SEVEN-MEMBERED HETEROCYCLIC DERIVATIVE ACTING AS KRAS G12C MUTANT PROTEIN INHIBITOR

Abstract

A class of KRAS G12C mutant protein inhibitors, specifically disclosing the compound shown in formula (I), and an isomer and a pharmaceutically acceptable salt thereof.

##STR00001##

Claims

1. A compound of formula (II), a pharmaceutically acceptable salt thereof or a stereoisomer or tautomer thereof, ##STR00028## wherein, R.sub.1 is H, F, Cl, Br, I, NH.sub.2 and C.sub.1-3 alkyl, wherein the C.sub.1-3 alkyl is optionally substituted with 1, 2 or 3 R.sub.a; R.sub.2, R.sub.3, R.sub.4 and R.sub.5 are each independently selected from the group consisting of H, F, Cl, Br, I and C.sub.1-3 alkyl, wherein the C.sub.1-3 alkyl is optionally substituted with 1, 2 or 3 R.sub.b; T is selected from the group consisting of C(R.sub.6) and N; R.sub.6 is selected from the group consisting of H, F, Cl, Br, I and C.sub.1-3 alkyl, wherein the C.sub.1-3 alkyl is optionally substituted with 1, 2 or 3 R.sub.c; R.sub.7 is selected from the group consisting of H and —CH.sub.2—CN; R.sub.8 is selected from the group consisting of H and F; L.sub.1 is selected from the group consisting of a single bond, ##STR00029## R.sub.9 is selected from the group consisting of H, Cl, pyrrolidinyl and C.sub.1-6 alkylamino, wherein the pyrrolidinyl and the C.sub.1-6 alkylamino are optionally substituted with 1, 2 or 3 R.sub.d; E is selected from the group consisting of —O— and —N-L.sub.2-L.sub.3-R.sub.10; L.sub.2 is selected from the group consisting of a single bond, —CH.sub.2— and —C(═O)—; L.sub.3 is selected from the group consisting of a single bond, —CH.sub.2— and ##STR00030## R.sub.10 is selected from the group consisting of H and C.sub.1-6 alkylamino; R.sub.a, R.sub.b, R.sub.c and R.sub.d are each independently selected from the group consisting of H, F, Cl, Br, I and CH.sub.3; and n is selected from 2.

2-21. (canceled)

22. The compound, the pharmaceutically acceptable salt thereof or the stereoisomer or tautomer thereof according to claim 1, wherein R.sub.1 is selected from NH.sub.2.

23. The compound, the pharmaceutically acceptable salt thereof or the stereoisomer or tautomer thereof according to claim 1, wherein R.sub.2 is selected from the group consisting of Cl and Br.

24. The compound, the pharmaceutically acceptable salt thereof or the stereoisomer or tautomer thereof according to claim 1, wherein R.sub.3 is selected from H.

25. The compound, the pharmaceutically acceptable salt thereof or the stereoisomer or tautomer thereof according to claim 1, wherein R.sub.4 is selected from the group consisting of Cl and Br.

26. The compound, the pharmaceutically acceptable salt thereof or the stereoisomer or tautomer thereof according to claim 1, wherein R.sub.5 is selected from F.

27. The compound, the pharmaceutically acceptable salt thereof or the stereoisomer or tautomer thereof according to claim 26, wherein the structural unit ##STR00031## is selected from the group consisting of ##STR00032##

28. The compound, the pharmaceutically acceptable salt thereof or the stereoisomer or tautomer thereof according to claim 1, wherein R.sub.6 is selected from the group consisting of H, F, Cl, Br, I, CH.sub.3, CF.sub.3, CHF.sub.2 and CH.sub.2F.

29. The compound, the pharmaceutically acceptable salt thereof or the stereoisomer or tautomer thereof according to claim 28, wherein R.sub.6 is selected from the group consisting of CF.sub.3 and CH.sub.3.

30. The compound, the pharmaceutically acceptable salt thereof or the stereoisomer or tautomer thereof according to claim 1, wherein R.sub.9 is selected from the group consisting of H, Cl, ##STR00033## and C.sub.1-4 alkylamino.

31. The compound, the pharmaceutically acceptable salt thereof or the stereoisomer or tautomer thereof according to claim 30, wherein R.sub.9 is selected from the group consisting of H, Cl, ##STR00034##

32. The compound, the pharmaceutically acceptable salt thereof or the stereoisomer or tautomer thereof according to claim 1, wherein the structural unit ##STR00035## is selected from the group consisting of H, Cl, ##STR00036##

33. The compound, the pharmaceutically acceptable salt thereof or the stereoisomer or tautomer thereof according to claim 1, wherein R.sub.10 is selected from the group consisting of H and ##STR00037##

34. The compound, the pharmaceutically acceptable salt thereof or the stereoisomer or tautomer thereof according to claim 1, wherein the structural unit -L.sub.2-L.sub.3-R.sub.10 is selected from the group consisting of H, ##STR00038##

35. The compound, the pharmaceutically acceptable salt thereof or the stereoisomer or tautomer thereof according to claim 1, wherein the compound is selected from formula (I): ##STR00039##

36. The compound, the pharmaceutically acceptable salt thereof or the stereoisomer or tautomer thereof according to claim 1, wherein the compound is selected from any one of: ##STR00040## ##STR00041## ##STR00042## ##STR00043##

37. The compound, the pharmaceutically acceptable salt thereof or the stereoisomer or tautomer thereof according to claim 36, selected from the group consisting of: ##STR00044## ##STR00045##

38. A pharmaceutical composition, comprising a therapeutically effective amount of the compound or the pharmaceutically acceptable salt, or the stereoisomer or tautomer thereof according to claim 1 as an active ingredient, and a pharmaceutically acceptable carrier.

39. A method for treating KRAS G12C-associated cancer, comprising administering to a subject in need thereof a compound, a pharmaceutically acceptable salt, or a stereoisomer or tautomer thereof according to claim 1.

Description

DETAILED DESCRIPTION

[0090] The present invention is described in detail below by way of examples. However, this is by no means disadvantageously limiting the scope of the present invention. The compounds disclosed herein can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific examples listed below, examples formed by combinations thereof with other chemical synthetic methods, and equivalents thereof known to those skilled in the art. Preferred examples include, but are not limited to, the examples of the present invention. It will be apparent to those skilled in the art that various changes and modifications can be made to the specific examples of the present invention without departing from the spirit and scope of the present invention.

Example 1. Preparation of Compounds 1-11A, 1-11B, 1-11C and 1-11D

[0091] ##STR00027##

[0092] Step 1.

[0093] A solution of compound 1-1 (5.0 g, 13.54 mmol, 1.0 eq) in propionic acid (50 mL) was heated to 125° C., followed by the slow addition of fuming nitric acid (2.56 g, 40.63 mmol, 1.83 mL, 3.0 eq). The reaction solution was reacted at 125° C. for 2 h. The reaction solution was slowly poured into 10 wt % ice brine (100 mL), and filtered. The filter cake was dried to give compound 1-2. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 8.86 (s, 1H), 8.35 (d, J=8.31 Hz, 1H), 8.11-8.19 (m, 1H), 8.02-8.10 (m, 1H), 7.61 (s, 1H). LCMS (ESI) m/z: 415.0 (m+1).sup.+.

[0094] Step 2.

[0095] A solution of compound 1-2 (2.5 g, 6.04 mmol, 1.0 eq) in phosphorus oxychloride (41.25 g, 269.02 mmol, 25 mL, 44.57 eq) was heated to 100° C., followed by the addition of N,N-dimethylaniline (731.37 mg, 6.04 mmol, 765.03 μL, 1.0 eq). The reaction mixture was stirred at 100° C. for 30 min, and distilled under reduced pressure to remove the solvent. The resulting residue was diluted with ethyl acetate (20 mL) and slowly added to ice water (50 mL), and the mixture was extracted with ethyl acetate (50 mL×2). The organic phases were combined, washed with saturated brine (50 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give compound 1-3. LCMS (ESI) m/z: 433.0 (m+1).sup.+.

[0096] Step 3.

[0097] To a solution of compound 1-3 (500 mg, 1.16 mmol, 1.0 eq) in dimethyl sulfoxide (5 mL) were added diisopropylethylamine (448.05 mg, 3.47 mmol, 603.85 μL, 3.0 eq) and compound 1-4 (532.28 mg, 2.31 mmol, 2.0 eq). The reaction solution was purged three times with nitrogen and reacted at 25° C. for 16 h. To the reaction solution was added room-temperature water (50 mL), and the resulting mixture was extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with saturated brine (50 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by silica gel chromatography (petroleum ether:ethyl acetate=2:1 to dichloromethane:methanol=20:1) to give compound 1-5. LCMS (ESI) m/z: 627.2 (m+1).sup.+.

[0098] Step 4.

[0099] To a solution of compound 1-5 (360 mg, 574.61 mmol, 1.0 eq) in N,N-dimethylformamide (36 mL) was added potassium carbonate (158.83 mg, 1.15 mmol, 2.0 eq). The reaction solution was reacted in an oil bath at 120° C. for 14 h. To the reaction solution was added room-temperature water (50 mL), and the resulting mixture was extracted with ethyl acetate (50 mL×2). The organic phases were combined, washed with saturated brine (50 mL×2) and concentrated under reduced pressure. The resulting residue was purified by silica gel chromatography (petroleum ether:ethyl acetate=1:1) to give compound 1-6. .sup.1H NMR (400 MHz, CDCl.sub.3-d) δ 8.52 (d, J=1.00 Hz, 1H), 8.13-8.23 (m, 1H), 7.73 (dt, J.sub.1=8.41 Hz, J.sub.2=5.27 Hz, 1H), 7.57-7.67 (m, 1H), 7.20 (d, J=5.52 Hz, 1H), 4.57 (br d, J=0.90 Hz, 1H), 4.10-4.19 (m, 2H), 3.80 (br s, 1H), 3.22-3.49 (m, 4H), 2.78-2.93 (m, 1H), 2.18-2.46 (m, 2H), 1.51 (s, 9H). LCMS (ESI) m/z: 580.3 (m+1).sup.+.

[0100] Step 5.

[0101] To a mixed solution of compound 1-6 (200 mg, 342.30 mmol, 1.0 eq) in ethanol (5 mL) and water (2.5 mL) were added iron powder (95.58 mg, 1.71 mmol, 5.0 eq) and ammonium chloride (91.55 mg, 1.71 mmol, 5.0 eq). The reaction solution was reacted in an oil bath at 80° C. for 2 h. The reaction solution was filtered and the filter cake was washed with ethanol (20 mL). The filtrate was concentrated under reduced pressure. The resulting residue was diluted with ethyl acetate (20 mL), washed with room-temperature water (20 mL) and saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give compound 1-7. .sup.1H NMR (400 MHz, CDCl.sub.3-d) δ 8.48-8.55 (m, 1H), 7.15-7.26 (m, 2H), 6.55-6.68 (m, 2H), 4.50-4.63 (m, 1H), 4.27-4.37 (m, 1H), 3.71-3.88 (m, 3H), 3.22-3.49 (m, 4H), 2.33-2.45 (m, 1H), 2.14-2.26 (m, 1H), 1.51 (s, 9H). LCMS (ESI) m/z: 550.3 (m+3). LCMS (ESI) m/z: 550.3 (m+1).sup.+.

[0102] Step 6.

[0103] To a solution of compound 1-7 (300 mg, 545.93 mmol, 1.0 eq) in acetonitrile (6 mL) was added chlorosuccinimide (138.51 mg, 1.04 mmol, 1.9 eq) at 0° C. The reaction solution was reacted in an oil bath at 60° C. for 2 h. The reaction solution was slowly added to a saturated sodium sulfite solution (20 mL), and the resulting mixture was extracted with ethyl acetate (20 mL×2). The organic phases were combined, washed with saturated brine (20 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by silica gel chromatography (petroleum ether:ethyl acetate=2:1) to give compounds 1-8A and 1-8B. LCMS (ESI) m/z: 618.2 (m+1).sup.+.

[0104] Step 7.

[0105] To a solution of compound 1-8A (Rf=0.5) (70 mg, 113.19 mmol, 1.0 eq) in ethyl acetate (0.5 mL) was added hydrogen chloride in ethyl acetate (4 mol/L, 0.5 mL, 17.67 eq). The reaction solution was reacted at 25° C. for 1 h. The reaction solution was concentrated under reduced pressure to give compound 1-9A hydrochloride. LCMS (ESI) m/z: 518.1 (m+1).sup.+.

[0106] Step 8.

[0107] To a solution of compound 1-9A (62 mg, 111.76 mmol, 1.0 eq, hydrochloride) in dichloromethane (2 mL) were added diisopropylethylamine (72.22 mg, 558.81 mmol, 97.33 μL, 5.0 eq) and compound 1-10 (11.13 mg, 122.94 mmol, 10.02 μL, 1.1 eq) at 0° C. The reaction solution was reacted at 0° C. for 0.5 h. To the reaction solution was added saturated ammonium chloride (10 mL), and the resulting mixture was extracted with ethyl acetate (10 mL×2). The organic phases were combined and concentrated under reduced pressure. The resulting residue was purified by preparative HPLC (mobile phase: water (10 mmol ammonium bicarbonate)-acetonitrile, gradient: acetonitrile 31%-61%, 10 min). The resulting product was purified by preparative SFC (column model: DAICEL CHIRALPAK AD (250 mm×30 mm, 10 μm), mobile phase: isopropanol (0.1% ammonium hydroxide), gradient: carbon dioxide critical fluid 40%-40%, 4.8 min, 35 min) to give compounds 1-11A (rt=0.953 min) and 1-11B (rt=1.264 min). 1-11A: .sup.1H NMR (400 MHz, MeOH-d.sub.4) δ 8.41 (s, 1H), 7.53 (d, J=7.38 Hz, 1H), 7.33 (s, 1H), 6.74-6.96 (m, 1H), 6.28 (br d, J=17.26 Hz, 1H), 5.82 (dd, J.sub.1=10.63 Hz, J.sub.2=1.75 Hz, 1H), 4.55 (br d, J=3.13 Hz, 1H), 4.24 (br s, 2H), 3.82-4.07 (m, 2H), 3.72 (br s, 1H), 3.45-3.51 (m, 1H), 3.38 (br d, J=10.76 Hz, 2H), 2.17-2.37 (m, 2H); LCMS (ESI) m/z: 572.0 (m+1).sup.+. 1-11B: .sup.1H NMR (400 MHz, MeOH-d.sub.4) δ 8.41 (s, 1H), 7.53 (d, J=7.38 Hz, 1H), 7.33 (s, 1H), 6.75-6.97 (m, 1H), 6.28 (br d, J=17.13 Hz, 1H), 5.82 (dd, J.sub.1=10.63 Hz, J.sub.2=1.75 Hz, 1H), 4.54 (br d, J=6.13 Hz, 1H), 4.17-4.39 (m, 2H), 3.83-4.08 (m, 2H), 3.72 (br s, 1H), 3.45-3.51 (m, 1H), 3.38 (br d, J=11.63 Hz, 2H), 2.16-2.35 (m, 2H); LCMS (ESI) m/z: 572.0 (m+1).sup.+.

[0108] Step 9.

[0109] To a solution of compound 1-8B (Rf=0.4) (80 mg, 129.36 mmol, 1.0 eq) in ethyl acetate (0.5 mL) was added hydrogen chloride in ethyl acetate (4 mol/L, 0.5 mL, 15.46 eq). The reaction solution was reacted at 25° C. for 1 h. The reaction solution was concentrated under reduced pressure to give compound 1-9B hydrochloride.

[0110] LCMS (ESI) m/z: 518.1 (m+1).sup.+.

[0111] Step 10.

[0112] To a solution of compound 1-9B (71 mg, 127.99 mmol, 1.0 eq, hydrochloride) in dichloromethane (2 mL) were added diisopropylethylamine (82.71 mg, 639.93 mmol, 111.46 μL, 5.0 eq) and compound 1-10 (12.74 mg, 140.78 mmol, 11.48 μL, 1.1 eq) at 0° C. The reaction solution was reacted at 0° C. for 0.5 h. To the reaction solution was added saturated ammonium chloride (10 mL), and the resulting mixture was extracted with ethyl acetate (10 mL×2). The organic phases were combined and concentrated under reduced pressure. The resulting residue was purified by preparative HPLC (mobile phase: water (10 mmol ammonium bicarbonate)-acetonitrile, gradient: acetonitrile 30%-63%, 11 min). The resulting product was purified by preparative SFC (DAICEL CHIRALPAK AD (250 mm×30 mm, 10 m), mobile phase: isopropanol (0.1% ammonium hydroxide), gradient: carbon dioxide critical fluid 50%-50%, 3.2 min, 40 min) to give compounds 1-11C (rt=0.632 min) and 1-11D (rt=1.519 min).

[0113] 1-11C: .sup.1H NMR (400 MHz, MeOH-d.sub.4) δ 8.41 (s, 1H), 7.53 (d, J=7.38 Hz, 1H), 7.35 (br s, 1H), 6.75-6.98 (m, 1H), 6.29 (br dd, J.sub.1=17.26 Hz, J.sub.2=6.50 Hz, 1H), 5.82 (br d, J=10.63 Hz, 1H), 4.56 (br d, J=12.51 Hz, 1H), 4.01-4.29 (m, 2H), 3.95 (br d, J=6.88 Hz, 2H), 3.76 (br s, 1H), 3.48 (br d, J=12.51 Hz, 1H), 3.34-3.42 (m, 2H), 2.18-2.38 (m, 2H); LCMS (ESI) m/z: 572.0 (m+1).sup.+.

[0114] 1-11D: .sup.1H NMR (400 MHz, MeOH-d.sub.4) δ 8.40 (s, 1H), 7.53 (d, J=7.34 Hz, 1H), 7.34 (s, 1H), 6.72-6.99 (m, 1H), 6.29 (br dd, J.sub.1=16.81 Hz, J.sub.2=5.20 Hz, 1H), 5.82 (br d, J=10.64 Hz, 1H), 4.32-4.71 (m, 3H), 4.21 (br s, 1H), 3.95 (br s, 1H), 3.74 (br d, J=11.49 Hz, 1H), 3.47 (br d, J=12.72 Hz, 1H), 3.35 (br s, 2H), 2.26 (br d, J=7.09 Hz, 2H); LCMS (ESI) m/z: 572.0 (m+1).sup.+.

Experimental Example 1. Cell Assay

[0115] Objective:

[0116] This experiment was conducted to verify the proliferation inhibitory effect of the compounds disclosed herein on KRAS G12C mutant NCI-H358 human non-small cell lung cancer cells and wild-type A375 human malignant melanoma cells.

[0117] Materials:

[0118] Cell line NCI-H358 (purchased from the Cell Bank of the Chinese Academy of Sciences), cell line A375 (purchased from the Cell Bank of the Chinese Academy of Sciences), DMEM medium, penicillin/streptomycin antibiotics purchased from Wisent, and fetal calf serum purchased from Biosera. CellTiter-Glo (chemiluminescence detection reagent for cell viability) reagent purchased from Promega.

[0119] Method:

[0120] 1) Anti-Proliferation Experiment on NCI-H358 Cells:

[0121] NCI-H358 cells were seeded in a white 96-well plate, with 80 μL of cell suspension containing 4000 NCI-H358 cells added to each well. The cell plate was incubated in a CO.sub.2 incubator overnight. The test compounds were serially diluted in a 3-fold gradient with a pipettor to 9th concentration, i.e. from 2 mM to 304 nM, and the duplicate wells were set up. 78 μL of medium was added to an intermediate plate, and the serially diluted compound was transferred to corresponding wells of the intermediate plate at 2 μL/well. After mixing, the mixture was transferred to the cell plate at 20 μL per well. The concentration of the compound transferred to the cell plate ranged from 10 μM to 1.52 nM. The cell plate was incubated in a CO.sub.2 incubator for 5 days. Another cell plate was read for signal values on the day of compound addition, and these values were used as the maximum values (the Max value in the equation below) in data analysis. To the cell plate was added the chemiluminescence detection reagent for cell viability at 25 μL/well, and the resulting plate was incubated at room temperature for 10 min to stabilize the luminescence signals. Readings were taken using a multi-marker analyzer. To the cell plate was added the chemiluminescence detection reagent for cell viability at 25 μL/well, and the resulting plate was incubated at room temperature for 10 min to stabilize the luminescence signals. Readings were taken using a multi-marker analyzer.

[0122] 2) Anti-Proliferation Experiment on A375 Cells:

[0123] A375 cells were seeded in a white 96-well plate, with 80 μL of cell suspension containing 2000 A375 cells added to each well. The cell plate was incubated in a CO.sub.2 incubator overnight. The test compounds were serially diluted in a 3-fold gradient with a pipettor to 9th concentration, i.e. from 2 mM to 304 nM, and the duplicate wells were set up. 78 μL of medium was added to an intermediate plate, and the serially diluted compound was transferred to corresponding wells of the intermediate plate at 2 μL/well. After mixing, the mixture was transferred to the cell plate at 20 μL per well. The concentration of the compound transferred to the cell plate ranged from 10 μM to 1.52 nM. The cell plate was incubated in a CO.sub.2 incubator for 5 days. Another cell plate was read for signal values on the day of compound addition, and these values were used as the maximum values (the Max value in the equation below) in data analysis. To the cell plate was added the chemiluminescence detection reagent for cell viability at 25 Lt/well, and the resulting plate was incubated at room temperature for 10 min to stabilize the luminescence signals. Readings were taken using a multi-marker analyzer. To the cell plate was added the chemiluminescence detection reagent for cell viability at 25 Lt/well, and the resulting plate was incubated at room temperature for 10 min to stabilize the luminescence signals. Readings were taken using a multi-marker analyzer.

[0124] Data analysis: the original data were converted to inhibition rate using the equation (Sample−Min)/(Max−Min)×100%, and the IC.sub.50 value was then curve fitted using four parameters (obtained from the “log(inhibitor) vs. response—Variable slope” model in GraphPad Prism).

[0125] Results: IC.sub.50 data for the anti-proliferative activity of the compounds disclosed herein against NCI-H358 (G12C mutant) and A375 (wild-type) cells are shown in Table 1.

[0126] Conclusion: compounds 1-11A and 1-11D disclosed herein show higher cell anti-proliferative activity against KRAS G12C mutant cells NCI-H358, and weaker anti-proliferative activity against wild type A375 cells, thereby showing high selectivity.

TABLE-US-00001 TABLE 1 Test compound NCI-H358 IC.sub.50 (μM) A375 IC.sub.50 (μM) 1-11A 0.10 6.12 1-11B 5.03 >10 1-11C 3.58 4.78 1-11D 0.33 >10

Experimental Example 2. Pharmacokinetic Evaluation on Mice

[0127] Objective:

[0128] To determine the drug concentration in the plasma of the test animals (male CD-1 mice) at different time points after intravenous and intragastric administration of the test compounds by an LC/MS/MS method. To investigate the pharmacokinetic performance of the test compounds in mice and to evaluate the pharmacokinetic characteristics.

[0129] Experimental scheme: test animals: 8 healthy adult male SD rats, divided into 4 groups, 2 in IV group (two groups) and 2 in PO group (two groups), according to the principle of similar body weight. The mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0130] Drug Preparation:

[0131] IV group: an appropriate amount of sample was weighed, and an appropriate amount of DMSO, PEG400 and water was sequentially added in the volume ratio of 10:60:30, and the mixture was stirred and ultrasonicated to a clear state (1.5 mg/mL).

[0132] PO group: an appropriate amount of sample was weighed, and an appropriate amount of DMSO, PEG400 and water was sequentially added in the volume ratio of 10:60:30, and the mixture was stirred and ultrasonicated to a clear state (1.0 mg/mL).

[0133] Administration:

[0134] The mice in IV group were each subjected to intravenous administration at a volume of 2 mL/kg and a dose of 3 mg/kg; after fasting overnight, the mice in PO group were each subjected to intragastric administration at a volume of 10 mL/kg and a dose of 10 mg/kg.

[0135] Procedures:

[0136] For IV group, 30 μL of blood was collected from the male SD rats at 0.0833 h, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 8 h and 24 h post-dose, and placed in a commercial anticoagulation tube with EDTA-K.sub.2 added in advance. For PO group, 200 μL of blood was collected from the male SD rats at 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 8 h, 12 h and 24 h post-dose, and placed in a commercial anticoagulation tube with EDTA-K.sub.2 added in advance. The tubes were centrifuged for 15 min to separate the plasma and stored at −60° C. The animals were given free access to food 2 h after the administration. LC/MS/MS method was used to determine the content of the test compounds in the plasma of rats after intravenous and intragastric administration. The linear range of the method was 2.00-6000 nM; plasma samples were analyzed after treatment with acetonitrile to precipitate proteins.

[0137] Results: see Table 2.

[0138] Conclusion: in the pharmacokinetic evaluation experiment of mice, compound 1-11A disclosed herein shows high exposure and oral bioavailability.

TABLE-US-00002 TABLE 2 Groups I-11A IV (3 mg/kg) Cl (mL/Kg/min) 15.2 V.sub.d (L/kg) 1.45 AUC (nM .Math. h) 5699 T.sub.1/2 (h) 1.15 PO (10 mg/kg) C.sub.max (nM) 1700 T.sub.max (h) 0.50 AUC (nM .Math. h) 8481 F (%) 47.5 Note: Cl: clearance rate; V.sub.d: volume of distribution; AUC: exposure; t.sub.1/2: half-life; C.sub.max: maximum compound concentration after oral administration; T.sub.max: time to C.sub.max; F: bioavailability.

Experimental Example 3. In Vivo Efficacy Study

[0139] Objective:

[0140] To evaluate the in vivo efficacy of test compounds in an NCI-H358 subcutaneous xenograft tumor model of human non-small cell lung cancer.

[0141] Procedures:

[0142] BALB/c nude mice, female, 6-8 weeks old, weighing 18-21 g, 96 in total, provided by Shanghai Lingchang Biotechnology Co., Ltd. NCI-H358 tumor cells were resuspended in PBS to obtain 0.1 mL (5×10.sup.6 cells) of cell suspension and inoculated subcutaneously in the right back (5×10.sup.6/mouse) of each mouse for tumor growth. The mice were randomly grouped and subjected to intragastric administration once daily when the mean tumor volume reached approximately 150-200 mm.sup.3, and the dosages were shown in Table 3. Tumor diameters were measured twice weekly using a vernier caliper. The tumor volume was calculated according to the formula: V=0.5a×b.sup.2, where a and b represent the long diameter and short diameter of the tumor, respectively. The efficacy of compounds against tumor was evaluated by TGI (%). TGI (%) refers to the tumor growth inhibition rate. TGI (%)=[(1−(mean tumor volume at the end of administration of a treatment group−mean tumor volume at the start of administration of the treatment group))/(mean tumor volume at the end of treatment of the solvent control group−mean tumor volume at the start of treatment of the solvent control group)]×100%.

[0143] Results: see Table 3.

TABLE-US-00003 TABLE 3 Tumor volume Groups (mm.sup.3) (day 20) TGI (%) Solvent control group 634 — 1-11A 3 mg/kg (0-20 days) 332 64.1 1-11A 10 mg/kg (0-20 days) 240 83.9

[0144] Conclusion:

[0145] Compound 1-11A disclosed herein exhibits good in vivo efficacy in an NCI-H358 subcutaneous xenograft tumor model of human non-small cell lung cancer. At the dose of 3 mg/kg, 1-11A has a strong tumor growth inhibitory effect, while at the increased dose of 10 mg/kg, 1-11A shows a stronger tumor inhibitory effect than that at 3 mg/kg, with the tumor growth inhibition rate up to 83.9%.