IMIDAZOLIDINONE COMPOUND, PREPARATION METHOD THEREFOR AND USE THEREOF
20230053342 · 2023-02-23
Assignee
Inventors
- Houxian ZU (Beijing, CN)
- Xizhen SONG (Beijing, CN)
- Kai CHEN (Beijing, CN)
- Xiangyong Liu (Beijing, CN)
- Jie Chen (Beijing, CN)
- Yajing BIAN (Beijing, CN)
- Lieming Ding (Hangzhou, CN)
- Jiabing Wang (Beijing, CN)
Cpc classification
International classification
Abstract
An imidazolidinone compound represented by formula (I), or a stereoisomer thereof, a geometric isomer thereof, or a tautomer thereof, or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing the compound, a synthesis method therefor and use thereof.
##STR00001##
Claims
1. A compound of Formula (I) or a stereoisomer, geometric isomer or tautomer, or a pharmaceutically acceptable salt, solvate, chelate, non-covalent complex or prodrug thereof, ##STR00079## wherein, X is selected from O and S; R.sub.1 is selected from H, CN, C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.3-6 cycloalkyl, C.sub.3-6 heterocyclyl, C.sub.6-8 aryl, C.sub.5-8 heteroaryl, OR.sub.a and —NR.sub.aR.sub.b; wherein C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.3-6 cycloalkyl, C.sub.3-6 heterocyclyl, C.sub.6-8 aryl and C.sub.5-8 heteroaryl are each optionally substituted with one or more substituents selected from halogen, CN, OR.sub.a, oxo, C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.3-6 cycloalkyl and C.sub.3-6 heterocyclyl; R.sub.2 is selected from H, C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.3-6 cycloalkyl, C.sub.3-6 heterocyclyl, C.sub.6-8 aryl, C.sub.5-8 heteroaryl; wherein C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.3-6 cycloalkyl, C.sub.3-6 heterocyclyl, C.sub.6-8 aryl and C.sub.5-8 heteroaryl are each optionally substituted with one or more substituents selected from halogen, CN, —OH, —NO.sub.2, C.sub.1-6 alkyl, C.sub.1-6 haloalkyl, C.sub.2-6 alkenyl, C.sub.2-6 haloalkenyl, C.sub.2-6 alkynyl, C.sub.2-6 haloalkynyl, C.sub.3-6 cycloalkyl, C.sub.3-6 halocycloalkyl, C.sub.3-6 heterocyclyl, C.sub.3-6 haloheterocyclyl, C.sub.6-8 aryl, C.sub.6-8 haloaryl, C.sub.5-8 heteroaryl, C.sub.5-8 haloheteroaryl, oxo, —OR.sub.a, —NR.sub.aR.sub.b, —C(O)R.sub.a, —C(O)O R.sub.a, —C(O)NR.sub.aR.sub.b, —S(O)R.sub.a and —S(O).sub.2R.sub.a; R.sub.3 is selected from H, halogen, CN, C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, —OR.sub.a and —NR.sub.aR.sub.b; wherein C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl are each optionally substituted with one or more substituents selected from halogen, CN, —OR.sub.a, —NR.sub.aR.sub.b, —C(O)R.sub.a, —C(O)OR.sub.a, —C(O)NR.sub.aR.sub.b, —S(O)R.sub.a and —S(O).sub.2R.sub.a; R.sub.4 is selected from H, halogen, CN, C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, oxo, C.sub.1-6 haloalkyl, C.sub.2-6 haloalkenyl, C.sub.2-6 haloalkynyl, C.sub.1-6 alkoxy, C.sub.1-6 halo alkoxy, —OR.sub.a, —NR.sub.aR.sub.b, —C(O)R.sub.a, —C(O)OR.sub.a, —C(O)NR.sub.aR.sub.b, —S(O)R.sub.a and —S(O).sub.2R.sub.a; R.sub.5 is selected from H, halogen, CN, C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.1-6 haloalkyl, C.sub.2-6 haloalkenyl, C.sub.2-6 haloalkynyl, C.sub.1-6 alkoxy, C.sub.1-6 haloalkoxy, —OR.sub.a, —NR.sub.aR.sub.b, —S(O)R.sub.a and —S(O).sub.2R.sub.a; R.sub.6 is selected from H, halogen, CN, oxo, C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.1-6 alkoxy, C.sub.3-6 cycloalkyl, C.sub.3-6 heterocyclyl, C.sub.6-8 aryl, C.sub.5-8 heteroaryl, —OR.sub.a, —NR.sub.aR.sub.b, —C(O)R.sub.a, —C(O)O R.sub.a, —C(O) NR.sub.aR.sub.b, —S(O)R.sub.a and —S(O).sub.2R.sub.a; wherein C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.1-6 alkoxy, C.sub.3-6 cycloalkyl, C.sub.3-6 heterocyclyl, C.sub.6-8 aryl and C.sub.5-8 heteroaryl are each optionally substituted with one or more substituents selected from halogen, CN, oxo, —NO.sub.2, C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, —OR.sub.a, —NR.sub.aR.sub.b, —C(O)R.sub.a, —C(O)O R.sub.a, —C(O)NR.sub.aR.sub.b, —S(O)R.sub.a and —S(O).sub.2R.sub.a; or, two R.sub.6 take with the C atoms which they are attached to form C.sub.3-6 cycloalkyl or C.sub.3-6 heterocyclyl; R.sub.a and R.sub.b are independently selected from H, C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.1-6 alkoxy, C.sub.3-6 cycloalkyl, C.sub.3-6 heterocyclyl, C.sub.6-8 aryl, C.sub.5-8 heteroaryl; wherein C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.1-6 alkoxy, C.sub.3-6 cycloalkyl, C.sub.3-6 heterocyclyl, C.sub.6-8 aryl, C.sub.5-8 heteroaryl are optionally substituted with halogen, CN, —OH, —NH.sub.2, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 haloalkyl, C.sub.1-6 haloalkoxy; m is selected from 0, 1, 2, 3 and 4; n is selected from 0, 1, 2 and 3; and y is selected from 0, 1, 2, 3, 4, 5 and 6.
2. The compound of claim 1, or a stereoisomer, geometric isomer or tautomer, or a pharmaceutically acceptable salt, solvate, chelate, non-covalent complex or prodrug thereof, wherein R.sub.1 is C.sub.1-6 alkyl or C.sub.3-6 cycloalkyl, said C.sub.1-6 alkyl and C.sub.3-6 cycloalkyl can be substituted optionally and independently with halogen.
3. The compound of claim 1, or a stereoisomer, geometric isomer or tautomer, or a pharmaceutically acceptable salt, solvate, chelate, non-covalent complex or prodrug thereof, wherein R.sub.2 is selected from H, C.sub.1-6 alkyl, C.sub.3-6 cycloalkyl, C.sub.3-6 heterocyclyl, C.sub.6-8 aryl and C.sub.5-8 heteroaryl; said C.sub.1-6 alkyl, C.sub.3-6 cycloalkyl, C.sub.3-6 heterocyclyl, C.sub.6-8 aryl and C.sub.5-8 heteroaryl are each optionally substituted with one or more substituents selected from halogen, —CN, —OH, —NR.sub.aR.sub.b, C.sub.1-6 alkyl and C.sub.1-6 haloalkyl; said R.sub.a and R.sub.b are independently selected from H and C.sub.1-6 alkyl.
4. The compound of claim 1, or a stereoisomer, geometric isomer or tautomer, or a pharmaceutically acceptable salt, solvate, chelate, non-covalent complex or prodrug thereof, wherein R.sub.3 is selected from H, halogen, CN, C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.1-6 haloalkyl, C.sub.2-6 haloalkenyl and C.sub.2-6 haloalkynyl.
5. The compound of claim 1, or a stereoisomer, geometric isomer or tautomer, or a pharmaceutically acceptable salt, solvate, chelate, non-covalent complex or prodrug thereof, wherein R.sub.6 is selected from H, halogen, CN, —NH.sub.2, oxo, —OH, C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.1-6 alkoxy, C.sub.1-6 haloalkyl, C.sub.2-6 haloalkenyl, C.sub.2-6 haloalkynyl, C.sub.3-6 cycloalkyl, C.sub.3-6 heterocyclyl, C.sub.6-8 aryl and C.sub.5-8 heteroaryl.
6. The compound of claim 1, or a stereoisomer, geometric isomer or tautomer, or a pharmaceutically acceptable salt, solvate, chelate, non-covalent complex or prodrug thereof, wherein the compound is of Formula (II): ##STR00080##
7. (canceled)
8. (canceled)
9. The compound of claim 1, or a stereoisomer, geometric isomer or tautomer, or a pharmaceutically acceptable salt, solvate, chelate, non-covalent complex or prodrug thereof, wherein R.sub.3 is selected from H, halogen, C.sub.1-6 alkyl and C.sub.2-6 alkenyl.
10. The compound of claim 1, or a stereoisomer, geometric isomer or tautomer, or a pharmaceutically acceptable salt, solvate, chelate, non-covalent complex or prodrug thereof, wherein R.sub.4 and R.sub.5 are both H.
11. The compound of claim 1, or a stereoisomer, geometric isomer or tautomer, or a pharmaceutically acceptable salt, solvate, chelate, non-covalent complex or prodrug thereof, wherein the compound is of Formula (III): ##STR00081## wherein: R.sub.1 is C.sub.1-6 alkyl or C.sub.1-6 haloalkyl; R.sub.2 is selected from H, C.sub.1-6 alkyl, C.sub.3-6 cycloalkyl, C.sub.3-6 heterocyclyl, C.sub.6-8 aryl and C.sub.5-8 heteroaryl; wherein C.sub.1-6 alkyl, C.sub.3-6 cycloalkyl, C.sub.3-6 heterocyclyl, C.sub.6-8 aryl and C.sub.5-8 heteroaryl are each optionally substituted with one or more substituents selected from halogen, —CN, —OH, —NR.sub.aR.sub.b, C.sub.1-6 alkyl, C.sub.1-6 haloalkyl, C.sub.1-6 alkoxy and C.sub.1-6 halo alkoxy; R.sub.3 is selected from H, halogen, C.sub.1-6 alkyl and C.sub.2-6 alkenyl; R.sub.6 is selected from H, halogen, —NH.sub.2, oxo, —OH, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 haloalkyl, C.sub.1-6 halo alkoxy, C.sub.3-6 cycloalkyl and C.sub.6-8 aryl; R.sub.a and R.sub.b are independently selected from H and C.sub.1-6 alkyl; and y is selected from 0, 1, 2, 3, 4, 5 and 6.
12. The compound of claim 11, or a stereoisomer, geometric isomer or tautomer, or a pharmaceutically acceptable salt, solvate, chelate, non-covalent complex or prodrug thereof, wherein X is O.
13. The compound of claim 1, or a stereoisomer, geometric isomer or tautomer, or a pharmaceutically acceptable salt, solvate, chelate, non-covalent complex or prodrug thereof, wherein the compound is of Formula (IV): ##STR00082##
14. The compound of claim 1, or a stereoisomer, geometric isomer or tautomer, or a pharmaceutically acceptable salt, solvate, chelate, non-covalent complex or prodrug thereof, wherein R.sub.2 is selected from H, C.sub.1-6 alkyl, C.sub.3-6 cycloalkyl, C.sub.3-6 heterocyclyl, C.sub.6 aryl and C.sub.5-6 heteroaryl; wherein C.sub.1-6 alkyl, C.sub.3-6 cycloalkyl, C.sub.3-6 heterocyclyl, C.sub.6 aryl and C.sub.5-6 heteroaryl are each optionally substituted with 1 or more substituents selected from halogen, —CN, —OH, —N—(CH.sub.3).sub.2, C.sub.1-6 alkyl and C.sub.1-6 haloalkyl.
15. The compound of claim 1, or a stereoisomer, geometric isomer or tautomer, or a pharmaceutically acceptable salt, solvate, chelate, non-covalent complex or prodrug thereof, wherein R.sub.6 is selected from H, halogen, —NH.sub.2, oxo, —OH, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.3-6 cycloalkyl and C.sub.6 aryl.
16. The compound of claim 1, or a stereoisomer, geometric isomer or tautomer, or a pharmaceutically acceptable salt, solvate, chelate, non-covalent complex or prodrug thereof, wherein the compound is of Formula (V): ##STR00083## wherein, R.sub.1 is selected from C.sub.1-6 alkyl and C.sub.1-6 haloalkyl; R.sub.2 is selected from H, C.sub.1-6 alkyl, C.sub.3-6 cycloalkyl, C.sub.3-6 heterocyclyl, C.sub.6 aryl and C.sub.5-6 heteroaryl; wherein C.sub.1-6 alkyl, C.sub.3-6 cycloalkyl, C.sub.3-6 heterocyclyl, C.sub.6 aryl and C.sub.5-6 heteroaryl are each optionally substituted with one or more substituents selected from halogen, —CN, —OH, —N—(CH.sub.3).sub.2, C.sub.1-6 alkyl and C.sub.1-6 haloalkyl.
17. The compound of claim 1, or a stereoisomer, geometric isomer or tautomer, or a pharmaceutically acceptable salt, solvate, chelate, non-covalent complex or prodrug thereof, wherein R.sub.1 is C.sub.1-4 alkyl, wherein R.sub.1 is substituted optionally with halogen.
18. The compound of claim 1, or a stereoisomer, geometric isomer or tautomer, or a pharmaceutically acceptable salt, solvate, chelate, non-covalent complex or prodrug thereof, wherein R.sub.1 is selected from —CH(CH.sub.3).sub.2, —C(CH.sub.3).sub.3, —CF.sub.3 and —CHF.sub.2.
19. The compound of claim 1, or a stereoisomer, geometric isomer or tautomer, or a pharmaceutically acceptable salt, solvate, chelate, non-covalent complex or prodrug thereof, wherein R.sub.2 is selected from H, C.sub.1-6 alkyl, C.sub.3-6 cycloalkyl, C.sub.3-6 heterocyclyl, C.sub.6 aryl and C.sub.5-6 heteroaryl, wherein R.sub.2 is substituted optionally with halogen.
20. (canceled)
21. (canceled)
22. The compound of claim 1, or a stereoisomer, geometric isomer or tautomer, or a pharmaceutically acceptable salt, solvate, chelate, non-covalent complex or prodrug thereof, wherein R.sub.2 is selected from —CH.sub.3, —CH.sub.2CH.sub.3, —CH(CH.sub.3).sub.2, —CH.sub.2CF.sub.3, cyclopropyl, cyclobutyl, phenyl and pyridyl; wherein —CH.sub.3, —CH.sub.2CH.sub.3, —CH(CH.sub.3).sub.2, —CH.sub.2CF.sub.3, cyclopropyl, cyclobutyl, phenyl and pyridyl are each optionally substituted with halogen.
23. The compound of claim 1, or a stereoisomer, geometric isomer or tautomer, or a pharmaceutically acceptable salt, solvate, chelate, non-covalent complex or prodrug thereof, wherein R.sub.2 is selected from —CH.sub.3, —CH(CH.sub.3).sub.2, cyclopropyl, phenyl and phenyl substituted with halogen.
24. The compound of claim 1, or a stereoisomer, geometric isomer or tautomer, or a pharmaceutically acceptable salt, solvate, chelate, non-covalent complex or prodrug thereof, wherein the compound is selected from: 1) (2S)-1-(2-(5-isopropyl-3-methyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4] oxazepin-9-yl)pyrrolidine-2-carboxamide; 2) (2S)-1-(2-(3-cyclopropyl-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4] oxazepin-9-yl)pyrrolidine-2-carboxamide; 3) (2S)-1-(2-(5-isopropyl-2,4-dioxo-3-(2,2,2-trifluoroethyl)imidazolidine-1-yl)-5,6-dihydrobenzo[f] imidazo [1,2-d] [1,4]oxazepin-9-yl)pyrrolidine-2-carboxamide; 4) (2S)-1-(2-(3-(4-fluorophenyl)-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo [1,2-d] [1,4] oxazepin-9-yl)pyrrolidine-2-carboxamide; 5) (2S)-1-(2-(5-cyclopropyl-3-methyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4] oxazepin-9-yl)pyrrolidine-2-carboxamide; 6) (2S)-1-(2-(3-cyclobutyl-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4] oxazepin-9-yl)pyrrolidine-2-carboxamide; 7) (2S)-1-(2-(3-ethyl-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4] oxazepin-9-yl)pyrrolidine-2-carboxamide; 8) (2S)-1-(2-(5-difluoromethyl-3-methyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4] oxazepin-9-yl)pyrrolidine-2-carboxamide; 9) (2S)-1-(2-(5-isopropyl-2,4-dioxo-3-phenylimidazolidine-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4] oxazepin-9-yl)pyrrolidine-2-carboxamide; 10) (S)-1-(2-((S)-5-isopropyl-2,4-dioxo-3-phenylimidazolidine-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4]oxazepin-9-yl)pyrrolidine-2-carboxamide; 11) (S)-1-(2-((R)-5-isopropyl-2,4-dioxo-3-phenylimidazolidine-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4]oxazepin-9-yl)pyrrolidine-2-carboxamide; 12) (2S)-1-(2-(5-isopropyl-2,4-dioxo-3-(4-(trifluoromethyl)phenyl)imidazolidine-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4] oxazepin-9-yl)pyrrolidine-2-carboxamide; 13) (2S)-1-(2-(5-isopropyl-2,4-dioxo-3-propylimidazolidine-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4] oxazepin-9-yl)pyrrolidine-2-carboxamide; 14) (2S)-1-(2-(3-cyclopropyl-5-isopropyl)-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4] oxazepin-9-yl)-2-methylpyrrolidine-2-carboxamide; 15) (2S)-1-(2-(3-cyclopropyl-5-isopropyl)-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4] thiazepin-9-yl)-4,4-difluoropyrrolidine-2-carboxamide; 16) (2S)-1-(2-(5-isopropyl-3-(1-methyl-1H-1,2,4-triazol-5-yl)-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4]oxazepin-9-yl)pyrrolidine-2-carboxamide; 17) (2S,4S)-4-amino-1-(2-(3-cyclopropyl-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f] imidazo [1,2-d] [1,4]oxazepin-9-yl)pyrrolidine-2-carboxamide; 18) (2S)-1-(2-(3-(2-fluorophenyl)-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4]oxazepin-9-yl)pyrrolidine-2-carboxamide; 19) (2S)-1-(2-(3-(4-chlorophenyl)-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4] oxazepin-9-yl)pyrrolidine-2-carboxamide; 20) (2S)-4-(tertbutoxy)-1-(2-(3-cyclopropyl-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f] imidazo [1,2]-d] [1,4]oxazepin-9-yl)pyrrolidine-2-carboxamide; 21) (2S)-1-(2-(3-cyclopropyl-5-isopropyl)-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4]oxazepin-9-yl)-4,4-dimethylpyrrolidine-2-carboxamide; 22) (2S)-1-(2-(3-(2,2-difluoroethyl)-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4]oxazepin-9-yl)pyrrolidine-2-carboxamide; 23) (2S)-1-(2-(3,5-diisopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4] oxazepin-9-yl)pyrrolidine-2-carboxamide; 24) (2S)-1-(2-(3-cyclopropyl-5-isopropyl)-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4]oxazepin-9-yl)-4-phenylpyrrolidine-2-carboxamide; 25) (2S)-1-(2-(3-(3-chloro5-cyanophenyl)-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4] thiazepin-9-yl)pyrrolidine-2-carboxamide; 26) (2S)-1-(2-(3,5-diisopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4]thiazepin-9-yl)pyrrolidine-2-carboxamide; 27) (2S)-1-(2-(3-azetidin-3-yl)-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4] oxazepin-9-yl)pyrrolidine-2-carboxamide; 28) (2S)-1-(2-(3-methyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4] oxazepin-9-yl)pyrrolidine-2-carboxamide; 29) (2S)-1-(2-(5-tertbutyl-3-methyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4] oxazepin-9-yl)pyrrolidine-2-carboxamide; 30) (S)-1-(2-((R)-3-(4-fluorophenyl)-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2]-d] [1,4]oxazepin-9-yl)pyrrolidine-2-carboxamide; 31) (S)-1-(2-((S)-3-(4-fluorophenyl)-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2]-d] [1,4]oxazepin-9-yl)pyrrolidine-2-carboxamide; 32) (2S)-1-(2-((5R)-3-cyclopropyl-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4]oxazepin-9-yl)pyrrolidine-2-carboxamide; 33) (2S)-1-(2-((5S)-3-cyclopropyl-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4]oxazepin-9-yl)pyrrolidine-2-carboxamide; 34) (2S)-1-(2-(3-(2-hydroxyethyl)-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4]oxazepin-9-yl)pyrrolidine-2-carboxamide; 35) (2S)-1-(2-(3-(2-fluoroethyl)-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4] oxazepin-9-yl)pyrrolidine-2-carboxamide; 36) (2S)-1-(2-(3-(2-(dimethylamino)ethyl)-5-)isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4]oxazepin-9-yl)pyrrolidine-2-carboxamide; 37) (2S)-1-(2-(3-cyclopropyl-5-isopropyl)-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4] thiazepin-9-yl)pyrrolidine-2-carboxamide; 38) (2S)-1-(2-(3-(6-fluoropyridin-3-yl)-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4]oxazepin-9-yl)pyrrolidine-2-carboxamide; 39) (2S)-1-(2-(3-(5-fluoropyridin-3-yl)-5-isopropyl-2,4-dioxoimidazolidin1-yl)-5,6-dihydrobenzo[f]imidazo[1,2]-d] [1,4]oxazepin-9-yl)pyrrolidine-2-carboxamide; 40) (2S)-1-(2-(3-(3-cyano-5-fluorophenyl)-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4]oxazepin-9-yl)-4-fluorinepyrrolidine-2-carboxamide; 41) (2S)-1-(2-(3-(3-chloropyrimidin-2-yl)-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4]oxazepin-9-yl)pyrrolidine-2-carboxamide; 42) (2S)-1-(2-(3-cyclopentyl-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4] oxazepin-9-yl)pyrrolidine-2-carboxamide; 43) (2S)-1-(2-(3-(cyanomethyl)-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4]oxazepin-9-yl)pyrrolidine-2-carboxamide; 44) (2S)-1-(2-(5-isopropyl-3-methyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4] thiazepin-9-yl)pyrrolidine-2-carboxamide; 45) (2S)-1-(2-(3-(2,2-difluoroethyl)-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4]thiazepin-9-yl)pyrrolidine-2-carboxamide; 46) (2S)-1-(2-(3-cyclopropyl-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-3-vinyl-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4]oxazepin-9-yl)pyrrolidine-2-carboxamide; 47) (2S)-1-(3-chloro-2-(3-cyclopropyl-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4]oxazepin-9-yl)pyrrolidine-2-carboxamide; 48) (2S)-1-(2-(3-cyclopropyl-5-(difluoromethyl)-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4]thiazepin-9-yl)pyrrolidine-2-carboxamide; 49) (2S,3S)-1-(2-(3-cyclopropyl-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4] oxazepin-9-yl)-3-hydroxypyrrolidine-2-carboxamide; 50) (2S)-1-(2-(3-cyclopropyl-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-3-methyl-5,6-dihydrobenzo[f]imidazo[1,2-d] [1,4]oxazepin-9-yl)-4-oxopyrrolidine-2-carboxamide; 51) (2S)-4-cyclohexyl-1-(2-(3-cyclopropyl-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo [1,2-d] [1,4]oxazepin-9-yl)pyrrolidine-2-carboxamide; and 52) (2S)-1-(2-(3-cyclopropyl-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f] imidazo [1,2-d] [1,4] thiazepin-9-yl)-5-oxopyrrolidine-2-carboxamide.
25. A pharmaceutical composition, which comprises a) a therapeutically effective amount of at least one compound of claim 1 or a stereoisomer, geometric isomer or tautomer, or a pharmaceutically acceptable salt, solvate, chelate, non-covalent complex or prodrug thereof and b) at least one pharmaceutical acceptable excipient, wherein the weight ratio of a) and b) is in the range from 0.0001 to 10.
26. (canceled)
27. (canceled)
28. (canceled)
29. (canceled)
30. (canceled)
31. (canceled)
32. (canceled)
33. (canceled)
34. (canceled)
35. A method for the treatment or prevention of a disease mediated by PI3K, wherein the method comprising administering to said subject in need thereof a therapeutically amount of the compound of claim 1 or a pharmaceutical composition comprising a therapeutically effective amount of at least one compound of claim 1 or a stereoisomer, geometric isomer or tautomer, or a pharmaceutically acceptable salt, solvate, chelate, non-covalent complex or prodrug thereof and at least one pharmaceutical acceptable excipient, wherein the disease is cancer.
36. The method of claim 35, wherein the PI3K include PI3Kα, PI3Kβ, PI3Kδ and/or PI3Kγ.
37. The method of claim 35, wherein PI3K is PI3Kα.
38. (canceled)
39. The method of claim 38, wherein the cancer is sarcoma, prostate cancer, breast cancer, pancreatic cancer, gastrointestinal cancer, colorectal cancer, thyroid cancer, liver cancer, adrenal cancer, glioma, endometrial cancer, melanoma, kidney cancer, bladder cancer, uterine cancer, vagina cancer, ovarian cancer, multiple myeloma, esophageal cancer, leukemia, brain cancer, oral and pharyngeal cancer, laryngeal cancer, lymphoma, basal cell carcinoma, polycythemia vera, or essential thrombocythemia.
40. A method for the treatment of cancer, comprising administering to a subject in need thereof a therapeutically effective amount of the compound of claim 1 or a pharmaceutical composition comprising a therapeutically effective amount of at least one compound of claim 1 or a stereoisomer, geometric isomer or tautomer, or a pharmaceutically acceptable salt, solvate, chelate, non-covalent complex or prodrug thereof and at least one pharmaceutical acceptable excipient, wherein the cancer is sarcoma, prostate cancer, breast cancer, pancreatic cancer, gastrointestinal cancer, colorectal cancer, thyroid cancer, liver cancer, adrenal cancer, glioma, endometrial cancer, melanoma, kidney cancer, bladder cancer, uterine cancer, vagina cancer, ovarian cancer, multiple myeloma, esophageal cancer, leukemia, brain cancer, oral and pharyngeal cancer, laryngeal cancer, lymphoma, basal cell carcinoma, polycythemia vera, or essential thrombocythemia.
41. The method of claim 40, wherein the subject is human.
Description
EXAMPLES
[0168] The present invention will use the following examples to illustrate the preparation of the compound of Formula (I), but there's no limit to the present invention.
[0169] The following examples are offered for illustrative purposes, so that those skilled in the art can understand the present invention, but are not intended to limit the invention in any manner. Unless otherwise indicated, the technical solutions or the methods of the examples in the present inventions are conventional. Unless otherwise indicated, all parts and percentages are by weight, all temperatures are in degrees Celsius of the present invention.
[0170] The following abbreviations have been used in the examples: [0171] DCM: Dichloromethane [0172] DMF: N, N-dimethylcarboxamide [0173] DIEA: N, N-diisopropylethylamine [0174] PE: Petroleum ether [0175] EA: Ethyl acetate [0176] NIS: N-Iodosuccinimide [0177] LCMS or LC-MS: Liquid chromatography-mass spectrometry [0178] THF: Tetrahydrofuran [0179] DMSO: Dimethyl sulfoxide [0180] Et.sub.3N or TEA: Triethylamine [0181] HATU: 2-(7-Azobenzotriazole)-N, N, N′, N′-tetramethylurea hexafluorophosphate [0182] Hex: n-hexane [0183] h, hr or hrs: hour [0184] LiHMDS: Lithium bis (trimethylsilyl) amide [0185] [PdCl.sub.2 (dppf)]CH.sub.2Cl.sub.2: [1,1′-bis (diphenylphosphine) ferrocene] dichloropalladium (II), complex with dichloromethane [0186] Boc: Tert-Butoxycarbonyl [0187] min: minute [0188] rt or RT: room temperature.
General Route
[0189] Compound of Formula (VI) was prepared by the following route:
##STR00011## ##STR00012## ##STR00013##
Preparation of Intermediate M-5
Step 1: Synthesis of Compound M-2
[0190] 4-bromo-2-hydroxybenzaldehyde (40 g) and MeOH (400 mL) were added into a 1000 mL single-neck flask, ammonia (136.50 g) was added with stirring in an ice bath, and then reacted in oil bath at 35° C. When LC-MS analysis indicated completion of the reaction, the reaction mixture was concentrated and diluted with water, extracted with EA for four times, and then the organic layers were combined, dried and concentrated. The residue was purified by column chromatography (PE:EA=3:1), and concentrated to give compound M-2 (34.55 g).
[0191] LC-MS [M+H.sup.+]: 239.
Step 2: Synthesis of Compound M-3
[0192] Compound M-2 (34.55 g), Cs.sub.2CO.sub.3 (133 g) and DMF (300 mL) were added into a 1000 mL single-neck flask, stirred at room temperature for 20 mins and then 1,2-dibromoethane (54.30 g) was added dropwised, after the addition was finished, the mixture was put into a 80° C. oil bath for a reflux reaction. When LC-MS analysis indicated the completion of the reaction, the reaction mixture was concentrated and diluted with water, extracted with EA for 3 times, and then the organic layers were combined, dried, concentrated under reduced pressure. The residue was purified by column chromatography (PE:EA=70:30), concentrated to give compound M-3 (21.37 g).
[0193] LC-MS [M+H.sup.+]: 265.
[0194] .sup.1H NMR (500 MHz, Chloroform-d) δ 8.38 (d, J=8.5 Hz, 1H), 7.30-7.15 (m, 3H), 6.99 (s, 1H), 4.47-4.43 (m, 2H), 4.41-4.35 (m, 2H).
Step 3: Synthesis of Compound M-4
[0195] Compound M-3 (21.37 g) and DMF (100 mL) were added into a 1000 mL single-neck flask, stirred to dissolve, and NIS (50.78 g) dissolved in DMF (100 mL) was added dropwised, after the addition was finished, the reaction was put into a 60° C. oil bath and reacted overnight. When LC-MS analysis indicated the completion of the reaction, water was added into the reaction mixture, the resulting precipitate was collected by vacuum filtration, and the filter cake was dried to give compound M-4 (35 g).
[0196] LC-MS [M+H.sup.+]: 517.
[0197] .sup.1H NMR (500 MHz, Chloroform-d) δ 8.30 (d, J=8.6 Hz, 1H), 7.25-7.16 (m, 2H), 4.46-4.41 (m, 2H), 4.36-4.32 (m, 2H).
Step 4: Synthesis of Compound M-5
[0198] Compound M-4 (35 g) and THF (150 mL) were added into a 500 mL three-neck flask, 100 mL ethylmagnesium bromide (1 M THF solution) was added dropwised at −40° C. under nitrogen protection, after the addition was finished, the reaction was stirred at −40° C. When LC-MS analysis indicated the completion of the reaction, the reaction mixture was quenched with saturated ammonium chloride solution in an ice bath, extracted with EA for 3 times, and then the organic layers were combined, dried, concentrated to provide a residue, the residue was slurried in methyltertbutyl ether, the suspension was filtered off with suction and the filter cake was dried to give compound M-5 (19.8 g).
[0199] LC-MS [M+H.sup.+]: 391.
[0200] .sup.1H NMR (500 MHz, DMSO-d6) δ 8.22 (d, J=8.6 Hz, 1H), 7.55 (s, 1H), 7.31-7.23 (m, 2H), 4.47-4.39 (m, 4H).
Example 1 Synthesis of (2S)-1-(2-(5-isopropyl-3-methyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f] imidazo [1,2-d][1,4]oxazepin-9-yl)pyrrolidine-2-carboxamide (Compound 1)
[0201] ##STR00014## ##STR00015##
Step 1: Synthesis of Compound 1-3
[0202] Compound 1-1 (9 g), compound 1-2 (16.8 g), DCM (200 mL) and HATU (31.25 g) were added into a 500 mL three-neck flask, TEA (50.21 g) was added dropwised in an ice bath, after the addition was finished, reacted at room temperature. When LC-MS analysis indicated the completion of the reaction, quenched the reaction with the addition of water, the organic layers were separated, rinsed with water, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromatography (PE:EA=50:50) to give compound 1-3 (9.2 g).
[0203] LC-MS [M-Boc+H].sup.+: 131.
Step 2: Synthesis of Compound 1-4
[0204] HCl/Dioxane (10 mL, 4.0 mol/L) was added to the solution of compound 1-3 (1.3 g) in dichloromethane (10 mL), stirred at room temperature for 3 hours. After the reaction was completed, the reaction mixture was concentrated to give compound 1-4 (0.9 g), which was used in the next reaction directly without further purification.
[0205] LC-MS [M+H].sup.+: 131.
Step 3: Synthesis of Compound 1-5
[0206] Compound 1-4 (900 mg) and TEA (4.19 g) were dissolved in THF (50 mL). Triphosgene (802 mg) in THF (10 mL) was added slowly to the reaction in an ice bath, the reaction mixture was naturally heated to room temperature and stirred for 12 h. When LC-MS analysis indicated the completion of the reaction, the reaction was quenched with the addition of water (5 mL), the reaction mixture was concentrated, extracted and separated with dichloromethane/water. The organic layers were dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromatography (PE:EA=1:1) to give compound 1-5 (732 mg).
[0207] LC-MS [M+H].sup.+: 157.
Step 4: Synthesis of Compound 1-6
[0208] Compound 1-5 (177 mg), compound M-5 (443 mg), CuI (64.75 mg), trans-N,N′-dimethylcyclohexane-1,2-diamine (48.36 mg) and K.sub.3PO.sub.4 (721.68 mg) were dissolved in DMF (5 mL), the atmosphere was replaced with nitrogen for three times, the reaction mixture was heated to 110° C. and reacted for 2 h. When LC-MS analysis indicated the completion of the reaction, the reaction mixture was diluted with EA (100 mL), washed with water. The organic layers were combined, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromatography (PE:EA=70:30) to give compound 1-6 (350 mg).
[0209] LC-MS [M+H].sup.+: 419.
Step 5: Synthesis of Compound 1-7
[0210] Compound 1-6 (350 mg), L-proline (290 mg), K.sub.3PO.sub.4 (848 mg), CuI (79.8 mg) and DMSO (5 mL) were added to a 100 mL single-neck flask, the reaction was stirred at 120° C. for 3 h under nitrogen protection. When LC-MS analysis indicated the completion of the reaction, the reaction mixture was filtered, the filter cake was washed with DMSO (3 mL), the filtrate was used in the next step directly.
[0211] LC-MS [M+H].sup.+: 454.
Step 6: Synthesis of Compound 1
[0212] DCM (12 mL), NH.sub.4Cl (445 mg) and DIEA (2.17 g) were added to the filtrate in Step 5 under nitrogen protection, cooled to 0° C., HATU (1.27 g) was added to the reaction system in an ice bath, stirred at 0° C. for 20 mins. When LC-MS analysis indicated the completion of the reaction, the reaction mixture was diluted with DCM, washed with water, the organic layers was dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The resulting crude product was purified by column chromatography (PE:EA=100:0-0:100) to give compound 1 (300 mg).
[0213] LC-MS [M+H].sup.+: 453.
[0214] .sup.1H NMR (500 MHz, DMSO-d6) δ 8.04 (d, J=8.9 Hz, 1H), 7.41 (s, 1H), 7.24 (s, 1H), 7.05 (s, 1H), 6.32 (d, J=8.7 Hz, 1H), 6.03 (s, 1H), 4.62-4.51 (m, 1H), 4.45-4.30 (m, 4H), 3.94 (d, J=8.7 Hz, 1H), 3.55 (t, J=7.3 Hz, 1H), 3.23 (q, J=7.2 Hz, 1H), 2.92 (s, 3H), 2.68 (m, 1H), 2.26-2.12 (m, 1H), 1.96 (m, 3H), 1.16 (d, J=7.0 Hz, 3H), 0.74 (d, J=6.8 Hz, 3H).
Example 2 Synthesis of (2S)-1-(2-(3-cyclopropyl-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f]imidazo[1,2-d][1,4]oxazepin-9-yl)pyrrolidine-2-carboxamide (Compound 2)
[0215] ##STR00016## ##STR00017##
Step 1: Synthesis of Compound 2-2
[0216] Compound 1-1 (5.0 g), DCM (100 mL) and HATU (9.55 g) were added to a 250 mL single-neck flask under nitrogen protection, TEA (8.15 g) and compound 2-1 (1.45 g) were added to the reaction in an ice bath, reacted at room temperature for 2 h. When LC-MS analysis indicated the completion of the reaction, the reaction mixture was concentrated, diluted with EA, washed with water, dried over anhydrous Na.sub.2SO.sub.4 and concentrated to give compound 2-2 (5.0 g), which was used in the next step directly.
[0217] LC-MS [M-Boc+H].sup.+: 157.
[0218] .sup.1H NMR (500 MHz, DMSO-d6) δ 7.91 (s, 1H), 6.54 (d, J=9.0 Hz, 1H), 3.69-3.58 (m, 1H), 2.65-2.55 (m, 1H), 1.88-1.80 (m, 1H), 1.37 (s, 9H), 0.79 (d, J=6.0 Hz, 6H), 0.62-0.58 (m, 2H), 0.42-0.30 (m, 2H).
Step 2: Synthesis of Compound 2-3
[0219] Compound 2-2 (5 g), DCM (20 mL) and HCl/Dioxane (20 mL, 4.0 mol/L) were added to a 250 mL single-neck flask, and reacted at room temperature for 2 h. When LC-MS analysis indicated the completion of the reaction, the reaction mixture was concentrated to give compound 2-3 (3.75 g).
[0220] LC-MS [M+H].sup.+: 157.
Step 3: Synthesis of Compound 2-4
[0221] Compound 2-3 (2.0 g), DCM (50 mL) and TEA (4.20 g) were added to a 250 mL single-neck flask, under nitrogen protection, triphosgene (1.54 g) dissolved in DCM (50 mL) was added to the reaction system in an ice bath, the mixture was stirred in an ice bath for 2 h. When LC-MS analysis indicated the completion of the reaction, the reaction was quenched with ice water in an ice bath, the reaction mixture was concentrated, extracted with EA, dried over anhydrous Na.sub.2SO.sub.4, concentrated to give a crude product. The crude product was purified by column chromatography (PE:EA=100:0-50:50), to give compound 2-4 (810 mg).
[0222] LC-MS [M+H].sup.+: 183/185.
[0223] .sup.1H NMR (500 MHz, Chloroform-d) δ 6.28 (s, 1H), 3.85 (d, J=3.6 Hz, 1H), 2.66-2.48 (m, J=3.7 Hz, 1H), 2.25-2.15 (m, 1H), 1.03 (d, J=6.5 Hz, 3H), 0.98-0.91 (m, 4H), 0.88 (d, J=7.0 Hz, 3H).
Step 4: Synthesis of Compound 2-5
[0224] Compound 2-4 (431 mg), compound M-5 (700 mg), DMF (10 mL), CuI (102 mg), trans-N,N′-dimethylcyclohexane-1,2-diamine (77 mg) and K.sub.3PO.sub.4 (760 mg) were added to a 50 mL single-neck flask, under nitrogen protection, the reaction mixture was heated to 120° C. and stirred for 2 h. When LC-MS analysis indicated the completion of the reaction, the reaction mixture was diluted with EA, washed with water, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The crude product was purified by column chromatography (PE:EA=100:0-60:40) to give compound 2-5 (642 mg).
[0225] LC-MS [M+H].sup.+: 445/447.
[0226] .sup.1H NMR (500 MHz, Chloroform-d) δ 8.23 (d, J=8.5 Hz, 1H), 7.27 (s, 1H), 7.22 (d, J=8.7 Hz, 1H), 7.20 (s, 1H), 4.63-4.59 (m, 1H), 4.51-4.38 (m, 2H), 4.36 (t, J=4.3 Hz, 2H), 2.79-2.71 (m, 1H), 2.68-2.60 (m, 1H), 1.24 (d, J=7.1 Hz, 3H), 1.02-0.94 (m, 4H), 0.81 (d, J=6.9 Hz, 3H).
Step 5: Synthesis of Compound 2-6
[0227] Compound 2-5 (642 mg), compound M-12 (415 mg), K.sub.3PO.sub.4 (919 mg) and DMSO (10 mL) were added to 30 mL microwave vial, purged with nitrogen, CuI (83 mg) was added, the reaction mixture was heated to 120° C. and reacted in a microwave reactor for 1 h. When LC-MS analysis indicated the completion of the reaction, the reaction mixture was used in the next step directly.
[0228] LC-MS [M+H].sup.+: 480.
Step 6: Synthesis of Compound 2
[0229] The reaction mixture in Step 5 was added to a 50 mL single-neck flask, the atmosphere was replaced with nitrogen, DCM (10 mL), NH.sub.4Cl (462 mg) and TEA (1.46 g) were added, the reaction system was cooled to 0° C., HATU (3.28 g) was added in an ice bath, the reaction mixture was stirred at 0° C. for 1 h. When LC-MS analysis indicated the completion of the reaction, the reaction mixture was diluted with DCM, washed with water, the organic layers were dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The crude product was purified by Pre-HPLC (C18 column, H.sub.2O:MeOH=95:5-50:50), to give compound 2 (85 mg).
[0230] LC-MS [M+H].sup.+: 479.
[0231] .sup.1H NMR (500 MHz, DMSO-d6) δ 8.03 (d, J=9.0 Hz, 1H), 7.40 (s, 1H), 7.24 (s, 1H), 7.05 (s, 1H), 6.32 (d, J=8.9 Hz, 1H), 6.03 (d, J=2.6 Hz, 1H), 4.48 (s, 1H), 4.41-4.29 (m, 4H), 3.94 (d, J=8.8 Hz, 1H), 3.55 (t, J=8.1 Hz, 1H), 3.25-3.19 (m, 1H), 2.74-2.55 (m, 2H), 2.25-2.14 (m, 1H), 2.07-1.85 (m, 3H), 1.13 (d, J=7.0 Hz, 3H), 0.88 (d, J=7.2 Hz, 2H), 0.81 (d, J=3.9 Hz, 2H), 0.71 (d, J=6.8 Hz, 3H).
Step 7: Preparation of Compound 32 and Compound 33
[0232] In the present examples, compound 32 (front peak) and compound 33 (back peak) were prepared by the separation of the compound 2 through the the following chiral column.
[0233] Chiral column HPLC conditions:
TABLE-US-00001 Column CHIRALPAK IA Column Specifications 3 cm × 25 cm, 5 um Injection volume 4.0 mL Mobile phase (Hex:DCM = 3:1):EtOH = 50:50 (v/v) Flow rates 35 mL/min Wavelength UV 220 nm Temperature 25° C. Sample solution EtOH:DCM = 3:1(12.3 mg/mL) Pre-HPLC Prep-HPLC-flash
[0234] Compound 32: LC-MS [M+H].sup.+: 479.
[0235] .sup.1H NMR (500 MHz, DMSO-d6) δ 8.03 (d, J=9.0 Hz, 1H), 7.40 (s, 1H), 7.24 (s, 1H), 7.05 (s, 1H), 6.32 (d, J=8.9 Hz, 1H), 6.03 (d, J=2.6 Hz, 1H), 4.48 (s, 1H), 4.41-4.29 (m, 4H), 3.94 (d, J=8.8 Hz, 1H), 3.55 (t, J=8.1 Hz, 1H), 3.25-3.19 (m, 1H), 2.74-2.55 (m, 2H), 2.25-2.14 (m, 1H), 2.07-1.85 (m, 3H), 1.13 (d, J=7.0 Hz, 3H), 0.88 (d, J=7.2 Hz, 2H), 0.81 (d, J=3.9 Hz, 2H), 0.71 (d, J=6.8 Hz, 3H).
[0236] Compound 33: LC-MS [M+H].sup.+: 479.
[0237] .sup.1H NMR (500 MHz, DMSO-d6) δ 8.03 (d, J=9.0 Hz, 1H), 7.40 (s, 1H), 7.24 (s, 1H), 7.05 (s, 1H), 6.32 (d, J=8.9 Hz, 1H), 6.03 (d, J=2.6 Hz, 1H), 4.48 (s, 1H), 4.41-4.29 (m, 4H), 3.94 (d, J=8.8 Hz, 1H), 3.55 (t, J=8.1 Hz, 1H), 3.25-3.19 (m, 1H), 2.74-2.55 (m, 2H), 2.25-2.14 (m, 1H), 2.07-1.85 (m, 3H), 1.13 (d, J=7.0 Hz, 3H), 0.88 (d, J=7.2 Hz, 2H), 0.81 (d, J=3.9 Hz, 2H), 0.71 (d, J=6.8 Hz, 3H).
Example 3 Synthesis of (2S)-1-(2-(5-isopropyl-2,4-dioxo-3-(2,2,2-trifluoroethyl)imidazolidine-1-yl)-5,6-dihydrobenzo[f]imdazo[1,2-d] [1,4]oxazepin-9-yl)pyrrolidine-2-carboxamide (Compound 3)
[0238] ##STR00018## ##STR00019##
Step 1: Synthesis of Compound 3-2
[0239] To a solution of compound 3-0 (2 g) and compound 3-1 (583 mg) in DCM (100 ml), HATU (2.67 g) and DIEA (2.28 g) were added successively, the mixture was stirred at room temperature for 12 h. Water was added, and the reaction mixture was separated, concentrated to give a residue. EA (25 mL) was added to the residue, and then the residue was washed with water and saturated brine. The organic layers were dried over anhydrous Na.sub.2SO.sub.4, concentrated, to give 1.8 g product.
[0240] LC-MS [M+H].sup.+: 421.
Step 2: Synthesis of Compound 3-3
[0241] Diethylamine (10 mL) was added to the solution of compound 3-2 (1.8 g) dissolved in DCM (10 ml), stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure to give a residue, and the residue was purified by column chromatography to give compound 3-3 (0.70 g).
[0242] LC-MS [M+H].sup.+: 199.
Step 3: Synthesis of Compound 3-4
[0243] Compound 3-3 (200 mg), CH.sub.3CN (10 mL), NaHCO.sub.3 (254 mg) were added to a 50 mL three-neck flask, the atmosphere was replaced with nitrogen for 3 times, p-Nitrophenyl chloroformate (203 mg) was added and the mixture was stirred at room temperature for 2 h. Water (6 mL) was added to the reaction system, and stirred at room temperature for 3 h. When the reaction was completed, the reaction mixture was concentrated under reduced pressure to provide the residue, and the residue was diluted with EA, the organic layers were washed with water, potassium carbonate aqueous solution and saturated brine successively, and dried over anhydrous Na.sub.2SO.sub.4, concentrated to provide the crude product. The crude product was purified by the column chromatography (PE:EA=60:40) to give compound 3-4 (195 mg).
[0244] LC-MS [M+H].sup.+: 225.
Step 4: Synthesis of Compound 3-5
[0245] Compound 3-4 (138 mg), compound M-5 (200 mg), CuI (39 mg), trans-N,N′-dimethylcyclohexane-1,2-diamine (29 mg), K.sub.3PO.sub.4 (326 mg) were dissolved in DMF (5 mL), the atmosphere was replaced with nitrogen for 3 times, the reaction mixture was heated to 110° C. for 2 h. When the reaction was completed, the reaction mixture was diluted with EA, washed with water for one time and saturated brine for three times, and the organic layers were dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromatography (PE:EA=100:0-70:30) to give compound 3-5 (180 mg).
[0246] LC-MS [M+H.sup.+]: 487/489.
Step 5: Synthesis of Compound 3-6
[0247] Compound 3-5 (117 mg), compound M-12 (138 mg), K.sub.3PO.sub.4 (356 mg), CuI (46 mg) and DMSO (3 mL) were added to a 50 mL single-neck flask, the atmosphere was replaced with nitrogen for 3 times, the reaction mixture was heated to 120° C. and stirred for 3 h. When the reaction was completed, the reaction mixture was filtered, the filter cake was washed with DMSO (3 mL), the filtrate was used in the next step directly.
[0248] LC-MS [M+H].sup.+: 522.
Step 6: Synthesis of Compound 3
[0249] DCM (6 mL), NH.sub.4Cl (127 mg), DIEA (613 mg) were added to the filtrate in Step 5 under nitrogen protection, the reaction mixture was cooled to 0° C., HATU (447 mg) was added in portions in an ice bath, stirred at 0° C. for 20 mins. When the reaction was completed, the reaction mixture was diluted with dichloromethane, washed with water and saturated brine, the organic layers were dried over anhydrous Na.sub.2SO.sub.4, concentrated under reduced pressure to provide crude product. The crude product was purified by Pre-HPLC to give compound 3 (85 mg).
[0250] LC-MS [M+H].sup.+: 521.
[0251] .sup.1H NMR (500 MHz, DMSO-d6) δ 8.05 (d, J=8.5 Hz, 1H), 7.41 (s, 1H), 7.29 (s, 1H), 7.06 (s, 1H), 6.33 (d, J=9 Hz, 1H), 6.04 (s, 1H), 4.71-4.70 (m, 1H), 4.38-4.24 (m, 6H), 3.95 (d, J=8.5 Hz, 1H), 3.55 (t, J=7.3 Hz, 1H), 3.28-3.20 (m, 1H), 2.70 (brs, 1H), 2.25-2.15 (m, 1H), 2.00-1.94 (m, 3H), 1.17 (d, J=7.0 Hz, 3H), 0.75 (d, J=7 Hz, 3H).
Example 4 Synthesis of (2S)-1-(2-(3-(4-fluorophenyl)-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f] imidazo [1,2-d][1,4]oxazepin-9-yl)pyrrolidine-2-carboxamide (Compound 4)
[0252] ##STR00020## ##STR00021##
Step 1: Synthesis of Compound 4-1
[0253] HATU (3.39 g) and DIEA (2.68 g) were added to the solution of compound 1-1 (1.5 g) and p-Fluoroaniline (921 mg) in DCM (50 ml). The reaction mixture was stirred at room temperature for 12 h. When the reaction was completed, the reaction mixture was diluted with water, separated, the organic layers were washed with 1 N HCl, NaHCO.sub.3 saturated aqueous solution and saturated brine successively, dried over anhydrous Na.sub.2SO.sub.4, concentrated to provide crude product. The crude product was purified by column chromatography (EA/PE=0-30%) to give compound 4-1 (2.1 g).
[0254] LC-MS [M-Boc+H].sup.+: 211.
Step 2: Synthesis of Compound 4-2
[0255] Compound 4-1 (2.1 g) was added to the solution of HCl/dioxane (10 mL, 4 M), stirred at room temperature for 12 h. The reaction mixture was concentrated under reduced pressure to give compound 4-2 (1.64 g), which was used in the next step directly without the further purification.
[0256] LC-MS [M+H].sup.+: 211.
Step 3: Synthesis of Compound 4-3
[0257] Compound 4-2 (500 mg), CH.sub.3CN (30 mL), NaHCO.sub.3 (681 mg) were added to a 50 mL three-neck flask, the atmosphere was replaced with nitrogen for 3 times, p-Nitrophenyl chloroformate (449 mg) was added, stirred at room temperature for 2 h. Water (18 mL) was added to the reaction mixture, and stirred at room temperature for 3 h.
[0258] When the reaction was completed, the reaction mixture was concentrated under reduced pressure to provide the residue, the residue was diluted with EA (100 mL), the organic layers were washed with water (50 mL), 5% K.sub.2CO.sub.3 aqueous solution (50 mL) and saturated brine respectively, seperated, the organic layers were dried over anhydrous Na.sub.2SO.sub.4, concentrated to provide the crude product. The crude product was purified by column chromatography (EA/PE=0-40%) to give compound 4-3 (447 mg).
[0259] LC-MS [M+H].sup.+: 237.
Step 4: Synthesis of Compound 4-4
[0260] Compound 4-3 (144 mg), compound M-5 (200 mg), CuI (39 mg), trans-N,N′-dimethylcyclohexane-1,2-diamine (29 mg), K.sub.3PO.sub.4 (326 mg) were dissolved in DMF (5 mL), the atmosphere was replaced with nitrogen for 3 times, the reaction mixture was heated to 110° C. and reacted for 2 h. When the reaction was completed, EA (100 mL) was added to dilute the reaction mixture. The organic layers were washed with water (60 mL) and saturated brine (60 mL), and dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromatography (PE:EA=100:0-70:30) to give compound 4-4 (215 mg).
[0261] LC-MS [M+H].sup.+: 499.
Step 5: Synthesis of Compound 4-5
[0262] 4-4 (120 mg), compound M-12 (138 mg), K.sub.3PO.sub.4 (356 mg), CuI (46 mg) and DMSO (3 mL) were added to a 100 mL single-neck flask, the atmosphere was replaced with nitrogen for 3 times, the reaction mixture was heated to 120° C., stirred and reacted for 3 h. When the reaction was completed, the reaction mixture was filtered, filter cake was washed with 3 mL of DMSO, the filtrate was used in the next step directly.
[0263] LC-MS [M+H.sup.+]: 534.
Step 6: Synthesis of Compound 4
[0264] DCM (12 mL), NH.sub.4Cl (445 mg) and DIEA (2.17 g) were added to the filtrate in Step 5 under nitrogen protection, the reaction mixture was cooled to 0° C., HATU (1.27 g) was added in portions in an ice bath, and stirred at 0° C. for 20 mins. When the reaction was completed, the reaction mixture was diluted with DCM (50 mL), the organic layers were washed with water (40 mL) and saturated brine (40 mL), dried over anhydrous Na.sub.2SO.sub.4, concentrated to provide the crude product. The crude product was purified by column chromatography (PE:EA=100:0-0:100) to give compound 4 (89 mg).
[0265] LC-MS [M+H].sup.+: 533.
[0266] .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 8.08 (d, J=8.9 Hz, 1H), 7.45-7.41 (m, 3H), 7.38-7.35 (m, 2H), 7.28 (s, 1H), 7.06 (s, 1H), 6.34 (d, J=8.9 Hz, 1H), 6.04 (s, 1H), 4.76 (s, 1H), 4.40-4.37 (m, 4H), 3.95 (d, J=8.8 Hz, 1H), 3.60-3.52 (m, 1H), 3.24 (q, J=8.5, 7.7 Hz, 1H), 2.75 (s, 1H), 2.26-2.14 (m, 1H), 2.00-1.93 (m, 3H), 1.22 (d, J=7.1 Hz, 3H), 0.88 (d, J=6.8 Hz, 3H).
[0267] Compound 30 and compound 31 were prepared following similar procedures as described in the Step 7 in Example 2.
Example 5 Synthesis of (2S)-1-(2-(5-cyclopropyl-3-methyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f] imidazo [1,2-d] [1,4] oxazepin-9-yl) pyrrolidine-2-carboxamide (Compound 5)
[0268] ##STR00022## ##STR00023##
[0269] Replacing compound 1-1 with compound 5-0, and p-fluoroaniline with methylamine hydrochloride, compound 5 was prepared following similar procedures as described in Example 4.
[0270] LC-MS [M+H].sup.+: 451.
Example 6 Synthesis of (2S)-1-(2-(3-cyclobutyl-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f] imidazo [1,2-d] [1,4] oxazepin-9-yl)pyrrolidine-2-carboxamide (Compound 6)
[0271] ##STR00024## ##STR00025##
[0272] Replacing p-fluoroaniline with cyclobutylamine, compound 6 was prepared following similar procedures as described in Example 4.
[0273] LC-MS [M+H].sup.+: 493.
[0274] .sup.1H NMR (500 MHz, DMSO-d6) δ 8.04 (d, J=9 Hz, 1H), 7.41 (s, 1H), 7.25 (s, 1H), 7.05 (s, 1H), 6.32 (d, J=9 Hz, 1H), 6.03 (s, 1H), 4.52-4.48 (m, 1H), 4.37-4.35 (m, 4H), 3.94 (d, J=8.5 Hz, 1H), 3.55 (t, J=7 Hz, 1H), 3.23 (q, J=7.2 Hz, 1H), 2.80-2.65 (m, 3H), 2.14-2.12 (m, 3H), 1.96-1.94 (m, 3H), 1.79-1.71 (m, 3H), 1.15 (d, J=7 Hz, 3H) 0.74 (d, J=6.5 Hz, 3H).
Example 7 Synthesis of (2S)-1-(2-(3-ethyl-5-isopropyl-2,4-dioxoimidazolidin-1-yl)-5,6-dihydrobenzo[f] imidazo [1,2-d] [1,4] oxazepin-9-yl) pyrrolidine-2-carboxamide (Compound 7)
[0275] ##STR00026## ##STR00027##
[0276] Replacing the compound 3-1 with compound 7-1, compound 7 was prepared following similar procedures as described in Example 3.
[0277] LC-MS [M+H].sup.+: 467.
[0278] The compounds in Table 1 were prepared in accordance with the synthetic protocols set forth in Example 1-7, using the appropriate starting materials and suitable reagents.
TABLE-US-00002 TABLE 1 EX. LCMS NO. Structure Chemical Name [M + H].sup.+ 8
[0279] All the racemic compounds in Table 1 can be synthesized using the corresponding chiral raw materials to obtain the corresponding enantiomers, or the enantiomers were separated on a chiral column basically according to the method described in Step 7 in Example 2.
Control Compound
[0280]
TABLE-US-00003 TABLE 2 EX. Compound name Structure 1 (2S)-1-(2-((4R)-4-methyl-2- oxooxazolidin-3-yl)-5,6- dihydrobenzo[f]imidazo[1,2-d] [1,4] oxazepin-9-yl)pyrrolidine-2-carboxamide
[0281] Control compound 1 (Example 102 in WO2017001658) and control compound 2 were prepared in accordance with the synthetic protocols set forth in WO2017001658, using the appropriate starting materials and intermediate and agents; The method to prepare the control compound 3 are as follows.
Preparation of Control Compound 3
[0282] ##STR00076## ##STR00077## ##STR00078##
Step 1: Synthesis of Compound D3-2
[0283] Compound D3-1 (2000 g) and oxalyl chloride (2019 g) were dissolved in dichloromethane (8 L). DMF (5 mL) was added to the reaction slowly, refluxed for 6 hours. The reaction system was concentrated to give 2245 g suspension containing yellow solid, which was used in the next step directly.
Step 2: Synthesis of Compound D3-3
[0284] To a solution of 4-methoxy-3-buten-2-one (642 g) in anhydrous THF (16 L), 6.42 L LiHMDS (1 mol/L) dissolved in THF was added dropwise at −78° C. After stirred at −78° C. for 2 h, suspension in step 1 dissolved in THF (1 L) was added to the reaction system slowly, stirred at −78° C. for 1-2 h, the reaction mixture was heated to room temperature and stirred for 16 h. The resulting reaction mixture was used in the next step directly.
[0285] LC-MS [M+H].sup.+: 239.
Step 3: Synthesis of Compound D3-4
[0286] The reaction mixture in Step 2 was quenched with the addition of water (500 mL) in an ice bath, and then trifluoroacetic acid (50 mL) was added, stirred at room temperature for 3 h. When the reaction was completed, the reaction mixture was concentrated and extracted with EA. The organic layers were washed with brine, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromatography (Hex/EA 100:1-10:1) to give brown solid. The brown solid was washed with a mixed solvent of HEX and EA (Volume ratio 50:1), the reaction mixture was filtered off with suction, the filtrate was collected and concentrated, to give compound D3-4 (395 g).
[0287] LC-MS [M+H].sup.+: 207.
[0288] .sup.1H NMR (MeOD-d6): 1.59 (s, 6H), 6.39-6.41 (m, 1H), 6.53-6.54 (m, 1H), 8.12-8.14 (m, 1H).
Step 4: Synthesis of Compound D3-5
[0289] Compound D3-4 (395 g) was added to 2 L of ammonia (30%), stirred and refluxed for 2 h. The reaction mixture was cooled, concentrated, to give 343.00 g compound D3-5, which was used in the next step directly.
[0290] LC-MS [M+H].sup.+: 206.
Step 5: Synthesis of Compound D3-6
[0291] The mixture of compound D3-5 (343 g) and phosphorus oxybromide (966.68 g) was heated to 120° C. and stirred for 1 h. The reaction mixture was poured into the ice water whilst hot. Sodium bicarbonate was added to adjust pH to neutral, the reaction mixture was extracted with EA. The organic layers were dried and concentrated. The residue was purified by column chromatography (Hex/EA=100:1) to give compound D3-6 (305 g).
[0292] LC-MS [M+H].sup.+: 269.
[0293] .sup.1H NMR (MeOD-d6): 1.59 (s, 6H), 7.52-7.54 (m, 1H), 7.78 (s, 1H), 8.41-8.42 (m, 1H).
Step 6: Synthesis of Compound D3-7
[0294] Bis(Pinacolato)diborane (710.45 mg), compound D3-6 (500 mg), potassium acetate (1.88 g), [PdCl.sub.2(dppf)]CH.sub.2Cl.sub.2 (152.32 mg) and dioxane (20 mL) were added to a single-neck flask under nitrogen protection, reacted in a 90° C. oil bath for 4 h. The reaction mixture was concentrated. The residue was purified by column chromatography (Hex/EA=3:1) to give compound D3-7 (450 mg).
[0295] LC-MS [M+H].sup.+: 316.
Step 7: Synthesis of Compound D3-8
[0296] [PdCl.sub.2(dppf)]CH.sub.2Cl.sub.2 (233.22 mg), potassium carbonate (394.70 mg), compound D3-7 (450 mg), compound M-5 (586.25 mg), dixone (3 mL) and H.sub.2O (0.5 mL) were added to a single-neck flask under nitrogen protection, refluxed in a 70° C. oil bath for 15 h. The reaction mixture was concentrated. The residue was purified by column chromatography (PE:EA=50:50) to give compound D3-8 (130 mg).
[0297] LC-MS [M+H].sup.+: 452.
Step 8: Synthesis of Compound D3-9
[0298] The compound D3-9 was prepared in accordance with the synthetic protocols set forth in step 5 in example 2.
[0299] LC-MS [M+H].sup.+: 487.
Step 9: Synthesis of Control Compound 3
[0300] Control compound 3 was prepared in accordance with the synthetic protocols set forth in step 6 in example 2.
[0301] LC-MS [M+H].sup.+: 486.
Pharmacological Experiment
[0302] The following experiments show that the preferred compounds of the present invention inhibits kinase activities of PI3Kα effectively in vitro. The anti-proliferative activities against tumor cells harboring PI3Kα mutants and the pharmacokinetic profile of the preferred compound in the present invention are better than the control compound with significant superiority.
Example A: Kinase Experiment
[0303] PI3Kα, PI3Kβ, PI3Kγ kinases undergo enzymatic reactions with their substrates ATP and PIP2:3PS, the amount of the product detected by ADP-Glo agent and luminescence method were used to reflect the enzymatic activities of PI3Kα, PI3Kβ, PI3Kγ (the final concentration of ATP is 10 μM). The above methods were used to test the inhibitory activity of certain compound of the present invention on PI3Kα, PI3Kβ and PI3Kγ kinase.
Method
[0304] Agent: Basic kinase buffer (pH 7.5); PI3Kα, PI3Kβ, PI3Kγ kinase solutions; PIP2:3PS and ATP solutions; ADP-Glo kit (containing 10 mM MgCl.sub.2).
[0305] Wherein, the buffer composition: 50 mM Hepes (pH 7.2-7.5), 3 mM MgCl.sub.2, 1 mM EGTA, 0.03% CHAPS, 100 mM NaCl, 2 mM DTT; [0306] Compound preparation: The tested compound was diluted to a specific concentration with 100% DMSO. [0307] Process: 1) PI3Kα, PI3Kβ, or PI3Kγ protein solution were dispensed to a 384-well assay plate (6008280, PerkinElmer), and centrifuged at 1000 rpm for 1 minute. 2) Tested compound, negative control DMSO or positive control BYL719 was dispensed to the above-mentioned 384-well assay plate with the kinase, centrifuged at 1000 rpm for 1 min, and incubated at 25° C. for 15 mins. 3) PIP2:3PS&ATP solution was added to the above-mentioned 384-well assay plate, centrifuged at 1000 rpm for 1 min, incubated at 25° C. for 60 mins. 4) 5 μL of ADP-Glo agent (10 mM MgCl.sub.2 contained) was transferred to the 384-well assay plate, centrifuged at 1000 rpm for 1 min, incubated at 25° C. for 40 mins. 5) 10 μL of Detection agent was transferred to the 384-well assay plate, centrifuged at 1000 rpm for 1 min, incubated at 25° C. for 40 mins. 6) RLU (Relative luminescence unit) value was measured by Envision multifunctional plate reader. The RLU value was used to characterize the degree of reaction between the enzyme and the substrate, and calculate the IC.sub.50 value. 7) Data analysis:
Compound inhibition rate(% inh)=(negative control RLU-tested compound RLU)/(negative control RLU-positive control RLU)*100%
[0308] The IC.sub.50 (half inhibitory concentration) of the compound was obtained using the following nonlinear fitting formula:
Y=minimum inhibition rate+(max inhibition rate-minimum inhibition rate)/(1+10{circumflex over ( )}((Log IC.sub.50−X)*slope)); wherein, X is the log value of the concentration of the tested compound, Y is the inhibition rate of tested compound(% inh).
[0309] The results of some examples of the present invention are shown in Table 3. Wherein, A represents the IC.sub.50 value5 nM; B represents the IC.sub.50 value is 5˜20 nM; C represents the IC.sub.50 value is 300˜600 nM; D represents the IC.sub.50 value>600 nM.
TABLE-US-00004 TABLE 3 IC.sub.50 of compound to PI3K (nM) Example α β γ Example 1 A D D Example 2 A D C Example 3 A / / Example 4 B D D Example 5 B / / Example 6 B / / Example 7 B / / Example 9 B / / Example 23 A / / Example 32 A C C Example 33 A D D Note: “/” represents not tested.
[0310] As Table 3 shows, the compound provided by the present invention have favorable PI3Kα kinase inhibitory activities, and good selectivity from PI3Kβ and PI3Kγ isoforms, which can avoid the potential adverse effects caused by multi-target inhibition.
Example B: Cell Proliferation Assay
[0311] Method: The anti-proliferative effects on human tumor cells MCF-7 and HGC27 of certain compounds of the present invention were evaluated by CellTiter Glo assay.
[0312] Detection method: MCF-7 cells were suspended in DMEM culture medium, the cell concentration of which was adjusted to 25000 cells/mL; HGC27 cells were suspended in RPMI-1640 culture medium, the cell concentration of which was adjusted to 5000 cells/mL. 100 μL of cell suspension was dispensed to a 96-well assay plate and placed in a CO.sub.2 incubator overnight. The tested compounds were dissolved in DMSO, and 3-fold derail dilution was performed to obtain 10-point concentrations. the 10-point concentrations of the tested compounds or or negative control was transferred to the well containing 100 μL of culture medium respectively, incubated at 37° C., 5% CO.sub.2, the HGC27 cells were incubated for 96 h, MCF-7 cells were incubated for 120 h. Then 100 μL of CellTiter-Glo agent was added to the above-mentioned 96-well assay plate, incubated at room temperature for 10 minutes to allow stable luminescence signal. VICTOR™ X5 instrument was used to record RLU (relative luminescence unit), and then calculate the IC.sub.50 values were calculated.
Compound inhibition rate(% inh)=100% −(tested compound RLU-blank control RLU)/(negative control RLU-blank control RLU)*100%
[0313] Negative control: DMSO;
[0314] Blank control: blank culture medium, with no compound or cell;
[0315] The IC.sub.50 (half inhibitory concentration) of the compound was obtained using the following nonlinear fitting formula:
Y=minimum inhibition rate+(max inhibition rate-minimum inhibition rate)/(1+10{circumflex over ( )}((Log IC.sub.50−X)*slope)); wherein, X: the log value of the concentration of the tested compound; Y: the inhibition rate of the tested compound(% inh).
[0316] Experiment data are shown in Table 4 and Table 5.
TABLE-US-00005 TABLE 4 IC.sub.50 value of Example compounds on MCF cell IC.sub.50(μM) of compound Example on MCF cell Control compound 1 0.444 Control compound 2 0.549 Control compound 3 0.519 Example 1 0.176 Example 2 0.070 Example 5 0.373 Example 9 0.142 Example 23 0.074 Example 28 0.468 Example 32 0.107 Example 33 0.047
TABLE-US-00006 TABLE 5 IC.sub.50 value of Example compounds on HGC-27 cell IC.sub.50(μM) of ompound Example on HGC-27cell Control compound 1 1.579 Control compound 3 2.067 Example 2 0.361 Example 5 4.071 Example 28 3.921 Example 32 0.415 Example 33 0.267
[0317] From the above table, the compounds provided by the present invention have a better cell proliferation inhibitory activities for cell lines carring PI3Kα point mutations than the control compounds, and have better anti-tumor effects at the same concentration. Therefore, the compound provided by the present invention is expected to be a PI3Kα kinase inhibitor with better anti-tumor effects.
Example C: Pharmacokinetic Test
[0318] Method: 42 male SD rats, weight: 150-300 g. Divided into 7 groups randomly, with 6 rats in each group. Among the 6 rats in each group, three of them were administrated a single intravenous injection of 2 mg/mL of the example compound, the other three were given intragastric administration of 10 mg/mL of the example compound, blood was collected from the orbital venous plexus at designated time points, plasma was separated, and stored in a −80° C. refrigerator for later use.
[0319] The protein of the resultant plasma was precipitated by acetonitrile, the supernatant was extracted, and mixed with water at the ratio of 1:1, took 10 μL of which for LC-MS/MS detection, and calculated the average was calculated. The results are shown in Table 6.
TABLE-US-00007 TABLE 6 Pharmacokinetic Test Results of Example Compounds Intravenous injection Gavage Compound Dosage CL Dosage C.sub.max AUC No. (mg/kg) (mL/min/kg) (mg/kg) (ng/mL) (hr*ng/mL) Control 2 16.6 10 1963 7882 compound 1 Example 2 2 1.98 10 4340 52993 Example 4 2 4.75 10 2690 29080 Example 23 2 8.76 10 1833 18563 Example 32 2 1.5 10 5033 66414 Example 33 2 11.1 10 1597 19832
[0320] According to relevant statistics (Kola I, Landis J. Nat Rev Drug Discov. 2004 August; 3(8):711-5.), about 40% of drug candidates failed clinical development due to poor PK/bioavailability in the early 1990s, therefore PK/bioavailability plays a very critical role in the clinical development of candidate drugs.
[0321] The above table shows that compounds provided by the present invention have unexpectedly higher oral exposure and lower in vivo clearance compared to the control compounds, which leads to higher systemic exposure at the same dose. The compound of the present invention achieves the same anti-tumor effects at a lower dose level, thereby reduces the probability of toxicity and safety risks, and increase the druggability of the compound as well.