Spiro ring-containing quinazoline compound

Abstract

The present invention relates to a compound of general formula (1) and a preparation method therefor, and use of the compound of formula (1) and isomers, crystalline forms and pharmaceutically acceptable salts thereof as an irreversible inhibitor for a K-Ras G12C mutant protein in preparing a medicament for resisting Ras-related diseases such as tumors. ##STR00001##

Claims

1. A compound of general formula (1) or pharmaceutically acceptable salts thereof: ##STR00204## wherein in the general formula (1): m is an integer of 1 or 2; n is an integer of 1 or 2; R.sup.1 is H, halogen, C1-C3 alkyl, C2-C4 alkenyl, C2-C4 alkynyl or C3-C6 cycloalkyl; R.sup.2 is C1-C3 alkyl or halogenated C1-C3 alkyl; R.sup.3 is ##STR00205## wherein R.sup.a is H or F, R.sup.b is H, F, Cl or Me, R.sup.c is H, F, Cl or Me, R.sup.d is H, F, Cl, NH.sub.2, Me or cyclopropyl, and R.sup.e, R.sup.f, R.sup.g, R.sup.h, R.sup.i, R.sup.j and R.sup.k are independently H, F, Cl, OH, OMe, NH.sub.2, CF.sub.3, C1-C3 alkyl or C3-C6 cycloalkyl; R.sup.4 is ##STR00206## ##STR00207## wherein n.sub.1 and n.sub.2 are independently integers of 1 or 2, m.sub.1, m.sub.2, m.sub.3 and m.sub.4 are independently integers of 0, 1, 2, 3 or 4, and m.sub.5 is an integer of 1, 2 or 3; A is CH.sub.2, O, S, SO, SO.sub.2 or N(C1-C3 alkyl)-, V is CH.sub.2, SO.sub.2 or CO, and L is O, S, SO.sub.2, SO or CO; X is a 5- to 7-membered heteroaryl or a partially saturated 5- to 7-membered heterocycloalkyl; Y is C3-C6 cycloalkyl, heterocycloalkyl, (C3-C6) cycloalkyl-(C1-C3) alkyl- or heterocycloalkyl-(C1-C3) alkyl-; R.sup.l and R.sup.m are independently C1-C3 alkyl, halogenated C1-C3 alkyl, hydroxyl-substituted C1-C3 alkyl, cyano-substituted C1-C3 alkyl, C3-C6 cycloalkyl, (C1-C3) alkoxy-(C2-C3) alkyl-, (halogenated C1-C3) alkoxy-(C2-C3) alkyl-, (C3-C6) cycloalkyl-(C1-C3) alkyl-; or R.sup.l and R.sup.m, together with a N atom, form 3- to 8-membered heterocycloalkyl, wherein the 3- to 8-membered heterocycloalkyl can be substituted with 1-3 substituents selected from OH, halogen, cyano, C1-C3 alkyl, C3-C6 cycloalkyl, heterocycloalkyl, (C1-C3) alkoxy and (halogenated C1-C3) alkoxy; R.sup.n is C1-C3 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, C3-C6 cycloalkyl, heterocycloalkyl, halogenated C1-C3 alkyl, hydroxyl-substituted C1-C3 alkyl, cyano-substituted C1-C3 alkyl, (C1-C3) alkoxy-(C2-C3) alkyl-, (halogenated C1-C3) alkoxy-(C2-C3) alkyl-, (C3-C6) cycloalkyl-(C1-C3) alkyl- or heterocycloalkyl-(C1-C3) alkyl-; and R.sup.o is OH, halogen, cyano, C1-C3 alkyl, C1-C3 alkoxy or C3-C6 cycloalkyl.

2. The compound according to claim 1, wherein in the general formula (1), R.sup.1 is H, F, Cl, Me, Et, isopropyl, vinyl, ethynyl or cyclopropyl.

3. The compound according to claim 1, wherein in the general formula (1), R.sup.2 is CH.sub.3, CH.sub.3CH.sub.2, CF.sub.3CH.sub.2, CHF.sub.2CH.sub.2 or CF.sub.3(CH.sub.3)CH.

4. The compound according to claim 1, wherein in the general formula (1), R.sup.3 is ##STR00208## ##STR00209## ##STR00210##

5. The compound according to claim 1, wherein in the general formula (1), R.sup.4 is: ##STR00211## ##STR00212## ##STR00213## ##STR00214## ##STR00215## ##STR00216## ##STR00217## ##STR00218## ##STR00219## ##STR00220## ##STR00221## ##STR00222## ##STR00223## ##STR00224##

6. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound has one of the following structures: ##STR00225## ##STR00226## ##STR00227## ##STR00228## ##STR00229## ##STR00230## ##STR00231## ##STR00232## ##STR00233## ##STR00234## ##STR00235## ##STR00236## ##STR00237## ##STR00238## ##STR00239## ##STR00240## ##STR00241## ##STR00242## ##STR00243## ##STR00244## ##STR00245## ##STR00246## ##STR00247## ##STR00248## ##STR00249## ##STR00250## ##STR00251## ##STR00252## ##STR00253## ##STR00254## ##STR00255## ##STR00256## ##STR00257## ##STR00258## ##STR00259## ##STR00260## ##STR00261## ##STR00262## ##STR00263## ##STR00264## ##STR00265## ##STR00266## ##STR00267## ##STR00268##

7. A pharmaceutical composition comprising a pharmaceutically acceptable excipient or carrier, and the compound or the pharmaceutically acceptable salts thereof according to claim 1 as active ingredients.

Description

DETAILED DESCRIPTION

(1) Various specific aspects, features and advantages of the compounds, methods and pharmaceutical compositions described above are set forth in detail in the following description, which makes the present invention clear. It should be understood that the detailed description and examples below describe specific embodiments for reference only. After reading the description of the present invention, those skilled in the art can make various changes or modifications to the present invention, and such equivalents also fall within the scope of the present invention defined herein.

(2) In all examples, 1H-NMR spectra were recorded with a Vian Mercury 400 nuclear magnetic resonance spectrometer, and chemical shifts are expressed in (ppm); silica gel for separation was 200-300 mesh silica gel if not specified, and the ratio of the eluents was volume ratio.

(3) In the present invention, the following abbreviations are used: MeCN represents acetonitrile; DCM represents dichloromethane; DIPEA represents diisopropylethylamine; dioxane represents 1,4-dioxane; DMF represents dimethylformamide; h represents hour; K.sub.3PO.sub.4 represents potassium phosphate; min represents minute; MS represents mass spectroscopy; NaH represents sodium hydride; NMR represents nuclear magnetic resonance; Pd.sub.2(dba).sub.3 represents tris(dibenzylideneacetone)dipalladium; Pd(dppf)Cl.sub.2 represents [1,1-bis(diphenylphosphino)ferrocene]palladium dichloride; TFA (CF.sub.3COOH) represents trifluoroacetic acid; TLC represents thin layer chromatography; THF represents tetrahydrofuran; and Xantphos represents 4,5-bis(diphenylphosphane)-9,9-dimethylxanthene.

Example 1 Synthesis of 1-(7-(6-cyclopropyl-2-(3-((dimethylamino)methyl)azetidin-1-yl)-7-(5-methyl-1H-indazol-4-yl)-8-(2,2,2-trifluoroethoxy)quinazolin-4-yl)-2,7-diazaspiro[3.5]non-2-yl)prop-2-en-1-one (Compound 1)

(4) ##STR00070## ##STR00071##

Step 1: Synthesis of Compound 1-3

(5) Compound 1-1 (5.53 g, 13.1 mmol) was suspended in dioxane (80 mL). DIPEA (10.1 g, 78.6 mmol) was added in an ice bath, followed by 1-2 (2.96 g, 13.1 mmol). The mixture was stirred for 30 min, and then warmed to room temperature and stirred at room temperature for 1 h. After the reaction was completed as detected by TLC, the reaction mixture was added with water and extracted with EA. The organic phase was dried and concentrated, and the residue was slurried with EA to obtain a yellow solid 1-3 (4.52 g, 56.3% yield).

(6) .sup.1H NMR (400 MHz, DMSO-d.sub.6) : 8.26 (d, J=1.5 Hz, 1H), 3.79 (s, 4H), 3.65 (s, 4H), 1.86 (t, J=5.3 Hz, 4H), 1.39 (s, 9H); MS(ESI): MS (ESI): 611.2 [M+1].sup.+.

Step 2: Synthesis of Compound 1-4

(7) Compound 1-3 (4.28 g, 7.0 mmol) was dissolved in a mixed solution of DMF (40 mL) and THF (40 mL). 1-(azetidin-3-yl)-N,N-dimethylmethylamine (1.60 g, 14.0 mmol) and DABCO (155 mg, 1.4 mmol) were added. The mixture was stirred at room temperature overnight. After the reaction was completed, the reaction mixture was added with water and extracted with EA. The organic phase was dried and concentrated, and the residue was purified by column chromatography to obtain compound 1-4 (3.62 g, 75.1% yield). MS(ESI): 689.2 [M+1].sup.+.

Step 3: Synthesis of Compound 1-5

(8) Trifluoroethanol (0.75 g, 7.5 mmol) was dissolved in anhydrous DMF (10 mL). NaH (60%, 0.60 g, 15.0 mmol) was added in an ice bath. The mixture was stirred at room temperature for 5 min to obtain sodium trifluoroethoxide. Compound 1-4 (3.45 g, 5.0 mmol) was dissolved in anhydrous THF (40 mL). The solution of sodium trifluoroethoxide in DMF prepared above was added. The mixture was stirred at room temperature overnight. After the reaction was completed, the reaction mixture was added with water and extracted with EA. The organic phase was dried and concentrated, and the residue was purified by column chromatography to obtain compound 1-5 (3.54 g, 92.1% yield). MS (ESI): 769.2 [M+1].sup.+.

Step 4: Synthesis of Compound 1-6

(9) To a single-necked flask were added compound 1-5 (3.08 g, 4.0 mmol), cyclopropylboronic acid (0.43 g, 5.0 mmol), Pd(dppf)Cl.sub.2 (0.59 g, 0.8 mmol) and K.sub.3PO.sub.4 (0.85 g, 4.0 mmol), followed sequentially by MeCN (40 mL), dioxane (40 mL) and H.sub.2O (16.5 mL). The mixture was stirred under nitrogen atmosphere at 100 C. for 5 h. After the reaction was completed, the reaction mixture was purified by column chromatography to obtain compound 1-6 (1.78 g, 65.2% yield). MS (ESI): 683.3 [M+1].sup.+.

Step 5: Synthesis of Compound 1-7

(10) To a single-necked flask were added compound 1-6 (1.37 g, 2.0 mmol), 5-methyl-1H-indazole-4-boronic acid (0.53 g, 3.0 mmol), Pd.sub.2(dba).sub.3 (0.27 g, 0.3 mmol), Xatphos (0.35 g, 0.6 mmol) and K.sub.3PO.sub.4 (0.85 g, 4.0 mmol), followed by dioxane (30 mL) and H.sub.2O (3 mL). The mixture was stirred under nitrogen at 120 C. overnight. After the reaction was completed, the mixture was purified by column chromatography to obtain compound 1-7 (557 mg, 26.4% yield). MS (ESI): 735.4 [M+1].sup.+.

Step 6: Synthesis of Compound 1-8

(11) Compound 1-7 (515 mg, 0.7 mmol) was dissolved in DCM (10 mL). TFA (3 mL) was added. The mixture was stirred at room temperature for 2 h. After the reaction was completed, the reaction mixture was concentrated, basified with saturated sodium carbonate, and extracted with EA. The organic phase was dried and concentrated to obtain compound 1-8 (445 mg, 100% yield). MS(ESI): 635.4 [M+1].sup.+.

Step 7: Synthesis of Compound 1

(12) Compound 1-8 (318 mg, 0.4 mmol) was dissolved in dry DCM (15 mL). DIPEA (65 mg, 0.5 mmol) was added in an ice salt bath, followed by slowly addition of acryloyl chloride (43 mg, 0.48 mmol). The mixture was reacted in an ice bath for 2 h. The reaction mixture was washed with saturated brine. The organic phase was dried and concentrated, and the residue was purified by column chromatography to obtain compound 1 (181 mg, yield 65.8%).

(13) .sup.1H NMR (400 MHz, DMSO-d.sub.6) 12.93 (s, 1H), 7.43-7.45 (m, 2H), 7.31 (d, J=8.4 Hz, 1H), 7.10 (s, 1H), 6.34 (dd, J=16.9, 10.3 Hz, 1H), 6.09-6.14 (m, 1H), 5.67-5.70 (m, 1H), 4.80-4.90 (m, 1H), 4.49-4.59 (m, 1H), 4.02-4.10 (m, 4H), 3.83-3.87 (m, 2H), 3.74 (s, 2H), 3.57 (s, 4H), 2.10-2.15 (m, 12H), 1.92-2.02 (m, 4H), 1.26-1.29 (m, 1H), 0.48-0.61 (m, 4H), MS (ESI): 689.4 [M+1].sup.+

(14) By the separation and purification on a chiral column, two axially chiral isomer of the compound 1 may be obtained:

(15) ##STR00072##

Example 2-132. Synthesis of Compound 2-132

(16) The target compound 2-132 was obtained using different starting materials according to a synthesis method similar to that in Example 1.

(17) TABLE-US-00001 TABLE 1 Compound structure [M + H].sup.+ 2 648.3 3 662.3 4 719.4 5 733.4 6 701.4 7 715.4 8 731.4 9 0 676.3 10 702.3 11 731.4 12 688.3 13 702.3 14 717.4 15 745.4 16 745.4 17 702.3 18 718.3 19 0 731.4 20 700.3 21 705.3 22 719.4 23 719.4 24 733.4 25 733.3 26 749.3 27 765.3 28 713.4 29 00 727.4 30 01 729.3 31 02 745.3 32 03 742.4 33 04 757.4 34 05 713.4 35 06 727.4 36 07 741.4 37 08 727.4 38 09 741.4 39 0 697.3 40 698.3 41 698.3 42 699.3 43 713.4 44 703.3 45 717.4 46 758.4 47 758.4 48 770.4 49 0 794.3 50 794.3 51 714.3 52 744.4 53 758.4 54 726.3 55 740.4 56 756.4 57 737.3 58 754.4 59 0 689.4 60 701.4 61 715.4 62 703.4 63 715.4 64 729.4 65 658.3 66 672.3 67 687.3 68 755.3 69 0 727.4 70 727.4 71 729.3 72 731.4 73 701.4 74 745.4 75 715.4 76 759.4 77 701.4 78 745.4 79 0 729.4 80 773.4 81 717.3 82 761.4 83 717.3 84 761.4 85 690.3 86 690.3 87 702.3 88 702.3 89 0 716.4 90 716.4 91 752.3 92 752.3 93 728.4 94 728.4 95 742.4 96 742.4 97 659.4 98 685.4 99 0 727.4 100 699.3 101 685.3 102 713.4 103 717.3 104 731.3 105 727.4 106 727.4 107 713.4 108 717.3 109 0 713.4 110 717.3 111 725.3 112 693.3 113 695.3 114 715.4 115 727.4 116 715.4 117 729.4 118 733.4 119 0 727.4 120 727.4 121 717.3 122 731.4 123 731.4 124 745.4 125 713.4 126 727.4 127 699.3 128 713.4 129 00 737.3 130 01 751.3 131 02 687.3 132 03 701.4

Example 133: Chiral Resolution of Compound 73

(18) The compounds of the present application may have axial chirality. The compounds with axial chirality can be resolved to obtain two chiral isomers.

(19) The sample was dissolved in ethanol to reach a concentration of 25 mg/mL, and the injection volume was 500 L. Conditions for preparative chromatography: CHIRALPAK AD-H (20250 mm, 5 m) chromatography column; mobile phase: ethanol-n-hexane (40:60); flow rate: 12 mL/min; wavelength of detection: 254 nm. The stepwise eluate was concentrated by rotary evaporation and dried to obtain products 73-a and 73-b: a first axially chiral isomer: 73-a; retention time on the chromatographic column: 8.532 min; and a second axially chiral isomer: 73-b; retention time on the chromatographic column: 10.126 min.

(20) The compound 67 was chirally resolved by a similar resolution method to obtain their two chiral isomers 67-a and 67-b, and the retention time on the chromatographic column was as follows: a first axially chiral isomer: 67-a; retention time on the chromatographic column: 5.413 min; and a second axially chiral isomer: 67-b; retention time on the chromatographic column: 7.938 min. Other compounds in the present application can also be chirally resolved using a similar method.

Example 134. pERK and ERK Protein Content Assay in 11358 Cells by Compounds

(21) H358 cells were seeded in a 24-well plate. After one day of growth, a test compound (at a concentration of 1 M) was added. After 24 h of action of the compound, the cells were lysed, and the cell lysate was transferred to a 96-well ELISA plate. The levels of pERK and ERK in the lysate were measured using an ELISA kit (abcam 176660). The ratio of pERK to ERK was calculated and compared with that of the DMSO group, and the percentage of inhibition of pERK activity by the compound was calculated. The results are shown in Table 2 below.

(22) TABLE-US-00002 TABLE 2 Inhibitory activity of the compounds of the present invention against the pERK level in H358 cells Com- Inhibition Com- Inhibition Com- Inhibition pound rate (%) pound rate (%) pound rate (%) 1 +++ 2 +++ 3 ++ 4 +++ 5 +++ 6 +++ 7 +++ 8 ++ 9 +++ 10 +++ 11 +++ 12 +++ 13 +++ 14 +++ 15 +++ 16 +++ 17 +++ 18 +++ 19 +++ 20 ++ 21 +++ 22 +++ 23 +++ 24 +++ 25 +++ 26 ++ 27 ++ 28 +++ 29 +++ 30 +++ 31 +++ 32 +++ 33 ++ 34 +++ 35 +++ 36 ++ 37 +++ 38 +++ 39 ++ 40 ++ 41 ++ 42 ++ 43 ++ 44 +++ 45 +++ 46 +++ 47 +++ 48 ++ 49 +++ 50 ++ 51 +++ 52 +++ 53 +++ 54 +++ 55 +++ 56 +++ 57 ++ 58 +++ 59 +++ 60 +++ 61 +++ 62 +++ 63 +++ 64 +++ 65 ++ 66 ++ 67 +++ 68 +++ 69 +++ 70 +++ 71 +++ 72 +++ 73 +++ 74 +++ 75 ++ 76 +++ 77 +++ 78 +++ 79 +++ 80 ++ 81 +++ 82 +++ 83 ++ 84 ++ 85 +++ 86 ++ 87 +++ 88 ++ 89 +++ 90 ++ 91 +++ 92 ++ 93 +++ 94 ++ 95 +++ 96 +++ 97 +++ 98 +++ 99 +++ 100 +++ 101 +++ 102 +++ 103 +++ 104 +++ 105 +++ 106 +++ 107 +++ 108 +++ 109 +++ 110 +++ 111 +++ 112 +++ 113 ++ 114 +++ 115 +++ 116 +++ 117 +++ 118 +++ 119 +++ 120 +++ 121 +++ 122 +++ 123 +++ 124 +++ 125 +++ 126 +++ 127 +++ 128 +++ 129 +++ 130 +++ 131 +++ 132 +++ 1-a +++ 1-b +++ 67-a +++ 67-b +++ 73-a +++ 73-b +++ B +++ + indicates an inhibition rate less than or equal to 50% ++ indicates an inhibition rate from 50% to 90% +++ indicates an inhibition rate greater than 90%.

Example 135. Antiproliferative Activity of Compounds Against 11358 Cells

(23) 2500 H358 cells were seeded in a 96-well ultra-low attachment plate (corning, 7007). After one day of growth, a serially diluted compound (a maximum concentration of 5 M, 5-fold dilution, a total of five doses) was added. Three days after the addition of the compound, Cell Titer Glow (Promega, G9681) was added to evaluate pellet growth, and the IC.sub.50 value was calculated. The results are shown in Table 3 below.

(24) TABLE-US-00003 TABLE 3 Antiproliferative activity of the compounds of the present invention against H358 cells Compound IC.sub.50 Compound IC.sub.50 Compound IC.sub.50 1 +++ 2 +++ 3 ++ 4 +++ 5 +++ 6 +++ 7 +++ 8 ++ 9 +++ 10 +++ 11 +++ 12 +++ 13 +++ 14 +++ 15 +++ 16 +++ 17 +++ 18 +++ 19 +++ 20 ++ 21 +++ 22 +++ 23 +++ 24 +++ 25 ++ 26 ++ 27 ++ 28 +++ 29 +++ 30 +++ 31 +++ 32 +++ 33 ++ 34 +++ 35 +++ 36 ++ 37 +++ 38 ++ 39 ++ 40 + 41 ++ 42 + 43 + 44 +++ 45 +++ 46 +++ 47 +++ 48 ++ 49 +++ 50 ++ 51 +++ 52 +++ 53 +++ 54 +++ 55 +++ 56 +++ 57 ++ 58 +++ 59 +++ 60 +++ 61 +++ 62 +++ 63 +++ 64 +++ 65 ++ 66 ++ 67 +++ 68 +++ 69 +++ 70 +++ 71 +++ 72 +++ 73 +++ 74 +++ 75 ++ 76 +++ 77 +++ 78 +++ 79 +++ 80 ++ 81 +++ 82 +++ 83 ++ 84 ++ 85 +++ 86 ++ 87 +++ 88 ++ 89 +++ 90 ++ 91 +++ 92 + 93 +++ 94 ++ 95 +++ 96 ++ 97 +++ 98 +++ 99 +++ 100 +++ 101 +++ 102 +++ 103 +++ 104 +++ 105 +++ 106 +++ 107 +++ 108 +++ 109 +++ 110 +++ 111 ++ 112 +++ 113 ++ 114 +++ 115 +++ 116 +++ 117 +++ 118 +++ 119 +++ 120 +++ 121 +++ 122 +++ 123 +++ 124 +++ 125 +++ 126 +++ 127 +++ 128 +++ 129 +++ 130 +++ 131 +++ 132 +++ 1-a +++ 1-b +++ 67-a +++ 67-b +++ 73-a +++ 73-b +++ B +++ + indicates the IC50 of the compound is greater than 1 M ++ indicates the IC50 of the compound is from 0.3 to 1 M +++ indicates the IC50 of the compound is less than 0.3 M.

(25) As can be seen from the data in Tables 2 and 3, most of the compounds of the present invention have antiproliferative activity against H358 cells less than 0.3 M, and when R.sup.4 is a non-aromatic heterocyclic ring or a spiro ring, the compounds have strong inhibitory activity against the ERK phosphorylation of the RAS pathway, and the compounds have strong inhibitory activity against the proliferation of H358 tumor cells carrying a K-RAS G12C mutation.

Example 136. Evaluation of Antitumor Activity in Mice

(26) Human pancreatic cancer Mia PaCa-2 cells were cultured conventionally in 1640 medium containing 10% fetal bovine serum in a 37 C./5% CO.sub.2 incubator. After being subcultured, the cells were collected when they reached the desired amount. 110.sup.7 Mia PaCa-2 cells were injected into the left dorsal side of each nude mouse, and the animals were randomly grouped for administration after tumors grew to 150 mm.sup.3. The groups were as follows: 1) solvent control group, 8 mice; 2) compound 6 group, compound 14 group, compound 28 group, compound 44 group, compound 60 group, compound 103 group and control drug B (Example 35 in WO2018/143315), 8 mice in each group. The mice in the solvent control group was intragastrically administered with 0.5% CMC-Na once a day; and the mice in the compound groups were intragastrically administered with 0.5% CMC-Na suspension once a day. On Tuesday and Thursday each week, tumor volumes and body weight of the mice were measured, and the nude mice were sacrificed on day 21 during the treatment period. The test results are shown in Table 4 below.

(27) TABLE-US-00004 TABLE 4 Experimental therapeutic effects of compounds on human pancreatic cancer Mia PaCa-2 xenograft tumors in nude mice Dose Administration Compound (mg/kg) regimen Anti-tumor effect 6 10 qd*21 35% regression 14 10 qd*21 37% regression 28 10 qd*21 42% regression 44 10 qd*21 23% regression 60 10 qd*21 30% regression 103 10 qd*21 39% regression B 10 qd*21 28% regression

(28) As can be seen from the data in the table above, the compounds of the present invention have strong in vivo antitumor activity, and can cause tumor regression after 21 days of continuous administration at a dosage of 10 mg/kg/day. The compounds 6, 14, 28, 60 and 103 have stronger in vivo activity than the control drug B.