SUBSTITUTED HETEROCYCLIC FUSED CYCLIC COMPOUND, PREPARATION METHOD THEREFOR AND PHARMACEUTICAL USE THEREOF
20230084095 · 2023-03-16
Assignee
- GENFLEET THERAPEUTICS (SHANGHAI) INC. (Shanghai, CN)
- ZHEJIANG GENFLEET THERAPEUTICS CO., LTD. (Shaoxing, CN)
Inventors
- Fusheng Zhou (Shanghai, CN)
- Tao Jiang (Shanghai, CN)
- Chonglan LIN (Shanghai, CN)
- Lijian CAI (Shanghai, CN)
- Wan He (Shanghai, CN)
- Jiong Lan (Shanghai, CN)
Cpc classification
C07D471/22
CHEMISTRY; METALLURGY
C07D498/22
CHEMISTRY; METALLURGY
International classification
C07D498/22
CHEMISTRY; METALLURGY
Abstract
A substituted heterocyclic fused cyclic compound as represented by formula (I) or formula (IA) and having a selective inhibitory effect on KRAS gene mutation, or a pharmaceutically-acceptable salt, a stereoisomer, a solvate or a prodrug thereof, a pharmaceutical composition containing the compound, and an application thereof in preparation of cancer drugs are provided.
##STR00001##
Claims
1. A compound of Formula (I), or a pharmaceutically acceptable salt, a stereoisomer, a solvate or a prodrug thereof, ##STR01338## wherein, Z is N—C(O)—CR.sub.3═CR.sub.1R.sub.2 or N—C(O)—C≡CR.sub.4; R.sub.1 and R.sub.2 are each independently hydrogen, halogen, cyano, —NR.sup.aR.sup.b, —C.sub.1-3 alkyl, -halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-3 alkoxy, —C.sub.1-3 alkyl-NR.sup.aR.sup.b, —C.sub.1-3 alkyl-3- to 6-membered heterocycloalkyl or —C.sub.1-3 alkyl-5- or 6-membered monocyclic heteroaryl, wherein the 3- to 6-membered heterocycloalkyl or the 5- or 6-membered monocyclic heteroaryl has 1, 2 or 3 heteroatoms selected from the group consisting of N, O, and S as ring atoms; R.sub.3 is hydrogen, halogen, —C.sub.1-3 alkyl, or —C.sub.1-3 alkoxy; R.sub.4 is hydrogen, -halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, or —C.sub.1-3 alkyl-C.sub.1-3 alkoxy; R.sub.11 and R.sub.12 are either identical or different and are each independently hydrogen, halogen, —C.sub.1-3 alkyl, -halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-6 alkoxy, —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkyl, or —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkoxy; R.sub.21 and R.sub.22 are either identical or different and are each independently hydrogen, halogen, —C.sub.1-3 alkyl, -halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-6 alkoxy, —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkyl, or —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkoxy; R.sub.31 and R.sub.32 are either identical or different and are each independently hydrogen, halogen, —C.sub.1-3 alkyl, -halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-6 alkoxy, —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkyl, or —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkoxy; R.sub.41 is hydrogen, halogen, —C.sub.1-3 alkyl, -halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-6 alkoxy, —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkyl, or —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkoxy; when the dashed line in ##STR01339## is a single bond, P is O, NH or NR.sup.m; R.sup.m is —C.sub.1-6 alkyl, -halogenated C.sub.1-6 alkyl, —C.sub.1-6 alkyl-hydroxyl, —C.sub.1-6 alkyl-cyano, —C.sub.1-6 alkyl-C.sub.1-6 alkoxy, —C.sub.1-6 alkyl-halogenated C.sub.1-6 alkoxy, —C.sub.1-6 alkyl-C.sub.3-6 cycloalkyl, or —C.sub.1-6 alkyl-3- to 6-membered heterocycloalkyl; R.sub.42 is —(C═O)—, —C.sub.1-3 alkyl-, —C.sub.1-3 alkyl (hydroxy)-, —C.sub.1-3 alkyl (cyano)-, —C.sub.1-3 alkyl (C.sub.1-6 alkyl)-, —C.sub.1-3 alkyl (halogenated C.sub.1-6 alkyl)-, —C.sub.1-3 alkyl (C.sub.1-6 alkyl-hydroxy)-, —C.sub.1-3 alkyl (C.sub.1-6 alkyl-cyano)-, —C.sub.1-3 alkyl (C.sub.1-6 alkoxy)-, or —C.sub.1-3 alkyl (halogenated C.sub.1-6 alkoxy)-; or when the dashed line in ##STR01340## is absent, P is hydrogen, halogen; R.sub.42 is hydrogen, halogen, —C.sub.1-3 alkyl, -halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-6 alkoxy, —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkyl, or —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkoxy; when Y.sub.1 is C, X.sub.1 is hydrogen, halogen, cyano, hydroxyl, amino, nitro, -substituted or unsubstituted C.sub.1-6 alkyl, -substituted or unsubstituted C.sub.3-6 cycloalkyl, -substituted or unsubstituted 3- to 6-membered heterocycloalkyl, —O-substituted or unsubstituted C.sub.1-6 alkyl, —O-substituted or unsubstituted C.sub.3-6 cycloalkyl, —O-substituted or unsubstituted 3- to 6-membered heterocycloalkyl, —NH-substituted or unsubstituted C.sub.1-6 alkyl, —N(substituted or unsubstituted C.sub.1-6 alkyl).sub.2, —NH-substituted or unsubstituted C.sub.3-6 cycloalkyl, —NH-substituted or unsubstituted 3- to 6-membered heterocycloalkyl, —NH(C═O)-substituted or unsubstituted C.sub.1-6 alkyl, —NH(C═O)—C.sub.3-6 cycloalkyl, —NH (SO.sub.2)-substituted or unsubstituted C.sub.1-6 alkyl, —NH(SO.sub.2)-substituted or unsubstituted C.sub.3-6 cycloalkyl, —SO.sub.2-substituted or unsubstituted C.sub.1-6 alkyl, —SO.sub.2-substituted or unsubstituted C.sub.3-6 cycloalkyl, —(C═O)—NR.sup.jR.sup.k—, —(C═O)—O-substituted or unsubstituted C.sub.1-6 alkyl, or —(C═O)—O-substituted or unsubstituted C.sub.3-6 cycloalkyl; wherein R.sup.j and R.sup.k are each independently hydrogen or C.sub.1-3 alkyl; or R.sup.j and R.sup.k form together with a nitrogen atom adjacent thereto a substituted or unsubstituted 3- to 6-membered N-containing heterocycloalkyl; the 3- to 6-membered heterocycloalkyl has 1, 2 or 3 heteroatoms selected from the group consisting of N, O, and S as ring atoms; the substituted or unsubstituted 3- to 6-membered N-containing heterocycloalkyl has 3 to 6 ring atoms, and one of the 3 to 6 ring atoms is nitrogen atom, while 0, 1 or 2 ring atoms among a rest of the 3 to 6 ring atoms are optionally heteroatoms selected from the group consisting of N, O, and S; the “substituted” means 1, 2, 3 or 4 hydrogen atoms in a group being each independently substituted by a group-S substituent; or when Y.sub.1 is N, X.sub.1 is absent; the group-S substituent is selected from the group consisting of hydroxyl, halogen, nitro, oxo, —C.sub.1-6 alkyl, -halogenated C.sub.1-6 alkyl, hydroxy-substituted C.sub.1-6 alkyl, benzyl, —(CH.sub.2).sub.u-cyano, —(CH.sub.2).sub.u—C.sub.1-6alkoxy, —(CH.sub.2).sub.u-halogenated C.sub.1-6 alkoxy, —(CH.sub.2).sub.u-halogenated C.sub.1-6 alkyl, —(CH.sub.2).sub.u-3- to 6-membered heterocycloalkyl, —(CH.sub.2).sub.u-5- or 6-membered monocyclic heteroaryl, —(CH.sub.2).sub.u—C.sub.3-8 cycloalkyl, —(CH.sub.2).sub.u—O—(CH.sub.2).sub.v—C.sub.3-8 cycloalkyl, —(CH.sub.2).sub.u—O—(CH.sub.2).sub.v—C.sub.1-6 alkoxy, —(CH.sub.2).sub.u—O—(CH.sub.2).sub.vOH, —(CH.sub.2).sub.u—SO.sub.2C.sub.1-6 alkyl, —(CH.sub.2).sub.u—NR.sub.a0R.sub.b0, —(CH.sub.2).sub.u—C(O)NR.sub.a0R.sub.b0, —(CH.sub.2).sub.u—C(O)C.sub.1-6 alkyl, —C(O)OC.sub.1-6 alkyl, NR.sub.a0C(O)—(CH.sub.2).sub.u—NR.sub.a0R.sub.b0, NR.sub.a0C(O)—(CH.sub.2).sub.uOH, and NR.sub.a0C(O)-halogenated C.sub.1-6 alkyl, wherein the 3- to 6-membered heterocycloalkyl or the 5- or 6-membered monocyclic heteroaryl each independently has 1, 2 or 3 heteroatoms selected from the group consisting of N, O, and S as ring atoms; the 3- to 6-membered heterocycloalkyl and the 5- or 6-membered monocyclic heteroaryl are each optionally substituted by 1, 2, or 3 substituents selected from the group consisting of halogen, cyano, —C.sub.1-3 alkyl, —C.sub.1-3 alkoxy, and —C.sub.3-6 cycloalkyl; u and v are each independently 0, 1, 2, 3 or 4; R.sub.a0 and R.sub.b0 are each independently hydrogen or C.sub.1-3 alkyl; E.sub.1 is N or CR.sub.5, wherein R.sub.5 is hydrogen, halogen, cyano, —C.sub.1-6 alkyl, —C.sub.1-6 alkoxy, -halogenated C.sub.1-6 alkyl, -halogenated C.sub.1-6 alkoxy, —C.sub.3-6 cycloalkyl, —NR.sup.hR.sup.i, —C.sub.1-4 alkyl-hydroxyl, —C.sub.1-4 alkyl-cyano, —C.sub.1-4 alkyl-C.sub.1-6 alkoxy, —C.sub.1-4 alkyl-halogenated C.sub.1-6 alkyl, or —C.sub.1-4alkyl-halogenated C.sub.1-6 alkoxy; E.sub.2 is N or CR.sub.6, wherein R.sub.6 is hydrogen, halogen, cyano, —C.sub.1-6 alkyl, —C.sub.1-6 alkoxy, -halogenated C.sub.1-6 alkyl, -halogenated C.sub.1-6 alkoxy, —C.sub.3-6 cycloalkyl, —NR.sup.hR.sup.i, —C.sub.1-4 alkyl-hydroxyl, —C.sub.1-4 alkyl-cyano, —C.sub.1-4 alkyl-C.sub.1-6 alkoxy, —C.sub.1-4 alkyl-halogenated C.sub.1-6 alkyl, or —C.sub.1-4 alkyl-halogenated C.sub.1-6 alkoxy, provided that Y.sub.1, E.sub.1 and E.sub.2 are not simultaneously N; Ar is C.sub.6-10 aryl, 5- or 6-membered monocyclic heteroaryl or 8- to 10-membered bicyclic heteroaryl, wherein the 5- or 6-membered monocyclic heteroaryl has 1, 2 or 3 heteroatoms selected from the group consisting of N, O, and S as ring atoms; the 8- to 10-membered bicyclic heteroaryl has 1, 2, 3, 4, or 5 heteroatoms selected from the group consisting of N, O, and S as ring atoms; and the C.sub.6-10 aryl, the 5- or 6-membered monocyclic heteroaryl or the 8- to 10-membered bicyclic heteroaryl is unsubstituted or substituted by 1, 2, 3, or 4 groups each independently selected from R.sub.s1; or, Ar has a structure of Formula (B): ##STR01341## wherein the ring B1 is a benzene ring or a 5- or 6-membered monocyclic heteroaryl ring; the ring B2 is a fused 5- or 6-membered monocyclic heterocycloalkyl ring or a fused 5- or 6-membered monocyclic cycloalkyl ring, wherein the 5- or 6-membered monocyclic heteroaryl ring or the fused 5- or 6-membered monocyclic heterocycloalkyl ring has 1, 2 or 3 heteroatoms selected from the group consisting of N, O, and S as ring atoms; (R.sub.s1).sub.p represents that hydrogens on the ring B1 is substituted by p R.sub.s1 groups, p being 0, 1, 2 or 3 and each R.sub.s1 being either identical or different; (R.sub.s2).sub.q represents that hydrogens on the ring B2 is substituted by q R.sub.s2 groups, q being 0, 1, 2 or 3 and each R.sub.s2 being either identical or different; R.sub.s1 and R.sub.s2 are each independently halogen, cyano, nitro, hydroxyl, —C.sub.1-6 alkyl, —C.sub.1-6 alkoxy, -halogenated C.sub.1-6 alkyl, -halogenated C.sub.1-6 alkoxy, —C.sub.3-6 cycloalkyl, —NR.sup.cR.sup.d, —C(O)NR.sup.eR.sup.f, —SO.sub.2C.sub.1-3 alkyl, —SO.sub.2halogenated C.sub.1-3 alkyl, —SO.sub.2NR.sup.eR.sup.f, —C.sub.1-4 alkyl-hydroxyl, —C.sub.1-4 alkyl-cyano, —C.sub.1-4 alkyl-C.sub.1-6 alkoxy, —C.sub.1-4 alkyl-halogenated C.sub.1-6 alkyl, —C.sub.1-4alkyl-halogenated C.sub.1-6 alkoxy, —C.sub.1-4 alkyl-3- to 6-membered heterocycloalkyl, —C.sub.1-4 alkyl-NR.sup.eR.sup.f, —C.sub.1-4 alkyl-C(O)NR.sup.eR.sup.f, —C.sub.1-4 alkyl-SO.sub.2C.sub.1-3 alkyl, or C.sub.2-4 alkynyl, wherein the 3- to 6-membered heterocycloalkyl has 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S as ring atoms; R.sub.0 is —C.sub.1-6 alkyl, —C.sub.3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, C.sub.6-10 aryl, 5- or 6-membered monocyclic heteroaryl, 8- to 10-membered bicyclic heteroaryl, 7- to 11-membered spirocycloalkyl, —C.sub.1-3 alkyl-C.sub.6-10 aryl, —C.sub.1-3 alkyl-5- or 6-membered monocyclic heteroaryl, —NR.sup.g—C.sub.6-10 aryl, —O—C.sub.6-10 aryl, —C.sub.1-3 alkyl-3- to 6-membered heterocycloalkyl, or —C.sub.1-3 alkyl-C.sub.3-6 cycloalkyl, wherein the 3- to 6-membered heterocycloalkyl, the 5- or 6-membered monocyclic heteroaryl or the 8- to 10-membered bicyclic heteroaryl has 1, 2 or 3 heteroatoms selected from the group consisting of N, O, and S as ring atoms; the C.sub.1-6 alkyl, the C.sub.3-6 cycloalkyl, the 3- to 6-membered heterocycloalkyl, the C.sub.6-10 aryl, the 5- or 6-membered monocyclic heteroaryl, the 8- to 10-membered bicyclic heteroaryl, and the 7- to 11-membered spirocycloalkyl are unsubstituted or each substituted by 1, 2, 3, or 4 groups each independently selected from R.sub.s3; and the —C.sub.1-3 alkyl- is unsubstituted or substituted by 1, 2, 3, or 4 groups each independently selected from C.sub.1-3 alkyl; or, R.sub.0 has a structure of Formula (A-1) or Formula (A-2): ##STR01342## wherein the ring A1 is a benzene ring or a 5- or 6-membered monocyclic heteroaryl ring; the ring A2 is a fused 5- or 6-membered monocyclic heterocycloalkyl ring or a fused 5- or 6-membered monocyclic cycloalkyl ring, wherein the 5- or 6-membered monocyclic heteroaryl ring or the fused 5- or 6-membered monocyclic heterocycloalkyl ring has 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S as ring atoms; (R.sub.s3).sub.t represents that hydrogens on the ring A1 is substituted by t R.sub.s3 groups, t being 0, 1, 2, or 3 and each R.sub.s3 being either identical or different; (R.sub.s4).sub.s represents that hydrogens on the ring A2 is substituted by s R.sub.s4 groups, s being 0, 1, 2, or 3 and each R.sub.s4 being either identical or different; R.sub.s3 and R.sub.s4 are each independently halogen, cyano, hydroxyl, —C.sub.1-6 alkyl, —C.sub.1-6 alkoxy, -halogenated C.sub.1-6 alkyl, -halogenated C.sub.1-6 alkoxy, —C.sub.3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, —NR.sup.hR.sup.i, —C(O)NR.sup.eR.sup.f, —SO.sub.2C.sub.1-3 alkyl, —SO.sub.2halogenated C.sub.1-3 alkyl, —SO.sub.2NR.sup.eR.sup.f, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-C.sub.2-4 alkynyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-6 alkoxy, —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkyl, —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkoxy, —C.sub.1-3 alkyl-3- to 6-membered heterocycloalkyl, —C.sub.1-3 alkyl-C.sub.3-6 cycloalkyl, —C.sub.1-3 alkyl-NR.sup.eR.sup.f, —C.sub.1-3 alkyl-C(O)NR.sup.eR.sup.f, —C.sub.1-3 alkyl-SO.sub.2C.sub.1-3 alkyl, or C.sub.2-4 alkynyl, wherein the 3- to 6-membered heterocycloalkyl has 1, 2 or 3 heteroatoms selected from the group consisting of N, O, and S as ring atoms; and the C.sub.1-6 alkyl, the —C.sub.1-6 alkoxy, the —C.sub.1-3 alkyl-, the —C.sub.3-6 cycloalkyl, and the 3- to 6-membered heterocycloalkyl are each optionally substituted by 1, 2 or 3 substituents each independently selected from the group consisting of halogen, methyl, ethyl, propyl, isopropyl, trifluoromethyl, amino, N(CH.sub.3).sub.2, hydroxyl, and carboxyl; R.sup.a, R.sup.b, R.sup.e, R.sup.f, and R.sup.g are each independently hydrogen or C.sub.1-3 alkyl; and R.sup.c, R.sup.d, R.sup.h, and R.sup.i are each independently hydrogen, —C.sub.1-3 alkyl, —C(O)C.sub.1-3 alkyl, or —CO.sub.2C.sub.1-3 alkyl.
2. A compound of Formula (II), or a pharmaceutically acceptable salt, a stereoisomer, a solvate or a prodrug thereof, ##STR01343## wherein: Z is N—C(O)—CR.sub.3═CR.sub.1R.sub.2 or N—C(O)—C≡CR.sub.4; R.sub.1 and R.sub.2 are each independently hydrogen, halogen, cyano, NR.sup.aR.sup.b, —C.sub.1-3 alkyl, halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-3 alkoxy, —C.sub.1-3 alkyl-NR.sup.aR.sup.b, —C.sub.1-3 alkyl-3- to 6-membered heterocycloalkyl, or —C.sub.1-3 alkyl-5- or 6-membered monocyclic heteroaryl, wherein the 3- to 6-membered heterocycloalkyl or the 5- or 6-membered monocyclic heteroaryl has 1, 2 or 3 heteroatoms selected from the group consisting of N, O, and S as ring atoms; R.sub.3 is hydrogen, halogen, —C.sub.1-3 alkyl, or —C.sub.1-3 alkoxy; R.sub.4 is hydrogen, halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, or —C.sub.1-3 alkyl-C.sub.1-3 alkoxy; R.sub.11 and R.sub.12 are either identical or different and are each independently hydrogen, halogen, —C.sub.1-3 alkyl, -halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-6 alkoxy, —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkyl, or —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkoxy; R.sub.21 and R.sub.22 are either identical or different and are each independently hydrogen, halogen, —C.sub.1-3 alkyl, -halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-6 alkoxy, —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkyl, or —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkoxy; R.sub.31 and R.sub.32 are either identical or different and are each independently hydrogen, halogen, —C.sub.1-3 alkyl, -halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-6 alkoxy, —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkyl, or —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkoxy; R.sub.41 is hydrogen, halogen, —C.sub.1-3 alkyl, -halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-6 alkoxy, —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkyl, or —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkoxy; P is O, NH, or NR.sup.m; R.sup.m is —C.sub.1-6 alkyl, -halogenated C.sub.1-6 alkyl, —C.sub.1-6 alkyl-hydroxyl, —C.sub.1-6alkyl-cyano, —C.sub.1-6 alkyl-C.sub.1-6 alkoxy, —C.sub.1-6 alkyl-halogenated C.sub.1-6 alkoxy, —C.sub.1-6 alkyl-C.sub.3-6 cycloalkyl, or —C.sub.1-6 alkyl-3- to 6-membered heterocycloalkyl; R.sub.42 is —(C═O)—, —C.sub.1-3 alkyl-, —C.sub.1-3 alkyl (hydroxy)-, —C.sub.1-3 alkyl (cyano)-, —C.sub.1-3 alkyl (C.sub.1-6 alkyl)-, —C.sub.1-3 alkyl (halogenated C.sub.1-6 alkyl)-, —C.sub.1-3 alkyl (C.sub.1-6 alkyl-hydroxy)-, —C.sub.1-3 alkyl (C.sub.1-6 alkyl-cyano)-, —C.sub.1-3 alkyl (C.sub.1-6 alkoxy)-, or —C.sub.1-3 alkyl (halogenated C.sub.1-6 alkoxy)-; X.sub.2 and Y.sub.2 are either identical or different and are each independently hydrogen, halogen, —C.sub.1-3 alkyl, -halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-6 alkoxy, —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkyl, or —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkoxy; or X.sub.2 and Y.sub.2 form together with a carbon atom adjacent thereto substituted or unsubstituted C.sub.3-6 cycloalkyl or substituted or unsubstituted 3- to 6-membered heterocycloalkyl; the 3- to 6-membered heterocycloalkyl has 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S as ring atoms; the “substituted” means 1, 2, 3, or 4 hydrogen atoms in a group being each independently substituted by a group-S substituent; E.sub.3 is N or C-L-R.sub.5, wherein: L is a bond, —CR.sub.L1R.sub.L2—, —O—(CR.sub.L1R.sub.L2).sub.t1—, or —NH—(CR.sub.L3R.sub.L4).sub.t2—, wherein R.sub.L1, R.sub.L2, R.sub.L3, and R.sub.L4 are either identical or different and are each independently hydrogen, halogen, hydroxyl, hydroxymethyl, hydroxyethyl, —C.sub.1-3 alkyl, or oxo; t1 and t2 are each independently 0, 1, 2, 3, or 4; when between R.sub.L1 and R.sub.L2 or between R.sub.L3 and R.sub.L4, when one is oxo, the other one is absent; R.sub.5 is hydrogen, halogen, hydroxyl, -substituted or unsubstituted C.sub.1-6 alkyl, -substituted or unsubstituted C.sub.3-6 cycloalkyl, -substituted or unsubstituted 3- to 6-membered heterocycloalkyl, —O-substituted or unsubstituted C.sub.1-6 alkyl, —O-substituted or unsubstituted C.sub.3-6 cycloalkyl, —O-substituted or unsubstituted 3- to 6-membered heterocycloalkyl, —SO.sub.2-substituted or unsubstituted C.sub.1-6 alkyl, —SO.sub.2-substituted or unsubstituted C.sub.3-6 cycloalkyl, —SO.sub.2-substituted or unsubstituted 3- to 6-membered heterocycloalkyl, -substituted or unsubstituted 5- or 6-membered monocyclic heteroaryl, or NR.sub.51R.sub.52, wherein R.sub.51 and R.sub.52 are each independently hydrogen, substituted or unsubstituted C.sub.1-6 alkyl, —SO.sub.2C.sub.1-6 alkyl, —SO.sub.2C.sub.3-6 cycloalkyl, —C(O)C.sub.1-6 alkyl, or —C(O)halogenated C.sub.1-6 alkyl; or R.sub.51 and R.sub.52 form together with a nitrogen atom adjacent thereto substituted or unsubstituted 3- to 6-membered N-containing heterocycloalkyl; wherein the 3- to 6-membered heterocycloalkyl and the 5- or 6-membered monocyclic heteroaryl each independently have 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S as ring atoms; the substituted or unsubstituted 3- to 6-membered N-containing heterocycloalkyl has 3 to 6 ring atoms, and one of the 3 to 6 ring atoms is nitrogen atom, while 0, 1, or 2 ring atoms among a rest of the 3 to 6 ring atoms are optionally heteroatoms selected from the group consisting of N, O, and S; the “substituted” means 1, 2, 3, or 4 hydrogen atoms in a group being each independently substituted by a group-S substituent; the group-S substituent is selected from the group consisting of hydroxyl, halogen, nitro, oxo, —C.sub.1-6 alkyl, -halogenated C.sub.1-6 alkyl, hydroxy-substituted C.sub.1-6 alkyl, benzyl, —(CH.sub.2).sub.u-cyano, —(CH.sub.2).sub.u—C.sub.1-6 alkoxy, —(CH.sub.2).sub.u-halogenated C.sub.1-6 alkoxy, —(CH.sub.2).sub.u-halogenated C.sub.1-6 alkyl, —(CH.sub.2).sub.u-3- to 6-membered heterocycloalkyl, —(CH.sub.2).sub.u-5- or 6-membered monocyclic heteroaryl, —(CH.sub.2).sub.u—C.sub.3-8 cycloalkyl, —(CH.sub.2).sub.u—O—(CH.sub.2).sub.v—C.sub.3-8 cycloalkyl, —(CH.sub.2).sub.u—O—(CH.sub.2).sub.v—C.sub.1-6 alkoxy, —(CH.sub.2).sub.u—O—(CH.sub.2).sub.vOH, —(CH.sub.2).sub.u—SO.sub.2C.sub.1-6 alkyl, —(CH.sub.2).sub.u—NR.sub.a0R.sub.b0, —(CH.sub.2).sub.u—C(O)NR.sub.a0R.sub.b0, —(CH.sub.2).sub.u—C(O)C.sub.1-6 alkyl, —C(O)OC.sub.1-6 alkyl, NR.sub.a0C(O)—(CH.sub.2).sub.u—NR.sub.a0R.sub.b0, NR.sub.a0C(O)—(CH.sub.2).sub.uOH, and NR.sub.a0C(O)-halogenated C.sub.1-6 alkyl, wherein the 3- to 6-membered heterocycloalkyl or the 5- or 6-membered monocyclic heteroaryl each independently has 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S as ring atoms; the 3- to 6-membered heterocycloalkyl and the 5- or 6-membered monocyclic heteroaryl are each optionally substituted by 1, 2, or 3 substituents selected from the group consisting of halogen, cyano, —C.sub.1-3 alkyl, —C.sub.1-3 alkoxy, and C.sub.3-6 cycloalkyl; u and v are each independently 0, 1, 2, 3, or 4; R.sub.a0 and R.sub.b0 are each independently hydrogen or C.sub.1-3 alkyl; E.sub.4 is N or CH; Ar is C.sub.6-10 aryl, 5- or 6-membered monocyclic heteroaryl, or 8- to 10-membered bicyclic heteroaryl, wherein the 5- or 6-membered monocyclic heteroaryl has 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S as ring atoms; the 8- to 10-membered bicyclic heteroaryl has 1, 2, 3, 4, or 5 heteroatoms selected from the group consisting of N, O, and S as ring atoms; and the C.sub.6-10 aryl, the 5- or 6-membered monocyclic heteroaryl, or the 8- to 10-membered bicyclic heteroaryl is unsubstituted or substituted by 1, 2, 3, or 4 groups each independently selected from R.sub.s1; or, Ar has a structure of Formula (B): ##STR01344## wherein the ring B1 is a benzene ring or a 5- or 6-membered monocyclic heteroaryl ring; the ring B2 is a fused 5- or 6-membered monocyclic heterocycloalkyl ring or a fused 5- or 6-membered monocyclic cycloalkyl ring, wherein the 5- or 6-membered monocyclic heteroaryl ring or the fused 5- or 6-membered monocyclic heterocycloalkyl ring has 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S as ring atoms; (R.sub.s1).sub.p represents that hydrogens on the ring B1 is substituted by p R.sub.s1 groups, p being 0, 1, 2, or 3 and each R.sub.s1 being either identical or different; (R.sub.s2).sub.q represents that hydrogens on the ring B2 is substituted by q R.sub.s2 groups, q being 0, 1, 2, or 3 and each R.sub.s2 being either identical or different; R.sub.s1, R.sub.s2 are each independently halogen, cyano, nitro, hydroxyl, —C.sub.1-6 alkyl, —C.sub.1-6 alkoxy, -halogenated C.sub.1-6 alkyl, -halogenated C.sub.1-6 alkoxy, —C.sub.3-6 cycloalkyl, —NR.sup.cR.sup.d, —C(O)NR.sup.eR.sup.f, —SO.sub.2C.sub.1-3 alkyl, —SO.sub.2halogenated C.sub.1-3 alkyl, —SO.sub.2NR.sup.eR.sup.f, —C.sub.1-4 alkyl-hydroxyl, —C.sub.1-4 alkyl-cyano, —C.sub.1-4 alkyl-C.sub.1-6 alkoxy, —C.sub.1-4 alkyl-halogenated C.sub.1-6 alkyl, —C.sub.1-4 alkyl-halogenated C.sub.1-6 alkoxy, —C.sub.1-4 alkyl-3- to 6-membered heterocycloalkyl, —C.sub.1-4 alkyl-NR.sup.eR.sup.f, —C.sub.1-4 alkyl-C(O)NR.sup.eR.sup.f, —C.sub.1-4 alkyl-SO.sub.2C.sub.1-3 alkyl, or —C.sub.2-4 alkynyl, wherein the 3- to 6-membered heterocycloalkyl has 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S as ring atoms; R.sup.a, R.sup.b, R.sup.e, and R.sup.f are each independently hydrogen or —C.sub.1-3 alkyl; and R.sup.c and R.sup.d are each independently hydrogen, —C.sub.1-3 alkyl, —C(O)C.sub.1-3 alkyl, or —CO.sub.2C.sub.1-3 alkyl.
3. A compound of Formula (IA), or a tautomer, a cis-trans isomer, a mesomer, a raceme, an enantiomer, a diastereoisomer, an atropisomer or a mixture thereof, or a pharmaceutically acceptable salt, a solvate or a prodrug thereof: ##STR01345## wherein: Z is N—C(O)—CR.sub.3═CR.sub.1R.sub.2 or N—C(O)—C≡CR.sub.4; R.sub.1 and R.sub.2 are each independently hydrogen, halogen, cyano, NR.sup.aR.sup.b, —C.sub.1-3 alkyl, halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-3 alkoxy, —C.sub.1-3 alkyl-NR.sup.aR.sup.b, —C.sub.1-3 alkyl-3- to 6-membered heterocycloalkyl, or —C.sub.1-3 alkyl-5- or 6-membered monocyclic heteroaryl, wherein the 3- to 6-membered heterocycloalkyl or the 5- or 6-membered monocyclic heteroaryl has 1, 2 or 3 heteroatoms selected from the group consisting of N, O, and S as ring atoms; R.sub.3 is hydrogen, halogen, —C.sub.1-3 alkyl, or —C.sub.1-3 alkoxy; R.sub.4 is hydrogen, halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, or —C.sub.1-3 alkyl-C.sub.1-3 alkoxy; R.sub.11 and R.sub.12 are either identical or different and are each independently hydrogen, halogen, —C.sub.1-3 alkyl, -halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-6 alkoxy, —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkyl, or —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkoxy; R.sub.21 and R.sub.22 are either identical or different and are each independently hydrogen, halogen, —C.sub.1-3 alkyl, -halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-6 alkoxy, —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkyl, or —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkoxy; R.sub.31 and R.sub.32 are either identical or different and are each independently hydrogen, halogen, —C.sub.1-3 alkyl, -halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-6 alkoxy, —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkyl, or —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkoxy; R.sub.41 is hydrogen, halogen, —C.sub.1-3 alkyl, -halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-6 alkoxy, —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkyl, or —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkoxy; when the dashed line in ##STR01346## is a single bond, P′ is O, NH, or NR.sup.m′; R.sup.m′ is —C.sub.1-6 deuteroalkyl, —C.sub.1-6 alkyl, -halogenated C.sub.1-6 alkyl, —C.sub.1-6 alkyl-hydroxyl, —C.sub.1-6 alkyl-cyano, —C.sub.1-6 alkyl-C.sub.1-6 alkoxy, —C.sub.1-6 alkyl-halogenated C.sub.1-6 alkoxy, —C.sub.1-6 alkyl-C.sub.3-6 cycloalkyl, or —C.sub.1-6 alkyl-3- to 6-membered heterocycloalkyl; R.sub.42′ is —C.sub.1-3 alkyl-(C═O)—, —(C═O)—, —C.sub.1-3 alkyl-, —C.sub.1-3 alkyl (hydroxy)-, —C.sub.1-3 alkyl (cyano)-, —C.sub.1-3 alkyl (C.sub.1-6 alkyl)-, —C.sub.1-3 alkyl (halogenated C.sub.1-6 alkyl)-, —C.sub.1-3 alkyl (C.sub.1-6 alkyl-hydroxy)-, —C.sub.1-3 alkyl (C.sub.1-6 alkyl-cyano)-, —C.sub.1-3 alkyl (C.sub.1-6 alkoxy)-, or —C.sub.1-3 alkyl (halogenated C.sub.1-6 alkoxy)-; or when the dashed line in ##STR01347## is absent, P′ is hydrogen or halogen; R.sub.42′ is hydrogen, halogen, —C.sub.1-3 alkyl, -halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-6 alkoxy, —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkyl, or —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkoxy; when Y.sub.1 is C, X.sub.1 is hydrogen, halogen, cyano, hydroxyl, amino, nitro, -substituted or unsubstituted C.sub.1-6 alkyl, -substituted or unsubstituted C.sub.3-6 cycloalkyl, -substituted or unsubstituted 3- to 6-membered heterocycloalkyl, —O-substituted or unsubstituted C.sub.1-6 alkyl, —O-substituted or unsubstituted C.sub.3-6cycloalkyl, —O-substituted or unsubstituted 3- to 6-membered heterocycloalkyl, —NH-substituted or unsubstituted C.sub.1-6 alkyl, —N(substituted or unsubstituted C.sub.1-6alkyl).sub.2, —NH-substituted or unsubstituted C.sub.3-6 cycloalkyl, —NH-substituted or unsubstituted 3- to 6-membered heterocycloalkyl, —NH(C═O)-substituted or unsubstituted C.sub.1-6 alkyl, —NH(C═O)—C.sub.3-6 cycloalkyl, —NH(SO.sub.2)-substituted or unsubstituted C.sub.1-6 alkyl, —NH(SO.sub.2)-substituted or unsubstituted C.sub.3-6 cycloalkyl, —SO.sub.2-substituted or unsubstituted C.sub.1-6 alkyl, —SO.sub.2-substituted or unsubstituted C.sub.3-6 cycloalkyl, —(C═O)—NR.sup.jR.sup.k—, —(C═O)—O-substituted or unsubstituted C.sub.1-6 alkyl, or —(C═O)—O-substituted or unsubstituted C.sub.3-6 cycloalkyl, wherein R.sup.j and R.sup.k are each independently hydrogen or C.sub.1-3 alkyl; or R.sup.j and R.sup.k form together with a nitrogen atom adjacent thereto substituted or unsubstituted 3- to 6-membered N-containing heterocycloalkyl; the 3- to 6-membered heterocycloalkyl has 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S as ring atoms; the substituted or unsubstituted 3- to 6-membered N-containing heterocycloalkyl has 3 to 6 ring atoms, and one of the 3 to 6 ring atoms is nitrogen atom, while 0, 1 or 2 ring atoms among a rest of the 3 to 6 ring atoms are optionally heteroatoms selected from the group consisting of N, O, and S; the “substituted” means 1, 2, 3, or 4 hydrogen atoms in a group being each independently substituted by a group-S substituent; or when Y.sub.1 is N, X.sub.1 is absent; the group-S substituent is selected from hydroxyl, halogen, nitro, oxo, —C.sub.1-6 alkyl, -halogenated C.sub.1-6 alkyl, hydroxyl-substituted C.sub.1-6 alkyl, benzyl, —(CH.sub.2).sub.u-cyano, —(CH.sub.2)—C.sub.1-6 alkoxy, —(CH.sub.2).sub.u-halogenated C.sub.1-6 alkoxy, —(CH.sub.2).sub.u-halogenated C.sub.1-6 alkyl, —(CH.sub.2).sub.u-3- to 6-membered heterocycloalkyl, —(CH.sub.2)-5- or 6-membered monocyclic heteroaryl, —(CH.sub.2).sub.u—C.sub.3-8 cycloalkyl, —(CH.sub.2).sub.u—O—(CH.sub.2).sub.v—C.sub.3-8 cycloalkyl, —(CH.sub.2).sub.u—O—(CH.sub.2).sub.v—C.sub.1-6 alkoxy, —(CH.sub.2).sub.u—O—(CH.sub.2).sub.vOH, —(CH.sub.2).sub.u—SO.sub.2C.sub.1-6 alkyl, —(CH.sub.2).sub.u—NR.sub.a0R.sub.b0, —(CH.sub.2).sub.u—C(O)NR.sub.a0R.sub.b0, —(CH.sub.2).sub.u—C(O)C.sub.1-6 alkyl, —C(O)OC.sub.1-6 alkyl, NR.sub.a0C(O)—(CH.sub.2).sub.u—NR.sub.a0R.sub.b0, NR.sub.a0C(O)—(CH.sub.2).sub.uOH, and NR.sub.a0C(O)-halogenated C.sub.1-6 alkyl, wherein the 3- to 6-membered heterocycloalkyl or the 5- or 6-membered monocyclic heteroaryl each independently has 1, 2 or 3 heteroatoms selected from the group consisting of N, O, and S as ring atoms; the 3- to 6-membered heterocycloalkyl and the 5- or 6-membered monocyclic heteroaryl are each optionally substituted by 1, 2, or 3 substituents selected from the group consisting of halogen, cyano, —C.sub.1-3 alkyl, —C.sub.1-3 alkoxy, and C.sub.3-6 cycloalkyl; u and v are each independently 0, 1, 2, 3, or 4; R.sub.a0 and R.sub.b0 are each independently hydrogen or C.sub.1-3 alkyl; E.sub.1′ is N or CR.sub.5′, wherein R.sub.5′ is hydrogen, halogen, cyano, —C.sub.1-6 alkyl, —C.sub.1-6 alkoxy, -halogenated C.sub.1-6 alkyl, -halogenated C.sub.1-6 alkoxy, —C.sub.3-6 cycloalkyl, —O—C.sub.3-6 cycloalkyl, —NR.sup.hR.sup.i, —C.sub.1-4 alkyl-hydroxyl, —C.sub.1-4 alkyl-cyano, —C.sub.1-4 alkyl-C.sub.1-6 alkoxy, —C.sub.1-4 alkyl-halogenated C.sub.1-6 alkyl, or —C.sub.1-4 alkyl-halogenated C.sub.1-6 alkoxy; E.sub.2′ is N or CR.sub.6′, wherein R.sub.6′ is hydrogen, halogen, cyano, —C.sub.1-6 alkyl, —C.sub.1-6 alkoxy, -halogenated C.sub.1-6 alkyl, -halogenated C.sub.1-6 alkoxy, —C.sub.3-6 cycloalkyl, —O—C.sub.3-6 cycloalkyl, —NR.sup.hR.sup.i, —C.sub.1-4 alkyl-hydroxyl, —C.sub.1-4 alkyl-cyano, —C.sub.1-4 alkyl-C.sub.1-6 alkoxy, —C.sub.1-4 alkyl-halogenated C.sub.1-6 alkyl, or —C.sub.1-4 alkyl-halogenated C.sub.1-6 alkoxy, provided that Y.sub.1, E.sub.1′ and E.sub.2′ are not simultaneously N; Ar′ is C.sub.6-10 aryl, 5- or 6-membered monocyclic heteroaryl, 8- to 10-membered bicyclic heteroaryl, or pyridonyl, wherein the 5- or 6-membered monocyclic heteroaryl has 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S as ring atoms; the 8- to 10-membered bicyclic heteroaryl has 1, 2, 3, 4, or 5 heteroatoms selected from the group consisting of N, O, and S as ring atoms; and the C.sub.6-10 aryl, the 5- or 6-membered monocyclic heteroaryl, the 8- to 10-membered bicyclic heteroaryl, and the pyridonyl are unsubstituted or each substituted by 1, 2, 3, or 4 groups each independently selected from R.sub.s1; or Ar′ has a structure of Formula (B): ##STR01348## wherein the ring B1 is a benzene ring or a 5- or 6-membered monocyclic heteroaryl ring; the ring B2 is a fused 5- or 6-membered monocyclic heterocycloalkyl ring or a fused 5- or 6-membered monocyclic cycloalkyl ring, wherein the 5- or 6-membered monocyclic heteroaryl ring or the fused 5- or 6-membered monocyclic heterocycloalkyl ring has 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S as ring atoms; (R.sub.s1).sub.p represents that hydrogens on the ring B1 is substituted by p R.sub.s1 groups, p being 0, 1, 2, or 3 and each R.sub.s1 being either identical or different; (R.sub.s2).sub.q represents that hydrogens on the ring B2 is substituted by q R.sub.s2 groups, q being 0, 1, 2, or 3 and each R.sub.s2 being either identical or different; R.sub.s1 and R.sub.s2 are each independently halogen, cyano, nitro, hydroxyl, —C.sub.1-6 alkyl, —C.sub.1-6 alkoxy, -halogenated C.sub.1-6 alkyl, -halogenated C.sub.1-6 alkoxy, —C.sub.3-6 cycloalkyl, —NR.sup.cR.sup.d, —C(O)NR.sup.eR.sup.f, —SO.sub.2C.sub.1-3 alkyl, —SO.sub.2halogenated C.sub.1-3 alkyl, —SO.sub.2NR.sup.eR.sup.f, —C.sub.1-4 alkyl-hydroxyl, —C.sub.1-4 alkyl-cyano, —C.sub.1-4 alkyl-C.sub.1-6 alkoxy, —C.sub.1-4 alkyl-halogenated C.sub.1-6 alkyl, —C.sub.1-4alkyl-halogenated C.sub.1-6 alkoxy, —C.sub.1-4 alkyl-3- to 6-membered heterocycloalkyl, —C.sub.1-4 alkyl-NR.sup.eR.sup.f, —C.sub.1-4 alkyl-C(O)NR.sup.eR.sup.f, —C.sub.1-4 alkyl-SO.sub.2C.sub.1-3 alkyl, or C.sub.2-4 alkynyl, wherein the 3- to 6-membered heterocycloalkyl has 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S as ring atoms; R.sub.0′ is —C.sub.1-6 alkyl, —C.sub.3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, C.sub.6-10 aryl, 5- or 6-membered monocyclic heteroaryl, 8- to 10-membered bicyclic heteroaryl, 7- to 11-membered spirocycloalkyl, —C.sub.1-3 alkyl-C.sub.6-10 aryl, —C.sub.1-3 alkyl-5- or 6-membered monocyclic heteroaryl, —NR.sup.g—C.sub.6-10 aryl, —O—C.sub.6-10 aryl, —C.sub.1-3 alkyl-3- to 6-membered heterocycloalkyl, —C.sub.1-3 alkyl-C.sub.3-6 cycloalkyl, or pyridonyl, wherein the 3- to 6-membered heterocycloalkyl, the 5- or 6-membered monocyclic heteroaryl, or the 8- to 10-membered bicyclic heteroaryl has 1, 2 or 3 heteroatoms selected from the group consisting of N, O, and S as ring atoms; the C.sub.1-6 alkyl, the C.sub.3-6 cycloalkyl, the 3- to 6-membered heterocycloalkyl, the C.sub.6-10 aryl, the 5- or 6-membered monocyclic heteroaryl, the 8- to 10-membered bicyclic heteroaryl, the 7- to 11-membered spirocycloalkyl, and the pyridonyl are unsubstituted or each substituted by 1, 2, 3, or 4 groups each independently selected from R.sub.s3; and the —C.sub.1-3 alkyl- is unsubstituted or substituted by 1, 2, 3, or 4 groups each independently selected from C.sub.1-3 alkyl; or R.sub.0′ has a structure of Formula (A-1) or Formula (A-2): ##STR01349## wherein the ring A1 is a benzene ring or a 5- or 6-membered monocyclic heteroaryl ring; the ring A2 is a fused 5- or 6-membered monocyclic heterocycloalkyl ring or a fused 5- or 6-membered monocyclic cycloalkyl ring, wherein the 5- or 6-membered monocyclic heteroaryl ring or the fused 5- or 6-membered monocyclic heterocycloalkyl ring has 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S as ring atoms; (R.sub.s3).sub.t represents that hydrogens on the ring A1 is substituted by t R.sub.s3 groups, t being 0, 1, 2, or 3 and each R.sub.s3 being either identical or different; (R.sub.s4).sub.s represents that hydrogens on the ring A2 is substituted by s R.sub.s4 groups, s being 0, 1, 2, or 3 and each R.sub.s4 being either identical or different; R.sub.s3 and R.sub.s4 are each independently halogen, cyano, hydroxyl, —C.sub.1-6 alkyl, —C.sub.1-6 alkoxy, -halogenated C.sub.1-6 alkyl, -halogenated C.sub.1-6 alkoxy, —C.sub.3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, —NR.sup.hR.sup.i, —C(O)NR.sup.eR.sup.f, —SO.sub.2C.sub.1-3 alkyl, —SO.sub.2halogenated C.sub.1-3 alkyl, —SO.sub.2NR.sup.eR.sup.f, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-C.sub.2-4 alkynyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-6 alkoxy, —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkyl, —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkoxy, —C.sub.1-3 alkyl-3- to 6-membered heterocycloalkyl, —C.sub.1-3 alkyl-C.sub.3-6 cycloalkyl, —C.sub.1-3 alkyl-NR.sup.eR.sup.f, —C.sub.1-3 alkyl-C(O)NR.sup.eR.sup.f, —C.sub.1-3 alkyl-SO.sub.2C.sub.1-3 alkyl, or C.sub.2-4 alkynyl, wherein the 3- to 6-membered heterocycloalkyl has 1, 2 or 3 heteroatoms selected from the group consisting of N, O, and S as ring atoms; and the C.sub.1-6 alkyl, the —C.sub.1-6 alkoxy, the —C.sub.1-3 alkyl-, the —C.sub.3-6 cycloalkyl, and the 3- to 6-membered heterocycloalkyl are each optionally substituted by 1, 2, or 3 substituents each independently selected from the group consisting of halogen, methyl, ethyl, propyl, isopropyl, trifluoromethyl, amino, N(CH.sub.3).sub.2, hydroxyl, and carboxyl; R.sup.a, R.sup.b, R.sup.e, R.sup.f, and R.sup.g are each independently hydrogen or C.sub.1-3 alkyl; and R.sup.c, R.sup.d, R.sup.h, and R.sup.i are each independently hydrogen, —C.sub.1-3 alkyl, —C(O)C.sub.1-3 alkyl, or —CO.sub.2C.sub.1-3 alkyl.
4. The compound according to claim 3, or a tautomer, a cis-trans isomer, a mesomer, a raceme, an enantiomer, a diastereoisomer, an atropisomer or a mixture thereof, or a pharmaceutically acceptable salt, a solvate or a prodrug thereof, wherein the compound of Formula (IA) is a compound of Formula (IB) or a compound of Formula (IC): ##STR01350## in Formula IB, P′ is O, NH, or NR.sup.m′; R.sup.m′ is-C.sub.1-6 deuteroalkyl, —C.sub.1-6 alkyl, -halogenated C.sub.1-6 alkyl, —C.sub.1-6 alkyl-hydroxyl, —C.sub.1-6 alkyl-cyano, —C.sub.1-6 alkyl-C.sub.1-6 alkoxy, —C.sub.1-6alkyl-halogenated C.sub.1-6 alkoxy, —C.sub.1-6 alkyl-C.sub.3-6 cycloalkyl, or —C.sub.1-6 alkyl-3- to 6-membered heterocycloalkyl; R.sub.42′ is —C.sub.1-3 alkyl-(C═O)—, —(C═O)—, —C.sub.1-3 alkyl-, —C.sub.1-3 alkyl (hydroxy)-, —C.sub.1-3 alkyl (cyano)-, —C.sub.1-3 alkyl (C.sub.1-6 alkyl)-, —C.sub.1-3 alkyl (halogenated C.sub.1-6 alkyl)-, —C.sub.1-3 alkyl (C.sub.1-6 alkyl-hydroxy)-, —C.sub.1-3 alkyl (C.sub.1-6 alkyl-cyano)-, —C.sub.1-3 alkyl (C.sub.1-6 alkoxy)-, or —C.sub.1-3 alkyl (halogenated C.sub.1-6 alkoxy)-; and R.sub.11, R.sub.12, R.sub.21, R.sub.22, R.sub.31, R.sub.32, R.sub.41, Z, R.sub.0′, Ar′, E.sub.1′, E.sub.2′, X.sub.1, and Y.sub.1 are as defined in claim 3; in Formula IC, P′ is hydrogen or halogen; R.sub.42′ is hydrogen, halogen, —C.sub.1-3 alkyl, -halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-6 alkoxy, —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkyl, or —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkoxy; and R.sub.11, R.sub.12, R.sub.21, R.sub.22, R.sub.31, R.sub.32, R.sub.41, Z, R.sub.0′, Ar′, E.sub.1′, E.sub.2′, X.sub.1, and Y.sub.1 are as defined in claim 3.
5. The compound according to claim 4, or a tautomer, a cis-trans isomer, a mesomer, a raceme, an enantiomer, a diastereoisomer, an atropisomer or a mixture thereof, or a pharmaceutically acceptable salt, a solvate or a prodrug thereof, wherein the compound of Formula (IB) is a compound of Formula (IB-1) or a compound of Formula (IB-2): ##STR01351## in Formula (IB-1) and Formula (IB-2), R.sub.21, R.sub.22, R.sub.11, R.sub.12, R.sub.31, R.sub.32, R.sub.41, R.sub.42′, Z, P′, R.sub.0′, Ar′, E.sub.1′, E.sub.2′, X.sub.1, and Y.sub.1 are as defined in claim 4.
6. The compound according to claim 4, or a tautomer, a cis-trans isomer, a mesomer, a raceme, an enantiomer, a diastereoisomer, an atropisomer or a mixture thereof, or a pharmaceutically acceptable salt, a solvate or a prodrug thereof, wherein the compound of Formula (IB) is a compound of Formula (IB-1a) or a compound of Formula (IB-2a): ##STR01352## wherein R.sub.1, R.sub.2, R.sub.3, R.sub.21, R.sub.22, R.sub.12, R.sub.11, R.sub.31, R.sub.32, P′, R.sub.0′, Ar′, E.sub.1′, and X.sub.1 are as defined in claim 4.
7. The compound according to claim 6, or a tautomer, a cis-trans isomer, a mesomer, a raceme, an enantiomer, a diastereoisomer, an atropisomer or a mixture thereof, or a pharmaceutically acceptable salt, a solvate or a prodrug thereof, wherein the compound of Formula (IB-1a) is a compound of Formula (IB-1aa), a compound of Formula (IB-1ab), a compound of Formula (IB-1ac), or a compound of Formula (IB-1ad): ##STR01353## in Formula (IB-1aa) and Formula (IB-1ab), R.sub.21′ is independently halogen, —C.sub.1-3 alkyl, -halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-6 alkoxy, —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkyl, or —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkoxy; and R.sub.1, R.sub.2, R.sub.3, R.sub.12, R.sub.11, R.sub.31, R.sub.32, P′, R.sub.0′, Ar′, E.sub.1′, and X.sub.1 are as defined in claim 6; in Formula (IB-1ac) and Formula (IB-1ad), R.sub.12′ is independently halogen, —C.sub.1-3 alkyl, -halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-6 alkoxy, —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkyl, or —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkoxy; and R.sub.1, R.sub.2, R.sub.3, R.sub.21, R.sub.22, R.sub.31, R.sub.32, P′, R.sub.0′, Ar′, E.sub.1′, and X.sub.1 are as defined in claim 6.
8. The compound according to claim 4, or a tautomer, a cis-trans isomer, a mesomer, a raceme, an enantiomer, a diastereoisomer, an atropisomer or a mixture thereof, or a pharmaceutically acceptable salt, a solvate or a prodrug thereof, wherein the compound of Formula (IB) is a compound of Formula (IB-1c) or a compound of Formula (IB-2c): ##STR01354## wherein R.sub.1, R.sub.2, R.sub.3, R.sub.21, R.sub.22, R.sub.12, R.sub.11, R.sub.31, R.sub.32, P′, R.sub.0′, Ar′, E.sub.1′, and X.sub.1 are as defined in claim 4.
9. The compound according to claim 8, or a tautomer, a cis-trans isomer, a mesomer, a raceme, an enantiomer, a diastereoisomer, an atropisomer or a mixture thereof, or a pharmaceutically acceptable salt, a solvate or a prodrug thereof, wherein the compound of Formula (IB-1c) is a compound of Formula (IB-1ca), a compound of Formula (IB-1cb), a compound of Formula (IB-1cc), or a compound of Formula (IB-1cd): ##STR01355## in Formula (IB-1ca) and Formula (IB-1cb), R.sub.21′ is independently halogen, —C.sub.1-3alkyl, -halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-6 alkoxy, —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkyl, or —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkoxy; and R.sub.1, R.sub.2, R.sub.3, R.sub.12, R.sub.11, R.sub.31, R.sub.32, P′, R.sub.0′, Ar′, E.sub.1′, and X.sub.1 are as defined in claim 8; in Formula (IB-1cc) and Formula (IB-1cd), R.sub.12′ is independently halogen, —C.sub.1-3 alkyl, -halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-6alkoxy, —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkyl, or —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkoxy; and R.sub.1, R.sub.2, R.sub.3, R.sub.21, R.sub.22, R.sub.31, R.sub.32, P′, R.sub.0′, Ar′, E.sub.1′, and X.sub.1 are as defined in claim 8.
10. The compound according to claim 4, or a tautomer, a cis-trans isomer, a mesomer, a raceme, an enantiomer, a diastereoisomer, an atropisomer or a mixture thereof, or a pharmaceutically acceptable salt, a solvate or a prodrug thereof, wherein the compound of Formula (IB) is a compound of Formula (IB-1b) or a compound of Formula (IB-2b): ##STR01356## wherein R.sub.1, R.sub.2, R.sub.3, R.sub.21, R.sub.22, R.sub.12, R.sub.11, R.sub.31, R.sub.32, P′, R.sub.0′, Ar′, E.sub.1′, and X.sub.1 are as defined in claim 4.
11. The compound according to claim 10, or a tautomer, a cis-trans isomer, a mesomer, a raceme, an enantiomer, a diastereoisomer, an atropisomer or a mixture thereof, or a pharmaceutically acceptable salt, a solvate or a prodrug thereof, wherein the compound of Formula (IB-1b) is a compound of Formula (IB-1ba), a compound of Formula (IB-1bb), a compound of Formula (IB-1bc), or a compound of Formula (IB-1bd): ##STR01357## in Formula (IB-1ba) and Formula (IB-1bb), R.sub.21′ is independently halogen, —C.sub.1-3 alkyl, -halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-6 alkoxy, —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkyl, or —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkoxy; and R.sub.1, R.sub.2, R.sub.3, R.sub.12, R.sub.11, R.sub.31, R.sub.32, P′, R.sub.0′, Ar′, E.sub.1′, and X.sub.1 are as defined in claim 10; in Formula (IB-1bc) and Formula (IB-1bd), R.sub.12′ is independently halogen, —C.sub.1-3 alkyl, -halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-6 alkoxy, —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkyl, or —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkoxy; and R.sub.1, R.sub.2, R.sub.3, R.sub.21, R.sub.22, R.sub.31, R.sub.32, P′, R.sub.0′, Ar′, E.sub.1′, and X.sub.1 are as defined in claim 10.
12. The compound according to claim 4, or a tautomer, a cis-trans isomer, a mesomer, a raceme, an enantiomer, a diastereoisomer, an atropisomer or a mixture thereof, or a pharmaceutically acceptable salt, a solvate or a prodrug thereof, wherein the compound of Formula (IB) is a compound of Formula (IB-1d) or a compound of Formula (IB-2d): ##STR01358## wherein R.sub.1, R.sub.2, R.sub.3, R.sub.21, R.sub.22, R.sub.12, R.sub.11, R.sub.31, R.sub.32, P′, R.sub.0′, Ar′, E.sub.1′, and X.sub.1 are as defined in claim 4.
13. The compound according to claim 12, or a tautomer, a cis-trans isomer, a mesomer, a raceme, an enantiomer, a diastereoisomer, an atropisomer or a mixture thereof, or a pharmaceutically acceptable salt, a solvate or a prodrug thereof, wherein the compound of Formula (IB-1d) is a compound of Formula (IB-1da), a compound of Formula (IB-1db), a compound of Formula (IB-1dc), or a compound of Formula (IB-1dd): ##STR01359## in Formula (IB-1da) and Formula (IB-1db), R.sub.21′ is independently halogen, —C.sub.1-3 alkyl, -halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-6 alkoxy, —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkyl, or —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkoxy; and R.sub.1, R.sub.2, R.sub.3, R.sub.12, R.sub.11, R.sub.31, R.sub.32, P′, R.sub.0′, Ar′, E.sub.1′, and X.sub.1 are as defined in claim 12; in Formula (IB-1dc) and Formula (IB-1dd), R.sub.12′ is independently halogen, —C.sub.1-3 alkyl, -halogenated C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, —C.sub.1-3 alkyl-C.sub.1-6 alkoxy, —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkyl, or —C.sub.1-3 alkyl-halogenated C.sub.1-6 alkoxy; and R.sub.1, R.sub.2, R.sub.3, R.sub.21, R.sub.22, R.sub.31, R.sub.32, P′, R.sub.0′, Ar′, E.sub.1′, and X.sub.1 are as defined in claim 12.
14. The compound according to claim 7, or a tautomer, a cis-trans isomer, a mesomer, a raceme, an enantiomer, a diastereoisomer, an atropisomer or a mixture thereof, or a pharmaceutically acceptable salt, a solvate or a prodrug thereof, wherein R.sub.21′ and R.sub.12′ are each independently —C.sub.1-3 alkyl, —C.sub.1-3 alkyl-hydroxyl, —C.sub.1-3 alkyl-cyano, or —C.sub.1-3 alkyl-C.sub.1-6 alkoxy.
15. The compound according to claim 3, or a tautomer, a cis-trans isomer, a mesomer, a raceme, an enantiomer, a diastereoisomer, an atropisomer or a mixture thereof, or a pharmaceutically acceptable salt, a solvate or a prodrug thereof, wherein X.sub.1 is hydrogen, halogen, -substituted or unsubstituted C.sub.1-6 alkyl, -substituted or unsubstituted C.sub.3-6 cycloalkyl, or —O-substituted or unsubstituted C.sub.1-6 alkyl; and the “substituted” means 1, 2, 3 or 4 hydrogen atoms in a group being each independently substituted by a group S substituent.
16. The compound according to claim 3, or a tautomer, a cis-trans isomer, a mesomer, a raceme, an enantiomer, a diastereoisomer, an atropisomer or a mixture thereof, or a pharmaceutically acceptable salt, a solvate or a prodrug thereof, wherein Ar′ is phenyl, 5- or 6-membered monocyclic heteroaryl, or pyridonyl; and the phenyl, the 5- or 6-membered monocyclic heteroaryl, and the pyridonyl are unsubstituted or each substituted by 1, 2, 3, or 4 groups each independently selected from the group consisting of halogen, cyano, hydroxyl, —C.sub.1-6 alkyl, —C.sub.1-6 alkoxy, —NR.sup.cR.sup.d, and —C.sub.1-4 alkyl-NR.sup.eR.sup.f, wherein R.sup.e and R.sup.f are each independently hydrogen or C.sub.1-3 alkyl; and R.sup.c and R.sup.d are each independently hydrogen, —C.sub.1-3 alkyl, —C(O)C.sub.1-3 alkyl, or —CO.sub.2C.sub.1-3 alkyl.
17. The compound according to claim 3, or a tautomer, a cis-trans isomer, a mesomer, a raceme, an enantiomer, a diastereoisomer, an atropisomer or a mixture thereof, or a pharmaceutically acceptable salt, a solvate or a prodrug thereof, wherein Ar′ has a structure selected from: ##STR01360## wherein R.sub.s1 and R.sub.s2 are as defined in claim 3.
18. The compound according to claim 3, or a tautomer, a cis-trans isomer, a mesomer, a raceme, an enantiomer, a diastereoisomer, an atropisomer or a mixture thereof, or a pharmaceutically acceptable salt, a solvate or a prodrug thereof, wherein R.sub.0′ is phenyl, 5- or 6-membered monocyclic heteroaryl, or pyridonyl, wherein the 5- or 6-membered monocyclic heteroaryl has 1, 2 or 3 heteroatoms selected from the group consisting of N, O, and S as ring atoms; and the phenyl, the 5- or 6-membered monocyclic heteroaryl, and the pyridonyl are unsubstituted or each substituted by 1, 2, 3, or 4 groups each independently selected from R.sub.s3.
19. The compound according to claim 3, or a tautomer, a cis-trans isomer, a mesomer, a raceme, an enantiomer, a diastereoisomer, an atropisomer or a mixture thereof, or a pharmaceutically acceptable salt, a solvate or a prodrug thereof, wherein R.sub.1 and R.sub.2 are each independently hydrogen, halogen, cyano, amino, NHCH.sub.3, N(CH.sub.3).sub.2, methyl, ethyl, n-propyl, isopropyl, monochloromethyl, dichloromethyl, trichloromethyl, monochloroethyl, 1,2-dichloroethyl, trichloroethyl, monobromoethyl, monofluoromethyl, difluoromethyl, trifluoromethyl, monofluoroethyl, difluoroethyl, trifluoroethyl, —CH.sub.2-hydroxyl, —CH.sub.2-cyano, —CH.sub.2-methoxy, —CH.sub.2-ethoxy, —CH.sub.2-propoxy, —CH.sub.2-isopropoxy, —CH.sub.2—NH.sub.2, —CH.sub.2—NHCH.sub.3, —CH.sub.2—N(CH.sub.3).sub.2, —CH.sub.2-3- to 6-membered heterocycloalkyl, or —CH.sub.2-5- or 6-membered monocyclic heteroaryl; the 3- to 6-membered heterocycloalkyl is selected from the group consisting of aziridine, ethylene oxide, azetidine, oxetane, tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, piperidine, piperazine, morpholine, thiomorpholine, thiomorpholin-1,1-dioxide, and tetrahydropyrane; the 5- or 6-membered monocyclic heteroaryl is selected from the group consisting of thiophene, N-alkylcyclopyrrole, furan, thiazole, isothiazole, imidazole, oxazole, pyrrole, pyrazole, triazole, 1,2,3-triazole, 1,2,4-triazole, 1,2,5-triazole, 1,3,4-triazole, tetrazole, isoxazole, oxadiazole, 1,2,3-oxadiazole, 1,2,4-oxadiazole, 1,2,5-oxadiazole, 1,3,4-oxadiazole, thiadiazole, pyridine, pyridazine, pyrimidine, and pyrazine; and the 3- to 6-membered heterocycloalkyl and the 5- or 6-membered monocyclic heteroaryl are each optionally substituted by 1 or 2 halogens or C.sub.1-3 alkyl.
20. The compound according to claim 3, or a tautomer, a cis-trans isomer, a mesomer, a raceme, an enantiomer, a diastereoisomer, an atropisomer or a mixture thereof, or a pharmaceutically acceptable salt, a solvate or a prodrug thereof, wherein R.sub.3 is hydrogen, halogen, methoxy, ethoxy, propoxy, or isopropoxy.
21. The compound according to claim 3, or a tautomer, a cis-trans isomer, a mesomer, a raceme, an enantiomer, a diastereoisomer, an atropisomer or a mixture thereof, or a pharmaceutically acceptable salt, a solvate or a prodrug thereof, wherein R.sub.4 is hydrogen, monochloromethyl, dichloromethyl, trichloromethyl, monochloroethyl, 1,2-dichloroethyl, trichloroethyl, monobromoethyl, monofluoromethyl, difluoromethyl, trifluoromethyl, monofluoroethyl, difluoroethyl, trifluoroethyl, —CH.sub.2-hydroxyl, —CH.sub.2-cyano, —CH.sub.2-methoxy, —CH.sub.2-ethoxy, —CH.sub.2-propoxy, or —CH.sub.2-isopropoxy.
22. The compound according to claim 3, or a tautomer, a cis-trans isomer, a mesomer, a raceme, an enantiomer, a diastereoisomer, an atropisomer or a mixture thereof, or a pharmaceutically acceptable salt, a solvate or a prodrug thereof, wherein P′ is O, NH, or NR.sup.m′; R.sup.m′ is —C.sub.1-6 deuteroalkyl or —C.sub.1-6 alkyl; R.sub.42′ is —C.sub.1-3 alkyl-(C═O)—, —(C═O)—, or —C.sub.1-3 alkyl-.
23. The compound according to claim 3, or a tautomer, a cis-trans isomer, a mesomer, a raceme, an enantiomer, a diastereoisomer, an atropisomer or a mixture thereof, or a pharmaceutically acceptable salt, a solvate or a prodrug thereof, wherein the compound of Formula (IA) is selected from the following: ##STR01361## ##STR01362## ##STR01363## ##STR01364## ##STR01365## ##STR01366## ##STR01367## ##STR01368## ##STR01369## ##STR01370## ##STR01371## ##STR01372## ##STR01373## ##STR01374## ##STR01375## ##STR01376## ##STR01377## ##STR01378## ##STR01379## ##STR01380## ##STR01381## ##STR01382## ##STR01383## ##STR01384## ##STR01385## ##STR01386## ##STR01387## ##STR01388## ##STR01389## ##STR01390## ##STR01391## ##STR01392## ##STR01393## ##STR01394## ##STR01395## ##STR01396## ##STR01397## ##STR01398## ##STR01399## ##STR01400## ##STR01401## ##STR01402## ##STR01403## ##STR01404## ##STR01405## ##STR01406## ##STR01407## ##STR01408## ##STR01409## ##STR01410## ##STR01411## ##STR01412## ##STR01413## ##STR01414## ##STR01415## ##STR01416## ##STR01417## ##STR01418## ##STR01419## ##STR01420## ##STR01421## ##STR01422## ##STR01423## ##STR01424## ##STR01425## ##STR01426## ##STR01427## ##STR01428## ##STR01429## ##STR01430## ##STR01431## ##STR01432## ##STR01433## ##STR01434## ##STR01435## ##STR01436## ##STR01437## ##STR01438## ##STR01439## ##STR01440## ##STR01441## ##STR01442## ##STR01443## ##STR01444## ##STR01445## ##STR01446## ##STR01447## ##STR01448## ##STR01449## ##STR01450## ##STR01451## ##STR01452## ##STR01453## ##STR01454## ##STR01455## ##STR01456## ##STR01457## ##STR01458## ##STR01459## ##STR01460## ##STR01461## ##STR01462## ##STR01463## ##STR01464## ##STR01465## ##STR01466## ##STR01467## ##STR01468## ##STR01469##
24. The compound according to claim 3, or a tautomer, a cis-trans isomer, a mesomer, a raceme, an enantiomer, a diastereoisomer, an atropisomer or a mixture thereof, or a pharmaceutically acceptable salt, a solvate or a prodrug thereof, wherein the compound of Formula (IA) is selected from the following: ##STR01470## ##STR01471## ##STR01472## ##STR01473## ##STR01474## ##STR01475## ##STR01476## ##STR01477## ##STR01478## ##STR01479## ##STR01480## ##STR01481##
25. A pharmaceutical composition, comprising the compound according to claim 1, or a tautomer, a cis-trans isomer, a mesomer, a raceme, an enantiomer, a diastereoisomer, an atropisomer or a mixture thereof, or a pharmaceutically acceptable salt, a solvate or a prodrug thereof, and a pharmaceutically acceptable carrier.
26. A method of use of the compound according to claim 1, or a tautomer, a cis-trans isomer, a mesomer, a raceme, an enantiomer, a diastereoisomer, an atropisomer or a mixture thereof, or a pharmaceutically acceptable salt, a solvate or a prodrug thereof, or a pharmaceutical composition comprising the compound according to claim 1, or the tautomer, the cis-trans isomer, the mesomer, the raceme, the enantiomer, the diastereoisomer, the atropisomer or the mixture thereof, or the pharmaceutically acceptable salt, the solvate or the prodrug thereof, and a pharmaceutically acceptable carrier in preparing a drug for preventing and/or treating a KRAS G12C mutation-induced disease.
27. A method of use of the compound according to claim 1, or a tautomer, a cis-trans isomer, a mesomer, a raceme, an enantiomer, a diastereoisomer, an atropisomer or a mixture thereof, or a pharmaceutically acceptable salt, a solvate or a prodrug thereof, or a pharmaceutical composition comprising the compound according to claim 1, or the tautomer, the cis-trans isomer, the mesomer, the raceme, the enantiomer, the diastereoisomer, the atropisomer or the mixture thereof, or the pharmaceutically acceptable salt, the solvate or the prodrug thereof, and a pharmaceutically acceptable carrier in preparing an inhibitor for a KRAS mutation, wherein the KRAS mutation is KRAS G12C mutation.
28. A pharmaceutical composition, comprising the compound according to claim 3, or a tautomer, a cis-trans isomer, a mesomer, a raceme, an enantiomer, a diastereoisomer, an atropisomer or a mixture thereof, or a pharmaceutically acceptable salt, a solvate or a prodrug thereof, and a pharmaceutically acceptable carrier.
29. A method of use of the compound according to claim 3, or a tautomer, a cis-trans isomer, a mesomer, a raceme, an enantiomer, a diastereoisomer, an atropisomer or a mixture thereof, or a pharmaceutically acceptable salt, a solvate or a prodrug thereof, or a pharmaceutical composition comprising the compound according to claim 3, or the tautomer, the cis-trans isomer, the mesomer, the raceme, the enantiomer, the diastereoisomer, the atropisomer or the mixture thereof, or the pharmaceutically acceptable salt, the solvate or the prodrug thereof, and a pharmaceutically acceptable carrier in preparing a drug for preventing and/or treating a KRAS G12C mutation-induced disease.
30. A method of use of the compound according to claim 3, or a tautomer, a cis-trans isomer, a mesomer, a raceme, an enantiomer, a diastereoisomer, an atropisomer or a mixture thereof, or a pharmaceutically acceptable salt, a solvate or a prodrug thereof, or a pharmaceutical composition comprising the compound according to claim 3, or the tautomer, the cis-trans isomer, the mesomer, the raceme, the enantiomer, the diastereoisomer, the atropisomer or the mixture thereof, or the pharmaceutically acceptable salt, the solvate or the prodrug thereof, and a pharmaceutically acceptable carrier in preparing an inhibitor for a KRAS mutation, wherein the KRAS mutation is KRAS G12C mutation.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
[0257]
[0258]
DETAILED DESCRIPTION OF THE EMBODIMENTS
[0259] The compounds of the present invention can be prepared by a plurality of synthesis methods well known to those skilled in the art, including specific embodiments listed below, embodiments derived therefrom in combination with other chemical synthesis methods, and equivalent replacements well known to those skilled in the art. Preferred embodiments include but are not limited to the Examples of the present invention. The present invention will be described in detail below with reference to Examples, which, however, do not constitute any unfavorable limitation to the present invention. The present invention has been described in detail herein, and the specific embodiments thereof are also disclosed. It will be obvious for those skilled in the art that various changes and improvements can be made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention. Where specific conditions are not indicated in Examples, conventional conditions or the conditions suggested by the manufacturer are adopted. For the used reagents or instruments that are not marked with the manufacturers, they are all conventional products that are commercially available.
[0260] List of abbreviations of reagents used in the following Examples: THF: tetrahydrofuran; DMSO: dimethyl sulfoxide; PE: petroleum ether; EtOAc: ethyl acetate; DCM: dichloromethane; MeOH: methanol; ACN: acetonitrile; IPA: isopropylamine; DMA: dimethylamine; TFA: trifluoroacetic acid; NH.sub.4Cl: ammonium chloride; SPhos: 2-dicyclohexylphosphino-2′,6′-dimethoxy-1,1′-biphenyl; SPhos-Pd-G2: chloro(2-dicyclohexylphosphino-2′,6′-dimethoxy-1,1′-biphenyl)[2-(2′-amino-1,1′-biphenyl)]palladium (II); NaHMDS: sodium bis(trimethylsilyl)amide; and LiHMDS: lithium bis(trimethylsilyl)amide.
[0261] For the preparative high-performance liquid chromatography (HPLC) used in the following Examples, the following conditions may be adopted: column type: Waters XBridge C18, 190*250 mm, 5 μm; mobile phase system: A: aqueous solution of 0.1% ammonium bicarbonate; B: preparative grade ACN; flow rate: 15 ml/min; B %=20%-100%; and column temperature: Room temperature.
[0262] If isomer compounds are tested by analytical scale HPLC, the following conditions may be adopted: column type: XBridge C18, 3.5 μm 4.6*150 mm; mobile phase: A: purified water (0.05% TFA); B: preparative grade ACN (0.05% TFA), gradient: 5%-95% B; run time: 15 min; flow rate: 1 ml/min; and column temperature=40° C.
Example 1 Preparation of Compounds Z1, Z1-1, and Z1-2
[0263] ##STR00476##
[0264] Step 1: 2-isopropyl-4-methylpyridin-3-amine (582 mg, 3.88 mmol) was dissolved in THF (20 mL), cooled to 0° C., added dropwise with NaHMDS (5.8 mL, 11.60 mmol, 2M in THF), stirred for 15 minutes to react, and then added dropwise with a solution of 2,5-difluoro-6-(2-fluoro-6-methoxyphenyl)nicotinic acid (1.0 g, 3.53 mmol) in THF (6 mL). The resulting mixture was stirred at room temperature for 3 hours to react. The resulting reaction liquid was poured into 30 mL of saturated NH.sub.4Cl. The reaction liquid was extracted with 40 mL of EtOAc for 3 times. The resulting organic phase was dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (0-5% MeOH/DCM) to obtain product 5-fluoro-6-(2-fluoro-6-methoxyphenyl)-2-((2-isopropyl-4-methylpyridin-3-yl)amino)nicotinic acid (850 mg, Y:58.2%), which was yellow solid. ES-API: [M+H].sup.+=414.1.
[0265] Step 2: 5-fluoro-6-(2-fluoro-6-methoxyphenyl)-2-((2-isopropyl-4-methylpyridin-3-yl)amino)nicotinic acid (700 mg, 1.69 mmol) was dissolved in 1,2-dichloroethane (15 mL), added with SOCl.sub.2 (2.0 g, 16.90 mmol), and stirred at 80° C. for 2 hours to react. By concentration after the reaction, product 5-fluoro-6-(2-fluoro-6-methoxyphenyl)-2-((2-isopropyl-4-methylpyridin-3-yl)amino)nicotinoyl chloride (721 mg, Y:100%) was obtained, which was directly used in next step without being purified.
[0266] Step 3: a solution of ethyl nitroacetate (449 mg, 3.38 mmol) in THF (2 ml) was added dropwise to a THF (25 mL) suspension containing NaH (608 mg, 15.21 mmol) at 0° C., stirred at the temperature of 0° C. for half an hour to react, and then added dropwise with a solution of 5-fluoro-6-(2-fluoro-6-methoxyphenyl)-2-((2-isopropyl-4-methylpyridin-3-yl)amino)nicotinoyl chloride (721 mg, 1.69 mmol) in THF (15 mL). The ice bath was removed. The resulting reaction liquid was stirred at 70° C. overnight. The reaction liquid was poured into ice water, mixed with 3.0 M diluted hydrochloric acid such that the pH was adjusted to 3, and extracted with EtOAc for 3 times. The resulting organic phase was dried and concentrated to obtain product 6-fluoro-7-(2-fluoro-6-methoxyphenyl)-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2 (1H)-one (1.05 g, crude), which was directed used in next step. ES-API: [M+H].sup.+=483.1.
[0267] Step 4: 6-fluoro-7-(2-fluoro-6-methoxyphenyl)-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2 (1H)-one (1.05 g, 1.69 mmol) was dissolved in ACN (25 mL), then orderly added with POCl.sub.3 (1.30 g, 8.45 mmol) and N,N-diisopropylethylamine (1.74 g, 13.52 mmol), and stirred at 80° C. for 1 hour to react. The resulting reaction liquid was concentrated, added with EtOAc, and washed orderly with ice water, water, and saturated salt water. After the resulting organic phase was dried and concentrated, the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-50%) to obtain product 4-chloro-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (185 mg, Y:21.9%), which was yellow solid. ES-API: [M+H].sup.+=500.1.
[0268] Step 5: 4-chloro-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (175 mg, 0.35 mmol) was dissolved in DMF (6 mL), added with tert-butyl (R)-3-(hydroxymethyl)piperazin-1-carboxylate (454 mg, 2.10 mmol), and stirred at 80° C. for 18 hours to react. The resulting reaction liquid was poured into 30 mL of water. The reaction liquid was extracted with 20 mL of EtOAc for 3 times. The resulting organic phase was washed with saturated salt solution for 3 times, dried and concentrated, and the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-70%) to obtain product tert-butyl (3R)-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-3-(hydroxymethyl)piperazin-1-carboxylate (85 mg, Y:35.7%), which was yellow solid. ES-API: [M+H].sup.+=681.3.
[0269] Step 6: the tert-butyl (3R)-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-3-(hydroxymethyl)piperazin-1-carboxylate (73 mg, 0.11 mmol) was dissolved in DMA (4 mL), added with NaH (22 mg, 0.55 mmol), and stirred at 145° C. for 10 hours to react. Cooled reaction liquid was poured into 15 mL of water. The reaction liquid was extracted with 30 mL of EtOAc for 3 times. The resulting organic phase was washed with saturated salt solution for 3 times, dried and concentrated, and the resulting crude product was subjected to thin-layer chromatography on a think plate (DCM/MeOH=20:1) to obtain product tert-butyl (4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-7-oxo-1,2,4a,5,7,8-hexahydropyrazino[1′,2′:4,5][1,4]oxo-[2,3-c][1,8]naphthyridin-3(4H)-formate (35 mg, Y:51.5%), which was yellow solid. ES-API: [M+H].sup.+=634.2.
[0270] Step 7: the tert-butyl (4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-7-oxo-1,2,4a,5,7,8-hexahydropyrazino[1′,2′:4,5][1,4]oxo[−2,3-c][1,8]naphthyridin-3(4H)-formate (35 mg, 0.055 mmol) was dissolved in DCM (2.5 mL), and added with TFA (0.5 mL). The resulting mixture was stirred at room temperature for 0.5 hour, and the resulting reaction liquid was concentrated to obtain product (4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-1,2,3,4,4a,5-hexahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,8]naphthyridin-7(8H)-one (40 mg, crude), which was directly used in next step. ES-API: [M+H].sup.+=534.3.
[0271] Step 8: the (4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-1,2,3,4,4a,5-hexahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,8]naphthyridin-7(8H)-one (40 mg, 0.055 mmol) was dissolved in DCM (4 mL), and added with triethylamine (28 mg, 0.28 mmol). The resulting reaction liquid was cooled to 0° C., and added dropwise with a solution (0.5 mL) of acrylic anhydride (6 mg, 0.05 mmol) in DCM. The resulting mixture was stirred at 0° C. for 15 minutes to react. The resulting reaction liquid was added with 10 mL of saturated solution of NaHCO.sub.3 and extracted with 10 mL of DCM for 3 times. The resulting organic phase was dried and concentrated, and the resulting crude product was subjected to thin-layer chromatography on a plate (DCM/MeOH=10:1) to obtain product (4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-1,2,3,4,4a,5-hexahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,8]naphthyridin-7(8H)-one (17 mg, Y:52.4%), which was faint yellow solid. ES-API: [M+H].sup.+=588.2.
[0272] Step 9: the (4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-1,2,3,4,4a,5-hexahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,8]naphthyridin-7(8H)-one (15 mg, 0.025 mmol) was dissolved in DCM (1.5 mL). The resulting solution was cooled to 0° C., and then added dropwise with a solution (1 mL) of 17% boron tribromide in DCM. The resulting mixture was stirred at room temperature for 3 hours to react. The resulting reaction liquid was poured into 20 mL of saturated solution of NaHCO.sub.3 and extracted with 20 mL of DCM for 3 times. The resulting organic phase was dried and concentrated, and the resulting crude product was purified by preparative scale HPLC to obtain product (4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-hydroxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-1,2,3,4,4a,5-hexahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,8]naphthyridin-7(8H)-one (Z1, 10 mg, Y: 68.3%), which was white solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 9.96 (s, 1H), 8.35 (d, J=4.8 Hz, 1H), 7.19-7.14 (m, 2H), 6.87-6.75 (m, 1H), 6.64 (d, J=8.5 Hz, 1H), 6.59 (t, J=8.5 Hz, 1H), 6.12 (d, J=15.9 Hz, 1H), 5.71-5.67 (m, 1H), 4.39-3.95 (m, 4H), 3.79-3.30 (m, 4H), 3.06-2.98 (m, 1H), 2.56-2.29 (m, 1H), 1.80-1.73 (m, 3H), 0.99-0.95 (m, 3H), 0.85-0.80 (m, 3H). ES-API: [M+H].sup.+=574.2.
[0273] Step 10: the compound Z1 was resolved by preparative scale chiral HPLC (column type: Chiralpak IC: 10 μm, 20*250 mm; mobile phase: ACN:isopropanol:aminomethanol=70:30:0.2; flow rate: 15 ml/min; and column temperature: room temperature) to obtain the following atropisomer compounds. One of the atropisomer compounds had a structure arbitrarily specified as Z1-1 (75 mg, peak 1, retention time: 3.94 min, Y: 15.4%), which was faint yellow solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.04 (s, 1H), 8.42 (d, J=4.9 Hz, 1H), 8.22 (d, J=8.3 Hz, 1H), 7.28-7.20 (m, 2H), 6.96-6.81 (m, 1H), 6.75-6.58 (m, 2H), 6.18 (d, J=17.1 Hz, 1H), 5.82-5.69 (m, 1H), 4.49-4.00 (m, 4H), 3.90-3.43 (m, 4H), 3.08 (t, J=11.0 Hz, 1H), 2.64-2.55 (m, 1H), 1.80 (s, 3H), 1.05 (d, J=6.7 Hz, 3H), 0.91 (d, J=6.7 Hz, 3H). ES-API: [M+H].sup.+=574.2. The other atropisomer compound had a structure arbitrarily specified as Z1-2 (115 mg, peak 2, retention time: 5.04 min, Y: 23.6%), which was faint yellow solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.05 (s, 1H), 8.42 (d, J=4.8 Hz, 1H), 8.22 (d, J=6.9 Hz, 1H), 7.28-7.20 (m, 2H), 6.96-6.81 (m, 1H), 6.74-6.59 (m, 2H), 6.19 (d, J=16.7 Hz, 1H), 5.83-5.68 (m, 1H), 4.49-4.00 (m, 4H), 3.94-3.44 (m, 4H), 3.08 (t, J=11.0 Hz, 1H), 2.49-2.41 (m, 1H), 1.87 (s, 3H), 1.03 (dd, J=6.3, 3.7 Hz, 3H), 0.88 (d, J=6.6 Hz, 3H). ES-API: [M+H].sup.+=574.2. The isomer compounds were detected by analytical scale chiral HPLC (column type: Chiralpak IC: 5 m, 4.6*250 mm; mobile phase: ACN:isopropanol:aminomethanol=70:30:0.2; flow rate: 1 ml/min; and column temperature=30° C.).
Example 2 Preparation of Compound Z2
[0274] ##STR00477##
[0275] Step 1: at room temperature, 5-(methoxymethylene)-2,2-dimethyl-1,3-dioxan-4,6-dione (2.9 g, 15.62 mmol) and isopropanol (40 mL) were added to a 100 mL round-bottom flask, and 2-chloropyridin-3-amine (2.0 g, 15.62 mmol) was added thereto in batches. The resulting mixture was refluxed and stirred for 15 minutes to react. The resulting reaction liquid was cooled to room temperature. The precipitated solid was filtered. The filter cake was washed with a small amount of isopropanol and dried in vacuum to obtain product 5-((2-chloropyridin-3-yl)amino)methylene)-2,2-dimethyl-1,3-dioxan-4,6-dione (3.90 g, Y: 58.2%), which was white solid. ES-API: [M+H].sup.+=283.1.
[0276] Step 2: 200 mL of diphenyl ether was added to a 500 mL round-bottom flask, heated to 220° C., and the 5-((2-chloropyridin-3-yl)amino)methylene)-2,2-dimethyl-1,3-dioxan-4,6-dione (3.9 g, 13.83 mmol) was added thereto in batches, and stirred at 220° C. for 20 minutes to react. The resulting reaction liquid was cooled to room temperature and poured into PE. The precipitated solid was filtered. The filter cake was washed with PE and dried in vacuum to obtain product 8-chloro-1,7-naphthyridin-4-ol (1.5 g, Y: 60%), which was light brown solid. ES-API: [M+H].sup.+=181.0.
[0277] Step 3: the 8-chloro-1,7-naphthyridin-4-ol (500 mg, 2.78 mmol), sodium acetate (300 mg, 2.78 mmol), anhydrous ethanol (25 mL), and 5% Pd/C (250 mg) were added to a 50 mL round-bottom flask, and stirred at room temperature for 3 days to react with hydrogen supply from a hydrogen balloon. The resulting reaction liquid was filtered by using diatomite, and the filtrate was concentrated. The resulting crude product was purified by flash column chromatography on silica gel (DCM/MeOH: 0-10%) to obtain product 1,7-naphthyridin-4-ol (200 mg, Y: 49.3%), which was yellow solid. ES-API: [M+H].sup.+=147.1.
[0278] Step 4: the 1,7-naphthyridin-4-ol (550 mg, 3.77 mmol) was dissolved in concentrated sulfuric acid (4.5 mL), cooled to 0° C., slowly added dropwise with concentrated nitric acid (1.0 mL, 15.08 mmol), and stirred at 100° C. for 1 hour to react. Cooled reaction liquid was poured into ice water, and mixed with concentrated ammonia water such that the pH was adjusted to 6-7. The precipitated solid was filtered and dried in vacuum to obtain product 3-nitro-1,7-naphthyridin-4-ol (530 mg, Y: 73.7%), which was yellow solid. ES-API: [M+H].sup.+=192.1.
[0279] Step 5: the 3-nitro-1,7-naphthyridin-4-ol (480 mg, 2.51 mmol) and phosphorus oxychloride (4.68 mL, 50.20 mmol) were added to a 20 mL round-bottom flask, cooled to −15° C., slowly added dropwise with triethylamine (1.8 mL, 12.55 mmol), and stirred at room temperature for 1 hour to react. The resulting reaction liquid was poured into ice water, mixed with a cold saturated sodium bicarbonate solution such that the pH was adjusted to 8, and extracted with DCM for 3 times. The resulting organic phase was dried and concentrated to obtain product 4-chloro-3-nitro-1,7-naphthyridine (450 mg, Y: 85.7%), which was brown solid. ES-API: [M+H].sup.+=210.1.
[0280] Step 6: the 4-chloro-3-nitro-1,7-naphthyridine (450 mg, 2.15 mmol) was dissolved in 1,4-dioxane (15 mL), orderly added with tert-butyl (R)-3-(hydroxymethyl)piperazin-1-carboxylate (1.02 g, 4.73 mmol) and N,N-diisopropylethylamine (832 mg, 6.45 mmol), and stirred at 80° C. for 3 hours to react. The resulting reaction liquid was concentrated, and the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 50-100%) to obtain product tert-butyl (R)-3-(hydroxymethyl)-4-(3-nitro-1,7-naphthyridin-4-yl)piperazin-1-carboxylate (330 mg, Y: 39.4%), which was yellow solid. ES-API: [M+H].sup.+=390.2.
[0281] Step 7: the tert-butyl (R)-3-(hydroxymethyl)-4-(3-nitro-1,7-naphthyridin-4-yl)piperazin-1-carboxylate (310 mg, 0.80 mmol), DMF (18 mL), and NaH (96 mg, 2.40 mmol) were orderly added to a 50 mL sealing tube, and stirred at 95° C. for 3 days to react. Cooled reaction liquid was poured into water and extracted with EtOAc twice. The resulting organic phase was washed with saturated salt solution for 3 times, dried and concentrated, and the resulting crude product was subjected to thin-layer chromatography on a plate (DCM/MeOH=15:1) to obtain product tert-butyl (R)-8a,9,11,12-tetrahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,7]naphthyridin-10(8H)-carboxylate (175 mg, Y: 64%), which was yellow solid. ES-API: [M+H].sup.+=343.3.
[0282] Step 8: the tert-butyl (R)-8a,9,11,12-tetrahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,7]naphthyridin-10(8H)-carboxylate (100 mg, 0.29 mmol) was dissolved in acetic acid (4 mL), added with sodium cyanoborohydride (73 mg, 1.16 mmol), and stirred at room temperature overnight to react. The resulting reaction liquid was poured into ice water, mixed with a saturated sodium bicarbonate solution such that the pH was adjusted to 8, and extracted with DCM twice. The resulting organic phase was washed with saturated salt solution, dried and concentrated, and the resulting crude product was subjected to thin-layer chromatography on a plate (DCM/MeOH/ammonia water=100:8:1) to obtain product tert-butyl (R)-1,2,3,4,8a,9,11,12-octahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,7]naphthyridin-10(8H)-carboxylate (50 mg, Y: 49.4%), which was faint yellow solid. ES-API: [M+H].sup.+=390.2.
[0283] Step 9: the tert-butyl (R)-1,2,3,4,8a,9,11,12-octahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,7]naphthyridin-10(8H)-carboxylate (50 mg, 0.14 mmol), 4-bromo-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole (83 mg, 0.28 mmol), cesium carbonate (136 mg, 0.42 mmol), Pd.sub.2(dba).sub.3 (51 mg, 0.056 mmol), Ruphos (26 mg, 0.056 mmol) and toluene (6 mL) were added to a 5 mL microwave tube, subjected to nitrogen replacement, placed into a microwave reactor at 120° C., and stirred for 1 hour to react. The resulting reaction liquid was cooled to room temperature, and filtered. The filtrate was dried and concentrated. The resulting crude product was subjected to thin-layer chromatography on a plate (DCM/MeOH/ammonia water=100:5:1) to obtain tert-butyl (8aR)-3-(5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-1,2,3,4,8a,9,11,12-octahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,7]naphthyridin-10(8H)-carboxylate (60 mg, Y: 74.1%). ES-API: [M+H].sup.+=561.3.
[0284] Step 10: the tert-butyl (8aR)-3-(5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-1,2,3,4,8a,9,11,12-octahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,7]naphthyridin-10(8H)-carboxylate (60 mg, 0.11 mmol) was dissolved in DCM (3 mL), and added with TFA (0.8 mL). The resulting mixture was stirred at room temperature for 1 hour. The resulting reaction liquid was concentrated to obtain product (R)-3-(5-methyl-1H-indazol-4-yl)-1,2,3,4,8,8a,9,10,11,12-decahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,7]naphthyridine (60 mg, crude percent yield), which was directly used in next step without being purified. ES-API: [M+H].sup.+=377.1.
[0285] Step 11: the (R)-3-(5-methyl-1H-indazol-4-yl)-1,2,3,4,8,8a,9,10,11,12-decahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,7]naphthyridine (60 mg, 0.11 mmol) and N,N-diisopropylethylamine (71 mg, 0.55 mmol) were dissolved in DCM (5 mL), and cooled to 0° C. A solution (0.5 mL) of acrylic anhydride (13 mg, 0.10 mmol) in DCM was added dropwise to the resulting reaction liquid. The resulting mixture was stirred at 0° C. for 10 minutes to react. The resulting reaction liquid was added with 10 mL of saturated solution of NaHCO.sub.3 and extracted with 10 mL of DCM for 3 times. The resulting organic phase was dried and concentrated, and the resulting crude product was purified by preparative scale HPLC to obtain product (R)-1-(3-(5-methyl-1H-indazol-4-yl)-1,2,3,4,8a,9,11,12-octahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,7]naphthyridin-10(8H)-yl)propyl-2-en-1-one (Z2, 12 mg, Y: 26.0%), which was white solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 12.91 (s, 1H), 8.04 (s, 1H), 7.86 (s, 1H), 7.21-7.07 (m, 2H), 6.89-6.64 (m, 1H), 6.09 (d, J=16.6 Hz, 1H), 5.67 (m, 1H), 4.21-4.11 (m, 3H), 3.96 (t, J=10.0 Hz, 1H), 3.86-3.53 (m, 4H), 3.45-3.30 (m, 2H), 3.15-3.08 (m, 1H), 2.86-2.75 (m, 2H), 2.28 (s, 3H). ES-API: [M+H].sup.+=431.2.
Example 3 Preparation of Compounds Z3a and Z3
[0286] ##STR00478##
[0287] Step 1: 4-chloro-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (700 mg, 1.40 mmol) was dissolved in DMF (10 mL), added with tert-butyl (2R,5R)-5-(hydroxymethyl)-2-methylpiperazin-1-carboxylate (1.61 g, 7.0 mmol), and stirred at 80° C. for 1 hour to react. The resulting reaction liquid was poured into 30 mL of water. The reaction liquid was extracted with 20 mL of EtOAc for 3 times. The resulting organic phase was washed with saturated salt solution for 3 times, dried and concentrated, and the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-70%) to obtain product tert-butyl (2R,5R)-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-5-(hydroxymethyl)-2-methylpiperazin-1-carboxylate (325 mg, Y: 33.4%), which was yellow solid. ES-API: [M+H].sup.+=695.2.
[0288] Step 2: the tert-butyl (2R,5R)-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-5-(hydroxymethyl)-2-methylpyridin-1-carboxylate (300 mg, 0.44 mmol) was dissolved in DMA (20 mL), added with NaH (52 mg, 1.32 mmol), and stirred at 125° C. for 20 hours to react. Cooled reaction liquid was poured into 15 mL of water. The reaction liquid was extracted with 30 mL of EtOAc for 3 times. The resulting organic phase was washed with saturated salt solution for 8 times, dried and concentrated, and the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-100%) to obtain product tert-butyl (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2-methyl-7-oxo-1,2,4a,5,7,8-hexahydropyrazino[1′,2′:4,5][1,4]oxozino[2,3-c][1,8]naphthyridin-3(4H)-carboxylate (60 mg, Y:21.4%), which was yellow solid. ES-API: [M+H].sup.+=648.3.
[0289] Step 3: the tert-butyl (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2-methyl-7-oxo-1,2,4a,5,7,8-hexahydropyrazino[1′,2′:4,5][1,4]oxozino[2,3-c][1,8]naphthyridin-3(4H)-carboxylate (60 mg, 0.093 mmol) was dissolved in DCM (3 mL), and added with TFA (0.7 mL). The resulting mixture was stirred at room temperature for 1 hour, and the resulting reaction liquid was concentrated to obtain product (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2-methyl-1,2,3,4,4a,5-hexahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,8]naphthyridin-7(8H)-one (61 mg, crude), which was directly used in next step. ES-API: [M+H].sup.+=548.2.
[0290] Step 4: the (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2-methyl-1,2,3,4,4a,5-hexahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,8]naphthyridin-7(8H)-one (61 mg, 0.093 mmol) was dissolved in DCM (5 mL), and added with triethylamine (47 mg, 0.46 mmol). The resulting reaction liquid was cooled to 0° C., and added dropwise with a solution (1 mL) of acrylic anhydride (17 mg, 0.14 mmol) in DCM. The resulting mixture was stirred at 0° C. for 15 minutes to react. The resulting reaction liquid was added with 10 mL of saturated solution of NaHCO.sub.3 and extracted with 10 mL of DCM for 3 times. The resulting organic phase was dried and concentrated, and the resulting crude product was subjected to thin-layer chromatography on a plate (DCM/MeOH=10:1) to obtain product ((2R,4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2-methyl-1,2,3,4,4a,5-hexahydropyrazino[1′,2′:4,5][1,4]oxopyrazino[2,3-c][1,8]naphthyridin-7(8H)-one (Z3a, 32 mg, Y: 57.4%), which was white solid. ES-API: [M+H].sup.+=602.2. .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 8.42 (d, J=4.1 Hz, 1H), 7.87 (d, J=8.8 Hz, 1H), 7.43 (dd, J=15.4, 8.3 Hz, 1H), 7.21 (d, J=4.8 Hz, 1H), 6.99-6.78 (m, 3H), 6.17 (d, J=17.4 Hz, 1H), 5.75 (d, J=10.5 Hz, 1H), 4.80-4.15 (m, 4H), 3.94-3.35 (m, 6H), 3.13-2.97 (m, 1H), 2.62-2.40 (m, 1H), 1.90-1.73 (m, 3H), 1.66-1.48 (m, 3H), 1.08-0.95 (m, 3H), 0.91-0.77 (m, 3H).
[0291] Step 5: (6aR,9R)-8-acryloyl-3-fluoro-2-(2-fluoro-6-methoxyphenyl)-13-(2-isopropyl-4-methylpyridin-3-yl)-9-methyl-6,6a, 7,8,9,10-hexahydropyrazino[1′,2′:4,5][1,4]oxopyrazino[3,2-c][1,8]naphthyridin-12(13H)-one (32 mg, 0.053 mmol) was dissolved in DCM (1.5 mL). The resulting solution was cooled to 0° C., and then added dropwise with a solution (1 mL) of 17% boron tribromide in DCM. The resulting mixture was stirred at room temperature for 3 hours to react. The resulting reaction liquid was poured into 20 mL of saturated solution of NaHCO.sub.3 and extracted with 20 mL of DCM for 3 times. The resulting organic phase was dried and concentrated, and the resulting crude product was purified by preparative scale HPLC to obtain product (6aR,9R)-8-acryloyl-3-fluoro-2-(2-fluoro-6-hydroxyphenyl)-13-(2-isopropyl-4-methylpyridin-3-yl)-9-methyl-6,6a, 7,8,9,10-hexahydropyrazino[1′,2′:4,5][1,4]oxopyrazino[3,2-c][1,8]naphthyridin-12(13H)-one (Z3, 18 mg, Y: 57.6%), which was white solid. ES-API: [M+H].sup.+=588.3.
Example 6 Preparation of Compound Z6
[0292] ##STR00479##
[0293] Step 1: 4,6-dichloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (1.8 g, 3.48 mmol) was dissolved in DMF (15 mL), added with tert-butyl (R)-3-(hydroxymethyl)piperazin-1-carboxylate (3 g, 13.92 mmol), and stirred at 80° C. for 2 hours to react. The resulting reaction liquid was poured into 30 mL of water. The reaction liquid was extracted with 20 mL of EtOAc for 3 times. The resulting organic phase was washed with saturated salt solution for 3 times, dried and concentrated, and the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-70%) to obtain product tert-butyl (3R)-4-(6-chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-3-(hydroxymethyl)piperazin-1-formate (1.3 g, 54%), which was yellow solid. ES-API: [M+H].sup.+=697.2.
[0294] Step 2: the tert-butyl (3R)-4-(6-chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-3-(hydroxymethyl)piperazin-1-formate (1.3 g, 1.86 mmol) was dissolved in DMA (10 mL), added with LHMDS (5.6 mmol, 5.6 mmol, 1 M tetrahydrofuran solution), and stirred at 140° C. for 20 hours to react. Cooled reaction liquid was poured into 15 mL of water. The reaction liquid was extracted with 30 mL of EtOAc for 3 times. The resulting organic phase was washed with saturated salt solution for 3 times, dried and concentrated, and the resulting crude product was purified by flash column chromatography on silica gel (MeOH/DCM: 0-10%) to obtain tert-butyl (4aR)-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-7-oxo-1,2,4a,5,7,8-hexahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,8]naphthyridin-3(4H)-carboxylate (0.24 g, 20%), which was yellow solid. ES-API: [M+H].sup.+=650.2.
[0295] Step 3: the tert-butyl (4aR)-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-7-oxo-1,2,4a,5,7,8-hexahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,8]naphthyridin-3(4H)-carboxylate (240 mg, 0.37 mmol) was dissolved in DCM (2 mL), and added with TFA (2 mL). The resulting mixture was stirred at room temperature for 0.5 hour, and the resulting reaction liquid was concentrated to obtain product (4aR)-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-1,2,3,4,4a,5-hexahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,8]naphthyridin-7(8H)-one (203 mg, crude), which was directly used in next step. ES-API: [M+H].sup.+=550.1.
[0296] Step 4: the (4aR)-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-1,2,3,4,4a, 5-hexahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,8]naphthyridin-7(8H)-one (203 mg, 0.37 mmol) was dissolved in DCM (4 mL), and added with triethylamine (187 mg, 1.85 mmol). The resulting reaction liquid was cooled to 0° C., and added dropwise with a solution (0.5 mL) of acrylic anhydride (37 mg, 0.30 mmol) in DCM. The resulting mixture was stirred at 0° C. for 10 minutes to react. The resulting reaction liquid was added with 10 mL of saturated solution of NaHCO.sub.3 and extracted with 10 mL of DCM for 3 times. The resulting organic phase was dried and concentrated, and the resulting crude product was subjected to thin-layer chromatography on a plate (DCM/MeOH=10:1) to obtain product (4aR)-3-acryloyl-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-1,2,3,4,4a, 5-hexahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,8]naphthyridin-7(8H)-one (223 mg, crude), which was faint yellow solid. ES-API: [M+H].sup.+=604.2.
[0297] Step 5: the (4aR)-3-acryloyl-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-1,2,3,4,4a,5-hexahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,8]naphthyridin-7(8H)-one (223 mg, 0.37 mmol) was dissolved in DCM (1.5 mL). The resulting solution was cooled to 0° C., and then added dropwise with a solution (3 mL) of 17% boron tribromide in DCM. The resulting mixture was stirred at room temperature for 1 hour to react. The resulting reaction liquid was poured into 20 mL of saturated solution of NaHCO.sub.3 and extracted with 20 mL of DCM for 3 times. The resulting organic phase was dried and concentrated, and the resulting crude product was purified by preparative grade HPLC to obtain product (4aR)-3-acryloyl-11-chloro-10-(2-fluoro-6-hydroxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-1,2,3,4,4a,5-hexahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,8]naphthyridin-7(8H)-one (Z6, 26.28 mg, 11%), which was white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 8.42-8.36 (m, 2H), 7.22-7.18 (m, 2H), 6.68-6.62 (m, 3H), 6.22-6.17 (m, 1H), 5.78-5.77 (m, 1H), 4.46-3.55 (m, 8H), 3.12-3.10 (m, 1H), 2.52-2.51 (m, 1H), 1.88-1.80 (m, 3H), 1.06-1.04 (m, 3H), 0.89-0.86 (m, 3H). ES-API: [M+H].sup.+=590.2.
Example 9 Preparation of Compounds Z9, Z9-1, and Z9-2
[0298] ##STR00480##
[0299] Step 1: 4-chloro-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (500 mg, 1.00 mmol) was dissolved in N,N-dimethylacetamide (6 mL), orderly added with (R)-1-(tert-butyl)3-methyl-piperazin-1,3-dicarboxylate (732 mg, 3.00 mmol) and N,N-diisopropylethylamine (387 mg, 3.00 mmol), and stirred at 120° C. for 2 hours to react. The resulting reaction liquid was added with 100 mL of EtOAc, washed with 30 mL of dilute brine for 4 times and then washed with 30 mL of saturated salt solution, dried and concentrated, and the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-70%) to obtain product (3R)-1-(tert-butyl)-3-methyl-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazin-1,3-dicarboxylate (500 mg, Y: 70.6%), which was yellow solid. ES-API: [M+H].sup.+=709.2.
[0300] Step 2: the (3R)-1-(tert-butyl)-3-methyl-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazin-1,3-dicarboxylate (500 mg, 0.71 mmol) was dissolved in acetic acid (8 mL), added with iron powder (138 mg, 2.47 mmol), and stirred at 80° C. for 30 minutes to react. The resulting reaction liquid was concentrated, and orderly added with 50 mL of EtOAc and 30 mL of saturated sodium bicarbonate. The resulting suspension was filtered by using diatomite. The filter cake was washed with EtOAc. The resulting organic phase was separated, washed orderly with 30 mL of saturated solution of sodium bicarbonate and 30 mL of saturated salt solution, dried and concentrated to obtain product tert-butyl (4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (450 mg, Y: 98.6%), which was faint yellow solid. ES-API: [M+H].sup.+=647.2.
[0301] Step 3: the tert-butyl (4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (450 mg, 0.70 mmol), 12 mL of acetone, anhydrous potassium carbonate (290 mg, 2.10 mmol), and iodomethane (596 mg, 4.20 mmol) were orderly added to a 15 mL sealing tube. The sealing tube was sealed, and the resulting mixture was stirred at 50° C. for 20 hours to react. The resulting reaction liquid was concentrated, added with 60 mL of EtOAc, washed orderly with 30 mL of water and 30 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-70%) to obtain product tert-butyl (4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-6-methyl-5,7-dioxy-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (390 mg, Y: 84.8%), which was orange solid. ES-API: [M+H].sup.+=661.3.
[0302] Step 4: the tert-butyl (4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-6-methyl-5,7-dioxy-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (940 mg, 1.42 mmol) was dissolved in DCM (6 mL), and added with TFA (2 mL). After stirring at room temperature for 2 hours, the resulting reaction liquid was concentrated to obtain product (4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (1.1 g, crude), which was directly used in next step. ES-API: [M+H].sup.+=561.3.
[0303] Step 5: the (4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (1.1 g, crude) was dissolved in DCM (20 mL), and added with N,N-diisopropylethylamine (916 mg, 7.10 mmol). The resulting reaction liquid was cooled to 0° C., added with acryloyl chloride (256 mg, 2.84 mmol), and stirred at 0° C. for 15 minutes to react. The resulting reaction liquid was added with 30 mL of DCM, washed orderly with 15 mL of water, 15 mL of saturated solution of NaHCO.sub.3 and 15 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-100%) to obtain product (4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (780 mg, Y: 88.3%), which was faint yellow solid. ES-API: [M+H].sup.+=615.3.
[0304] Step 6: the (4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (390 mg, 0.64 mmol) was dissolved in DCM (9 mL). The resulting reaction liquid was cooled to 0° C., and then added dropwise with a solution (7 mL) of 17% boron tribromide in DCM. The resulting mixture was stirred at room temperature for 6 hours to react. The resulting reaction liquid was poured into 60 mL of saturated solution of NaHCO.sub.3 and extracted with 80 mL of DCM twice. The resulting organic phase was washed orderly with 50 mL of saturated solution of NaHCO.sub.3 and 80 mL of saturated salt solution, dried and concentrated to obtain product (4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-hydroxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8,8]naphthyridin-5,7-dione (Z9, 375 mg, Y: 98.4%), which was faint yellow solid. ES-API: [M+H].sup.+=601.2.
[0305] Step 7: the (4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-hydroxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8,8]naphthyridin-5,7-dione (750 mg, 1.25 mmol) was purified by preparative scale HPLC, and then resolved by preparative scale chiral HPLC (column type: IB:10 μm, 30*250 mm; mobile phase: hexane:EtOH=65:35; flow rate: 25 ml/min; and column temperature: room temperature) to obtain the following atropisomer compounds. One of the atropisomer compounds had a structure arbitrarily specified as Z9-1 (250 mg, peak 1, retention time: 6.463 min, Y: 33.3%), which was faint yellow solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.11 (d, J=1.3 Hz, 1H), 8.46-8.34 (m, 2H), 7.30-7.19 (m, 2H), 7.10-6.79 (m, 1H), 6.74-6.62 (m, 2H), 6.15 (d, J=16.9 Hz, 1H), 5.75 (d, J=12.0 Hz, 1H), 4.73 (d, J=13.3 Hz, 1H), 4.45 (d, J=12.7 Hz, 1H), 4.10-3.97 (m, 1H), 3.63-3.47 (m, 2H), 3.39-3.08 (m, 4H), 2.83-2.59 (m, 1H), 2.48-2.39 (m, 1H), 1.99 (s, 3H), 1.02 (d, J=6.7 Hz, 3H), 0.85 (d, J=6.7 Hz, 3H). ES-API: [M+H].sup.+=601.2. The other atropisomer compound had a structure arbitrarily specified as Z9-2 (350 mg, peak 2, retention time: 8.252 min, Y: 46.7%), which was faint yellow solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.14 (s, 1H), 8.44 (d, J=4.9 Hz, 1H), 8.38 (d, J=9.0 Hz, 1H), 7.29-7.20 (m, 2H), 7.10-6.79 (m, 1H), 6.76-6.59 (m, 2H), 6.15 (d, J=16.9 Hz, 1H), 5.75 (d, J=11.1 Hz, 1H), 4.73 (d, J=14.0 Hz, 1H), 4.45 (d, J=12.4 Hz, 1H), 4.02-3.89 (m, 1H), 3.62-3.50 (m, 2H), 3.33-3.09 (m, 4H), 2.88-2.61 (m, 2H), 1.79 (s, 3H), 1.10 (d, J=6.7 Hz, 3H), 0.98 (d, J=6.7 Hz, 3H). ES-API: [M+H].sup.+=601.2. The isomer compounds were detected by analytical scale chiral HPLC (column type: IB:5 m, 4.6*250 mm; mobile phase: hexane:EtOH=65:35; flow rate: 1 ml/min; and column temperature=30° C.).
Example 10 Preparation of Compounds Z10, Z10-1, and Z10-2
[0306] ##STR00481##
[0307] Step 1: tert-butyl (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2-methyl-5,7-dioxo-1,2, 4,4a,5,6,7,8-octyl-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (800 mg, 1.21 mmol), 20 mL of acetone, anhydrous potassium carbonate (500 mg, 3.63 mmol), and iodomethane (1.03 g, 7.26 mmol) were orderly added to a 50 mL sealing tube. The sealing tube was sealed, and the resulting mixture was stirred at 50° C. for 18 hours to react. The resulting reaction liquid was concentrated, added with 60 mL of EtOAc, washed orderly with 20 mL of water and 30 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-70%) to obtain product tert-butyl (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,6-dimethyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octyl-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (790 mg, Y: 96.7%), which was orange solid. ES-API: [M+H].sup.+=675.3.
[0308] Step 2: the tert-butyl (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,6-dimethyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octyl-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (790 mg, 1.42 mmol) was dissolved in DCM (6 mL), and added with TFA (2 mL). After stirring at room temperature for 2 hours, the resulting reaction liquid was concentrated to obtain product (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (810 mg, crude), which was directly used in next step. ES-API: [M+H].sup.+=575.2.
[0309] Step 3: the (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (810 mg, crude) was dissolved in DCM (15 mL), and added with N,N-diisopropylethylamine (755 mg, 5.85 mmol). The resulting reaction liquid was cooled to 0° C., added with acryloyl chloride (211 mg, 2.34 mmol), and stirred at 0° C. for 15 minutes to react. The resulting reaction liquid was added with 50 mL of DCM, washed orderly with 20 mL of water, 40 mL of saturated solution of NaHCO.sub.3 and 20 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-100%) to obtain product (2R,4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (670 mg, Y: 91.0%), which was faint yellow solid. ES-API: [M+H].sup.+=629.2.
[0310] Step 4: the (2R,4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (370 mg, 0.59 mmol) was dissolved in DCM (8 mL). The resulting reaction liquid was cooled to 0° C., and then added dropwise with a solution (7 mL) of 17% boron tribromide in DCM. The resulting mixture was stirred at room temperature for 3 hours to react. The resulting reaction liquid was poured into 60 mL of saturated solution of NaHCO.sub.3 and extracted with 80 mL of DCM twice. The resulting organic phase was washed orderly with 50 mL of saturated solution of NaHCO.sub.3 and 80 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by preparative scale HPLC to obtain product (2R,4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-hydroxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,6-dimethyl-2,3,4,4a,6-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8,8]naphthyridin-5-dione (Z10, 249 mg, Y: 68.7%), which was faint yellow solid. ES-API: [M+H].sup.+=615.2.
[0311] Step 5: the compound Z10 (450 mg, 1.06 mmol) was resolved by preparative scale chiral HPLC (column type: OD-H: 10 μm, 20*250 mm; mobile phase: hexane:EtOH=80:20; flow rate: 15 ml/min; and column temperature: room temperature) to obtain the following atropisomer compounds. One of the atropisomer compounds had a structure arbitrarily specified as Z10-1 (206 mg, peak 1, retention time: 8.321 min, Y: 45.7%), which was faint yellow solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.13 (d, J=1.3 Hz, 1H), 8.44 (d, J=4.9 Hz, 1H), 8.02-7.95 (m, 1H), 7.33-7.20 (m, 2H), 7.06-6.82 (m, 1H), 6.76-6.63 (m, 2H), 6.24-6.08 (m, 1H), 5.82-5.67 (m, 1H), 5.05-4.73 (m, 1H), 4.63-4.37 (m, 1H), 4.07-3.97 (m, 1H), 3.73 (dd, J=14.1, 4.2 Hz, 1H), 3.39-3.20 (m, 4H), 2.94-2.78 (m, 1H), 2.49-2.39 (m, 1H), 1.99 (s, 3H), 1.61-1.49 (m, 3H), 1.02 (d, J=6.7 Hz, 3H), 0.85 (d, J=6.7 Hz, 3H). ES-API: [M+H].sup.+=615.2. The other atropisomer compound had a structure arbitrarily specified as Z10-2 (209 mg, peak 2, retention time: 10.183 min, Y: 46.4%), which was faint yellow solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.15 (s, 1H), 8.45 (d, J=4.9 Hz, 1H), 8.03-7.95 (m, 1H), 7.30-7.18 (m, 2H), 7.06-6.82 (m, 1H), 6.75-6.61 (m, 2H), 6.21-6.09 (m, 1H), 5.80-5.65 (m, 1H), 5.05-4.72 (m, 1H), 4.63-4.37 (m, 1H), 4.01-3.92 (m, 1H), 3.74 (dd, J=14.2, 4.2 Hz, 1H), 3.43-3.21 (m, 4H), 2.95-2.82 (m, 1H), 2.80-2.72 (m, 1H), 1.80 (s, 3H), 1.60-1.48 (m, 3H), 1.11 (d, J=6.7 Hz, 3H), 0.98 (d, J=6.7 Hz, 3H). ES-API: [M+H].sup.+=615.2. The isomer compounds were detected by analytical scale chiral HPLC (column type: OD-H: 5 m, 4.6*250 mm; mobile phase: hexane:EtOH=80:20; flow rate: 1 ml/min; and column temperature=30° C.).
Examples 4-5, 7-8, and 11-20
[0312] Compounds Z4-Z5, Z7-Z8, and Z11-Z20 were prepared by similar methods to that for the compound Z1 or Z2, where starting materials for different compounds were commercially available or prepared by existing methods well known to those skilled in the art. Furthermore, it would be easy for a person skilled in the art to synthesize intermediates with reference to existing similar methods.
TABLE-US-00003 Ex- am- Com- ple pound MS No. No. Compound Structure [M + H] 4 Z4
Example 21 Preparation of Compounds Z21, Z21-1 and Z21-2
[0313] ##STR00496## ##STR00497## ##STR00498## ##STR00499##
[0314] Step 1: 7-chloro-6-fluoro-4-hydroxy-1-(4-isopropyl-6-methylpyrimidin-5-yl)-2-oxo-1,2-dihydro-1,8-naphthyridin-3-nitrile (2 g, 5.34 mmol), 12 mL of water, and 12 mL of dioxane were added to a round-bottom flask. The resulting system was cooled to 0° C., and 12 mL of concentrated sulfuric acid was added dropwise to the resulting reaction liquid. After the completion of dropwise addition, the resulting mixture was stirred at 120° C. for 18 hours to react. After the completion of the reaction, a large amount of solid was precipitated. The resulting reaction mixture was filtered, and the filter cake was washed with water for 3 times. The filter cake was dried to obtain 7-chloro-6-fluoro-4-hydroxy-1-(4-isopropyl-6-methylpyrimidin-5-yl)-1,8-naphthyridin-2(1H)-one (1.4 g, 75%), which was white solid. The resulting crude product was directly used in next step. ES-API: [M+H].sup.+=349.1.
[0315] Step 2: the 7-chloro-6-fluoro-4-hydroxy-1-(4-isopropyl-6-methylpyrimidin-5-yl)-1,8-naphthyridin-2(1H)-one (1.3 g, 3.72 mmol), sodium nitrite (26 mg, 0.37 mmol), and 8 mL of glacial acetic acid were added to a round-bottom flask. Concentrated nitric acid (700 mg, 11.1 mmol) was added dropwise to the resulting reaction liquid. The reaction liquid was placed into an oil bath at 30° C. to be heated for 2 hours. The reaction liquid was poured into ice water, and solid was precipitated. The resulting reaction mixture was filtered, and the filter cake was washed with water. The filter cake was collected and dried in vacuum to obtain 7-chloro-6-fluoro-4-hydroxy-1-(4-isopropyl-6-methylpyrimidin-5-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (1.2 g, purity: 76%), which was yellow solid. The resulting crude product was directly used in next step. ES-API: [M+H].sup.+=394.1.
[0316] Step 3: the 7-chloro-6-fluoro-4-hydroxy-1-(4-isopropyl-6-methylpyrimidin-5-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (1.2 g, 3 mmol), 2-fluoro-6-methoxyphenylboronic acid (2 g, 12 mmol), 2-dicyclohexylphosphino-2′,6′-dimethoxybiphenyl (123 mg, 0.3 mmol), SPhos-Pd-G2 (216 mg, 0.3 mmol), potassium phosphate (1.9 g, 9 mmol), 15 mL of dioxane, and 3 mL of water were added to a reaction flask. The resulting mixture was stirred in an oil bath at 110° C. for 1 hour to react under the protection of nitrogen, and then the reaction was terminated. The resulting reaction liquid was added with an aqueous solution (30 mL) of 1 M potassium carbonate, and extracted with 20 mL of EtOAc/PE (1:1) once to remove impurities. The water phase was then mixed with an aqueous solution of 6 M potassium carbonate such that the pH was adjusted to 4. Extraction was performed with EtOAc for 3 times. The resulting organic phase was dried by using sodium sulfate and concentrated to obtain 6-fluoro-7-(2-fluoro-6-methoxyphenyl)-4-hydroxy-1-(4-isopropyl-6-methylpyrimidin-5-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (1.1 g, 75%), which was yellow solid. ES-API: [M+H].sup.+=483.1.
[0317] Step 4: the 6-fluoro-7-(2-fluoro-6-methoxyphenyl)-4-hydroxy-1-(4-isopropyl-6-methylpyrimidin-5-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (1.2 g, 2.48 mmol), diisopropylethylamine (3 g, 23.1 mmol), and ACN (20 mL) were added to a round-bottom flask. Phosphorus oxychloride (2.2 g, 14.5 mmol) was added thereto. The resulting mixture was stirred at 85° C. for 1 hour to react. Whether the reaction was completed was detected by liquid chromatography-mass spectrograph (LC-MS). The resulting reaction liquid was poured into ice water and extracted with EtOAc. The resulting organic phase was dried by using sodium sulfate and concentrated to obtain 4-chloro-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(4-isopropyl-6-methylpyrimidin-5-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (1.1 g, purity: 83%). The resulting crude product was directly used in next step. ES-API: [M+H].sup.+=502.1.
[0318] Step 5: the 4-chloro-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(4-isopropyl-6-methylpyrimidin-5-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (1 g, 2 mmol), 1-(tert-butyl)3-methyl(R)-piperazin-1,3-dicarboxylate (1.94 g, 8 mmol), N,N-diisopropylethylamine (516 mg, 4 mmol), and N,N-dimethylacetamide (10 mL) were added to a round-bottom flask, and stirred at 120° C. for 2 hours to react. Whether the reaction was completed was detected by LC-MS. The resulting reaction liquid was poured into 30 mL of water. Extraction was performed with EtOAc for 3 times. The resulting organic phase was washed with saturated salt solution/water (v/v, 1:1) for 4 times, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-40%) to obtain 1-(tert-butyl)3-methyl(3R)-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(4-isopropyl-6-methylpyrimidin-5-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazin-1,3-dicarboxylate (1 g, purity: 82%). ES-API: [M+H].sup.+=710.2.
[0319] Step 6: the 1-(tert-butyl)3-methyl(3R)-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(4-isopropyl-6-methylpyrimidin-5-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazin-1,3-dicarboxylate (1 g, 1.4 mmol), iron powder (390 mg, 7 mmol), and 15 mL of glacial acetic acid were added to a reaction flask. The resulting mixture was stirred at 80° C. for 1 hour to react. Whether the reaction was completed was detected by LC-MS. The resulting reaction liquid was poured into 50 mL of aqueous solution of sodium bicarbonate and extracted with 30 mL of EtOAc for 3 times. The resulting organic phase was dried and concentrated to obtain tert-butyl (4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(4-isopropyl-6-methylpyrimidin-5-yl)-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (850 mg, 93%), which was yellow solid. ES-API: [M+H].sup.+=648.3.
[0320] Step 7: the tert-butyl (4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(4-isopropyl-6-methylpyrimidin-5-yl)-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (450 mg, 0.69 mmol), iodomethane (789 mg, 5.55 mmol), potassium carbonate (286 mg, 2.07 mmol), and 10 mL of acetone were added to a round-bottom flask. The resulting mixture was stirred at 50° C. for 16 hours to react in a sealing tube, and whether the reaction was completed was detected by LC-MS. The resulting reaction liquid was filtered by using diatomite. The filtrate was concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-60%) to obtain tert-butyl (4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(4-isopropyl-6-methylpyrimidin-5-yl)-6-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (260 mg, 57%), which was yellow solid. ES-API: [M+H].sup.+=662.2.
[0321] Step 8: the tert-butyl (4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(4-isopropyl-6-methylpyrimidin-5-yl)-6-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (260 mg, 0.39 mmol), 1 mL of DCM, and 3 mL of TFA were added to a round-bottom flask. The resulting mixture was stirred at room temperature for 1 hour, and whether the reaction was completed was detected by LC-MS. The resulting reaction liquid was concentrated to obtain (4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(4-isopropyl-6-methylpyrimidin-5-yl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (219 mg), which was yellow solid. The resulting crude product was directly used in next step. ES-API: [M+H].sup.+=562.2.
[0322] Step 9: the (4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(4-isopropyl-6-methylpyrimidin-5-yl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (219 mg, 0.39 mmol), 3 mL of DCM, and triethylamine (158 mg, 1.56 mmol) were added to a 50 mL round-bottom flask. The resulting reaction liquid was cooled to 0° C., and then added dropwise with a solution of acryloyl chloride in DCM (71 mg, 0.78 mmol, 0.5 mL). The resulting mixture was stirred at 0° C. for 10 minutes to react. The resulting reaction liquid was added with 40 mL of saturated solution of sodium bicarbonate and extracted with 20 mL of DCM for 3 times. The resulting organic phase was dried and concentrated to obtain (4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(4-isopropyl-6-methylpyrimidin-5-yl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (240 mg, purity: 87%), which was yellow solid. The resulting crude product was directly used in next step. ES-API: [M+H].sup.+=616.3.
[0323] Step 10: the (4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(4-isopropyl-6-methylpyrimidin-5-yl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (240 mg, 0.39 mmol) and 3 mL of DCM were added to a round-bottom flask. The resulting reaction liquid was cooled to 0° C., and then added dropwise with a solution (6 mL) of 17% boron tribromide in DCM. After the completion of dropwise addition, the resulting mixture was stirred at room temperature for 2 hours to react. The resulting reaction liquid was poured into 30 mL of glacial saturated solution of NaHCO.sub.3 and extracted with 20 mL of DCM for 3 times. The resulting organic phase was dried and concentrated, and the resulting crude product was purified by preparative scale HPLC to obtain product (4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-hydroxyphenyl)-8-(4-isopropyl-6-methylpyrimidin-5-yl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (Z21, 130 mg, 55%), which was yellow solid. ES-API: [M+H].sup.+=602.2.
[0324] Step 11: the compound (4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-hydroxyphenyl)-8-(4-isopropyl-6-methylpyrimidin-5-yl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (130 mg) was resolved by preparative scale chiral HPLC (column type: Chiralpak IE: 10 μm, 20*250 mm; mobile phase: hexane:EtOH:diethylamine=70:30:0.2; flow rate: 15 ml/min; and column temperature: room temperature) to obtain the following atropisomer compounds. One of the atropisomer compounds had a structure arbitrarily specified as Z21-1 (peak 2, retention time: 12.33 min, 47 mg), which was yellow solid. ES-API: [M+H].sup.+=602.2. .sup.1HNMR (500 MHz, DMSO-d.sub.6): 10.17 (s, 1H), 9.03 (s, 1H), 8.41 (d, J=9 Hz, 1H), 7.26-7.25 (m, 1H), 7.08-7.05 (m, 1H), 6.68-6.66 (m, 2H), 6.17-6.14 (m, 1H), 5.77-5.75 (m, 1H), 4.75-4.73 (m, 1H), 4.46-4.44 (m, 1H), 4.0-3.95 (m, 1H), 3.55-3.54 (m, 2H), 3.41 (s, 3H), 3.20-3.18 (m, 1H), 2.85-2.83 (m, 1H), 2.68-2.65 (m, 1H), 2.00 (s, 3H), 1.13 (d, J=6.5 Hz, 3H), 1.06 (d, J=6.5 Hz, 3H). The other atropisomer compound had a structure arbitrarily specified as Z21-2 (peak 1, retention time: 10.58 min, 48 mg), which was yellow solid. ES-API: [M+H].sup.+=602.2. .sup.1HNMR (500 MHz, DMSO-d.sub.6): 10.16 (s, 1H), 9.03 (s, 1H), 8.41 (d, J=9 Hz, 1H), 7.26-7.25 (m, 1H), 7.08-7.05 (m, 1H), 6.68-6.66 (m, 2H), 6.17-6.14 (m, 1H), 5.77-5.75 (m, 1H), 4.75-4.73 (m, 1H), 4.46-4.44 (m, 1H), 4.0-3.95 (m, 1H), 3.55-3.54 (m, 2H), 3.41 (s, 3H), 3.20-3.18 (m, 1H), 2.65-2.60 (m, 1H), 2.52-2.51 (m, 1H), 2.20 (s, 3H), 1.06 (d, J=6.5 Hz, 3H), 0.86 (d, J=6.5 Hz, 3H). The isomer compounds were detected by analytical scale chiral HPLC (column type: Chiralpak IE: 5 μm, 4.6*250 mm; mobile phase: hexane:EtOH:aminomethanol=70:30:0.2; flow rate: 1 ml/min; and column temperature=30° C.).
Example 22 Preparation of Compound Z22
[0325] ##STR00500## ##STR00501## ##STR00502##
[0326] Step 1: 7-chloro-6-fluoro-4-hydroxy-1-(3-isopropylpyrazin-2-yl)-1,8-naphthyridin-2(1H)-one (2 g, 6 mmol) was dissolved in acetic acid (5 mL), orderly added with sodium nitrite (41 mg, 0.6 mmol) and concentrated nitric acid (1.5 g, 24 mmol), and stirred at room temperature for 30 minutes to react. The resulting reaction liquid was slowly poured into 100 mL of ice water. The precipitated solid was filtered. The filter cake was washed with 20 ml of ice water and dried in vacuum to obtain product 7-chloro-6-fluoro-4-hydroxy-1-(3-isopropylpyrazin-2-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (1.5 g, 65%), which was yellow solid. ES-API: [M+H]=380.2.
[0327] Step 2: the 7-chloro-6-fluoro-4-hydroxy-1-(3-isopropylpyrazin-2-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (1.5 g, 3.94 mmol), (2-fluoro-6-methoxyphenyl)boric acid (2.04 g, 12 mmol), SPhos-Pd-G2 (288 mg, 0.4 mmol), 2-dicyclohexylphosphino-2′,6′-dimethoxybiphenyl (164 mg, 0.4 mmol), potassium phosphate (2.5 g, 12 mmol), 10 mL of water, and 40 mL of dioxane were added to a 100 mL three-necked round-bottom flask. The resulting mixture was stirred at 100° C. for 2-3 hours to react under the protection of nitrogen. After the completion of the reaction, the resulting reaction liquid was cooled to room temperature, added with 80 mL of water and 100 mL of methyl tert-butyl ether extracted once. The water phase was then mixed with 1M hydrochloric acid solution such that the pH was adjusted to a range of 3 to 5, and extracted with EtOAc (200 mL*2). The resulting combined EtOAc phase was dried by using anhydrous sodium sulfate, and filtered. The filtrate was dried in vacuum to obtain product 6-fluoro-7-(2-fluoro-6-methoxyphenyl)-4-hydroxy-1-(3-isopropylpyrazin-2-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (1.6 g, crude), which was faint yellow solid. ES-API: [M+H].sup.+=470.1.
[0328] Step 3: the 6-fluoro-7-(2-fluoro-6-methoxyphenyl)-4-hydroxy-1-(3-isopropylpyrazin-2-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (1.6 g, 3.4 mmol) was dissolved in ACN (30 mL), orderly added with phosphorus oxychloride (2.6 g, 17 mmol) and N,N-diisopropylethylamine (3 g, 23.8 mmol), and gradually heated to 80° C. and stirred for 30 minutes to react. The resulting reaction liquid was concentrated, added with 30 mL of cold ACN, then added dropwise to 150 mL of saturated sodium bicarbonate solution in an ice water bath, and extracted with EtOAc (200 mL*2). The resulting combined organic phase was washed with 200 mL of saturated salt solution once. The organic phase was then dried by using anhydrous sodium sulfate, filtered, dried and concentrated, and the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-50%) to obtain 4-chloro-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(3-isopropylpyrazin-2-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (340 mg, Y: 20%), which was yellow solid. ES-API: [M+H].sup.+=488.2.
[0329] Step 4: the 4-chloro-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(3-isopropylpyrazin-2-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (310 mg, 0.64 mmol) was dissolved in N,N-dimethylacetamide (5 mL), orderly added with 1-(tert-butyl)3-methyl(3R,6R)-6-methylpiperazin-1,3-dicarboxylic acid (247 mg, 0.96 mmol) and N,N-diisopropylethylamine (250 mg, 1.92 mmol), and stirred at 120° C. for 2 hours to react. The resulting reaction liquid was added with 50 mL of EtOAc, and washed with 30 mL of saturated salt solution for 3 times. The EtOAc phase was dried and concentrated, and the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-80%) to obtain product 1-(tert-butyl)3-methyl(3R,6R)-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(3-isopropylpyrazin-2-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-6-methylpiperazin-1,3-dicarboxylate (317 mg, Y: 70%), which was yellow solid. ES-API: [M+H].sup.+=710.2.
[0330] Step 5: the 1-(tert-butyl)3-methyl(3R,6R)-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(3-isopropylpyrazin-2-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-6-methylpiperazin-1,3-dicarboxylate (280 mg, 0.4 mmol) was dissolved in acetic acid (4 mL), added with iron powder (78 mg, 1.4 mmol), and stirred at 80° C. for 30 minutes to react. The resulting reaction liquid was orderly added with 50 mL of EtOAc and 30 mL of saturated sodium bicarbonate solution. The resulting suspension was filtered by using diatomite. The filter cake was washed with EtOAc. The resulting organic phase was separated, washed orderly with 100 mL of saturated sodium bicarbonate and 30 mL of saturated salt solution, dried and concentrated to obtain product tert-butyl (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(3-isopropylpyrazin-2-yl)-2-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (312 mg, crude), which was yellow solid. ES-API: [M+H].sup.+=648.1.
[0331] Step 6: the tert-butyl (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(3-isopropylpyrazin-2-yl)-2-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (295 mg, 0.46 mmol), 3 mL of acetone, anhydrous potassium carbonate (1 g, 6.9 mmol), and iodomethane (253 mg, 1.84 mmol) were added to a 15 mL sealing tube. The sealing tube was sealed, and the resulting mixture was stirred at 55° C. for 18 hours to react. The resulting reaction liquid was added with 50 mL of EtOAc, washed with 20 mL of saturated salt solution for 3 times, dried and concentrated to obtain product tert-butyl (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(3-isopropylpyrazin-2-yl)-2,6-dimethyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (356 mg, crude), which was yellow solid. ES-API: [M+H].sup.+=662.2.
[0332] Step 7: the tert-butyl (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(3-isopropylpyrazin-2-yl)-2,6-dimethyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (356 mg, 0.54 mmol) was dissolved in DCM (8 mL), and added with TFA (4 mL). The resulting mixture was stirred at room temperature for 2 hours. The resulting reaction liquid was concentrated to obtain product (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(3-isopropylpyrazin-2-yl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (415 mg, crude), which was directly used in next step. ES-API: [M+H].sup.+=562.2.
[0333] Step 8: the (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(3-isopropylpyrazin-2-yl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (415 mg, 0.74 mmol) was dissolved in DCM (15 mL), and added with triethylamine (3.0 mL, 21.62 mmol). The resulting reaction liquid was cooled to 0° C., and then added dropwise with acryloyl chloride (115 mg, 1.28 mmol). The resulting mixture was stirred at 0° C. for 5 minutes to react. The resulting reaction liquid was added with 50 mL of DCM, washed with 50 mL of saturated solution of NaHCO.sub.3 and 80 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-60%) to obtain product (2R,4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(3-isopropylpyrazin-2-yl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (201 mg, Y: 44%), which was yellow solid. ES-API: [M+H].sup.+=616.2.
[0334] Step 9: under the condition of an ice water bath, the (2R,4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(3-isopropylpyrazin-2-yl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (201 mg, 0.33 mmol) was added to dry DCM (3.0 mL), and then added with boron tribromide (5.0 mL) to react at room temperature for 30 minutes. Under the condition of the ice water bath, the above reaction liquid was added dropwise to saturated sodium bicarbonate solution, extracted with DCM (50 mL) twice, dried and concentrated. The resulting crude product was purified by preparative scale HPLC to obtain product (2R,4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-hydroxyphenyl)-8-(3-isopropylpyrazin-2-yl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (Z22, 65 mg, Y: 33%) ES-API: [M+H].sup.+=602.2. .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.17 (s, 1H), 8.75 (dd, J=4.0, 2.6 Hz, 1H), 8.55 (dd, J=15.5, 2.4 Hz, 1H), 8.05-7.98 (m, 1H), 7.26 (dd, J=15.0, 8.2 Hz, 1H), 7.03 (dd, J=16.8, 10.0 Hz, 1H), 6.73 (d, J=8.4 Hz, 1H), 6.67 (t, J=8.8 Hz, 1H), 6.16 (t, J=12.4 Hz, 1H), 5.74 (dd, J=20.0, 11.8 Hz, 1H), 4.78 (s, 1H), 4.65-4.56 (m, 1H), 4.00 (t, J=28.0 Hz, 1H), 3.80-3.70 (m, 1H), 3.36 (d, J=2.4 Hz, 3H), 3.05-2.62 (m, 2H), 1.63-1.48 (m, 3H), 1.18 (d, J=6.8 Hz, 2H), 1.10 (d, J=6.8 Hz, 3H), 1.00 (d, J=6.7 Hz, 2H).
Example 23 Preparation of Compound Z23
[0335] ##STR00503## ##STR00504##
[0336] Step 1: 4-chloro-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(4-isopropyl-6-methylpyrimidin-5-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (0.8 g, 1.46 mmol), 1-(tert-butyl)3-methyl(3R,6R)-6-methylpiperazin-1,3-dicarboxylate (567 mg, 2.2 mmol), N,N-diisopropylethylamine (565 mg, 4.38 mmol), and N,N-dimethylacetamide (10 mL) were added to a round-bottom flask, and stirred at 120° C. for 1 hour to react. Whether the reaction was completed was detected by LC-MS. The resulting reaction liquid was poured into 30 mL of water. Extraction was performed with EtOAc for 3 times. The resulting organic phase was washed with saturated salt solution/water (v/v, 1:1) for 4 times, dried and concentrated to obtain 1-(tert-butyl)3-methyl(3R,6R)-4-(6-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-7-(2-fluoro-6-methoxyphenyl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-6-methylpiperazin-1,3-dicarboxylate (1 g, yield: 89%). ES-API: [M+H].sup.+=768.3.
[0337] Step 2: the 1-(tert-butyl)3-methyl(3R,6R)-4-(6-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-7-(2-fluoro-6-methoxyphenyl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-6-methylpiperazin-1,3-dicarboxylate (1 g, 1.3 mmol), iron powder (300 mg, 5.3 mmol), and 8 mL of glacial acetic acid was added to a reaction flask. The resulting mixture was stirred at 80° C. for 0.5 hour to react. Whether the reaction was completed was detected by LC-MS. The resulting reaction liquid was poured into 50 mL of aqueous solution of sodium bicarbonate and extracted with 30 mL of EtOAc for 3 times. The resulting organic phase was dried and concentrated to obtain crude product tert-butyl (2R,4aR)-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-10-(2-fluoro-6-methoxyphenyl)-2-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (761 mg, 83%), which was yellow solid. ES-API:[M+H].sup.+=706.3.
[0338] Step 3: the tert-butyl (2R,4aR)-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-10-(2-fluoro-6-methoxyphenyl)-2-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (761 mg, 1.08 mmol), iodomethane (1.5 g, 10.79 mmol), potassium carbonate (596 mg, 4.32 mmol), and 15 mL of acetone were added to a round-bottom flask. The resulting mixture was stirred at 50° C. for 16 hours to react in a sealing tube, and whether the reaction was completed was detected by LC-MS. The resulting reaction liquid was filtered by using diatomite. The filtrate was concentrated to obtain crude product tert-butyl (2R,4aR)-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-10-(2-fluoro-6-methoxyphenyl)-2,6-dimethyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (738 mg, 95%), which was yellow solid. ES-API: [M+H].sup.+=720.3.
[0339] Step 4: the tert-butyl (2R,4aR)-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-10-(2-fluoro-6-methoxyphenyl)-2,6-dimethyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (738 mg, 1.02 mmol), 2 mL of DCM, and 5 mL of TFA were added to a round-bottom flask. The resulting mixture was stirred at room temperature for 1 hour, and whether the reaction was completed was detected by LC-MS. The resulting reaction liquid was concentrated to obtain (2R,4aR)-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-10-(2-fluoro-6-methoxyphenyl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (632 mg, 100%), which was yellow solid. The resulting crude product was directly used in next step. ES-API: [M+H].sup.+=620.3.
[0340] Step 5: the (2R,4aR)-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-10-(2-fluoro-6-methoxyphenyl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (632 mg, 1.02 mmol), 3 mL of DCM, and triethylamine (677 mg, 6.7 mmol) were added to a 50 mL round-bottom flask. The resulting reaction liquid was cooled to 0° C., and then added dropwise with a solution of acryloyl chloride in DCM (249 mg, 2.77 mmol, 0.5 mL). The resulting mixture was stirred at 0° C. for 10 minutes to react. The resulting reaction liquid was added with 40 mL of saturated solution of sodium bicarbonate and extracted with 20 mL of DCM for 3 times. The resulting organic phase was dried and concentrated, and the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-60%) to obtain (2R,4aR)-3-acryloyl-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-10-(2-fluoro-6-methoxyphenyl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (500 mg, 72%), which was yellow solid. The crude product was directly used in next step. ES-API: [M+H].sup.+=674.2.
[0341] Step 6: the (2R,4aR)-3-acryloyl-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-10-(2-fluoro-6-methoxyphenyl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (500 mg, 0.74 mmol) and 3 mL of DCM were added to a round-bottom flask. The resulting reaction liquid was cooled to 0° C., and then added dropwise with a solution (12 mL) of 17% boron tribromide in DCM. After the completion of dropwise addition, the resulting mixture was stirred at 25° C. for 25 hours to react. The resulting reaction liquid was poured into 30 mL of glacial saturated solution of NaHCO.sub.3 and extracted with 20 mL of DCM for 3 times. The resulting organic phase was dried and concentrated, and the resulting crude product was purified by preparative scale HPLC to obtain product (2R,4aR)-3-acryloyl-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-10-(2-fluoro-6-hydroxyphenyl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (Z23, 200 mg, 40%), which was yellow solid. .sup.1HNMR (500 MHz, DMSO-d.sub.6): δ 10.10-10.5 (m, 1H), 9.11 (s, 1H), 8.25-8.23 (m 1H), 7.22-7.21 (m, 1H), 6.86-6.74 (m, 1H), 6.67-6.64 (m, 2H), 6.17-6.14 (m, 1H), 5.75-5.71 (m, 1H), 5.04-5.01 (m, 1H), 4.62-4.42 (m, 1H), 4.03-3.98 (m, 1H), 3.74-3.72 (m, 1H), 3.42-3.33 (m, 5H), 2.77-2.64 (m, 2H), 1.56-1.52 (m, 3H), 1.05-0.97 (m, 9H), 0.86-0.84 (m, 3H). ES-API: [M+H].sup.+=660.3.
Example 24 Preparation of Compounds Z24, Z24-1, and Z24-2
[0342] ##STR00505## ##STR00506## ##STR00507##
[0343] Step 1: 6,7-dichloro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridin-3-carbonitrile (30.0 g, 77.319 mmol) was suspended in a mixed solution of 1,4-dioxane (120 mL) and water (120 mL), and slowly added with concentrated sulfuric acid (120 mL). The resulting mixture was stirred at 120° C. for 36 hours to react. Cooled reaction liquid was poured into 200 mL of ice water, mixed with sodium carbonate to adjust the pH to a range of 2 to 3, and extracted with EtOAc (1000 mL*2). The resulting combined EtOAc phase was dried by anhydrous sodium sulfate, and filtered. The filtrate was dried in vacuum to obtain product 6,7-dichloro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-1,8-naphthyridin-2(1H)-one (24 g, Y: 85.7%), which was light brown solid. ES-API: [M+H].sup.+=364.1.
[0344] Step 2: the 6,7-dichloro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-1,8-naphthyridin-2(1H)-one (3.16 g, 8.705 mmol) was dissolved in acetic acid (15 mL), orderly added with sodium nitrite (100 mg, 1.58 mmol) and concentrated nitric acid (5.0 mL, 74.52 mmol), and stirred at room temperature for 30 minutes to react. The resulting reaction liquid was slowly poured into 100 mL ice water. The precipitated solid was filtered. The filter cake was washed with 20 ml of ice water and dried in vacuum to obtain product 6,7-dichloro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (3.5 g, Y: 92%), which was yellow solid. ES-API: [M+H].sup.+=409.1.
[0345] Step 3: the 6,7-dichloro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (3.5 g, 8.570 mmol), (2-fluoro-6-methoxyphenyl)boric acid (5.8 g, 34.10 mmol), tetrakis(triphenylphosphine)palladium (1.15 g, 0.9956 mmol), sodium carbonate (3.5 g, 33.02 mmol), 10 mL of water, and 40 mL of dioxane were added to a 100 mL three-necked round-bottom flask. The resulting mixture was stirred at 100° C. for 2-3 hours to react under the protection of nitrogen. After the completion of the reaction, the resulting reaction liquid was cooled to room temperature, added with 80 mL of water and 100 mL of methyl tert-butyl ether, and extracted once. The water phase was then mixed with 1M hydrochloric acid solution to adjust the pH to a range of 3 to 5, and extracted with EtOAc (200 mL*2). The resulting combined EtOAc phase was dried by anhydrous sodium sulfate, and filtered. The filtrate was dried in vacuum to obtain product 6-chloro-7-(2-fluoro-6-methoxyphenyl)-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (4.5 g, crude), which was faint yellow solid. ES-API: [M+H].sup.+=499.1.
[0346] Step 4: the 6-chloro-7-(2-fluoro-6-methoxyphenyl)-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (4.6 g, 8.57 mmol) was dissolved in ACN (30 mL), orderly added with phosphorus oxychloride (7.5 g, 48.92 mmol) and N,N-diisopropylethylamine (10.5 g, 81.24 mmol), and gradually warmed to 80° C. and stirred for 30 minutes to react. The resulting reaction liquid was concentrated, added with 30 mL of cold ACN, added dropwise to 150 mL of saturated sodium bicarbonate solution in an ice water bath, and extracted with EtOAc (200 mL*2). The resulting combined EtOAc phase was washed with 200 mL of saturated salt solution once, dried by anhydrous sodium sulfate, and filtered. After the organic phase was dried and concentrated, the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-50%) to obtain 4,6-dichloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (3.05 g, Y: 76%), which was yellow solid. ES-API: [M+H].sup.+=517.2.
[0347] Step 5: the 4,6-dichloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (2.5 g, 4.843 mmol) was dissolved in N,N-dimethylacetamide (25 mL), orderly added with 1-(tert-butyl)3-methyl(R)-piperazin-1,3-dicarboxylate (3.5 g, 14.34 mmol) and N,N-diisopropylethylamine (2.0 g, 15.47 mmol), and stirred at 120° C. for 2 hours to react. The resulting reaction liquid was added with 80 mL of EtOAc, and washed with 80 mL of saturated salt solution for 3 times. The EtOAc phase was dried and concentrated, and the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-80%) to obtain product 1-(tert-butyl)3-methyl(3R)-4-(6-chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazin-1,3-dicarboxylate (2.7 g, Y: 77%), which was yellow solid. ES-API: [M+H].sup.+=725.2.
[0348] Step 6: the 1-(tert-butyl)3-methyl(3R)-4-(6-chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazin-1,3-dicarboxylate (2.7 g, 3.728 mmol) was dissolved in acetic acid (30 mL), added with iron powder (835 mg, 14.91 mmol), and stirred at 80° C. for 30 minutes to react. The resulting reaction liquid was concentrated, orderly added with 200 mL of EtOAc and 100 mL of saturated sodium bicarbonate solution. The resulting suspension was filtered by diatomite. The filter cake was washed with EtOAc. The resulting organic phase was separated, washed orderly with 100 mL of saturated sodium bicarbonate solution and 150 mL of saturated salt solution, dried and concentrated to obtain product tert-butyl (4aR)-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (2.70 g, crude), which was yellow solid. ES-API: [M+H]+=663.2.
[0349] Step 7: the tert-butyl (4aR)-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (250 mg, 0.3774 mmol), 4 mL of DCM, and 4 mL of TFA were orderly added to a 100 mL single-necked flask, stirred at room temperature for 2 hours. The resulting reaction liquid was concentrated to obtain product (4aR)-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (300 mg, crude), which was directly used in next step. ES-API: [M+H].sup.+=563.2.
[0350] Step 8: the (4aR)-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (300 mg, 0.3774 mmol) was dissolved in DCM (10 mL), and added with triethylamine (3.0 mL, 21.62 mmol). The resulting reaction liquid was cooled to 0° C., and added dropwise with acryloyl chloride (50 mg, 0.5524 mmol). The resulting mixture was stirred at 0° C. for 15 minutes to react. The resulting reaction liquid was added with 80 mL of DCM, washed with 100 mL of saturated solution of NaHCO.sub.3 and 80 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (MeOH/DCM: 0-20%) to obtain product (4aR)-3-acryloyl-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,3,4,4a,6,8-hexa hydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (243 mg, crude), which was yellow solid. ES-API: [M+H].sup.+=617.2.
[0351] Step 9: under the condition of an ice water bath, the (4aR)-3-acryloyl-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,3,4,4a,6,8-hexa hydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (243 mg, 0.3774 mmol) was added to dry DCM (6.0 mL), then added with boron tribromide (5.0 mL, 5.0 mmol), and warmed to room temperature to react overnight. Under the condition of the ice water bath, the above reaction liquid was added dropwise to saturated sodium bicarbonate solution, extracted with DCM (80 mL) twice, dried, concentrated, and purified by preparative scale HPLC to obtain (4aR)-3-acryloyl-11-chloro-10-(2-fluoro-6-hydroxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,3,4,4a,6,8-hexa hydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (Z24, 76 mg, Y: 32%). [M+H].sup.+=603.2.
[0352] Step 10: the compound Z24 (76.0 mg, 0.1262 mmol) was resolved by preparative scale chiral HPLC (column type: IA: 10 μm, 30*250 mm; mobile phase: hexane:EtOH=40:60; flow rate: 25 ml/min; and column temperature: room temperature) to obtain the following atropisomer compounds. One of the atropisomer compounds had a structure arbitrarily specified as Z24-1 (13.7 mg, peak 1, retention time: 2.612 min, Y: 18%), ES-API: [M+H].sup.+=603.2. The other atropisomer compound had a structure arbitrarily specified as Z24-2 (21.4 mg, peak 2, retention time: 3.985 min, Y: 28%), ES-API: [M+H].sup.+=603.2. The isomer compounds were detected by analytical scale chiral HPLC (column type IA: 5 μm, 4.6*150 mm; mobile phase: hexane:EtOH=40:60; flow rate: 1 ml/min; and column temperature=30° C.).
Example 25 Preparation of Compounds Z25, Z25-1, and Z25-2
[0353] ##STR00508## ##STR00509## ##STR00510## ##STR00511##
[0354] Step 1: 6,7-dichloro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridin-3-carbonitrile (30.0 g, 77.319 mmol) was suspended in a mixed solution of 1,4-dioxane (120 mL) and water (120 mL), and slowly added with concentrated sulfuric acid (120 mL). The resulting mixture was stirred at 120° C. for 36 hours to react. Cooled reaction liquid was poured into 200 mL of ice water, mixed with sodium carbonate to adjust the pH to a range of 2 to 3, and extracted with EtOAc (1000 mL*2). The resulting combined EtOAc phase was dried by anhydrous sodium sulfate, and filtered. The filtrate was dried in vacuum to obtain product 6,7-dichloro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-1,8-naphthyridin-2(1H)-one (24 g, Y: 85.7%), which was light brown solid. ES-API: [M+H].sup.+=364.1.
[0355] Step 2: the 6,7-dichloro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-1,8-naphthyridin-2(1H)-one (3.16 g, 8.705 mmol) was dissolved in acetic acid (15 mL), order added with sodium nitrite (100 mg, 1.58 mmol) and concentrated nitric acid (5.0 mL, 74.52 mmol), and stirred at room temperature for 30 minutes to react. The resulting reaction liquid was slowly poured into 100 mL ice water. The precipitated solid was filtered. The filter cake was washed with 20 ml of ice water and dried in vacuum to obtain product 6,7-dichloro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (3.5 g, Y: 92%), which was yellow solid. ES-API: [M+H].sup.+=409.1.
[0356] Step 3: the 6,7-dichloro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (3.5 g, 8.570 mmol), (2-fluoro-6-methoxyphenyl)boric acid (5.8 g, 34.10 mmol), tetrakis(triphenylphosphine)palladium (1.15 g, 0.9956 mmol), sodium carbonate (3.5 g, 33.02 mmol), 10 mL of water, and 40 mL of dioxane were added to a 100 mL three-necked round-bottom flask. The resulting mixture was stirred at 100° C. for 2-3 hours to react under the protection of nitrogen. After the completion of the reaction, the resulting reaction liquid was cooled to room temperature, added with 80 mL of water and 100 mL of methyl tert-butyl ether, and extracted once. The water phase was then mixed with 1M hydrochloric acid solution to adjust the pH to a range of 3 to 5, and extracted with EtOAc (200 mL*2). The resulting combined EtOAc phase was dried by anhydrous sodium sulfate, and filtered. The filtrate was dried in vacuum to obtain product 6-chloro-7-(2-fluoro-6-methoxyphenyl)-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (4.5 g, crude), which was faint yellow solid. ES-API: [M+H].sup.+=499.1.
[0357] Step 4: the 6-chloro-7-(2-fluoro-6-methoxyphenyl)-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (4.6 g, 8.57 mmol) was dissolved in ACN (30 mL), orderly added with phosphorus oxychloride (7.5 g, 48.92 mmol) and N,N-diisopropylethylamine (10.5 g, 81.24 mmol), and gradually heated to 80° C. and stirred for 30 minutes to react. The resulting reaction liquid was concentrated, added with 30 mL of cold ACN, added dropwise to 150 mL of saturated sodium bicarbonate solution in an ice water bath, and extracted with EtOAc (200 mL*2). The resulting combined EtOAc phase was washed with 200 mL of saturated salt solution once, dried by anhydrous sodium sulfate, and filtered. After the organic phase was dried and concentrated, the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-50%) to obtain 4,6-dichloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (3.05 g, Y: 76%), which was yellow solid. ES-API: [M+H].sup.+=517.2.
[0358] Step 5: the 4,6-dichloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (2.5 g, 4.843 mmol) was dissolved in N,N-dimethylacetamide (25 mL), orderly added with 1-(tert-butyl)3-methyl(R)-piperazin-1,3-dicarboxylate (3.5 g, 14.34 mmol) and N,N-diisopropylethylamine (2.0 g, 15.47 mmol), and stirred at 120° C. for 2 hours to react. The resulting reaction liquid was added with 80 mL of EtOAc, and washed with 80 mL of saturated salt solution for 3 times. The EtOAc phase was dried and concentrated, and the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-80%) to obtain product 1-(tert-butyl)3-methyl(3R)-4-(6-chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazin-1,3-dicarboxylate (2.7 g, Y: 77%), which was yellow solid. ES-API: [M+H].sup.+=725.2.
[0359] Step 6: the 1-(tert-butyl)3-methyl(3R)-4-(6-chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazin-1,3-dicarboxylate (2.7 g, 3.728 mmol) was dissolved in acetic acid (30 mL), added with iron powder (835 mg, 14.91 mmol), and stirred at 80° C. for 30 minutes to react. The resulting reaction liquid was concentrated, orderly added with 200 mL of EtOAc and 100 mL of saturated sodium bicarbonate solution. The resulting suspension was filtered by diatomite. The filter cake was washed with EtOAc. The resulting organic phase was separated, washed orderly with 100 mL of saturated sodium bicarbonate solution and 150 mL of saturated salt solution, dried and concentrated to obtain product tert-butyl (4aR)-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (2.70 g, crude), which was yellow solid. ES-API: [M+H].sup.+=663.2.
[0360] Step 7: the tert-butyl (4aR)-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (2.7 g, 3.728 mmol), 30 mL of acetone, anhydrous potassium carbonate (2.2 g, 15.94 mmol), and iodomethane (5.4 g, 38.03 mmol) were orderly added to a 150 mL sealing tube. The sealing tube was sealed, and the resulting mixture was stirred at 55° C. for 18 hours to react. The resulting reaction liquid was added with 150 mL of EtOAc, washed with 100 mL of saturated salt solution for 3 times, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-80%) to obtain product tert-butyl (4aR)-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-6-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (2.2 g, Y: 87%), which was yellow solid. ES-API: [M+H].sup.+=677.2.
[0361] Step 8: the tert-butyl (4aR)-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-6-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (517 mg, 0.7549 mmol) was dissolved in DCM (8 mL), and added with TFA (2 mL). The resulting mixture was stirred at room temperature for 2 hours. The resulting reaction liquid was concentrated to obtain product (4aR)-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (530 mg, crude), which was directly used in next step. ES-API: [M+H].sup.+=577.2.
[0362] Step 9: the (4aR)-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (530 mg, 0.7549 mmol) was dissolved in DCM (15 mL), and added with triethylamine (3.0 mL, 21.62 mmol). The resulting reaction liquid was cooled to 0° C., and added dropwise with acryloyl chloride (100 mg, 1.1048 mmol). The resulting mixture was stirred at 0° C. for 15 minutes to react. The resulting reaction liquid was added with 80 mL of DCM, washed with 100 mL of saturated solution of NaHCO.sub.3 and 80 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-60%) to obtain product (4aR)-3-acryloyl-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (280 mg, Y: 59%), which was yellow solid. ES-API: [M+H].sup.+=631.2.
[0363] Step 10: under the condition of an ice water bath, the (4aR)-3-acryloyl-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (280 mg, 0.444 mmol) was added to dry DCM (6.0 mL), then added with boron tribromide (5.0 mL, 5.0 mmol), and warmed to room temperature to react overnight. Under the condition of the ice water bath, the above reaction liquid was added dropwise to saturated sodium bicarbonate solution, extracted with DCM (80 mL) twice, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-60%) to obtain product (4aR)-3-acryloyl-11-chloro-10-(2-fluoro-6-hydroxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (Z25, 233 mg, Y: 85%).
[0364] Step 11: the compound Z25 was resolved by preparative scale chiral HPLC (column type: IA: 10 μm, 30*250 mm; mobile phase: hexane:EtOH=60:40; flow rate: 25 ml/min; and column temperature: room temperature) to obtain the following atropisomer compounds. One of the atropisomer compounds had a structure arbitrarily specified as Z25-1 (76.8 mg, peak 1, retention time: 2.531 min, Y: 34%). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.03 (d, J=18.4 Hz, 1H), 8.52 (d, J=7.3 Hz, 1H), 8.43 (d, J=4.7 Hz, 1H), 7.23 (d, J=9.6 Hz, 2H), 7.08 (dd, J=16.6, 10.5 Hz, 1H), 6.74-6.62 (m, 2H), 6.15 (d, J=16.8 Hz, 1H), 5.75 (d, J=10.7 Hz, 1H), 4.73 (d, J=14.2 Hz, 1H), 4.46 (d, J=12.9 Hz, 1H), 4.00 (s, 1H), 3.61 (d, J=10.0 Hz, 1H), 3.51 (s, 1H), 3.34 (s, 3H), 3.22 (s, 1H), 2.64 (t, J=11.5 Hz, 1H), 2.48-2.42 (m, 1H), 1.98 (d, J=5.1 Hz, 3H), 1.03 (t, J=6.9 Hz, 3H), 0.86 (t, J=7.9 Hz, 3H). ES-API: [M+H].sup.+=617.2. The other atropisomer compound had a structure arbitrarily specified as Z25-2 (70 mg, peak 2, retention time: 3.683 min, Y: 31%). .sup.1H NMR (500 MHz, CDCl.sub.3) δ 8.64-8.59 (m, 1H), 8.35 (s, 1H), 8.07 (s, 1H), 7.27-7.20 (m, 2H), 7.14-7.02 (m, 1H), 6.75-6.63 (m, 2H), 6.39 (dd, J=17.0, 2.0 Hz, 1H), 5.88-5.77 (m, 1H), 4.91 (d, J=14.0 Hz, 1H), 4.83 (d, J=13.0 Hz, 1H), 3.72-3.58 (m, 2H), 3.50 (s, 3H), 3.43 (d, J=12.0 Hz, 1H), 3.16 (t, J=13.0 Hz, 1H), 2.91 (t, J=12.0 Hz, 1H), 2.82-2.73 (m, 1H), 1.93 (s, 3H), 1.24 (d, J=7.0 Hz, 3H), 1.12 (d, J=7.0 Hz, 3H). ES-API: [M+H].sup.+=617.2. The isomer compounds were detected by analytical scale chiral HPLC (column type: IA: 5 m, 4.6*150 mm; mobile phase: hexane:EtOH=60:40; flow rate: 1 ml/min; and column temperature=30° C.).
Example 26 Preparation of Compounds Z26, Z26-1, and Z26-2
[0365] ##STR00512## ##STR00513##
[0366] Step 1: tert-butyl (4aR)-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-6-(methyl-d3)-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (511 mg, 0.7549 mmol) was dissolved in DCM (8 mL), and added with TFA (2 mL). The resulting mixture was stirred at room temperature for 2 hours. The resulting reaction liquid was concentrated to obtain product (4aR)-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-6-(methyl-d3)-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (520 mg, crude), which was directly used in next step. ES-API: [M+H].sup.+=580.3.
[0367] Step 2: the (4aR)-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-6-(methyl-d3)-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (520 mg, 0.7549 mmol) was dissolved in DCM (10 mL), and added with triethylamine (3.0 mL, 21.62 mmol). The resulting reaction liquid was cooled to 0° C., and added dropwise with acryloyl chloride (100 mg, 1.1048 mmol). The resulting mixture was stirred at 0° C. for 15 minutes to react. The resulting reaction liquid was added with 80 mL of DCM, washed with 100 mL of saturated solution of NaHCO.sub.3 and 80 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-60%) to obtain product (4aR)-3-acryloyl-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-6-(methyl-d3)-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (232 mg, Y:48%), which was yellow solid. ES-API: [M+H].sup.+=634.2.
[0368] Step 3: under the condition of an ice water bath, the (4aR)-3-acryloyl-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-6-(methyl-d3)-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (240 mg, 0.3791 mmol) was added to dry DCM (6.0 mL), then added with boron tribromide (5.0 mL, 5.0 mmol), and warmed to room temperature to react overnight. Under the condition of the ice water bath, the above reaction liquid was added dropwise to saturated sodium bicarbonate solution, extracted with DCM (80 mL) twice, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-60%) to obtain product (4aR)-3-acryloyl-11-chloro-10-(2-fluoro-6-hydroxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-6-(methyl-d3)-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (Z26, 187 mg, Y: 79%). [M+H].sup.+=620.3.
[0369] Step 4: the compound Z26 (187 mg, 0.302 mmol) was resolved by preparative scale chiral HPLC (column type: IA: 10 μm, 30*250 mm; mobile phase: hexane:EtOH=60:40; flow rate: 25 ml/min; and column temperature: room temperature) to obtain the following atropisomer compounds. One of the atropisomer compounds had a structure arbitrarily specified as Z26-1 (68.8 mg, peak 1, retention time: 2.525 min, Y: 36.7%). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.03 (d, J=17.9 Hz, 1H), 8.51 (d, J=7.4 Hz, 1H), 8.43 (d, J=4.7 Hz, 1H), 7.29-7.18 (m, 2H), 7.08 (dd, J=17.0, 10.6 Hz, 1H), 6.74-6.61 (m, 2H), 6.15 (d, J=16.6 Hz, 1H), 5.75 (d, J=11.5 Hz, 1H), 4.73 (d, J=13.5 Hz, 1H), 4.46 (d, J=12.3 Hz, 1H), 4.00 (s, 1H), 3.61 (d, J=10.5 Hz, 1H), 3.50 (s, 1H), 3.22 (s, 1H), 2.65 (t, J=12.5 Hz, 1H), 2.49-2.42 (m, 1H), 1.98 (d, J=5.0 Hz, 3H), 1.02 (d, J=7.0 Hz, 3H), 0.86 (t, J=7.9 Hz, 3H). ES-API: [M+H].sup.+=620.3. The other atropisomer compound had a structure arbitrarily specified as Z26-2 (63.2 mg, peak 2, retention time: 3.683 min, Y: 33.79%). .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.62 (d, J=4.8 Hz, 1H), 8.35 (s, 1H), 8.07 (s, 1H), 7.24-7.20 (m, 2H), 7.16-7.01 (m, 1H), 6.74-6.63 (m, 2H), 6.39 (dd, J=16.8, 2.0 Hz, 1H), 5.82 (dd, J=10.4, 2.0 Hz, 1H), 4.91 (d, J=13.6 Hz, 1H), 4.83 (d, J=13.6 Hz, 1H), 3.71-3.57 (m, 2H), 3.42 (d, J=12.0 Hz, 1H), 3.16 (t, J=12.8 Hz, 1H), 2.91 (t, J=12.0 Hz, 1H), 2.81-2.70 (m, 1H), 1.92 (s, 3H), 1.22 (d, J=6.8 Hz, 3H), 1.10 (d, J=6.8 Hz, 3H). ES-API: [M+H].sup.+=620.3. The isomer compounds were detected by analytical scale chiral HPLC (column type: IA: 5 μm, 4.6*150 mm; mobile phase: hexane:EtOH=60:40; flow rate: 1 ml/min; and column temperature=30° C.).
Example 27 Preparation of Compounds Z27, Z27-1 and Z27-2
[0370] ##STR00514## ##STR00515## ##STR00516##
[0371] Step 1: 4,6-dichloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (500 mg, 0.9686 mmol) was dissolved in N,N-dimethylacetamide (5 mL), orderly added with 1-(tert-butyl)3-methyl(3R,6R)-6-methylpiperazin-1,3-dicarboxylate (375 mg, 1.452 mmol) and N,N-diisopropylethylamine (375 mg, 2.907 mmol), and stirred at 120° C. for 2 hours to react. The resulting reaction liquid was added with 80 mL of EtOAc, and washed with 80 mL of saturated salt solution for 3 times. The EtOAc phase was dried and concentrated, and the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-80%) to obtain product 1-(tert-butyl)3-methyl(3R,6R)-4-(6-chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-6-methylpiperazin-1,3-dicarboxylate (535 mg, Y: 74.5%), which was yellow solid. ES-API: [M+H].sup.+=739.2.
[0372] Step 2: the 1-(tert-butyl)3-methyl(3R,6R)-4-(6-chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-6-methylpiperazin-1,3-dicarboxylate (530 mg, 0.7179 mmol) was dissolved in acetic acid (6 mL), added with iron powder (200 mg, 3.571 mmol), and stirred at 80° C. for 30 minutes to react. The resulting reaction liquid was concentrated and orderly added with 200 mL of EtOAc and 100 mL of saturated sodium bicarbonate. The resulting suspension was filtered by diatomite. The filter cake was washed with EtOAc. The resulting organic phase was separated, washed orderly with 100 mL of saturated sodium bicarbonate and 150 mL of saturated salt solution, dried and concentrated to obtain product tert-butyl (2R,4aR)-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (452 mg, Y: 92%), which was yellow solid. ES-API: [M+H].sup.+=677.2.
[0373] Step 3: the tert-butyl (2R,4aR)-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (445 mg, 0.6583 mmol), 10 mL of acetone, anhydrous potassium carbonate (500 mg, 2.633 mmol), and iodomethane (1.20 g, 6.5828 mmol) were orderly added to a 150 mL sealing tube. The sealing tube was sealed, and the resulting mixture was stirred at 55° C. for 18 hours to react. The resulting reaction liquid was added with 150 mL of EtOAc, washed with 100 mL of saturated salt solution for 3 times, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-80%) to obtain product tert-butyl (2R,4aR)-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,6-dimethyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (455 mg, crude), which was yellow solid. ES-API: [M+H].sup.+=691.3.
[0374] Step 4: the tert-butyl (2R,4aR)-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,6-dimethyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (511 mg, 0.7549 mmol) was dissolved in DCM (8 mL), and added with TFA (2 mL). After stirring at room temperature for 2 hours, the resulting reaction liquid was concentrated to obtain product (2R,4aR)-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (462 mg, crude), which was directly used in next step. ES-API: [M+H].sup.+=591.3.
[0375] Step 5: the (2R,4aR)-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (462 mg, 0.6283 mmol) was dissolved in DCM (8 mL), and added with triethylamine (2.0 mL, 14.41 mmol). The resulting reaction liquid was cooled to 0° C., and added dropwise with acryloyl chloride (100 mg, 1.1048 mmol). The resulting mixture was stirred at 0° C. for 15 minutes to react. The resulting reaction liquid was added with 80 mL of DCM, washed with 100 mL of saturated solution of NaHCO.sub.3 and 80 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-60%) to obtain product (2R,4aR)-3-acryloyl-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (290 mg, Y: 68%), which was yellow solid. ES-API: [M+H].sup.+=645.2.
[0376] Step 6: under the condition of an ice water bath, the (2R,4aR)-3-acryloyl-11-chloro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (290 mg, 0.4503 mmol) was added to dry DCM (6.0 mL), then added with boron tribromide (6.0 mL, 6.0 mmol), and warmed to room temperature to react overnight. Under the condition of an ice water bath, the above reaction liquid was added dropwise to saturated sodium bicarbonate solution, extracted with DCM (80 mL) twice, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-60%) to obtain product (2R,4aR)-3-acryloyl-11-chloro-10-(2-fluoro-6-hydroxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (Z27, 307 mg, crude). [M+H].sup.+=631.2.
[0377] Step 7: the compound Z27 was resolved by preparative scale chiral HPLC (column type: IA*:10 μm, 30*250 mm; mobile phase: hexane:EtOH=60:40; flow rate: 25 ml/min; and column temperature: room temperature) to obtain the following atropisomer compounds. One of the atropisomer compounds had a structure arbitrarily specified as Z27-1 (67.7 mg, peak 1, retention time: 2.394 min, Y: 23.4%). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.05 (d, J=17.8 Hz, 1H), 8.43 (d, J=4.8 Hz, 1H), 8.23 (d, J=9.9 Hz, 1H), 7.23 (d, J=9.9 Hz, 2H), 7.02 (dd, J=16.8, 10.6 Hz, 1H), 6.74-6.63 (m, 2H), 6.15 (dd, J=16.8, 2.3 Hz, 1H), 5.76 (dd, J=10.5, 2.3 Hz, 1H), 4.78 (s, 1H), 4.60 (d, J=13.8 Hz, 1H), 4.00 (d, J=3.5 Hz, 1H), 3.75 (dd, J=14.1, 3.9 Hz, 1H), 3.41-3.33 (m, 1H), 3.34 (s, 3H), 2.81 (d, J=12.1 Hz, 1H), 2.48-2.42 (m, 1H), 1.98 (s, 3H), 1.53 (d, J=6.7 Hz, 3H), 1.03 (d, J=5.5 Hz, 3H), 0.85 (t, J=6.2 Hz, 3H). ES-API: [M+H].sup.+=631.2. The other atropisomer compound had a structure arbitrarily specified as Z27-2 (64.6 mg, peak 2, retention time: 3.382 min, Y: 23.2%). .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.57 (d, J=4.8 Hz, 1H), 8.36 (s, 1H), 8.28 (s, 1H), 7.25-7.15 (m, 2H), 7.03 (dd, J=16.8, 10.8 Hz, 1H), 6.72-6.61 (m, 2H), 6.34 (dd, J=16.8, 2.0 Hz, 1H), 5.80 (dd, J=10.8, 2.0 Hz, 1H), 5.11-5.01 (m, 1H), 4.77 (d, J=14.0 Hz, 1H), 3.82 (dd, J=14.0, 4.4 Hz, 1H), 3.61 (d, J=4.4 Hz, 1H), 3.49 (s, 3H), 3.30-3.17 (m, 1H), 3.03 (dd, J=12.0, 3.6 Hz, 1H), 2.80-2.68 (m, 1H), 1.91 (s, 3H), 1.66 (d, J=6.8 Hz, 3H), 1.22 (d, J=6.8 Hz, 3H), 1.08 (d, J=6.8 Hz, 3H). ES-API: [M+H].sup.+=631.2. The isomer compounds were detected by analytical scale chiral HPLC (column type: IA: 5 μm, 4.6*150 mm; mobile phase: hexane:EtOH=60:40; flow rate: 1 ml/min; and column temperature=30° C.).
Example 28 Preparation of Compound Z28
[0378] ##STR00517## ##STR00518##
[0379] Step 1: 4-chloro-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (400 mg, 0.80 mmol) was dissolved in DMF (5 mL), added with tert-butyl (S)-3-(hydroxymethyl)piperazin-1-carboxylate (432 mg, 2.00 mmol) and N,N-diisopropylethylamine (310 mg, 2.40 mmol), and stirred at 75° C. for 2 hours to react. The resulting reaction liquid was diluted by 100 mL of EtOAc, washed with 40 mL of saturated salt solution for 5 times, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-70%) to obtain product tert-butyl (3S)-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-3-(hydroxymethyl)piperazin-1-carboxylate (420 mg, Y: 77.2%), which was yellow solid. ES-API: [M+H].sup.+=681.3.
[0380] Step 2: the tert-butyl (3S)-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-3-(hydroxymethyl)piperazin-1-carboxylate (420 mg, 0.62 mmol) was dissolved in DMA (20 mL), added with LiHMDS (1.55 mL, 1.55 mmol, 1.0 M in THF), and then slowly heated to 140° C. and stirred for 24 hours to react. The resulting reaction liquid was diluted by 100 mL of EtOAc, washed with 40 mL of dilute brine for 4 times and then with 40 mL of saturated salt solution once, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-100%) to obtain product tert-butyl (4aS)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-7-oxo-1,2,4a,5,7,8-hexahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,8]naphthyridin-3(4H)-carboxylate (180 mg, Y: 46.0%), which was yellow solid. ES-API: [M+H].sup.+=634.3.
[0381] Step 3: (4aS)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-7-oxo-1,2,4a,5,7,8-hexahydropyrazino[1′,2′:4,5][1,4]oxo[2,3-c][1,8]naphthyridin-3(4H)-carboxylate (35 mg, 0.055 mmol) was dissolved in DCM (4 mL), and added with TFA (1 mL). After stirring at room temperature for 1 hour, the resulting reaction liquid was concentrated to obtain product (4aS)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-1,2,3,4,4a, 5-hexahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,8]naphthyridin-7(8H)-one (185 mg, crude), which was directly used in next step. ES-API: [M+H].sup.+=534.3.
[0382] Step 4: the (4aS)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-1,2,3,4,4a, 5-hexahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,8]naphthyridin-7(8H)-one (185 mg, crude) was dissolved in DCM (6 mL), and added with N,N-diisopropylethylamine (180 mg, 1.40 mmol). The resulting liquid was cooled to 0° C., and then added dropwise with a solution (0.5 mL) of acryloyl chloride (50 mg, 0.56 mmol) in DCM. The resulting mixture was stirred at 0° C. for 15 minutes to react. The resulting reaction liquid was added with 50 mL of DCM, washed orderly with 15 mL of water, with 15 mL of saturated solution of NaHCO.sub.3 twice and then with 15 mL of saturated salt solution. The resulting organic phase was dried and concentrated to obtain product (4aS)-3-acryloyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-1,2,3,4,4a, 5-hexahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,8]naphthyridin-7(8H)-one (160 mg, Y: 95.8%), which was yellow solid. ES-API: [M+H].sup.+=588.3.
[0383] Step 5: the (4aS)-3-acryloyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-1,2,3,4,4a, 5-hexahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,8]naphthyridin-7(8H)-one (160 mg, 0.27 mmol) was dissolved in DCM (4 mL). The resulting solution was cooled to 0° C., and then added dropwise with a solution (3 mL) of 17% boron tribromide in DCM. The resulting mixture was stirred at room temperature for 3 hours to react. The resulting reaction liquid was poured into 25 mL of cold saturated solution of NaHCO.sub.3 and extracted with 50 mL of DCM. The resulting organic phase was washed orderly with 25 mL of saturated solution of NaHCO.sub.3 and 25 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by preparative scale HPLC to obtain product (4aS)-3-acryloyl-11-fluoro-10-(2-fluoro-6-hydroxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-1,2,3,4,4a,5-hexahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,8]naphthyridin-7(8H)-one (Z28, 90 mg, Y: 57.6%), which was faint yellow solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.04 (s, 1H), 8.42 (d, J=4.8 Hz, 1H), 8.22 (d, J=7.4 Hz, 1H), 7.27-7.17 (m, 2H), 7.01-6.78 (m, 1H), 6.77-6.58 (m, 2H), 6.18 (d, J=16.3 Hz, 1H), 5.87-5.66 (m, 1H), 4.51-3.97 (m, 4H), 3.91-3.39 (m, 4H), 3.14-3.01 (m, 1H), 2.62-2.41 (m, 1H), 1.91-1.76 (m, 3H), 1.12-0.98 (m, 3H), 0.94-0.83 (m, 3H). ES-API: [M+H].sup.+=574.2.
Example 29 Preparation of Compound Z29
[0384] ##STR00519## ##STR00520##
[0385] Step 1: 7-chloro-6-fluoro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-1,8-naphthyridin-2(1H)-one (4.0 g, 11.53 mmol) was dissolved in acetic acid (9 mL), orderly added with sodium nitrite (79 mg, 1.15 mmol) and concentrated nitric acid (2.3 mL, 34.6 mmol), and stirred at room temperature for 30 minutes to react. The resulting reaction liquid was slowly poured into 100 mL of ice water. The precipitated solid was filtered. The filter cake was washed with 20 ml of ice water and dried in vacuum to obtain product 7-chloro-6-fluoro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (3.1 g, Y: 80%), which was yellow solid. ES-API: [M+H].sup.+=393.1.
[0386] Step 2: the 7-chloro-6-fluoro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (1.0 g, 2.55 mmol), (5-methyl-1H-indazol-4-yl)boric acid (1.8 g, 10.2 mmol), tetrakis(triphenylphosphine)palladium (589 mg, 0.51 mmol), potassium carbonate (1.76 g, 12.75 mmol), 2 mL of water, and 8 mL of dioxane were added to a 100 mL three-necked round-bottom flask. The resulting mixture was stirred at 110° C. for 1 hour to react under the protection of nitrogen. After the completion of the reaction, the resulting reaction liquid was cooled to room temperature, added with 80 mL of water and 100 mL of methyl tert-butyl ether, and extracted once. The water phase was then mixed with 1 M hydrochloric acid solution to adjust the pH to a range of 3 to 4. Solid was precipitated and filtered to obtain a solid product which was dried in vacuum to obtain product 6-fluoro-7-(5-methyl-1H-indazol-4-yl)-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2 (1H)-one (0.8 g, 50%), which was faint yellow solid. ES-API: [M+H].sup.+=489.2.
[0387] Step 3: the 6-fluoro-7-(5-methyl-1H-indazol-4-yl)-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2 (1H)-one (0.6 g, 1.23 mmol) was dissolved in ACN (20 mL), orderly added with phosphorus oxychloride (0.94 g, 6.15 mmol) and N,N-diisopropylethylamine (1.27 g, 9.84 mmol), and gradually heated to 80° C. and stirred for 24 hours to react. The resulting reaction liquid was concentrated, added with 30 mL of cold ACN, added dropwise to 30 mL of saturated sodium bicarbonate solution under the condition of an ice water bath, and extracted with EtOAc, and then combine the ethyl acetate phase (200 mL*2). The resulting combined EtOAc phase was washed with 50 mL of saturated salt solution once, dried by anhydrous sodium sulfate and filtered. After the resulting organic phase was dried and concentrated, the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-80%) to obtain 4-chloro-6-fluoro-7-(5-methyl-1H-indazol-4-yl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (0.3 g, Y: 50%), which was yellow solid. ES-API: [M+H]½=507.0.
[0388] Step 4: the tert-butyl 4-chloro-6-fluoro-7-(5-methyl-1H-indazol-4-yl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (150 mg, 0.296 mmol) was dissolved in N,N-dimethylacetamide (25 mL), subsequently added with tert-butyl (R)-3-(hydroxymethyl)piperazin-1-formate (1.48 g, 320 mmol), and stirred at 80° C. for 1.5 hours to react. The resulting reaction liquid was added with 80 mL of EtOAc, and washed with 80 mL of saturated salt solution for 3 times. The EtOAc phase was dried and concentrated, and the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-80%) to obtain product tert-butyl (3R)-4-(6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)-7-(5-methyl-1H-indazol-4-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-3-(hydroxymethyl)piperazin-1-carboxylate (10 mg, Y: 40%), which was yellow solid. ES-API: [M+H].sup.+=787.3.
[0389] Step 5: the tert-butyl (3R)-4-(6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)-7-(5-methyl-1H-indazol-4-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-3-(hydroxymethyl)piperazin-1-carboxylate (90 mg, 0.13 mmol) was dissolved in was dissolved in N,N-dimethylacetamide (25 mL), added with sodium hydride (15.7 mg, 0.39 mmol), and stirred at 130° C. for 18 hours to react. The resulting reaction liquid was cooled to room temperature, poured into ice water, mixed with 3 M hydrochloric acid to adjust the pH to 7, and added with 30 mL of EtOAc. The organic phase was separated, washed orderly with 30 mL of water and 30 mL of saturated salt solution, dried and concentrated to obtain product tert-butyl (4aR)-11-fluoro-8-(2-isopropyl-4-methylpyridin-3-yl)-10-(5-methyl-1H-indazol-4-yl)-7-oxo-1,2,4a,5,7,8-hexahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,8]naphthyridin-3(4H)-carboxylate (60 mg, Y: 70%), which was yellow solid. ES-API: [M+H].sup.+=640.3.
[0390] Step 6: the tert-butyl (4aR)-11-fluoro-8-(2-isopropyl-4-methylpyridin-3-yl)-10-(5-methyl-1H-indazol-4-yl)-7-oxo-1,2,4a,5,7,8-hexahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,8]naphthyridin-3(4H)-carboxylate (60 mg, 0.094 mmol) was dissolved in DCM (4 mL), and added with TFA (2 mL). After stirring at room temperature for 2 hours, the resulting reaction liquid was concentrated to obtain product (4aR)-11-fluoro-8-(2-isopropyl-4-methylpyridin-3-yl)-10-(5-methyl-1H-indazol-4-yl)-1,2,3,4,4a, 5-hexahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,8]naphthyridin7(8H)-one (50 mg, crude), which was directly used in next step. ES-API: [M+H].sup.+=540.2.
[0391] Step 7: the (4aR)-11-fluoro-8-(2-isopropyl-4-methylpyridin-3-yl)-10-(5-methyl-1H-indazol-4-yl)-1,2,3,4,4a, 5-hexahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,8]naphthyridin7(8H)-one (50 mg, 0.093 mmol) was dissolved in DCM (5 mL), and added with triethylamine (60 mg, 0.465 mmol). The resulting reaction liquid was cooled to 0° C., and then added dropwise with acryloyl chloride (10.5 mg, 0.083 mmol). The resulting mixture was stirred at 0° C. for 15 minutes to react. The resulting reaction liquid was added with 20 mL of DCM, washed with 20 mL of saturated solution of NaHCO.sub.3 and 20 mL of saturated salt solution, dried and concentrated, and purified by preparative scale HPLC to obtain product (4aR)-3-acryloyl-11-fluoro-8-(2-isopropyl-4-methylpyridin-3-yl)-10-(5-methyl-1H-indazol-4-yl)-1,2,3,4,4a,5-hexahydropyrazino[1′,2′:4,5][1,4]oxazino[2,3-c][1,8]naphthyridin-7(8H)-one (Z29, 15 mg, Y: 28%). ES-API: [M+H].sup.+=594.2. .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 13.08 (s, 1H), 8.37 (d, J=4.8 Hz, 1H), 8.32 (d, J=9.4 Hz, 1H), 7.49 (d, J=8.1 Hz, 2H), 7.24 (d, J=8.5 Hz, 1H), 7.20 (d, J=5.0 Hz, 1H), 6.90 (s, 1H), 6.20 (d, J=16.7 Hz, 1H), 5.78 (s, 1H), 4.45 (d, J=46.5 Hz, 1H), 4.35-4.20 (m, 2H), 4.05 (s, 1H), 3.85 (d, J=51.4 Hz, 1H), 3.75-3.57 (m, 2H), 3.48 (s, 1H), 3.15 (s, 1H), 2.05 (s, 3H), 1.90 (d, J=33.7 Hz, 3H), 1.03 (t, J=6.7 Hz, 3H), 0.81 (dd, J=20.9, 6.4 Hz, 3H).
Example 30 Preparation of Compounds Z30, Z30-1, and Z30-2
[0392] ##STR00521## ##STR00522##
[0393] Step 1: tert-butyl (4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (310 mg, 0.48 mmol), 10 mL of acetone, anhydrous potassium carbonate (265 mg, 1.92 mmol), and iodoethane (599 mg, 3.84 mmol) were orderly added to a 15 mL sealing tube. The sealing tube was sealed, and the resulting mixture was stirred at 55° C. for 18 hours to react. The resulting reaction liquid was concentrated, added with 60 mL of EtOAc, washed orderly with 30 mL of water and 30 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-70%) to obtain product tert-butyl (4aR)-6-ethyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-5,7-dioxo-1,2,4,4a,5,6,7,8-octyl-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (290 mg, Y: 89.7%), which was orange solid. ES-API: [M+H].sup.+=675.3.
[0394] Step 2: the tert-butyl (4aR)-6-ethyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-5,7-dioxo-1,2,4,4a,5,6,7,8-octyl-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (290 mg, 0.43 mmol) was dissolved in DCM (4 mL), and added with TFA (1 mL). After stirring at room temperature for 2 hours, the resulting reaction liquid was concentrated to obtain product (4aR)-6-ethyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (300 mg, crude), which was directly used in next step. ES-API: [M+H].sup.+=575.2.
[0395] Step 3: the (4aR)-6-ethyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (300 mg, crude) was dissolved in DCM (15 mL), and added with N,N-diisopropylethylamine (464 mg, 3.60 mmol). The resulting reaction liquid was cooled to 0° C., added with acryloyl chloride (130 mg, 1.44 mmol), and stirred at 0° C. for 15 minutes to react. The reaction liquid was added with 45 mL of DCM, washed orderly with 25 mL of water, 25 mL of saturated solution of NaHCO.sub.3 and 25 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-90%) to obtain product (4aR)-3-acryloyl-6-ethyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,3,4,4a, 6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (2450 mg, Y: 90.7%), which was faint yellow solid. ES-API: [M+H].sup.+=615.3.
[0396] Step 4: the (4aR)-3-acryloyl-6-ethyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,3,4,4a, 6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (245 mg, 0.39 mmol) was dissolved in DCM (5 mL). The resulting solution was cooled to 0° C., and then added dropwise with a solution (5 mL) of 17% boron tribromide in DCM. The resulting mixture was stirred at room temperature for 3 hours to react. The resulting reaction liquid was poured into 60 mL of saturated solution of NaHCO.sub.3 and extracted with 50 mL of DCM twice. The resulting organic phase was washed orderly with 30 mL of saturated solution of NaHCO.sub.3 and mL of saturated salt solution, dried and concentrated to obtain product (4aR)-3-acryloyl-6-ethyl-11-fluoro-10-(2-fluoro-6-hydroxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (Z30, 240 mg, Y: 100%), which was faint yellow solid. ES-API:[M+H].sup.+=615.3.
[0397] Step 5: the compound Z30 (240 mg, 0.39 mmol) was purified by preparative scale HPLC and then resolved by preparative scale chiral HPLC (column type: IB: 10 m, 30*250 mm; mobile phase: hexane:EtOH=70:30; flow rate: 25 ml/min; and column temperature: room temperature) to obtain the following atropisomer compounds. One of the atropisomer compounds had a structure arbitrarily specified as Z30-1 (71 mg, peak 1, retention time: 6.342 min, Y: 29.6%), which was faint yellow solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.11 (d, J=1.1 Hz, 1H), 8.45 (d, J=4.9 Hz, 1H), 8.40 (d, J=8.8 Hz, 1H), 7.32-7.18 (m, 2H), 7.12-6.80 (m, 1H), 6.75-6.62 (m, 2H), 6.15 (dd, J=16.8, 2.0 Hz, 1H), 5.75 (d, J=12.2 Hz, 1H), 4.72 (d, J=13.5 Hz, 1H), 4.46 (d, J=11.9 Hz, 1H), 4.18-3.93 (m, 3H), 3.63-3.50 (m, 2H), 3.26-3.06 (m, 1H), 2.80-2.55 (m, 1H), 2.50-2.39 (m, 1H), 1.97 (s, 3H), 1.10-0.95 (m, 6H), 0.86 (d, J=6.7 Hz, 3H). ES-API: [M+H].sup.+=615.2. The other atropisomer compound had a structure arbitrarily specified as Z30-2 (73 mg, peak 2, retention time: 7.970 min, Y: 30.5%), which was faint yellow solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.13 (s, 1H), 8.45 (d, J=4.9 Hz, 1H), 8.39 (d, J=8.8 Hz, 1H), 7.31-7.19 (m, 2H), 7.12-6.80 (m, 1H), 6.77-6.62 (m, 2H), 6.15 (dd, J=16.8, 2.1 Hz, 1H), 5.75 (d, J=12.3 Hz, 1H), 4.73 (d, J=14.1 Hz, 1H), 4.46 (d, J=13.0 Hz, 1H), 4.20-4.02 (m, 2H), 4.00-3.91 (m, 1H), 3.65-3.53 (m, 2H), 3.26-3.06 (m, 1H), 2.82-2.58 (m, 2H), 1.80 (s, 3H), 1.15-0.93 (m, 9H). .sup.1ES-API: [M+H].sup.+=615.2. The isomer compounds were detected by analytical scale chiral HPLC (column type: IB: 5 μm, 4.6*250 mm; mobile phase: hexane:EtOH=70:30; flow rate: 1 ml/min; and column temperature=30° C.).
Example 31 Preparation of Compound Z31
[0398] ##STR00523## ##STR00524##
[0399] Step 1: 6-chloro-7-(2-fluoro-6-methoxyphenyl)-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (1.0 g, 2.0 mmol), potassium cypropyltrifluoroborate (1.48 g, 10.0 mmol), SPhos-Pd-G2 (144 mg, 0.20 mmol), SPhos (82 mg, 0.20 mmol), potassium carbonate (1.66 g, 12.0 mmol), 2 mL of water, and 20 mL of toluene were added to a 250 mL round-bottom flask. The resulting mixture was stirred at 125° C. for 18 hours to react under the protection of nitrogen. The resulting reaction liquid was concentrated, added with 50 mL of water, mixed with 3.0 M diluted hydrochloric acid to adjust the pH to 3.0, and extracted with 50 mL of DCM twice. The resulting organic phase was dried and concentrated to obtain product 6-cyclopropyl-7-(2-fluoro-6-methoxyphenyl)-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (850 mg, crude), which was brown solid. ES-API: [M+H].sup.+=505.2.
[0400] Step 2: the 6-cyclopropyl-7-(2-fluoro-6-methoxyphenyl)-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (1.6 g, crude) was dissolved in ACN (50 mL), orderly added with phosphorus oxychloride (2.43 g, 15.85 mmol) and N,N-diisopropylethylamine (3.27 g, 25.36 mmol), and stirred at 85° C. for 1 hour to react. The resulting reaction liquid was concentrated, added with 120 mL of EtOAc, and washed orderly with 60 mL of water, with 60 mL of saturated sodium bicarbonate solution twice and then with 60 mL saturated salt solution. After the resulting organic phase was dried and concentrated, the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-35%) to obtain product 4-chloro-6-cyclopropyl-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (520 mg, Y: 24.8%), which was faint yellow solid. ES-API: [M+H].sup.+=523.2.
[0401] Step 3: the 4-chloro-6-cyclopropyl-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2 (1H)-one (490 mg, 0.94 mmol) was dissolved in N,N-dimethylacetamide (6 mL), orderly added with methyl (3R,6R)-1-N-BOC-6-methylpiperazin-3-formate (485 mg, 1.88 mmol) and N,N-diisopropylethylamine (364 mg, 2.82 mmol), and stirred at 125° C. for 3 hours to react. The resulting reaction liquid was added with 100 mL of EtOAc, washed with 30 mL of dilute brine for 4 times and then with 30 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-50%) to obtain product methyl (3R,6R)-1-N-BOC-4-(6-cyclopropyl-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-6-methylpiperazin-3-formate (485 mg, Y: 69.4%), which was orange solid. ES-API: [M+H].sup.+=745.3.
[0402] Step 4: the methyl (3R,6R)-1-N-BOC-4-(6-cyclopropyl-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-6-methylpiperazin-3-formate (455 mg, 0.61 mmol) was dissolved in acetic acid (8 mL), added with iron powder (120 mg, 2.14 mmol), and stirred at 80° C. for 30 minutes to react. The resulting reaction liquid was concentrated, and orderly added with 80 mL of EtOAc and 50 mL of saturated sodium bicarbonate solution. The resulting suspension was filtered by diatomite. The filter cake was washed with EtOAc. The resulting organic phase was separated, washed orderly with 25 mL of saturated sodium bicarbonate solution and 25 mL of saturated salt solution, dried and concentrated to obtain product tert-butyl (2R,4aR)-11-cyclopropyl-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octyl-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (415 mg, Y: 99.5%), which was faint yellow solid. ES-API: [M+H].sup.+=683.3.
[0403] Step 5: the tert-butyl (2R,4aR)-11-cyclopropyl-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octyl-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (415 mg, 0.61 mmol), 12 mL of acetone, anhydrous potassium carbonate (337 mg, 2.44 mmol), and iodomethane (693 mg, 4.88 mmol) were orderly added to a 50 mL sealing tube. The sealing tube was sealed, and the resulting mixture was stirred at 50° C. for 18 hours to react. The resulting reaction liquid was added with 60 mL of EtOAc, washed orderly with 15 mL of water and 15 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified on a preparative thin-layer chromatographic plate (DCM/MeOH=25:1) to obtain product tert-butyl (2R,4aR)-11-cyclopropyl-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,6-dimethyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octyl-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (160 mg, Y: 37.8%), which was faint yellow solid. ES-API: [M+H].sup.+=697.3.
[0404] Step 6: the tert-butyl (2R,4aR)-11-cyclopropyl-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,6-dimethyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octyl-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (160 mg, 0.23 mmol) was dissolved in DCM (3.5 mL), and added with TFA (0.8 mL). After stirring at room temperature for 2 hours, the resulting reaction liquid was concentrated to obtain product (2R,4aR)-11-cyclopropyl-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,6-dimethyl-1,2,4,4a, 6,8-hexahydro-312-pyrazino[4′,3′: 4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (165 mg, crude), which was directly used in next step. ES-API: [M+H].sup.+=597.2.
[0405] Step 7: the (2R,4aR)-11-cyclopropyl-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,6-dimethyl-1,2,4,4a, 6,8-hexahydro-312-pyrazino[4′,3′: 4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (165 mg, crude) was dissolved in DCM (10 mL), and added with N,N-diisopropylethylamine (148 mg, 1.15 mmol). The resulting reaction liquid was cooled to 0° C., and added dropwise with acryloyl chloride (41 mg, 0.46 mmol). The resulting mixture was stirred at 0° C. for 15 minutes to react. The resulting reaction liquid was added with 30 mL of DCM, washed orderly with 15 mL of water, 15 mL of saturated solution of NaHCO.sub.3 and 15 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (DCM/MeOH: 0-5%) to obtain product (2R,4aR)-3-acryloyl-11-cyclopropyl-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,6-dim ethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (140 mg, Y:93.7%), which was faint yellow solid. ES-API: [M+H].sup.+=651.3.
[0406] Step 8: the (2R,4aR)-3-acryloyl-11-cyclopropyl-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,6-dim ethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (130 mg, 0.20 mmol) was dissolved in DCM (3 mL). The resulting solution was cooled to 0° C., and then added dropwise with a solution (3 mL) of 17% boron tribromide in DCM. The resulting mixture was stirred at room temperature for 3 hours to react. The resulting reaction liquid was poured into 40 mL of saturated solution of NaHCO.sub.3 and extracted with 25 mL of DCM twice. The resulting organic phase was dried and concentrated, and the resulting crude product was purified by preparative scale HPLC to obtain product (2R,4aR)-3-acryloyl-11-cyclopropyl-10-(2-fluoro-6-hydroxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,6-dim ethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (Z31, 65 mg, Y: 51.1%), which was white solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 9.90 (s, 1H), 8.42 (d, J=4.8 Hz, 1H), 7.75-7.73 (m, 1H), 7.22-7.17 (m, 2H), 7.03 (dd, J=16.8, 10.5 Hz, 1H), 6.74-6.60 (m, 2H), 6.22-6.08 (m, 1H), 5.81-5.69 (m, 1H), 5.05-4.81 (m, 1H), 4.62-4.41 (m, 1H), 4.03-3.90 (m, 1H), 3.75 (dd, J=14.1, 4.2 Hz, 1H), 3.39-3.25 (m, 4H), 2.83-2.67 (m, 1H), 2.48-2.37 (m, 1H), 2.01-1.75 (m, 3H), 1.70-1.46 (m, 4H), 1.14-0.55 (m, I0H). ES-API: [M+H].sup.+=637.3.
Example 32 Preparation of Compound Z32
[0407] ##STR00525## ##STR00526##
[0408] Step 1: 4,6-bicyclopropylpyrimidin-5-amine (742 mg, 4.24 mmol) was dissolved in dry tetrahydrofuran (20 mL), added with 2 M NaHMDS (8.48 mL, 16.96 mmol) under the condition of an ice water bath, and stirred for 20 minutes under the condition of the ice water bath. The resulting mixture was added with 2,5-difluoro-6-(2-fluoro-6-methoxyphenyl)nicotinic acid (1.2 g, 4.24 mmol), and stirred at room temperature for 3 hours. The resulting reaction liquid was slowly poured into 30 mL of ice water, mixed with diluted hydrochloric acid (3M) to adjust the pH to a range of 5 to 6, extracted with EtOAc, washed with 50 mL of saturated salt solution once, dried by anhydrous sodium sulfate, and filtered. After the resulting organic phase was dried and concentrated, the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 20-40%) to obtain product 2-((4,6-bicyclopropylpyrimidin-5-yl)amino)-5-fluoro-6-(2-fluoro-6-methoxyphenyl)nicotinic acid (1.8 g, Y: 98%), which was yellow solid. ES-API: [M+H].sup.+=439.1.
[0409] Step 2: the 2-((4,6-bicyclopropylpyrimidin-5-yl)amino)-5-fluoro-6-(2-fluoro-6-methoxyphenyl)nicotinic acid (1.5 g, 3.42 mmol) was dissolved in dichloroethane, and added with thionyl chloride (4.07 g, 34.2 mmol). The resulting mixture was stirred at 80° C. for 2 hours to react. After the completion of the reaction, the resulting reaction liquid was cooled to room temperature, concentrated, and dried in 50 degrees of vacuum for 4 hours to obtain product 2-((4,6-dicyclopropylpyrimidin-5-yl)amino)-5-fluoro-6-(2-fluoro-6-methoxyphenyl)nicotinoyl chloride (1.57 g, crude), which was faint yellow solid. MeOH was used to detect ES-API: [M+H].sup.+=453.2.
[0410] Step 3: under the condition of the ice water bath, sodium hydride (1.97 g, 49.35 mmol) was added to a solution of ethyl nitroacetate (1.31 g, 9.86 mmol) in tetrahydrofuran, stirred for 30 minutes, subsequently added with the 2-((4,6-bicyclopropylpyrimidin-5-yl)amino)-5-fluoro-6-(2-fluoro-6-methoxyphenyl)nicotinoyl chloride (1.57 g, 3.29 mmol), stirred at room temperature for 1 hour, and then heated to 80° C. to react for 2 hours. The resulting reaction liquid was poured into ice water, mixed with 3 M hydrochloric acid to adjust the pH to 34, extracted with EtOAc, dried by anhydrous sodium sulfate, and filtered. The resulting organic phase was dried and concentrated to obtain product 1-(4,6-bicyclopropylpyrimidin-5-yl)-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-4-hydroxy-3-nitro-1,8-naphthyridin-2(1H)-one (110 mg, Y: 20%). ES-API: [M+H].sup.+=508.1.
[0411] Step 4: the 1-(4,6-bicyclopropylpyrimidin-5-yl)-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-4-hydroxy-3-nitro-1,8-naphthyridin-2(1H)-one (110 mg, 0.20 mmol) was dissolved in ACN (10 mL), orderly added with phosphorus oxychloride (153 mg, 1.0 mmol) and N,N-diisopropylethylamine (77 g, 0.6 mmol), and gradually heated to 80° C. and stirred for 3 hours to react. The resulting reaction liquid was concentrated, added with 30 mL of cold ACN, added dropwise to 30 mL of saturated sodium bicarbonate solution under the condition of the ice water bath, and extracted with EtOAc (50 mL*2). The resulting combined organic phase was washed with 30 mL of saturated salt solution, dried by anhydrous sodium sulfate, and filtered. After the resulting organic phase was dried and concentrated, the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-80%) to obtain 4-chloro-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(4,6-bicyclopropylpyrimidin-5-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (110 mg, Y: 68%), which was yellow solid. ES-API: [M+H].sup.+=526.2.
[0412] Step 5: the 4-chloro-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(4,6-bicyclopropylpyrimidin-5-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (110 mg, 0.296 mmol) was dissolved in N,N-dimethylacetamide (3 mL), subsequently added with 1-(tert-butyl)3-methyl(R)-piperazin-1,3-dicarboxylate (54 mg, 0.22 mmol), and stirred at 120° C. for 2 hours to react. After the completion of the reaction, the resulting product was added with 30 mL of EtOAc and washed with 30 mL of saturated salt solution for 3 times. The EtOAc phase was dried and concentrated to obtain a crude product, namely target product (3R)-1-tert-butyl3-methyl4-(1-(4,6-bicyclopropylpyrimidin-5-yl)-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazin-1,3-dione (50 mg, Y: 46%), which was yellow solid. ES-API: [M+H].sup.+=734.3.
[0413] Step 6: the (3R)-1-tert-butyl3-methyl4-(1-(4,6-bicyclopropylpyrimidin-5-yl)-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazin-1,3-dione (50 mg, 0.068 mmol) was dissolved in acetic acid (25 mL), added with iron powder (11.5 mg, 0.204 mmol), and stirred at 80° C. for 30 minutes to react. The resulting reaction liquid was concentrated, and orderly added with 30 mL of EtOAc and 30 mL of saturated sodium bicarbonate solution. The resulting suspension was filtered by diatomite. The filter cake was washed with EtOAc. The resulting organic phase was separated, washed orderly with 30 mL of saturated sodium bicarbonate solution and 30 mL of saturated salt solution, dried and concentrated to obtain product tert-butyl (4aR)-8-(4,6-bicyclopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-5,7-dioxo-4,4a,5,6,7,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3(2H)-carboxylate (40 mg, crude), which was yellow solid. ES-API: [M+H].sup.+=672.2.
[0414] Step 7: the tert-butyl (4aR)-8-(4,6-bicyclopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-5,7-dioxo-4,4a,5,6,7,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3(2H)-carboxylate (40 mg, 0.059 mmol), 30 mL of acetone, anhydrous potassium carbonate (33 mg, 0.24 mmol), and iodomethane (85 mg, 0.59 mmol) were sealed in a sealing tube, and stirred at 50° C. for 18 hours to react. The resulting reaction liquid was added with 20 mL of EtOAc, washed with 20 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-80%) to obtain product (4aR)-tert-butyl8-(4,6-bicyclopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-6-methyl-5,7-dioxo-4,4a,5,6,7,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3(2H)-one (40 mg, Y: 90%), which was yellow solid. ES-API: [M+H].sup.+=686.2.
[0415] Step 8: the (4aR)-tert-butyl8-(4,6-bicyclopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-6-methyl-5,7-dioxo-4,4a,5,6,7,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3(2H)-one (44 mg) was dissolved in DCM (3 mL), and added with TFA (1 mL). After stirring at room temperature for 2 hours, the resulting reaction liquid was concentrated to obtain product (4aR)-8-(4,6-bicyclopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-6-methyl-2,3,4,4a-tetrahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7(6H, 8H)-dione (40 mg, crude), which was directly used in next step. ES-API: [M+H].sup.+=586.2.
[0416] Step 9: the (4aR)-8-(4,6-bicyclopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-6-methyl-2,3,4,4a-tetrahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7(6H, 8H)-dione (40 mg, 0.068 mmol) was dissolved in DCM (5 mL), and added with diisopropylethylamine (53 mL, 0.408 mmol). The resulting reaction liquid was cooled to 0° C., and added dropwise with acryloyl chloride (12.4 mg, 0.137 mmol). The resulting mixture was stirred at 0° C. for 15 minutes to react. The resulting reaction liquid was added with 20 mL of DCM, washed with 20 mL of saturated solution of NaHCO.sub.3 and 20 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by preparative scale HPLC to obtain product (4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(4,6-bicyclopropylpyrimidin-5-yl)-6-methyl-2,3,4,4a, 6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (Z32, 10 mg, Y: 22%), which was yellow solid. ES-API: [M+H].sup.+=640.2.
Example 33 Preparation of Compounds Z33, Z33-1, and Z33-2
[0417] ##STR00527## ##STR00528## ##STR00529##
[0418] Step 1: 7-chloro-6-fluoro-4-hydroxy-1-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-2-oxo-1,2-dihydro-1,8-naphthyridin-3-nitrile (3.6 g, 10.0 mmol) was suspended in a mixed solution of 1,4-dioxane (10 mL) and water (120 mL), and slowly added with concentrated sulfuric acid (10 mL). The resulting mixture was stirred at 120° C. for 18 hours to react. Cooled reaction liquid was poured into 20 mL of ice water, mixed with sodium carbonate to adjust the pH to a range of 2 to 3, and extracted with EtOAc (1000 mL*2). The resulting combined EtOAc phase was dried by anhydrous sodium sulfate, and filtered. The filtrate was dried in vacuum to obtain product 7-chloro-6-fluoro-4-hydroxy-1-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-1,8-naphthyridin-2(1H)-one (3.36 g, Y: 92%), which was light brown solid. ES-API: [M+H].sup.+=337.1.
[0419] Step 2: the 7-chloro-6-fluoro-4-hydroxy-1-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-1,8-naphthyridin-2(1H)-one (3.36 g, 10 mmol) was dissolved in acetic acid (7 mL), orderly added with sodium nitrite (69 mg, 1.0 mmol) and concentrated nitric acid (2.0 mL, 30 mmol), and stirred at room temperature for 30 minutes to react. The resulting reaction liquid was slowly added into 21 mL of ice water. The precipitated solid was filtered. The filter cake was washed with 10 ml of ice water and dried in vacuum to obtain product 7-chloro-6-fluoro-4-hydroxy-1-(1-isopropyl-4-methyl-1H-pyrazol-5-yl).sup.3-nitro-1,8-naphthyridin-2(1H)-one (3.0 g, Y: 90%), which was yellow solid. ES-API: [M+H].sup.+=382.1.
[0420] Step 3: the 7-chloro-6-fluoro-4-hydroxy-1-(1-isopropyl-4-methyl-1H-pyrazol-5-yl).sup.3-nitro-1,8-naphthyridin-2(1H)-one (1.5 g, 3.93 mmol), (2-fluoro-6-methoxyphenyl)boric acid (2.67 g, 15.72 mmol), tetrakis(triphenylphosphine)palladium (908 mg, 0.786 mmol), potassium carbonate (2.72 g, 19.65 mmol), 4 mL of water, and 20 mL of dioxane were added to a 100 mL three-necked round-bottom flask. The resulting mixture was stirred at 100° C. for 3 hours to react under the protection of nitrogen. After the completion of the reaction, the resulting reaction liquid was cooled to room temperature, added with 20 mL of water and 50 mL of methyl tert-butyl ether, and extracted once. The water phase was then mixed with 1 M hydrochloric acid solution to adjust the pH to a range of 3 to 5, and extracted with EtOAc (50 mL*2). The resulting combined EtOAc phase was dried by anhydrous sodium sulfate, and filtered. The filtrate was dried in vacuum to obtain product 6-fluoro-7-(2-fluoro-6-methoxyphenyl)-4-hydroxy-1-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (1.5 g, crude), which was faint yellow solid. ES-API: [M+H].sup.+=472.1.
[0421] Step 4: the 6-fluoro-7-(2-fluoro-6-methoxyphenyl)-4-hydroxy-1-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (1.5 g, 3.18 mmol) was dissolved in ACN (15 mL), orderly added with phosphorus oxychloride (2.4 ml, 25.5 mmol) and N,N-diisopropylethylamine (2.6 ml, 15.9 mmol), and gradually heated to 80° C. and stirred for 30 minutes to react. The resulting reaction liquid was concentrated, added with 10 mL of cold ACN, added dropwise to 20 mL of saturated sodium bicarbonate solution in an ice water bath, and extracted with EtOAc (20 mL*2). The resulting combined EtOAc phase was washed with 20 mL of saturated salt solution once, dried by anhydrous sodium sulfate, and filtered. After the resulting organic phase was dried and concentrated, the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-50%) to obtain 4-chloro-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (0.9 g, Y: 65%), which was yellow solid. ES-API: [M+H].sup.+=490.1.
[0422] Step 5: the 4-chloro-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (490 mg, 1.0 mmol) was dissolved in N,N-dimethylacetamide (5 mL), orderly added with (3R,6R)-1-tert-butyl3-methyl6-methylpiperazin-1,3-dicarboxylic acid (310 mg, 1.2 mmol) and N,N-diisopropylethylamine (390 mg, 3 mmol), and stirred at 120° C. for 2 hours to react. The resulting reaction liquid was added with 20 mL of EtOAc, and washed with 20 mL of saturated salt solution for 3 times. The EtOAc phase was dried and concentrated to obtain product (3R,6R)-1-tert-butyl-3-methyl4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazin-1,3-dione (620 mg, crude), which was yellow solid. ES-API: [M+H].sup.+=712.2.
[0423] Step 6: the (3R,6R)-1-tert-butyl-3-methyl4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazin-1,3-dione (620 mg, 0.872 mmol) was dissolved in acetic acid (8 mL), added with iron powder (146 mg, 2.62 mmol), and stirred at 80° C. for 30 minutes to react. The resulting reaction liquid was concentrated, and orderly added with 30 mL of EtOAc and 30 mL of saturated sodium bicarbonate solution. The resulting suspension was filtered by using diatomite. The filter cake was washed with EtOAc. The resulting organic phase was separated, washed orderly with 30 mL of saturated sodium bicarbonate solution and 30 mL of saturated salt solution, dried and concentrated to obtain product tert-butyl (2R,4aR)-tert-butyl11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-2-methyl-5,7-dioxo-4,4a,5,6,7,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3(2H)-carboxylate (300 mg, crude), which was yellow solid. ES-API: [M+H].sup.+=650.3.
[0424] Step 7: the tert-butyl (2R,4aR)-tert-butyl11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-2-methyl-5,7-dioxo-4,4a,5,6,7,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3(2H)-carboxylate (300 mg, 0.462 mmol), 6 mL of acetone, anhydrous potassium carbonate (255 mg, 1.84 mmol), and iodomethane (656 mg, 4.62 mmol) were sealed in a sealing tube, and stirred at 50° C. for 18 hours to react. The resulting reaction liquid was added with 20 mL of EtOAc, washed with 20 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-80%) to obtain product tert-butyl (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-2,6-dimethyl-5,7-di oxo-4,4a,5,6,7,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3(2H)-carboxylate (350 mg, Y: 95%), which was yellow solid. ES-API: [M+H].sup.+=664.3.
[0425] Step 8: the tert-butyl (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-2,6-dimethyl-5,7-di oxo-4,4a,5,6,7,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3(2H)-carboxylate (350 mg) was dissolved in DCM (4 mL), and added with TFA (2 mL). After stirring at room temperature for 2 hours, the resulting reaction liquid was concentrated to obtain product (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-2,6-dimethyl-2,3,4,4a-tetrahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7(6H, 8H)-dione (40 mg, crude), which was directly used in next step. ES-API: [M+H].sup.+=564.2.
[0426] Step 9: the (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-2,6-dimethyl-2,3,4,4a-tetrahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7(6H, 8H)-dione (350 mg, 0.62 mmol) was dissolved in DCM (6 mL), and added with diisopropylethylamine (480 mg, 3.72 mmol). The resulting reaction liquid was cooled to 0° C., and added dropwise with acryloyl chloride (112.5 mg, 1.24 mmol). The resulting mixture was stirred at 0° C. for 15 minutes to react. The resulting reaction liquid was added with 20 mL of DCM, washed with 20 mL of saturated solution of NaHCO.sub.3 and 20 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by preparative scale chromatography to obtain product (2R,4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-2,6-dimethyl-2,3,4,4a-tetrahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7(6H, 8H)-dione (250 mg, Y:60%), which was yellow solid. ES-API: [M+H].sup.+=618.3.
[0427] Step 10: under the condition of the ice water bath, the (2R,4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-2,6-dimethyl-2,3,4,4a-tetrahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7(6H, 8H)-dione (250 mg, 0.405 mmol) was added to dry DCM (6.0 mL), then added with boron tribromide (4.0 mL, 4.0 mmol), and warmed to room temperature to react for 1 hour. Under the condition of the ice water bath, the above reaction liquid was added dropwise to saturated sodium bicarbonate solution, extracted with DCM (30 mL) twice, dried and concentrated to obtain product (2R,4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-hydroxyphenyl)-8-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-2,6-dimethyl-2,3,4,4a-tetrahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7(6H, 8H)-dione (Z33).
[0428] Step 11: the compound Z33 was purified by preparative scale HPLC to obtain the following atropisomer compounds. One of the atropisomer compounds had a structure arbitrarily specified as Z33-1 (peak 1, 30 mg, retention time: 9.576 min, Y: 50%). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.17 (s, 1H), 7.98 (dd, J=8.4, 5.5 Hz, 1H), 7.40 (d, J=5.5 Hz, 1H), 7.29 (q, J=7.9 Hz, 1H), 7.02 (dd, J=16.8, 10.6 Hz, 1H), 6.81-6.68 (m, 2H), 6.20-6.11 (m, 1H), 5.81-5.69 (m, 1H), 4.77 (s, 1H), 4.61 (d, J=14.7 Hz, 1H), 4.01-3.83 (m, 2H), 3.73 (dd, J=14.2, 4.2 Hz, 1H), 3.35 (d, J=5.8 Hz, 3H), 2.86 (dd, J=48.2, 12.0 Hz, 1H), 1.76-1.59 (m, 3H), 1.55 (dd, J=16.6, 6.7 Hz, 3H), 1.26 (dd, J=32.6, 6.6 Hz, 3H), 1.21-1.10 (m, 3H). The other atropisomer compound had a structure arbitrarily specified as Z33-2 (peak 2, 15 mg, retention time: 9.663 min, Y: 25%). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.17 (s, 1H), 7.96 (m, 1H), 7.40 (d, J=5.5 Hz, 1H), 7.29 (q, J=7.9 Hz, 1H), 7.02 (m, 1H), 6.81-6.68 (m, 2H), 6.20-6.11 (m, 1H), 5.81-5.69 (m, 1H), 4.77 (s, 1H), 4.61 (d, J=14.7 Hz, 1H), 4.01-3.83 (m, 2H), 3.73 (dd, J=14.2, 4.2 Hz, 1H), 3.35 (d, J=5.8 Hz, 3H), 2.86 (m, 1H), 1.76-1.59 (m, 3H), 1.55 (m, 3H), 1.30 (dd, J=32.6, 6.6 Hz, 3H), 1.23-1.15 (m, 3H). The isomer compounds were detected by analytical scale HPLC.
Example 34 Preparation of Compounds Z34, Z34-1, and Z34-2
[0429] ##STR00530## ##STR00531##
[0430] Step 1: tert-butyl (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octyl-3H-pyrazion[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (115 mg, 0.17 mmol), 4 mL of acetone, anhydrous potassium carbonate (94 mg, 0.68 mmol), and deuteroiodomethane (246 mg, 1.70 mmol) were added to a 15 mL sealing tube. The sealing tube was sealed, and the resulting mixture was stirred at 50° C. for 18 hours to react. The resulting reaction liquid was added with 30 mL of EtOAc, washed orderly with 12 mL of water and 15 mL of saturated salt solution, dried and concentrated to obtain product tert-butyl (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2-methyl-6-(methyl-d3)-5,7-dioxo-1,2,4,4a,5,6,7,8-octyl-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (118 mg, Y: 100.0%), which was yellow solid. ES-API: [M+H].sup.+=678.3.
[0431] Step 2: the tert-butyl (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2-methyl-6-(methyl-d3)-5,7-dioxo-1,2,4,4a,5,6,7,8-octyl-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (118 mg, 0.17 mmol) was dissolved in DCM (3 mL), and added with TFA (0.7 mL). After stirring at room temperature for 2 hours, the resulting reaction liquid was concentrated to obtain product (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2-methyl-6-(methyl-d3)-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (120 mg, crude), which was directly used in next step. ES-API: [M+H].sup.+=578.2.
[0432] Step 3: the (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2-methyl-6-(methyl-d3)-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (120 mg, crude) was dissolved in DCM (5 mL), and added with N,N-diisopropylethylamine (110 mg, 0.85 mmol). The resulting reaction liquid was cooled to 0° C., and added dropwise with acryloyl chloride (31 mg, 0.34 mmol). The resulting mixture was stirred at 0° C. for 15 minutes to react. The resulting reaction liquid was added with 25 mL of DCM, washed orderly with 15 mL of water, 15 mL of saturated solution of NaHCO.sub.3 and 15 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-100%) to obtain product (2R,4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2-methyl-6-(methyl-d3)-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (85 mg, Y: 77.3%), which was faint yellow solid. ES-API: [M+H].sup.+=632.2.
[0433] Step 4: the (2R,4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2-methyl-6-(methyl-d3)-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (85 mg, 0.13 mmol) was dissolved in DCM (1.5 mL). The resulting solution was cooled to 0° C., and then added dropwise with a solution (1.5 mL) of 17% boron tribromide in DCM. The resulting mixture was stirred at room temperature for 4 hours to react. The resulting reaction liquid was poured into 40 mL of saturated solution of NaHCO.sub.3 and extracted with 25 mL of DCM twice. The resulting organic phase was dried and concentrated, and the resulting crude product was purified by preparative scale HPLC to obtain product (2R,4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2-methyl-6-(methyl-d3)-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (Z34).
[0434] Step 5: the compound Z34 was resolved by preparative scale chiral HPLC (column type: OD-H: 10 μm, 20*250 mm; mobile phase: hexane:EtOH=80:20; flow rate: 15 ml/min; and column temperature: room temperature) and purified to obtain the following atropisomer compounds. One of the atropisomer compounds had a structure arbitrarily specified as Z34-1 (23 mg, peak 1, retention time: 11.056 min, Y: 28.7%), which was faint yellow solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.14 (s, 1H), 8.44 (d, J=4.9 Hz, 1H), 8.01-7.97 (m, 1H), 7.28-7.23 (m, 2H), 7.05-6.84 (m 1H), 6.77-6.64 (m, 2H), 6.18-6.13 (m, 1H), 5.77-5.71 (m, 1H), 5.03-4.77 (m, 1H), 4.61-4.41 (m, 1H), 4.06-4.00 (m, 1H), 3.73 (dd, J=14.1, 4.2 Hz, 1H), 3.39-3.20 (m, 1H), 2.92-2.79 (m, 1H), 2.47-2.36 (m, 1H), 1.99 (s, 3H), 1.58-1.53 (m, 3H), 1.03 (d, J=6.7 Hz, 3H), 0.85 (d, J=6.7 Hz, 3H). ES-API: [M+H].sup.+=618.2. the other atropisomer compound had a structure arbitrarily specified as Z34-2 (25 mg, peak 2, retention time: 14.067 min, Y: 31.2%), which was faint yellow solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.15 (s, 1H), 8.45 (d, J=4.9 Hz, 1H), 8.01-7.97 (m, 1H), 7.28-7.23 (m, 2H), 7.05-6.85 (m 1H), 6.74-6.63 (m, 2H), 6.18-6.13 (m, 1H), 5.77-5.71 (m, 1H), 5.04-4.77 (m, 1H), 4.62-4.41 (m, 1H), 4.00-3.94 (m, 1H), 3.73 (dd, J=14.1, 4.2 Hz, 1H), 3.43-3.25 (m, 1H), 2.95-2.83 (m, 1H), 2.79-2.74 (m, 1H), 1.80 (s, 3H), 1.58-1.53 (m, 3H), 1.11 (d, J=6.7 Hz, 3H), 0.98 (d, J=6.7 Hz, 3H). ES-API: [M+H].sup.+=618.2. The isomer compounds were detected by analytical scale chiral HPLC (column type: OD-H: 5 μm, 4.6*250 mm; mobile phase: hexane:EtOH=80:20; flow rate: 1 ml/min; and column temperature=30° C.).
Example 35 Preparation of Compounds Z35, Z35-1, and Z35-2
[0435] ##STR00532## ##STR00533##
[0436] Step 1: tert-butyl (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octyl-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (200 mg, 0.30 mmol) was dissolved in DCM (4 mL), and added with TFA (1 mL). After stirring at room temperature for 2 hours, the resulting reaction liquid was concentrated to obtain product (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (210 mg, crude), which was directly used in next step. ES-API: [M+H].sup.+=561.3.
[0437] Step 2: the (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (210 mg, crude) was dissolved in DCM (10 mL), and added with N,N-diisopropylethylamine (194 mg, 1.50 mmol). The resulting reaction liquid was cooled to 0° C., and added dropwise with acryloyl chloride (54 mg, 0.60 mmol). The resulting mixture was stirred at 0° C. for 15 minutes to react. The resulting reaction liquid was added with 30 mL of DCM, washed orderly with 15 mL of water, 15 mL of saturated solution of NaHCO.sub.3 and 15 mL of saturated salt solution, dried and concentrated to obtain product (2R,4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (175 mg, Y: 95.0%), which was faint yellow solid. ES-API: [M+H].sup.+=615.3.
[0438] Step 3: the (2R,4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (175 mg, 0.28 mmol) was dissolved in DCM (4 mL). The resulting solution was cooled to 0° C., and then added dropwise with a solution (4 mL) of 17% boron tribromide in DCM. The resulting mixture was stirred at room temperature for 3 hours to react. The resulting reaction liquid was poured into 80 mL of saturated solution of NaHCO.sub.3 and extracted with 30 mL of DCM twice. The resulting organic phase was dried and concentrated to obtain product (2R,4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-hydroxyphenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4]pyrazino[2,3,3-c][1,8,8]naphthyridin-5-dione (Z35, 170 mg, Y: 99.4%), which was faint yellow solid. ES-API: [M+H].sup.+=601.2.
[0439] Step 4: the compound Z35 (170 mg, 0.28 mmol) was purified by preparative scale HPLC and then resolved by preparative scale chiral HPLC (column type: IA: 10 m, 30*250 mm; mobile phase: hexane:EtOH=40:60; flow rate: 25 ml/min; and column temperature: room temperature) to obtain the following atropisomer compounds. One of the atropisomer compounds had a structure arbitrarily specified as Z35-1 (19 mg, peak 1, retention time: 2.905 min, Y: 11.1%), which was faint yellow solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.60-10.46 (m, 1H), 10.12 (s, 1H), 8.45 (d, J=4.8 Hz, 1H), 8.01-7.91 (m, 1H), 7.31-7.18 (m, 2H), 7.03-6.82 (m, 1H), 6.75-6.63 (m, 2H), 6.20-6.10 (m, 1H), 5.79-5.70 (m, 1H), 5.08-4.70 (m, 1H), 4.67-4.39 (m, 1H), 4.09-3.97 (m, 1H), 3.72 (dd, J=14.1, 4.0 Hz, 1H), 3.31-3.18 (m, 1H), 3.02-2.87 (m, 1H), 2.57-2.50 (m, 1H), 1.89 (s, 3H), 1.58-1.44 (m, 3H), 1.05 (d, J=6.7 Hz, 3H), 0.89 (d, J=6.7 Hz, 3H). ES-API: [M+H].sup.+=601.2. The other atropisomer compound had a structure arbitrarily specified as Z35-2 (19 mg, peak 2, retention time: 8.769 min, Y: 11.1%), which was faint yellow solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.60-10.46 (m, 1H), 10.12 (s, 1H), 8.45 (d, J=4.9 Hz, 1H), 8.01-7.91 (m, 1H), 7.31-7.18 (m, 2H), 7.03-6.82 (m, 1H), 6.75-6.63 (m, 2H), 6.20-6.08 (m, 1H), 5.78-5.67 (m, 1H), 5.10-4.70 (m, 1H), 4.67-4.39 (m, 1H), 4.09-3.97 (m, 1H), 3.72 (dd, J=14.1, 3.8 Hz, 1H), 3.31-3.18 (m, 1H), 3.06-2.92 (m, 1H), 2.66-2.57 (m, 1H), 1.83 (s, 3H), 1.58-1.44 (m, 3H), 1.08 (d, J=6.7 Hz, 3H), 0.91 (d, J=6.7 Hz, 3H). ES-API: [M+H].sup.+=601.2. The isomer compounds were detected by analytical scale chiral HPLC (column type: IA: 5 m, 4.6*150 mm; mobile phase: hexane:EtOH=40:60; flow rate: 1 ml/min; and column temperature=30° C.).
Example 36 Preparation of Compounds Z36, Z36-1, and Z36-2
[0440] ##STR00534## ##STR00535## ##STR00536## ##STR00537##
[0441] Step 1: 7-chloro-6-fluoro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridin-3-carbonitrile (3.6 g, 10.0 mmol) was suspended in a mixed solution of 1,4-dioxane (10 mL) and water (120 mL), and slowly added with concentrated sulfuric acid (10 mL). The resulting mixture was stirred at 120° C. for 18 hours to react. Cooled reaction liquid was poured into 20 mL of ice water, mixed with sodium carbonate to adjust the pH to a range of 2 to 3, and extracted with EtOAc (1000 mL*2). The resulting combined EtOAc phase was dried by anhydrous sodium sulfate, and filtered. The filtrate was dried in vacuum to obtain product 7-chloro-6-fluoro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-1,8-naphthyridin-2(1H)-one (3.36 g, Y: 90%), which was light brown solid. ES-API: [M+H]=348.1.
[0442] Step 2: the 7-chloro-6-fluoro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-1,8-naphthyridin-2(1H)-one (3.36 g, 10 mmol) was dissolved in acetic acid (7 mL), orderly added with sodium nitrite (69 mg, 1.0 mmol) and concentrated nitric acid (2.0 mL, 30 mmol), and stirred at room temperature for 30 minutes to react. The resulting reaction liquid was slowly poured into 21 mL of ice water. The precipitated solid was filtered. The filter cake was washed with 10 ml of ice water and dried in vacuum to obtain product 7-chloro-6-fluoro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl).sup.3-nitro-1,8-naphthyridin-2(1H)-one (3.0 g, Y: 90%), which was yellow solid. ES-API: [M+H].sup.+=393.1.
[0443] Step 3: the 7-chloro-6-fluoro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl).sup.3-nitro-1,8-naphthyridin-2(1H)-one (1.5 g, 3.93 mmol), (2-fluorophenyl)boric acid (2.67 g, 15.72 mmol), tetrakis(triphenylphosphine)palladium (908 mg, 0.786 mmol), potassium carbonate (2.72 g, 19.65 mmol), 4 mL of water, and 20 mL of dioxane were added to a 100 mL three-necked round-bottom flask. The resulting mixture was stirred at 100° C. for 3 hours to react under the protection of nitrogen. After the completion of the reaction, the resulting reaction liquid was cooled to room temperature, added with 20 mL of water and 50 mL of methyl tert-butyl ether, and extracted once. The water phase was then mixed with 1 M hydrochloric acid solution to adjust the pH to a range of 3 to 5, and extracted with EtOAc (50 mL*2). The resulting combined EtOAc phase was dried by anhydrous sodium sulfate, and filtered. The filtrate was dried in vacuum to obtain product 6-fluoro-7-(2-fluorophenyl)-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (1.5 g, Y: 65%), which was faint yellow solid. ES-API: [M+H].sup.+=453.1.
[0444] Step 4: the 6-fluoro-7-(2-fluorophenyl)-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (1.5 g, 3.18 mmol) was dissolved in ACN (15 mL), orderly added with phosphorus oxychloride (2.4 ml, 25.5 mmol) and N,N-diisopropylethylamine (2.6 ml, 15.9 mmol), and gradually heated to 80° C. and stirred for 30 minutes to react. The resulting reaction liquid was concentrated, added with 10 mL of cold ACN, added dropwise to 20 mL of saturated sodium bicarbonate solution in an ice water bath, and extracted with EtOAc (20 mL*2). The resulting combined EtOAc phase was washed with 20 mL of saturated salt solution once, dried by anhydrous sodium sulfate, and filtered. After the resulting organic phase was dried and concentrated, the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-50%) to obtain 4-chloro-6-fluoro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (0.9 g, Y: 65%), which was yellow solid. ES-API: [M+H].sup.+=471.1.
[0445] Step 5: the 4-chloro-6-fluoro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (490 mg, 1.0 mmol) was dissolved in N,N-dimethylacetamide (5 mL), orderly added with (3R,6R)-1-tert-butyl3-methyl 6-methylpiperazin-1,3-dicarboxylic acid (310 mg, 1.2 mmol) and N,N-diisopropylethylamine (390 mg, 3 mmol), and stirred at 120° C. for 1 hour to react. The resulting reaction liquid was added with 20 mL of EtOAc, and washed with 20 mL of saturated salt solution for 3 times. The EtOAc phase was dried and concentrated to obtain product (3R,6R)-1-tert-butyl-3-methyl4-(6-fluoro-7-(2-fluorophenyl)-1-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazin-1,3-dione (620 mg, Y: 50%), which was yellow solid. ES-API: [M+H].sup.+=692.2.
[0446] Step 6: the (3R,6R)-1-tert-butyl-3-methyl4-(6-fluoro-7-(2-fluorophenyl)-1-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazin-1,3-dione (620 mg, 0.872 mmol) was dissolved in acetic acid (8 mL), added with iron powder (146 mg, 2.62 mmol), and stirred at 80° C. for 30 minutes to react. The resulting reaction liquid was concentrated, and orderly added with 30 mL of EtOAc and 30 mL of saturated sodium bicarbonate solution. The resulting suspension was filtered by diatomite. The filter cake was washed with EtOAc. The resulting organic phase was separated, washed orderly with 30 mL of saturated sodium bicarbonate and 30 mL of saturated salt solution, dried and concentrated to obtain product tert-butyl (2R,4aR)-11-fluoro-10-(2-fluorophenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2-methyl-5,7-dioxo-4,4a,5,6,7,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3(2H)-carboxylate (300 mg, Y: 80%), which was yellow solid. ES-API: [M+H].sup.+=631.3.
[0447] Step 7: the tert-butyl (2R,4aR)-11-fluoro-10-(2-fluorophenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2-methyl-5,7-dioxo-4,4a,5,6,7,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3(2H)-carboxylate (300 mg, 0.462 mmol), 6 mL of acetone, anhydrous potassium carbonate (255 mg, 1.84 mmol), and iodomethane (656 mg, 4.62 mmol) were sealed in a sealing tube, and stirred at 50° C. for 18 hours to react. The resulting reaction liquid was added with 20 mL of EtOAc, washed with 20 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-80%) to obtain product tert-butyl (2R,4aR)-11-fluoro-10-(2-fluorophenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,6-dimethyl-5,7-dioxo-4,4a,5,6,7,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3(2H)-carboxylate (350 mg, Y: 95%), which was yellow solid. ES-API: [M+H].sup.+=645.2.
[0448] Step 8: the tert-butyl (2R,4aR)-11-fluoro-10-(2-fluorophenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,6-dimethyl-5,7-dioxo-4,4a,5,6,7,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3(2H)-carboxylate (350 mg) was dissolved in DCM (4 mL), and added with TFA (2 mL). After stirring at room temperature for 2 hours, the resulting reaction liquid was concentrated to obtain product (2R,4aR)-11-fluoro-10-(2-fluorophenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,6-dimethyl-2,3,4,4a-tetrahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7(6H, 8H)-dione (40 mg, crude), which was directly used in next step. ES-API: [M+H].sup.+=545.2.
[0449] Step 9: the (2R,4aR)-11-fluoro-10-(2-fluorophenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,6-dimethyl-2,3,4,4a-tetrahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7(6H, 8H)-dione (350 mg, 0.62 mmol) was dissolved in DCM (6 mL), and added with diisopropylethylamine (480 mg, 3.72 mmol). The resulting reaction liquid was cooled to 0° C., and added dropwise with acryloyl chloride (112.5 mg, 1.24 mmol). The resulting mixture was stirred at 0° C. for 15 minutes to react. The resulting reaction liquid was added with 20 mL of DCM, washed with 20 mL of saturated solution of NaHCO.sub.3 and 20 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by preparative scale HPLC to obtain product (2R,4aR)-3-acryloyl-11-fluoro-10-(2-fluorophenyl)-8-(2-isopropyl-4-methylpyridin-3-yl)-2,6-dimethyl-2,3,4,4a-tetrahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7(6H, 8H)-dione (Z36, 60 mg, Y: 18%), which was yellow solid. ES-API: [M+H].sup.+=698.3. .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 8.49 (d, J=4.9 Hz, 1H), 8.04 (dd, J=14.3, 9.2 Hz, 1H), 7.52 (tdd, J=7.7, 5.3, 1.9 Hz, 1H), 7.36-7.26 (m, 3H), 7.23 (td, J=7.5, 2.0 Hz, 1H), 6.95-6.77 (m, 1H), 6.21-6.09 (m, 1H), 5.80-5.69 (m, 1H), 5.07-4.75 (m, 1H), 4.64-4.39 (m, 1H), 4.03 (dd, J=27.3, 4.3 Hz, 1H), 3.74 (dd, J=14.2, 4.3 Hz, 1H), 3.39 (d, J=2.1 Hz, 4H), 2.95-2.80 (m, 1H), 2.45 (q, J=6.7 Hz, 1H), 2.02 (d, J=2.5 Hz, 3H), 1.56 (dd, J=17.1, 6.7 Hz, 3H), 1.04 (d, J=6.6 Hz, 3H), 0.87 (d, J=6.7 Hz, 3H).
[0450] Step 10: the compound Z36 was resolved by preparative scale chiral HPLC (column: Chiralpak IB: 10 μm, 30*250 mm; mobile phase: hexane:EtOH:aminomethanol=50:50:0.2; flow rate: 25 ml/min; and column temperature: room temperature) to obtain the following atropisomer compounds. One of the atropisomer compounds had a structure arbitrarily specified as Z36-1 (peak 1, 22 mg, retention time: 10.211 min, Y: 38%), ES-API: [M+H].sup.+=599.2; .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 8.49 (d, J=4.9 Hz, 1H), 8.04 (dd, J=14.3, 9.2 Hz, 1H), 7.52 (t, J=7.7, 5.3, 1.9 Hz, 1H), 7.36-7.26 (m, 3H), 7.23 (t, J=7.5, 2.0 Hz, 1H), 6.95-6.70 (m, 1H), 6.21-6.09 (m, 1H), 5.80-5.69 (m, 1H), 5.07-4.75 (m, 1H), 4.64-4.39 (m, 1H), 4.03-3.70 (m, 1H), 3.74 (dd, J=14.2, 4.3 Hz, 1H), 3.39 (d, J=2.1 Hz, 4H), 2.95-2.80 (m, 1H), 2.45 (q, J=6.7 Hz, 1H), 2.02 (d, J=2.5 Hz, 3H), 1.56 (dd, J=17.1, 6.7 Hz, 3H), 1.04 (d, J=6.6 Hz, 3H), 0.87 (d, J=6.7 Hz, 3H). The other atropisomer compound had a structure arbitrarily specified as Z36-2 (peak 2, 20 mg, retention time: 12.534 min, Y: 34%), ES-API: [M+H].sup.+=599.2; .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 8.49 (d, J=4.9 Hz, 1H), 8.03 (dd, J=14.5, 9.2 Hz, 1H), 7.52-7.44 (m, 1H), 7.36-7.22 (m, 4H), 6.95-6.77 (m, 1H), 6.20-6.11 (m, 1H), 5.78-5.70 (m, 1H), 4.77 (s, 1H), 4.61 (d, J=14.1 Hz, 1H), 4.02-3.93 (m, 1H), 3.74 (dd, J=14.2, 4.2 Hz, 1H), 3.35 (s, 4H), 2.92-2.76 (m, 2H), 1.82 (s, 3H), 1.56-1.45 (m, 3H), 1.11 (d, J=6.9 Hz, 3H), 0.99 (dd, J=6.8, 2.9 Hz, 3H). The isomer compounds were detected by analytical scale chiral HPLC (column: Chiralpak IB: 5 μm, 4.6*250 mm; mobile phase: hexane:EtOH:aminomethanol=50:50:0.2; flow rate: 1 ml/min; column temperature=30° C.).
Example 37 Preparation of Compounds Z37, Z37-1 and Z37-2
[0451] ##STR00538## ##STR00539## ##STR00540## ##STR00541##
[0452] Step 1: 7-bromo-6-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-8-fluoro-4-hydroxy-2-oxo-1,2-dihydroquinolin-3-carbonitrile (2.0 g, 4.17 mmol) was suspended in 1,4-dioxane (10 mL), and slowly added with a mixed liquid of concentrated sulfuric acid (10 mL) and water (10 mL). The resulting mixture was stirred at 120° C. for 18 hours to react. Cooled reaction liquid was poured into 50 mL of ice water. The precipitated solid was filtered. The filter cake was washed with a small amount of water and dried in vacuum to obtain product 7-bromo-6-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-8-fluoro-4-hydroxylquinolin-2(1H)-one (1.5 g, Y: 79.1%), which was light brown solid. ES-API: [M+H].sup.+=454.0, 456.1.
[0453] Step 2: the 7-bromo-6-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-8-fluoro-4-hydroxylquinolin-2(1H)-one (1.4 g, 3.08 mmol) was dissolved in acetic acid (4 mL), orderly added with sodium nitrite (21 mg, 0.31 mmol) and concentrated nitric acid (0.62 mL, 9.24 mmol), and stirred at room temperature for 30 minutes to react. The resulting reaction liquid was poured into 10 mL of ice water. The precipitated solid was filtered. The filter cake was washed with 6 mL of water and dried in vacuum to obtain product 7-bromo-6-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-8-fluoro-4-hydroxy-3-nitroquinolin-2(1H)-one (1.25 g, Y:81.2%), which was yellow solid. ES-API: [M+H].sup.+=499.0, 501.0.
[0454] Step 3: the 7-bromo-6-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-8-fluoro-4-hydroxy-3-nitroquinolin-2(1H)-one (1.25 g, 2.50 mmol) was dissolved in ACN (25 mL), orderly added with phosphorus oxychloride (1.15 mL, 12.50 mmol) and N,N-diisopropylethylamine (3.48 mL, 20.0 mmol), and stirred at 85° C. for 30 minutes to react. The resulting reaction liquid was concentrated, added with 100 mL of EtOAc, and washed with 30 mL of water, with 30 mL saturated sodium bicarbonate solution twice and then with 30 mL of saturated salt solution. After the resulting organic phase was dried and concentrated, the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-15%) to obtain product 7-bromo-4,6-dichloro-1-(4,6-diisopropylpyrimidin-5-yl)-8-fluoro-3-nitroquinolin-2(1H)-one (650 mg, Y: 50.1%), which was yellow solid. ES-API: [M+H].sup.+=517.0, 519.0.
[0455] Step 4: the 7-bromo-4,6-dichloro-1-(4,6-diisopropylpyrimidin-5-yl)-8-fluoro-3-nitroquinolin-2(1H)-one (370 mg, 0.71 mmol) was dissolved in N,N-dimethylacetamide (5 mL), orderly added with (R)-1-(tert-butyl)3-methyl-piperazin-1,3-dicarboxylate (520 mg, 2.13 mmol) and N,N-diisopropylethylamine (275 mg, 2.13 mmol), and stirred at 120° C. for 2 hours to react. The resulting reaction liquid was added with 80 mL of EtOAc, washed with 30 mL of dilute brine for 4 times and then with 30 mL saturated salt solution, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-25%) to obtain product (R)-1-(tert-butyl)-3-methyl-4-(7-bromo-6-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-8-fluoro-3-nitro-2-oxo-1,2-dihydroquinolin-4-yl)piperazin-1,3-dicarboxylate (400 mg, Y: 77.1%), which was orange solid. ES-API: [M+H].sup.+=725.0, 727.2.
[0456] Step 5: the (R)-1-(tert-butyl)-3-methyl-4-(7-bromo-6-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-8-fluoro-3-nitro-2-oxo-1,2-dihydroquinolin-4-yl)piperazin-1,3-dicarboxylate (380 mg, 0.52 mmol) was dissolved in acetic acid (7 mL), added with iron powder (103 mg, 1.83 mmol), and stirred at 80° C. for 30 minutes to react. The resulting reaction liquid was concentrated, and orderly added with 100 mL of EtOAc and 60 mL of saturated sodium bicarbonate. The resulting suspension was filtered by using diatomite. The filter cake was washed with EtOAc. The resulting organic phase was separated, washed orderly with 40 mL of saturated sodium bicarbonate solution and 40 mL of saturated salt solution, then dried and concentrated to obtain product tert-butyl (R)-10-bromo-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-9-fluoro-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]quinolin-3-carboxylate (347 mg, Y: 100%), which was yellow solid. ES-API: [M+H].sup.+=663.2, 665.2.
[0457] Step 6: the tert-butyl (R)-10-bromo-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-9-fluoro-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]quinolin-3-carboxylate (317 mg, 0.48 mmol), 12 mL of acetone, anhydrous potassium carbonate (265 mg, 1.92 mmol), and iodomethane (678 mg, 4.80 mmol) were orderly added to a 50 mL sealing tube. The sealing tube was sealed, and the resulting mixture was stirred at 50° C. for 18 hours to react. The resulting reaction liquid was added with 60 mL of EtOAc, washed orderly with 25 mL of water and 25 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-30%) to obtain product tert-butyl (R)-10-bromo-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-9-fluoro-6-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octyl-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]quinolin-3-carboxylate (310 mg, Y: 95.8%), which was yellow solid. ES-API: [M+H].sup.+=677.1, 679.2.
[0458] Step 7: the tert-butyl (R)-10-bromo-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-9-fluoro-6-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octyl-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]quinolin-3-carboxylate (285 mg, 0.42 mmol), (2-fluoro-6-hydroxyphenyl)boric acid (262 mg, 1.68 mmol), SPhos-Pd-G2 (30 mg, 0.042 mmol), 2-dicyclohexylphosphino-2′,6′-dimethoxybiphenyl (17 mg, 0.042 mmol), potassium phosphate (356 mg, 1.68 mmol), 3 mL of water, and 15 mL of dioxane were added to a 100 mL round-bottom flask. The resulting mixture was stirred at 90° C. for 2 hours to react under the protection of nitrogen. The resulting reaction liquid was concentrated, and the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-50%) to obtain product tert-butyl (4aR)-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-9-fluoro-10-(2-fluoro-6-hydroxyphenyl)-6-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]quinolin-3-carboxylate (230 mg, Y: 77.2%), which was directly used in next step. ES-API: [M+H].sup.+=709.2.
[0459] Step 8: the tert-butyl (4aR)-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-9-fluoro-10-(2-fluoro-6-hydroxyphenyl)-6-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]quinolin-3-carboxylate (230 mg, 0.32 mmol) was dissolved in DCM (3 mL), and added with TFA (0.8 mL). After stirring at room temperature for 1 hour, the resulting reaction liquid was concentrated to obtain product (4aR)-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-9-fluoro-10-(2-fluoro-6-hydroxyphenyl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′,5]pyrazino[2,3-c]quinolin-5,7-dione (270 mg, crude), which was directly used in next step. ES-API: [M+H].sup.+=609.2.
[0460] Step 9: the (4aR)-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-9-fluoro-10-(2-fluoro-6-hydroxyphenyl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′,5]pyrazino[2,3-c]quinolin-5,7-dione (270 mg, crude) was dissolved in DCM (12 mL), and added with N,N-diisopropylethylamine (206 mg, 1.60 mmol). The resulting reaction liquid was cooled to 0° C., and added dropwise with acryloyl chloride (26 mg, 0.29 mmol). The resulting mixture was stirred at 0° C. for 15 minutes to react. The resulting reaction liquid was added with 30 mL of DCM, washed orderly with 10 mL of water, 10 mL of saturated solution of NaHCO.sub.3 and 10 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by preparative scale HPLC to obtain the following atropisomer compounds. One of the atropisomer compounds had a structure arbitrarily specified as Z37-1 (retention time: 10.095 min; 40 mg, Y: 18.6%), which was white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 9.13 (s, 1H), 8.02 (s, 1H), 7.27 (dd, J=15.4, 8.1 Hz, 1H), 7.07 (dd, J=16.4, 10.6 Hz, 1H), 6.77-6.67 (m, 2H), 6.15 (d, J=16.7 Hz, 1H), 5.76 (d, J=11.1 Hz, 1H), 4.72 (d, J=13.4 Hz, 1H), 4.48 (d, J=13.0 Hz, 1H), 4.07-3.97 (m, 1H), 3.67-3.41 (m, 2H), 3.31-3.11 (m, 4H), 3.00-2.89 (m, 1H), 2.74-2.56 (m, 2H), 1.30-0.72 (m, 12H). ES-API: [M+H].sup.+=663.2. The other atropisomer compound had a structure arbitrarily specified as Z37-2 (retention time: 10.424 min; 50 mg, Y:23.2%), which was white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.21 (s, 1H), 9.13 (s, 1H), 8.03 (s, 1H), 7.27 (dd, J=15.4, 8.3 Hz, 1H), 7.07 (dd, J=16.7, 10.7 Hz, 1H), 6.77-6.65 (m, 2H), 6.15 (dd, J=16.8, 2.2 Hz, 1H), 5.75 (d, J=10.2 Hz, 1H), 4.72 (d, J=13.9 Hz, 1H), 4.48 (d, J=13.8 Hz, 1H), 4.07-3.97 (m, 1H), 3.66-3.41 (m, 2H), 3.30-3.10 (m, 4H), 3.00-2.89 (m, 1H), 2.70-2.54 (m, 2H), 1.21-0.84 (m, 12H). ES-API: [M+H].sup.+=663.2. The isomer compounds were detected by analytical scale HPLC.
Example 38 Preparation of Compounds Z38, Z38-1 and Z38-2
[0461] ##STR00542## ##STR00543## ##STR00544## ##STR00545##
[0462] Step 1: 2-cyano-6-isopropylphenyl-3-amine (742 mg, 4.24 mmol) was dissolved in dry tetrahydrofuran (20 mL), added with 2 M NaHMDS (8.48 mL, 16.96 mmol) under the condition of an ice water bath, and stirred for 20 minutes under the condition of the ice water bath. The resulting mixture was added with 2,5-difluoro-6-(2-fluoro-6-methoxyphenyl)nicotinic acid (1.2 g, 4.24 mmol), and stirred at room temperature for 3 hours. The resulting reaction liquid was slowly poured into 30 mL of ice water, mixed with diluted hydrochloric acid (3 M) to adjust the pH to a range of 5 to 6, extracted with EtOAc, washed with 50 mL of saturated salt solution once, dried by anhydrous sodium sulfate, and filtered. After the resulting organic phase was dried and concentrated, the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 20-40%) to obtain product 2-((2-cyano-6-isopropylphenyl)amino)-5-fluoro-6-(2-fluoro-6-methoxyphenyl)nicotinic acid (1.8 g, Y: 98%), which was yellow solid. ES-API: [M+H].sup.+=424.1.
[0463] Step 2: the 2-((2-cyano-6-isopropylphenyl)amino)-5-fluoro-6-(2-fluoro-6-methoxyphenyl)nicotinic acid (1.5 g, 3.42 mmol) was dissolved in dichloroethane, and added with thionyl chloride (4.07 g, 34.2 mmol). The resulting mixture was stirred at 80° C. for 2 hours to react. After the completion of the reaction, the resulting reaction liquid was cooled to room temperature, concentrated, and dried in 50 degrees of vacuum for 4 hours to obtain product 2-((2-cyano-6-isopropylphen-3-yl)amino)-5-fluoro-6-(2-fluoro-6-methoxyphenyl)nicotinoyl chloride (1.57 g, crude), which was faint yellow solid. MeOH was used to detect ES-API: [M+H].sup.+=438.1 (MeOH).
[0464] Step 3: under the condition of the ice water bath, sodium hydride (1.97 g, 49.35 mmol) was added to a solution of ethyl nitroacetate (1.31 g, 9.86 mmol) in tetrahydrofuran, stirred for 30 minutes, subsequently added with the 2-((2-cyano-6-isopropylphen-3-yl)amino)-5-fluoro-6-(2-fluoro-6-methoxyphenyl)nicotinoyl chloride (1.57 g, 3.29 mmol), stirred at room temperature for 1 hour, and then heated to 80° C. to react for 2 hours. The resulting reaction liquid was poured into ice water, mixed with 3 M hydrochloric acid to adjust the pH to a range of 3 to 4, extracted with EtOAc, dried by anhydrous sodium sulfate, and filtered. The resulting organic phase was dried and concentrated to obtain product 1-(2-cyano-6-isopropylphen-3-yl)-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-4-hydroxy-3-nitro-1,8-naphthyridin-2(1H)-one (110 mg, Y: 20%). ES-API: [M+H].sup.+=493.1.
[0465] Step 4: the 1-(2-cyano-6-isopropylphen-3-yl)-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-4-hydroxy-3-nitro-1,8-naphthyridin-2(1H)-one (110 mg, 0.20 mmol) was dissolved in ACN (10 mL), orderly added with phosphorus oxychloride (153 mg, 1.0 mmol) and N,N-diisopropylethylamine (77 g, 0.6 mmol), and gradually heated to 80° C. and stirred for 3 hours to react. The resulting reaction liquid was concentrated, added with 30 mL of cold ACN, added dropwise to 30 mL of saturated sodium bicarbonate solution under the condition of the ice water bath, and extracted with EtOAc (50 mL*2). The resulting combined organic phase was washed with 30 mL of saturated salt solution, dried by anhydrous sodium sulfate, and filtered. After the resulting organic phase was dried and concentrated, the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-80%) to obtain 4-chloro-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(2-cyano-6-isopropylphen-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (110 mg, Y: 68%), which was yellow solid. ES-API: [M+H].sup.+=511.1.
[0466] Step 5: the 4-chloro-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(2-cyano-6-isopropylphen-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (110 mg, 0.296 mmol) was dissolved in N,N-dimethylacetamide (3 mL), subsequently added with (3R,6R)-1-tert-butyl3-methyl6-methylpiperazin-1,3-dicarboxylic acid (54 mg, 0.22 mmol), and stirred at 120° C. for 2 hours to react. After the completion of the reaction, the resulting product was added with 30 mL of EtOAc and washed with 30 mL of saturated salt solution for 3 times. The EtOAc phase was dried and concentrated to obtain a crude product, namely target product (3R,6R)-1-tert-butyl3-methyl4-(1-(2-cyano-6-isopropylphenyl)-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-6-methylpiperazin-1,3-dicarboxylic acid (50 mg, Y: 46%), which was yellow solid. ES-API: [M+H].sup.+=733.3.
[0467] Step 6: the (3R,6R)-1-tert-butyl3-methyl4-(1-(2-cyano-6-isopropylphenyl)-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-6-methylpiperazin-1,3-dicarboxylic acid (50 mg, 0.068 mmol) was dissolved in acetic acid (25 mL), added with iron powder (11.5 mg, 0.204 mmol), and stirred at 80° C. for 30 minutes to react. The resulting reaction liquid was concentrated, and orderly added with 30 mL of EtOAc and 30 mL of saturated sodium bicarbonate solution. The resulting suspension was filtered by diatomite. The filter cake was washed with EtOAc. The resulting organic phase was separated, washed orderly with 30 mL of saturated sodium bicarbonate and 30 mL of saturated salt solution, dried and concentrated to obtain product (2R,4aR)-tert-butyl8-(2-cyano-6-isopropylphenyl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-2-methyl-5,7-dioxo-4,4a, 5,6,7,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3(2H)-carboxylate (40 mg, crude), which was yellow solid. ES-API: [M+H].sup.+=671.2.
[0468] Step 7: the (2R,4aR)-tert-butyl8-(2-cyano-6-isopropylphenyl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-2-methyl-5,7-dioxo-4,4a, 5,6,7,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3(2H)-carboxylate (40 mg, 0.059 mmol), 30 mL of acetone, anhydrous potassium carbonate (33 mg, 0.24 mmol), and iodomethane (85 mg, 0.59 mmol) were sealed in a sealing tube, and stirred at 50° C. for 18 hours to react. The resulting reaction liquid was added with 20 mL of EtOAc, washed with 20 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-80%) to obtain product (2R,4aR)-tert-butyl8-(2-cyano-6-isopropylphenyl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-2,6-dimethyl-5,7-dioxo-4,4a,5,6,7,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3(2H)-carboxylate (40 mg, Y: 90%), which was yellow solid. ES-API: [M+H].sup.+=685.2.
[0469] Step 8: the (2R,4aR)-tert-butyl8-(2-cyano-6-isopropylphenyl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-2,6-dimethyl-5,7-dioxo-4,4a,5,6,7,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3(2H)-carboxylate (44 mg) was dissolved in DCM (3 mL), and added with TFA (1 mL). After stirring at room temperature for 2 hours, the resulting reaction liquid was concentrated to obtain product 2-((2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-2,6-dimethyl-5,7-dioxo-2,3,4,4a,5,6-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-8(7H)-yl)-3-isopropylbenzonitrile (40 mg, crude), which was directly used in next step. ES-API: [M+H].sup.+=585.2.
[0470] Step 9: the 2-((2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-2,6-dimethyl-5,7-dioxo-2,3,4,4a,5,6-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-8(7H)-yl)-3-isopropylbenzonitrile (40 mg, 0.068 mmol) was dissolved in DCM (5 mL), and added with diisopropylethylamine (53 mL, 0.408 mmol). The resulting reaction liquid was cooled to 0° C., and added dropwise with acryloyl chloride (12.4 mg, 0.137 mmol). The resulting mixture was stirred at 0° C. for 15 minutes to react. The resulting reaction liquid was added with 20 mL of DCM, washed with 20 mL of saturated solution of NaHCO.sub.3 and 20 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by preparative scale chromatography to obtain product 2-((2R,4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-2,6-dimethyl-5,7-dioxo-2,3,4,4a,5,6-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-8(7H)-yl)-3-isopropylbenzonitrile (32 mg, Y: 70%), which was yellow solid. ES-API: [M+H].sup.+=639.3.
[0471] Step 10: the 2-((2R,4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-2,6-dimethyl-5,7-dioxo-2,3,4,4a,5,6-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-8(7H)-yl)-3-isopropylbenzonitrile (32 mg, 0.068 mmol) was added to dry DCM (4.0 mL), then added with boron tribromide (4.0 mL, 4.0 mmol), and warmed to room temperature to react for 1 hour. Under the condition of the ice water bath, the above reaction liquid was added dropwise to saturated sodium bicarbonate solution, extracted with DCM (30 mL) twice, dried and concentrated. The resulting crude product was purified by preparative scale thin-layer chromatography (DCM/MeOH: 10/1) to obtain product 2-((2R,4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-hydroxyphenyl)-2,6-dimethyl-5,7-dioxo-2,3,4,4a,5,6-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-8(7H)-yl)-3-isopropylbenzonitrile (Z38, 25 mg, Y: 80%), which was yellow solid. ES-API: [M+H].sup.+=625.2.
[0472] Step 11: the compound Z38 was resolved by preparative scale chiral HPLC (column type: Chiralpak IB: 10 μm, 30*250 mm; mobile phase: hexane:EtOH:aminomethanol=50:50:0.2; flow rate: 25 ml/min; and column temperature: room temperature) to obtain the following atropisomer compounds. One of the atropisomer compounds had a structure arbitrarily specified as Z38-1 (peak 1, 7 mg, retention time: 10.117 min, Y: 28%), ES-API: [M+H].sup.+=625.2. The other atropisomer compound had a structure arbitrarily specified as Z38-2 (peak 2, 9 mg, retention time: 12.237 min, Y: 39%), ES-API: [M+H].sup.+=625.2. The isomer compounds were detected by analytical scale chiral HPLC (column: Chiralpak IB: 5 μm, 4.6*250 mm; mobile phase: hexane:EtOH:aminomethanol=50:50:0.2; flow rate: 1 ml/min; and column temperature=30° C.).
Example 39 Preparation of Compounds Z39, Z39-1 and Z39-2
[0473] ##STR00546## ##STR00547## ##STR00548##
[0474] Step 1: tert-butyl (R)-10-bromo-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-9-fluoro-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]quinolin-3-carboxylate (230 mg, 0.35 mmol), 8 mL of acetone, anhydrous potassium carbonate (193 mg, 1.40 mmol), and deuteroiodomethane (507 mg, 3.50 mmol) were orderly added to a 15 mL sealing tube. The sealing tube was sealed, and the resulting mixture was stirred at 50° C. for 18 hours to react. The resulting reaction liquid was added with 60 mL of EtOAc, washed orderly with 25 mL of water and 25 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-30%) to obtain product tert-butyl (R)-10-bromo-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-9-fluoro-6-(deuteromethyl-d.sub.3)-5,7-dioxo-1,2,4,4a,5,6,7,8-octyl-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]quinolin-3-carboxylate (185 mg, Y: 78.50%), which was faint yellow solid. ES-API: [M+H].sup.+=680.2, 682.2.
[0475] Step 2: tert-butyl (R)-10-bromo-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-9-fluoro-6-deuteromethyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octyl-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]quinolin-3-carboxylate (185 mg, 0.27 mmol), (2-fluoro-6-hydroxyphenyl)boric acid (262 mg, 1.35 mmol), SPhos-Pd-G2 (19 mg, 0.027 mmol), 2-dicyclohexylphosphino-2′,6′-dimethoxybiphenyl (11 mg, 0.027 mmol), potassium phosphate (229 mg, 1.08 mmol), 2 mL of water, and 10 mL of dioxane were added to a 100 mL round-bottom flask. The resulting mixture was stirred at 90° C. for 4 hours to react under the protection of nitrogen. The resulting reaction liquid was concentrated, and the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-50%) to obtain product tert-butyl (4aR)-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-9-fluoro-10-(2-fluoro-6-hydroxyphenyl)-6-(deuteromethyl-d.sub.3)-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]quinolin-3-carboxylate (140 mg, Y: 72.4%), which was yellow solid. ES-API: [M+H].sup.+=712.3.
[0476] Step 3: the tert-butyl (4aR)-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-9-fluoro-10-(2-fluoro-6-hydroxyphenyl)-6-(deuteromethyl-d.sub.3)-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]quinolin-3-carboxylate (140 mg, 0.20 mmol) was dissolved in DCM (3 mL), and added with TFA (0.8 mL). After stirring at room temperature for 1 hour, the resulting reaction liquid was concentrated to obtain product (4aR)-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-9-fluoro-10-(2-fluoro-6-hydroxyphenyl)-6-(deuteromethyl-d.sub.3)-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′,5]pyrazino[2,3-c]quinolin-5,7-dione (165 mg, crude), which was directly used in next step. ES-API: [M+H].sup.+=612.3.
[0477] Step 4: the (4aR)-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-9-fluoro-10-(2-fluoro-6-hydroxyphenyl)-6-(deuteromethyl-d.sub.3)-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′,5]pyrazino[2,3-c]quinolin-5,7-dione (165 mg, crude) was dissolved in DCM (8 mL), and added with N,N-diisopropylethylamine (129 mg, 1.0 mmol). The resulting reaction liquid was cooled to 0° C., and added dropwise with acryloyl chloride (16 mg, 0.18 mmol). The resulting mixture was stirred at 0° C. for 15 minutes to react. The resulting reaction liquid was added with 30 mL of DCM, washed orderly with 10 mL of water, 10 mL of saturated solution of NaHCO.sub.3 and 10 mL of saturated salt solution, dried and concentrated to obtain product (4aR)-3-acryloyl-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-9-fluoro-10-(2-fluoro-6-hydroxyphenyl)-6-(deuteromethyl-d3)-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′,5]pyrazino[2,3-c]quinolin-5,7-dione (Z39).
[0478] Step 5: the compound Z39 was purified by preparative scale HPLC to obtain the following atropisomer compounds. One of the atropisomer compounds had a structure arbitrarily specified as Z39-1 (retention time: 10.088 min; 23 mg, Y: 17.6%), which was white solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.19 (d, J=1.5 Hz, 1H), 9.13 (s, 1H), 8.02 (s, 1H), 7.27 (dd, J=15.5, 8.2 Hz, 1H), 7.07 (dd, J=16.7, 10.7 Hz, 1H), 6.77-6.69 (m, 2H), 6.15 (d, J=16.8 Hz, 1H), 5.75 (d, J=11.2 Hz, 1H), 4.72 (d, J=14.4 Hz, 1H), 4.48 (d, J=12.2 Hz, 1H), 3.99 (s, 1H), 3.62 (d, J=10.5 Hz, 1H), 3.46 (d, J=11.1 Hz, 1H), 3.19 (t, J=12.1 Hz, 1H), 3.00-2.90 (m, 1H), 2.70-2.54 (m, 2H), 1.14-0.95 (m, 12H). ES-API: [M+H].sup.+=666.2. The other atropisomer compound had a structure arbitrarily specified as Z39-2 (retention time: 10.420 min; 30 mg, Y: 22.9%), which was white solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.20 (d, J=1.4 Hz, 1H), 9.13 (s, 1H), 8.03 (s, 1H), 7.27 (dd, J=15.4, 8.2 Hz, 1H), 7.07 (dd, J=16.7, 10.2 Hz, 1H), 6.79-6.66 (m, 2H), 6.15 (d, J=16.7 Hz, 1H), 5.75 (d, J=10.9 Hz, 1H), 4.72 (d, J=13.7 Hz, 1H), 4.49 (d, J=13.3 Hz, 1H), 4.00 (s, 1H), 3.63-3.44 (m, 2H), 3.19 (t, J=12.1 Hz, 1H), 3.00-2.90 (m, 1H), 2.73-2.55 (m, 2H), 1.25-0.78 (m, 12H). ES-API: [M+H].sup.+=666.2. The isomer compounds were detected by analytical scale HPLC.
Example 40 Preparation of Compound Z40
[0479] ##STR00549## ##STR00550## ##STR00551##
[0480] Step 1 7-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-6-fluoro-4-hydroxy-2-oxo-1,2-dihydro-1,8-naphthyridin-3-carbonitrile (2 g, 5 mmol) was suspended in 1,4-dioxane (12 mL), and slowly added with a mixed liquid of concentrated sulfuric acid (12 mL) and water (12 mL). The resulting mixture was stirred at 120° C. to react overnight. The resulting reaction liquid was cooled and poured into ice water (50 mL), and extracted with EtOAc (50 mL*3). The resulting organic phase was dried and concentrated to obtain product 7-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-6-fluoro-4-hydroxy-1,8-naphthyridin-2(1H)-one (1.8 g, Y:96%), which was faint yellow solid. ES-API: [M+H].sup.+=377.2.
[0481] Step 2: the 7-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-6-fluoro-4-hydroxy-1,8-naphthyridin-2(1H)-one (1.56 g, 4.14 mmol) was dissolved in acetic acid (5.46 mL), orderly added with sodium nitrite (29 mg, 0.42 mmol) and concentrated nitric acid (780 mg, 12.42 mmol), and stirred at room temperature for 20 minutes. The resulting reaction liquid was poured into 6 mL of water. Yellow solid was precipitated and then filtered. The filter cake was washed with 6 mL of water and dried in vacuum to obtain product 7-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-6-fluoro-4-hydroxy-3-nitro-1,8-naphthyridin-2(1H)-one (1.6 g, Y: 91%), which was faint yellow solid. ES-API: [M+H].sup.+=422.0.
[0482] Step 3: a mixed solution of the 7-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-6-fluoro-4-hydroxy-3-nitro-1,8-naphthyridin-2(1H)-one (1.56 g, 3.70 mmol), (2-fluoro-6-methoxyphenyl)boric acid (3.14 g, 18.49 mmol), chloro(2-dicyclohexylphosphino-2′,6′-dimethoxy-1,1′-biphenyl)(2′-amino-1,1′-biphen-2-yl)palladium(II) (266 mg, 0.37 mmol), 2-dicyclohexylphosphino-2′,6′-dimethoxybiphenyl (152 mg, 0.37 mmol), and potassium phosphate (2.36 g, 11.09 mmol) in 8 mL of water and 40 mL of dioxane was replaced with nitrogen for 3 times and allowed to react at 100° C. for 2 hours. The resulting reaction liquid was poured into 50 mL of water, and washed with methyl tert-butyl ether (30 mL*2). The water phase was mixed with 1.0 M diluted hydrochloric acid to adjust the pH to 6.0, and extracted with EtOAc (30 mL*2). The resulting organic phase was dried and concentrated to obtain crude product 1-(4,6-diisopropylpyrimidin-5-yl)-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-4-hydroxy-3-nitro-1,8-naphthyridin-2(1H)-one (1.55 g, Y: 82%), which was yellow solid. ES-API: [M+H].sup.+=512.2.
[0483] Step 4: phosphorus oxychloride (2.29 g, 14.96 mmol) and N,N-diisopropylethylamine (3.09 g, 23.93 mmol) were orderly added to a solution of the 1-(4,6-diisopropylpyrimidin-5-yl)-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-4-hydroxy-3-nitro-1,8-naphthyridin-2(1H)-one (1.53 g, 2.99 mmol) in ACN (20 mL). The resulting mixture was stirred at 80° C. for 1 hour to react. The resulting reaction liquid was concentrated, added with 50 mL of EtOAc, and washed orderly with 30 mL of saturated sodium bicarbonate twice and then with 30 mL of water and 30 mL of saturated salt solution. After the resulting organic phase was dried and concentrated, the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-50%) to obtain product 4-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-3-nitro-1,8-naphthyridin-2(1H)-one (800 mg, Y: 50%), which was faint yellow solid. ES-API: [M+H].sup.+=530.2.
[0484] Step 5: the 4-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-3-nitro-1,8-naphthyridin-2(1H)-one (800 mg, 1.51 mmol) was dissolved in N,N-dimethylacetamide (8 mL), orderly added with 1-(tert-butyl)3-methyl(3R,6R)-6-methylpiperazin-1,3-dicarboxylic acid (390 mg, 1.51 mmol) and N,N-diisopropylethylamine (585 mg, 4.53 mmol), and stirred at 120° C. for 2 hours. Cooled reaction liquid was added with 30 mL of EtOAc, washed with 30 mL of sodium bicarbonate twice, with 30 mL of dilute brine twice, with 30 mL of water once and with 30 mL of saturated salt solution once, dried and concentrated, and purified by flash column chromatography on silica gel (EtOAc/PE: 0-100%) to obtain 1-(tert-butyl)3-methyl(3R,6R)-4-(1-(4,6-diisopropylpyrimidin-5-yl)-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-6-methylpiperazin-1,3-dicarboxylic acid (540 mg, Y: 47%). ES-API: [M+H].sup.+=752.2.
[0485] Step 6: the 1-(tert-butyl)3-methyl(3R,6R)-4-(1-(4,6-diisopropylpyrimidin-5-yl)-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-6-methylpiperazin-1,3-dicarboxylic acid (540 mg, 0.72 mmol) was dissolved in acetic acid (5 mL), added with iron powder (140 mg, 2.51 mmol), and stirred at 80° C. for 1 hour. The resulting reaction liquid was concentrated, added with 50 mL of EtOAc, and mixed with saturated sodium bicarbonate solution to adjust the pH to 8. The resulting suspension was filtered by diatomite. The filter cake was washed with EtOAc. The resulting organic phase was separated, washed orderly with 25 mL of saturated sodium bicarbonate and 25 mL of saturated salt solution, dried and concentrated to obtain crude product tert-butyl (2R,4aR)-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-2-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (540 mg), which was yellow solid. ES-API: [M+H].sup.+=690.3.
[0486] Step 7: the tert-butyl (2R,4aR)-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-2-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (540 mg, 0.78 mmol), acetone (10 mL), anhydrous potassium carbonate (433 mg, 3.13 mmol), and iodomethane (1.11 g, 7.83 mmol) were orderly added to a 50 mL sealing tube. The sealing tube was sealed, and the resulting mixture was stirred at 50° C. to react overnight. The resulting reaction liquid was filtered, concentrated, dissolved in DCM (10 mL), and washed with water (10 mL). The resulting organic phase was dried and concentrated to obtain crude product tert-butyl (2R,4aR)-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-2,6-dimethyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (420 mg), which was yellow solid. ES-API: [M+H].sup.+=704.3.
[0487] Step 8: the tert-butyl (2R,4aR)-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-2,6-dimethyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (420 mg, 0.60 mmol) was dissolved in DCM (20 mL), and added with TFA (4 mL). After stirring at room temperature for 2 hours, the resulting reaction liquid was concentrated to obtain crude product (2R,4aR)-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (360 mg), which was directly used in next step. ES-API: [M+H].sup.+=604.3.
[0488] Step 9: the (2R,4aR)-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (360 mg, 0.60 mmol) was dissolved in DCM (5 mL), and added with N,N-diisopropylethylamine (385 mg, 2.98 mmol). The resulting reaction liquid was cooled to 0° C., and added dropwise with acryloyl chloride (108 mg, 1.19 mmol). The reaction liquid was stirred at 0° C. for 5 minutes, and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (0-80% EtOAc/PE) to obtain product (2R,4aR)-3-acryloyl-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (220 mg, Y: 56%), which was faint yellow solid. ES-API: [M+H].sup.+=658.2.
[0489] Step 10: the (2R,4aR)-3-acryloyl-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (220 mg, 0.33 mmol) was dissolved in DCM (3 mL). The resulting solution was cooled to 0° C., added dropwise with a solution (3 mL) of 17% boron tribromide in DCM, and stirred at room temperature overnight. The resulting reaction liquid was poured into 40 mL of saturated solution of sodium bicarbonate and extracted with 25 mL of DCM twice. The resulting organic phase was dried and concentrated, and the resulting crude product was purified by preparative scale HPLC to obtain product (2R,4aR)-3-acryloyl-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-hydroxyphenyl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (Z40, 115 mg, Y: 53%), which was faint yellow solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.14 (s, 1H), 9.12 (s, 1H), 8.02 (dd, J=14.8, 8.5 Hz, 1H), 7.26 (dd, J=15.4, 8.2 Hz, 1H), 6.95 (ddd, J=75.5, 16.8, 10.6 Hz, 1H), 6.73 (d, J=8.3 Hz, 1H), 6.68 (t, J=8.8 Hz, 1H), 6.22-6.09 (m, 1H), 5.79-5.61 (m, 1H), 5.13-4.55 (m, 1H), 4.77-4.42 (m, 1H), 4.02 (dd, J=28.2, 3.9 Hz, 1H), 3.74 (dd, J=14.2, 4.2 Hz, 1H), 3.42 (t, J=15.3 Hz, 1H), 3.33-3.13 (m, 1H), 3.03-2.85 (m, 1H), 2.79 (dd, J=13.5, 6.8 Hz, 1H), 2.51-2.45 (m, 3H), 1.56 (dd, J=18.3, 6.7 Hz, 3H), 1.11 (d, J=6.7 Hz, 3H), 1.05 (d, J=6.6 Hz, 3H), 1.00 (d, J=6.6 Hz, 3H), 0.85 (d, J=6.7 Hz, 3H). ES-API: [M+H].sup.+=644.2.
Example 41 Preparation of Compound Z41
[0490] ##STR00552## ##STR00553##
[0491] Step 1: 4-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-3-nitro-1,8-naphthyridin-2(1H)-one (1 g, 1.89 mmol) was dissolved in N,N-dimethylacetamide (10 mL), orderly added with 1-(tert-butyl)3-methyl(R)-piperazin-1,3-dicarboxylate (1.39 g, 5.67 mmol) and N,N-diisopropylethylamine (733 mg, 5.67 mmol), and stirred at 120° C. for 2 hours. Cooled reaction liquid was added with 30 mL of EtOAc, washed with 30 mL of sodium bicarbonate twice, with 30 mL of dilute brine twice, 30 mL of water once and with 30 mL of saturated salt solution once, dried and concentrated, and purified by flash column chromatography on silica gel (0-100% EtOAc/PE) to obtain 1-(tert-butyl)3-methyl(3R)-4-(1-(4,6-diisopropylpyrimidin-5-yl)-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazin-1,3-dicarboxylate (1.3 g, Y: 80%). ES-API: [M+H].sup.+=738.3.
[0492] Step 2: the 1-(tert-butyl)3-methyl(3R)-4-(1-(4,6-diisopropylpyrimidin-5-yl)-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazin-1,3-dicarboxylate (1.23 g, 1.67 mmol) was dissolved in acetic acid (12 mL), added with iron powder (326 mg, 5.84 mmol), and stirred at 80° C. for 2 hours. The resulting reaction liquid was concentrated, added with 50 mL of EtOAc, and mixed with saturated sodium bicarbonate solution to adjust the pH to 8. The resulting suspension was filtered by diatomite. The filter cake was washed with EtOAc. The resulting organic phase was separated, washed orderly with 25 mL of saturated sodium bicarbonate and 25 mL of saturated salt solution, dried and concentrated to obtain crude product tert-butyl (4aR)-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-5,7-dioxo-1,2,4,4a,5,6,7,8-octa hydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylic acid (1.08 g, Y: 95%), which was grey-green solid. ES-API: [M+H].sup.+=676.2.
[0493] Step 3: the tert-butyl (4aR)-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-5,7-dioxo-1,2,4,4a,5,6,7,8-octa hydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (500 mg, 0.74 mmol), acetone (10 mL), anhydrous potassium carbonate (307 mg, 2.22 mmol), and iodomethane (630 mg, 4.44 mmol) were orderly added to a 15 mL sealing tube. The sealing tube was sealed, and the resulting mixture was stirred at 50° C. to react overnight. The resulting reaction liquid was filtered, concentrated, dissolved in DCM (10 mL), and washed with water (10 mL). The resulting organic phase was dried and concentrated to obtain crude product tert-butyl (4aR)-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-6-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (410 mg, Y: 80%), which was yellow solid. ES-API: [M+H].sup.+=690.2.
[0494] Step 4: the tert-butyl (4aR)-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-6-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (410 mg, 0.60 mmol) was dissolved in DCM (4 mL), and added with TFA (1 mL). After stirring at room temperature for 2 hours, the resulting reaction liquid was concentrated to obtain crude product (4aR)-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (350 mg), which was directly used in next step. ES-API: [M+H].sup.+=590.2.
[0495] Step 5: the (4aR)-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (350 mg, 0.59 mmol) was dissolved in DCM (5 mL), and added with N,N-diisopropylethylamine (230 mg, 1.78 mmol). The resulting reaction liquid was cooled to 0° C., and added dropwise with acryloyl chloride (107 mg, 1.19 mmol). The reaction liquid was stirred at 0° C. for 5 minutes, and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (0-100% EtOAc/PE) to obtain product (4aR)-3-acryloyl-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (300 mg, Y: 80%), which was faint yellow solid. ES-API: [M+H].sup.+=644.2.
[0496] Step 6: the (4aR)-3-acryloyl-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (300 mg, 0.47 mmol) was dissolved in DCM (5 mL). The resulting solution was cooled to 0° C., added dropwise with a solution (5 mL) of 17% boron tribromide in DCM, and stirred at room temperature overnight. The resulting reaction liquid was poured into 40 mL of saturated solution of sodium bicarbonate and extracted with 25 mL of DCM twice. The resulting organic phase was dried and concentrated, and the resulting crude product was purified by preparative scale HPLC to obtain product (4aR)-3-acryloyl-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-hydroxyphenyl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (Z41, 97 mg, Y: 35%), which was faint yellow solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.13 (s, 1H), 9.12 (s, 1H), 8.42 (d, J=8.7 Hz, 1H), 7.26 (dd, J=15.5, 8.2 Hz, 1H), 7.03 (dt, J=111.6, 55.9 Hz, 1H), 6.73 (d, J=8.3 Hz, 1H), 6.68 (t, J=8.8 Hz, 1H), 6.16 (d, J=16.9 Hz, 1H), 5.76 (d, J=11.0 Hz, 1H), 4.74 (d, J=13.9 Hz, 1H), 4.45 (d, J=12.3 Hz, 1H), 4.00 (s, 1H), 3.58 (t, J=12.7 Hz, 2H), 3.33 (s, 3H), 3.22 (t, J=11.5 Hz, 1H), 2.79 (dt, J=13.5, 6.7 Hz, 1H), 2.69 (t, J=10.7 Hz, 1H), 2.51-2.45 (m, 1H), 1.11 (d, J=6.7 Hz, 3H), 1.04 (d, J=6.6 Hz, 3H), 1.01 (d, J=6.6 Hz, 3H), 0.85 (d, J=6.7 Hz, 3H). ES-API: [M+H]+=630.2.
Example 42 Preparation of Compound Z42
[0497] ##STR00554## ##STR00555##
[0498] Step 1 4-chloro-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2 (1H)-one (1.2 g, 2.40 mmol) was dissolved in N,N-dimethylacetamide (10 mL), orderly added with tert-butyl (3-(2-methoxy-2-oxyethyl)piperazin-1-carboxylate (743 mg, 2.87 mmol) and N,N-diisopropylethylamine (930 mg, 7.20 mmol), and stirred at 120° C. for 2 hours to react. The resulting reaction liquid was added with 50 mL of EtOAc, washed with 25 mL of dilute brine for 4 times and then with 20 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-50%) to obtain yellow solid tert-butyl-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-3-(2-methoxy-2-oxyethyl)piperazin-1-carboxylic acid (580 mg, Y: 28%). ES-API: [M+H].sup.+=723.3.
[0499] Step 2: the tert-butyl 4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-3-(2-methoxy-2-oxyethyl)piperazin-1-carboxylate (550 mg, 0.76 mmol) was dissolved in acetic acid (5.5 mL), added with iron powder (149 mg, 2.66 mmol), and stirred at 80° C. for 2 hours to react. The resulting reaction liquid was concentrated, and orderly added with 50 mL of EtOAc and 30 mL of saturated sodium bicarbonate. The resulting suspension was filtered by using diatomite. The filter cake was washed with EtOAc. The resulting organic phase was separated, washed orderly with 25 mL of saturated sodium bicarbonate and 25 mL of saturated salt solution, dried and concentrated to obtain yellow solid tert-butyl 12-fluoro-11-(2-fluoro-6-methoxyphenyl)-9-(2-isopropyl-4-methylpyridin-3-yl)-6,8-dioxo-1,2,4a,5,6,7,8,9-octahydropyrazino[1′,2′:4,5][1,4]diaza[2,3-c][1,8]naphthyridin-3(4H)-carboxylate (520 mg). ES-API: [M+H].sup.+=661.3.
[0500] Step 3: the tert-butyl 12-fluoro-11-(2-fluoro-6-methoxyphenyl)-9-(2-isopropyl-4-methylpyridin-3-yl)-6,8-dioxo-1,2,4a,5,6,7,8,9-octahydropyrazino[1′,2′:4,5][1,4]diaza[2,3-c][1,8]naphthyridin-3(4H)-carboxylate (100 mg, 0.15 mmol) was dissolved in DCM (4 mL), and added with TFA (1 mL). After stirring at room temperature for 2 hours, the resulting reaction liquid was concentrated to obtain product 12-fluoro-11-(2-fluoro-6-methoxyphenyl)-9-(2-isopropyl-4-methylpyridin-3-yl)-1,2,3,4,4a,5,7,9-octahydropyrazino [1′,2′:4,5][1,4]diaza[2,3-c][1,8]naphthyridin-6,8-dione (85 mg, crude), which was directly used in next step. ES-API: [M+H].sup.+=561.3.
[0501] Step 4: the 12-fluoro-11-(2-fluoro-6-methoxyphenyl)-9-(2-isopropyl-4-methylpyridin-3-yl)-1,2,3,4,4a,5,7,9-octahydropyrazino [1′,2′:4,5][1,4]diaza[2,3-c][1,8]naphthyridin-6,8-dione (85 mg, crude) was dissolved in DCM (2 mL), and added with N,N-diisopropylethylamine (58 mg, 0.45 mmol). The resulting reaction liquid was cooled to 0° C., added dropwise with acryloyl chloride (27 mg, 0.30 mmol), and stirred at 0° C. for 10 minutes. The resulting reaction liquid was added with 25 mL of DCM, washed orderly with 15 mL of water, 15 mL of saturated solution of sodium bicarbonate and 15 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-100%) to obtain product 3-acryloyl-12-fluoro-11-(2-fluoro-6-methoxyphenyl)-9-(2-isopropyl-4-methylpyridin-3-yl)-1,2,3,4,4a,5,7,9-octahydropyrazino[1′,2′:4,5][1,4]diaza[2,3-c][1,8]naphthyridin-6,8-dione (60 mg, Y:65%), which was faint yellow solid. ES-API: [M+H].sup.+=615.3.
[0502] Step 5: the 3-acryloyl-12-fluoro-11-(2-fluoro-6-methoxyphenyl)-9-(2-isopropyl-4-methylpyridin-3-yl)-1,2,3,4,4a,5,7,9-octahydropyrazino[1′,2′:4,5][1,4]diaza[2,3-c][1,8]naphthyridin-6,8-dione (60 mg, 0.10 mmol) was dissolved in DCM (3 mL). The resulting reaction liquid was cooled to 0° C., and then added dropwise with a solution (3 mL) of 17% boron tribromide in DCM. The resulting mixture was stirred at room temperature for 2 hours to react. The resulting reaction liquid was poured into 40 mL of saturated solution of sodium bicarbonate and extracted with 25 mL of DCM twice. The resulting organic phase was dried and concentrated, and the resulting crude product was purified by preparative scale HPLC to obtain product 3-acryloyl-12-fluoro-11-(2-fluoro-6-hydroxyphenyl)-9-(2-isopropyl-4-methylpyridin-3-yl)-1,2,3,4,4a,5,7,9-octahydropyrazino[1′,2′:4,5][1,4]diaza[2,3-c][1,8]naphthyridin-6,8-dione (Z42, 27 mg, Y: 46%), which was faint yellow solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.11 (s, 1H), 8.62 (s, 1H), 8.44 (d, J=4.6 Hz, 1H), 8.24 (s, 1H), 7.23-7.28 (m, 2H), 6.85-7.00 (m, 1H), 6.78-6.62 (m, 2H), 6.24 (s, 1H), 5.77 (d, J=9.9 Hz, 1H), 3.93 (s, 7H), 3.09 (s, 1H), 2.83 (s, 1H), 2.60 (s, 1H), 1.86 (d, J=23.4 Hz, 3H), 1.06 (t, J=7.1 Hz, 3H), 0.91 (dd, J=9.7, 6.8 Hz, 3H). ES-API: [M+H].sup.+=601.2.
Example 43 Preparation of Compound Z43
[0503] ##STR00556## ##STR00557##
[0504] Step 1: under the condition of an ice water bath, sodium hydride (91 mg, 2.27 mmol) was added to a solution of tert-butyl 12-fluoro-11-(2-fluoro-6-methoxyphenyl)-9-(2-isopropyl-4-methylpyridin-3-yl)-6,8-dioxo-1,2,4a,5,6,7,8,9-octahydropyrazino[1′,2′:4,5][1,4]diaza[2,3-c][1,8]naphthyridin-3(4H)-carboxylate (300 mg, 0.45 mmol) in tetrahydrofuran (10 mL), and stirred for 30 minutes. Iodomethane (650 mg, 4.54 mmol) was then added to the resulting reaction liquid, and stirred at room temperature overnight. The reaction liquid was quenched by using saturated ammonium chloride solution (30 mL), and extracted with EtOAc (30 mL*3). The resulting organic phase was dried, concentrated, and purified by flash column chromatography on silica gel (0-10% MeOH/DCM) to obtain product tert-butyl 12-fluoro-11-(2-fluoro-6-methoxyphenyl)-9-(2-isopropyl-4-methylpyridin-3-yl)-7-methyl-6,8-dioxo-1,2,4a,5,6,7,8,9-octahydropyrazino[1′,2′:4,5][1,4]diaza[2,3-c][1,8]naphthyridin-3(4H)-carboxylate (190 mg, Y: 62%), which was yellow solid. ES-API: [M+H].sup.+=675.3.
[0505] Step 2: the tert-butyl 12-fluoro-11-(2-fluoro-6-methoxyphenyl)-9-(2-isopropyl-4-methylpyridin-3-yl)-7-methyl-6,8-dioxo-1,2,4a,5,6,7,8,9-octahydropyrazino[1′,2′:4,5][1,4]diaza[2,3-c][1,8]naphthyridin-3(4H)-carboxylate (188 mg, 0.28 mmol) was dissolved in DCM (8 mL), and added with TFA (2 mL). After stirring at room temperature for 2 hours, the resulting reaction liquid was concentrated to obtain crude product 12-fluoro-11-(2-fluoro-6-methoxyphenyl)-9-(2-isopropyl-4-methylpyridin-3-yl)-7-methyl-1,2,3,4,4a,5,7,9-octahydropyrazino[1′,2′:4,5][1,4]diaza[2,3-c][1,8]naphthyridin-6,8-dione (160 mg), which was directly used in next step. ES-API: [M+H].sup.+=575.2.
[0506] Step 3: the 12-fluoro-11-(2-fluoro-6-methoxyphenyl)-9-(2-isopropyl-4-methylpyridin-3-yl)-7-methyl-1,2,3,4,4a,5,7,9-octahydropyrazino[1′,2′:4,5][1,4]diaza[2,3-c][1,8]naphthyridin-6,8-dione (160 mg, 0.28 mmol) was dissolved in DCM (4 mL), and added with N,N-diisopropylethylamine (108 mg, 0.84 mmol). The resulting reaction liquid was cooled to 0° C., and added dropwise with acryloyl chloride (50 mg, 0.56 mmol). The reaction liquid was stirred at 0° C. for 5 minutes, and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (0-80% EtOAc/PE) to obtain product 3-acryloyl-12-fluoro-11-(2-fluoro-6-methoxyphenyl)-9-(2-isopropyl-4-methylpyridin-3-yl)-7-methyl-1,2,3,4,4a,5,7,9-octahydropyrazino[1′,2′:4,5][1,4]diaza[2,3-c][1,8]naphthyridin-6,8-dione (170 mg, Y: 97%), which was faint yellow solid. ES-API: [M+H].sup.+=629.2.
[0507] Step 4: 3-acryloyl-12-fluoro-11-(2-fluoro-6-methoxyphenyl)-9-(2-isopropyl-4-methylpyridin-3-yl)-7-methyl-1,2,3,4,4a,5,7,9-octahydropyrazino[1′,2′:4,5][1,4]diaza[2,3-c][1,8]naphthyridin-6,8-dione (170 mg, 0.27 mmol) was dissolved in DCM (3 mL). The resulting solution was cooled to 0° C., added dropwise with a solution (3 mL) of 17% boron tribromide in DCM, and stirred at room temperature for 2 hours. The resulting reaction liquid was poured into 40 mL of saturated solution of sodium bicarbonate and extracted with 25 mL of DCM twice. The resulting organic phase was dried and concentrated, and the resulting crude product was purified by preparative scale HPLC to obtain product 3-acryloyl-12-fluoro-11-(2-fluoro-6-hydroxyphenyl)-9-(2-isopropyl-4-methylpyridin-3-yl)-7-methyl-1,2,3,4,4a,5,7,9-octahydropyrazino[1′,2′:4,5][1,4]diaza[2,3-c][1,8]naphthyridin-6,8-dione (Z43, 95 mg, Y: 57%), which was faint yellow solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 8.47 (d, J=9.8 Hz, 1H), 8.42 (dd, J=4.8, 2.1 Hz, 1H), 7.26-7.19 (m, 2H), 7.04-6.82 (m, 1H), 6.72-6.67 (m, 1H), 6.66-6.62 (m, 1H), 6.19 (d, J=16.5 Hz, 1H), 5.76 (d, J=10.4 Hz, 1H), 4.60-4.45 (m, 1H), 4.36-4.10 (m, 1H), 4.02 (s, 1H), 3.93-3.78 (m, 1H), 3.65 (dd, J=21.9, 10.6 Hz, 1H), 3.29-3.22 (m, 2H), 3.09 (s, 3H), 3.17-2.99 (m, 1H), 2.76-2.63 (m, 2H), 2.37-2.28 (m, 1H), 1.90 (d, J=47.0 Hz, 3H), 1.05 (dd, J=31.1, 6.7 Hz, 3H), 0.93 (dd, J=42.5, 6.6 Hz, 3H). ES-API: [M+H].sup.+=615.2.
Example 44 Preparation of Compounds Z44a and Z44
[0508] ##STR00558## ##STR00559##
[0509] Step 1: tert-butyl (2R,4aR)-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-2-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (350 mg, 0.51 mmol), acetone (5 mL), anhydrous potassium carbonate (210 mg, 1.52 mmol), and deuteroiodomethane (735 mg, 5.07 mmol) were orderly added to a 15 mL sealing tube. The sealing tube was sealed, and the resulting mixture was stirred at 50° C. overnight. The resulting reaction liquid was filtered and concentrated to obtain product tert-butyl (2R,4aR)-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-2-methyl-6-(methyl-d3)-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (250 mg), which was yellow solid. ES-API: [M+H].sup.+=707.3.
[0510] Step 2: the tert-butyl (2R,4aR)-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-2-methyl-6-(methyl-d3)-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (250 mg, 0.35 mmol) was dissolved in DCM (8 mL), and added with TFA (2 mL). After stirring at room temperature for 2 hours, the resulting reaction liquid was concentrated to obtain crude product (2R,4aR)-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-2-methyl-6-(methyl-d3)-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (215 mg), which was directly used in next step. ES-API: [M+H].sup.+=607.3.
[0511] Step 3: the (2R,4aR)-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-2-methyl-6-(methyl-d3)-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (215 mg, 0.35 mmol) was dissolved in DCM (3 mL), and added with N,N-diisopropylethylamine (226 mg, 1.75 mmol). The resulting reaction liquid was cooled to 0° C., and added dropwise with acryloyl chloride (63 mg, 0.70 mmol). The reaction liquid was stirred at 0° C. for 10 minutes, and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-80%) to obtain product Z44a (122 mg, P: 100%, Y: 52%), which was faint yellow solid. ES-API: [M+H].sup.+=661.3.
[0512] Step 4: (2R,4aR)-3-acryloyl-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-2-methyl-6-(methyl-d3)-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (Z44a, 120 mg, 0.18 mmol) was dissolved in DCM (3 mL). The resulting solution was cooled to 0° C., added dropwise with a solution (3 mL) of 17% boron tribromide in DCM, and stirred at room temperature overnight. The resulting reaction liquid was poured into 40 mL of saturated solution of sodium bicarbonate and extracted with 25 mL of DCM twice. The resulting organic phase was dried and concentrated, and the resulting crude product was purified by preparative scale HPLC to obtain product (2R,4aR)-3-acryloyl-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-hydroxyphenyl)-2-methyl-6-(methyl-d3)-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (Z44, 69 mg, P: 100%, Y: 59%), which was faint yellow solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 9.12 (s, 1H), 8.00 (dd, J=14.2, 8.7 Hz, 1H), 7.24 (dd, J=15.3, 7.8 Hz, 1H), 6.95 (ddd, J=75.8, 16.7, 10.5 Hz, 1H), 6.71 (s, 1H), 6.65 (t, J=8.5 Hz, 1H), 6.24-6.01 (m, 1H), 5.82-5.66 (m, 1H), 5.04-4.60 (m, 1H), 4.77-4.42 (m, 1H), 4.02 (dd, J=28.4, 3.7 Hz, 1H), 3.74 (dd, J=14.1, 4.1 Hz, 1H), 3.44 (m, 1H), 3.25 (m, 1H), 2.99-2.74 (m, 2H), 2.48-2.44 (m, 1H), 1.55 (dd, J=18.5, 6.7 Hz, 3H), 1.11 (d, J=6.7 Hz, 3H), 1.04 (d, J=6.7 Hz, 3H), 1.00 (d, J=6.6 Hz, 3H), 0.84 (d, J=6.7 Hz, 3H). ES-API: [M+H].sup.+=647.3.
Example 45 Preparation of Compound Z45
[0513] ##STR00560## ##STR00561##
[0514] Step 1: tert-butyl (2R,4aR)-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-2-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (50 mg, 0.07 mmol) was dissolved in DCM (4 mL), and added with TFA (1 mL). After stirring at room temperature for 2 hours, the resulting reaction liquid was concentrated to obtain crude product (2R,4aR)-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-2-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (43 mg), which was directly used in next step. ES-API: [M+H].sup.+=590.2.
[0515] Step 2: the (2R,4aR)-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-2-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (43 mg, 0.07 mmol) was dissolved in DCM (2 mL), and added with N,N-diisopropylethylamine (45 mg, 0.35 mmol). The resulting reaction liquid was cooled to 0° C., and added dropwise with acryloyl chloride (13 mg, 0.14 mmol). The reaction liquid was stirred at 0° C. for 5 minutes, and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-80%) to obtain product (2R,4aR)-3-acryloyl-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-2-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (26 mg, Y: 56%), which was faint yellow solid. ES-API: [M+H].sup.+=644.2.
[0516] Step 3: the (2R,4aR)-3-acryloyl-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-2-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (26 mg, 0.04 mmol) was dissolved in DCM (0.5 mL). The resulting solution was cooled to 0° C., added dropwise with a solution (0.5 mL) of 17% boron tribromide in DCM, and stirred at room temperature overnight. The resulting reaction liquid was poured into 4 mL of saturated solution of sodium bicarbonate and extracted with 3 mL of DCM twice. The resulting organic phase was dried and concentrated, and the resulting crude product was purified by preparative scale HPLC to obtain product (2R,4aR)-3-acryloyl-8-(4,6-diisopropylpyrimidin-5-yl)-11-fluoro-10-(2-fluoro-6-hydroxyphenyl)-2-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (Z45, 8 mg, Y: 31%), which was faint yellow solid. ES-API: [M+H].sup.+=630.2.
Example 46 Preparation of Compound Z46
[0517] ##STR00562## ##STR00563## ##STR00564##
[0518] Step 1: 7-chloro-6-fluoro-4-hydroxy-1-(2-isopropyl-4,6-dimethylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridin-3-nitrile (8 g, 20.68 mmol) was suspended in 1,4-dioxane (40 mL), and slowly added with a mixed liquid of concentrated sulfuric acid (40 mL) and water (40 mL). The resulting mixture was stirred at 120° C. overnight to react. Cooled reaction liquid was poured into 150 mL of ice water, mixed with aqueous solution of potassium hydroxide to adjust the pH to 6, and extracted with EtOAc. The resulting organic phase was dried and concentrated to obtain product 7-chloro-6-fluoro-4-hydroxy-1-(2-isopropyl-4,6-dimethylpyridin-3-yl)-1,8-naphthyridin-2(1H)-one (7.6 g, Y: 96%), which was faint yellow solid. ES-API: [M+H].sup.+=362.2.
[0519] Step 2: the 7-chloro-6-fluoro-4-hydroxy-1-(2-isopropyl-4,6-dimethylpyridin-3-yl)-1,8-naphthyridin-2(1H)-one (7.6 g, 21.01 mmol) was dissolved in acetic acid (26 mL), orderly added with sodium nitrite (145 mg, 2.10 mmol) and concentrated nitric acid (3.97 g, 63.01 mmol), and stirred at room temperature for 20 minutes. The resulting reaction liquid was poured into 20 mL of water. Yellow solid was precipitated and filtered. The filter cake was washed with 6 mL of water and dried in vacuum to obtain product 7-chloro-6-fluoro-4-hydroxy-1-(2-isopropyl-4,6-dimethylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (4 g, Y: 47%), which was faint yellow solid. ES-API: [M+H].sup.+=407.1.
[0520] Step 3: a mixed solution of 7-chloro-6-fluoro-4-hydroxy-1-(2-isopropyl-4,6-dimethylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (4 g, 9.83 mmol), (2-fluoro-6-methoxyphenyl)boric acid (8.36 g, 49.16 mmol), chloro(2-dicyclohexylphosphino-2′,6′-dimethoxy-1,1′-biphenyl)(2′-amino-1,1′-biphen-2-yl)palladium (II) (705 mg, 0.98 mmol), 2-dicyclohexylphosphino-2′,6′-dimethoxybiphenyl (402 mg, 0.98 mmol), and potassium phosphate (6.26 g, 29.50 mmol) in 8 mL of water and 40 mL of dioxane was replaced with nitrogen for 3 times and allowed to react at 100° C. for 2 hours. The resulting reaction liquid was poured into 50 mL of water, and washed with methyl tert-butyl ether (30 mL*2). The water phase was mixed with 3.0 M diluted hydrochloric acid to adjust the pH to 6.0, and then extracted with EtOAc (30 mL*2). The resulting organic phase was dried and concentrated to obtain crude product 6-fluoro-7-(2-fluoro-6-methoxyphenyl)-4-hydroxy-1-(2-isopropyl-4,6-dimethylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (4 g, Y: 82%). ES-API: [M+H].sup.+=497.2.
[0521] Step 4: phosphorus oxychloride (6.18 g, 40.30 mmol) and N,N-diisopropylethylamine (8.33 g, 68.48 mmol) were orderly added to a solution of the 6-fluoro-7-(2-fluoro-6-methoxyphenyl)-4-hydroxy-1-(2-isopropyl-4,6-dimethylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (4 g, 8.06 mmol) in ACN (60 mL). The resulting mixture was stirred at 80° C. temperature for 1 hour to react. The resulting reaction liquid was concentrated, added with 150 mL of EtOAc, and washed orderly with 80 mL of saturated sodium bicarbonate twice and then with 80 mL of water and 80 mL of saturated salt solution. After the resulting organic phase was dried and concentrated, the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-50%) to obtain product 4-chloro-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4,6-dimethylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (3.5 g, Y: 85%), which was faint yellow solid. ES-API: [M+H].sup.+=515.2.
[0522] Step 5: the 4-chloro-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4,6-dimethylpyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (1 g, 1.94 mmol) was dissolved in N,N-dimethylacetamide (10 mL), orderly added with 1-(tert-butyl)3-methyl(3R,6R)-6-methylpiperazin-1,3-dicarboxylic acid (600 mg, 2.33 mmol) and N,N-diisopropylethylamine (750 mg, 5.82 mmol), and stirred at 120° C. for 2 hours. Cooled reaction liquid was added with 100 mL of EtOAc, washed with 50 mL of sodium bicarbonate twice, with 50 mL of dilute brine twice, with 50 mL of water once and with 50 mL of saturated salt solution once, dried and concentrated to obtain crude product 1-(tert-butyl)3-methyl(3R,6R)-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4,6-dimethylpyridin-3-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-6-methylpiperazin-1,3-dicarboxylate (1.6 g). ES-API: [M+H].sup.+=737.2.
[0523] Step 6: the 1-(tert-butyl)3-methyl(3R,6R)-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropyl-4,6-dimethylpyridin-3-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-6-methylpiperazin-1,3-dicarboxylate (1.6 g, 2.17 mmol) was dissolved in acetic acid (16 mL), added with iron powder (425 mg, 7.60 mmol), and stirred at 80° C. for 1 hour. The resulting reaction liquid was concentrated, added with 50 mL of EtOAc, and mixed with saturated sodium bicarbonate solution to adjust the pH to 6. The resulting suspension was filtered by diatomite. The filter cake was washed with EtOAc. The resulting organic phase was separated, washed orderly with 25 mL of saturated sodium bicarbonate and 25 mL of saturated salt solution, dried and concentrated to obtain crude product tert-butyl (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4,6-dimethylpyridin-3-yl)-2-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (1.4 g), which was yellow solid. ES-API: [M+H].sup.+=675.3.
[0524] Step 7: the tert-butyl (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4,6-dimethylpyridin-3-yl)-2-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (1.4 g, 2.07 mmol), acetone (15 mL), anhydrous potassium carbonate (860 mg, 6.21 mmol), and iodomethane (2.95 g, 20.7 mmol) were orderly added to a 50 mL sealing tube. The sealing tube was sealed, and the resulting mixture was stirred at 50° C. to react overnight. The resulting reaction liquid was filtered, concentrated, and purified by flash column chromatography on silica gel (EtOAc/PE: 0-80%) to obtain product tert-butyl (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4,6-dimethylpyridin-3-yl)-2,6-dimethyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (340 mg, Y: 24%), which was yellow solid. ES-API: [M+H].sup.+=706.3.
[0525] Step 8: the tert-butyl (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4,6-dimethylpyridin-3-yl)-2,6-dimethyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (340 mg, 0.49 mmol) was dissolved in DCM (4 mL), and added with TFA (1 mL). After stirring at room temperature for 2 hours, the resulting reaction liquid was concentrated to obtain crude product (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4,6-dimethylpyridin-3-yl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (317 mg), which was directly used in next step. ES-API: [M+H].sup.+=589.3.
[0526] Step 9: the (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4,6-dimethylpyridin-3-yl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (317 mg, 0.49 mmol) was dissolved in DCM (5 mL), and added with N,N-diisopropylethylamine (320 mg, 2.45 mmol). The resulting reaction liquid was cooled to 0° C., and added dropwise with acryloyl chloride (89 mg, 0.99 mmol). The reaction liquid was stirred at 0° C. for 5 minutes, and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-80%) to obtain product (2R,4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4,6-dimethylpyridin-3-yl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (280 mg, Y: 81%), which was faint yellow solid. ES-API: [M+H].sup.+=643.3.
[0527] Step 10: the (2R,4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4,6-dimethylpyridin-3-yl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (280 mg, 0.44 mmol) was dissolved in DCM (3 mL). The resulting solution was cooled to 0° C., added dropwise with a solution (3 mL) of 17% boron tribromide in DCM, and stirred at room temperature for 2 hours. The resulting reaction liquid was poured into 40 mL of saturated solution of sodium bicarbonate and extracted with 25 mL of DCM twice. The resulting organic phase was dried and concentrated, and the resulting crude product was purified by preparative scale HPLC to obtain product (2R,4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(2-isopropyl-4,6-dimethylpyridin-3-yl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (Z46, 108 mg, Y: 40%), which was faint yellow solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.14 (s, 1H), 7.99 (t, J=10.4 Hz, 1H), 7.27 (dd, J=15.5, 7.7 Hz, 1H), 7.13-6.83 (m, 2H), 6.78-6.65 (m, 2H), 6.16 (t, J=13.5 Hz, 1H), 5.81-5.69 (m, 1H), 5.08-4.72 (m, 1H), 4.54 (t, J=51.7 Hz, 1H), 3.99 (t, J=29.0 Hz, 1H), 3.73 (d, J=10.5 Hz, 1H), 3.44-3.23 (m, 1H), 3.35 (s, 3H), 2.95-2.80 (m, 1H), 2.75-2.68 (m, 1H), 2.45 (d, J=1.9 Hz, 3H), 2.41-2.37 (m, 1H), 1.84 (d, J=96 Hz, 1H), 1.55 (dd, J=16.5, 5.4 Hz, 3H), 1.05 (dd, J=39.3, 6.7 Hz, 3H), 0.90 (dd, J=65.2, 6.6 Hz, 3H). ES-API: [M+H].sup.+=629.2.
Example 47 Preparation of Compound Z47
[0528] ##STR00565## ##STR00566## ##STR00567##
[0529] Step 1: 7-chloro-6-fluoro-4-hydroxy-1-(3-isopropyl-1,5-dimethyl-1H-pyrazol-4-yl)-2-oxo-1,2-dihydro-1,8-naphthyridin-3-nitrile (3.5 g, 9.3 mmol) was suspended in 1,4-dioxane (40 mL), and slowly added with a mixed liquid of concentrated sulfuric acid (40 mL) and water (40 mL). The resulting mixture was stirred at 120° C. for 20 hours to react. Cooled reaction liquid was poured into 150 mL of ice water, mixed with 4.0 M aqueous potassium hydroxide solution to adjust the pH to 3. The precipitated solid was filtered. The filtrate was extracted with 200 mL of DCM. The resulting solid was dissolved in a mixed liquid of DCM/MeOH=10:1, and filtered. The resulting combined organic phase was dried and concentrated to obtain product 7-chloro-6-fluoro-4-hydroxy-1-(3-isopropyl-1,5-dimethyl-1H-pyrazol-4-yl)-1,8-naphthyridin-2(1H)-one (2.3 g, Y: 71.6%), which was light brown solid. ES-API: [M+H].sup.+=351.1.
[0530] Step 2: the 7-chloro-6-fluoro-4-hydroxy-1-(3-isopropyl-1,5-dimethyl-1H-pyrazol-4-yl)-1,8-naphthyridin-2(1H)-one (2.3 g, 6.5 mmol) was dissolved in acetic acid (8 mL), orderly added with sodium nitrite (45 mg, 0.65 mmol) and concentrated nitric acid (1.5 mL, 19.5 mmol), and stirred at room temperature for 1 hour to react. The resulting reaction liquid was poured into 30 mL of ice water. The precipitated solid was filtered. The filter cake was washed with 6 mL of water and dried in vacuum to obtain product 7-chloro-6-fluoro-4-hydroxy-1-(3-isopropyl-1,5-dimethyl-1H-pyrazol-4-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (1.5 g, Y: 60%), which was faint yellow solid. ES-API: [M+H].sup.+=396.1.
[0531] Step 3: the 7-chloro-6-fluoro-4-hydroxy-1-(3-isopropyl-1,5-dimethyl-1H-pyrazol-4-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (1.5 g, 3.8 mmol), (2-fluoro-6-methoxyphenyl)boric acid (0.96 g, 5.7 mmol), chloro(2-dicyclohexylphosphino-2′,6′-dimethoxy-1,1′-biphenyl)[2-(2′-amino-1,1′-biphenyl)]palladium(II)-G2-Pd (273 mg, 0.38 mmol), 2-dicyclohexylphosphino-2′,6′-dimethoxybiphenyl (155 mg, 0.38 mmol), potassium phosphate (2.41 g, 11.4 mmol), 4 mL of water, and 20 mL of dioxane were added to a 250 mL round-bottom flask. The resulting mixture was stirred at 85° C. for 4 hours to react under the protection of nitrogen. The resulting reaction liquid was concentrated, added with 100 mL of water, mixed with 3.0 M diluted hydrochloric acid to adjust the pH to 3.0, and extracted with 120 mL of DCM twice. The resulting organic phase was dried and concentrated to obtain product 6-fluoro-7-(2-fluoro-6-methoxyphenyl)-4-hydroxy-1-(3-isopropyl-1,5-dimethyl-1H-pyrazol-4-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (2.4 g, crude), which was directly used in next step. ES-API: [M+H].sup.+=486.1.
[0532] Step 4: the 6-fluoro-7-(2-fluoro-6-methoxyphenyl)-4-hydroxy-1-(3-isopropyl-1,5-dimethyl-1H-pyrazol-4-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (2.4 g, crude) was dissolved in ACN (50 mL), orderly added with phosphorus oxychloride (3.8 g, 24.7 mmol) and N,N-diisopropylethylamine (6.38 g, 49.4 mmol), and stirred at 85° C. for 1 hour to react. The resulting reaction liquid was concentrated, added with 150 mL of EtOAc, and washed orderly with 80 mL of water, with 80 mL saturated sodium bicarbonate solution twice and then with 80 mL of saturated salt solution. After the resulting organic phase was dried and concentrated, the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-40%) to obtain product 4-chloro-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(3-isopropyl-1,5-dimethyl-1H-pyrazol-4-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (0.29 g, Y:10%), which was faint yellow solid. ES-API: [M+H].sup.+=504.1.
[0533] Step 5: the 4-chloro-6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(3-isopropyl-1,5-dimethyl-1H-pyrazol-4-yl)-3-nitro-1,8-naphthyridin-2(1H)-one (290 mg, 0.57 mmol) was dissolved in N,N-dimethylacetamide (3 mL), orderly added with methyl (3R,6R)-1-N-BOC-6-methylpiperazin-3-formate (155 mg, 0.60 mmol) and N,N-diisopropylethylamine (116 mg, 0.90 mmol), and stirred at 120° C. for 2 hours to react. The resulting reaction liquid was added with 50 mL of EtOAc, washed with 25 mL of dilute brine for 4 times and then with 20 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-50%) to obtain product 1-(tert-butyl)3-methyl(3R,6R)-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(3-isopropyl-1,5-dimethyl-1H-pyrazol-4-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-6-methylpiperazin-1,3-dicarboxylic acid (280 mg, Y: 68.8%), which was orange solid. ES-API: [M+H].sup.+=726.3.
[0534] Step 6: the 1-(tert-butyl)3-methyl(3R,6R)-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-1-(3-isopropyl-1,5-dimethyl-1H-pyrazol-4-yl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-6-methylpiperazin-1,3-dicarboxylic acid (280 mg, 0.38 mmol) was dissolved in acetic acid (2.5 mL), added with iron powder (37 mg, 0.66 mmol), and stirred at 80° C. for 30 minutes to react. The resulting reaction liquid was concentrated, and orderly added with 50 mL of EtOAc and 30 mL of saturated sodium bicarbonate. The resulting suspension was filtered by diatomite. The filter cake was washed with EtOAc. The resulting organic phase was separated, washed orderly with 25 mL of saturated sodium bicarbonate and 25 mL of saturated salt solution, dried and concentrated to obtain product tert-butyl (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(3-isopropyl-1,5-dimethyl-1H-pyrazol-4-yl)-2-methyl-5,7-di oxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (250 mg, Y: 97.6%), which was yellow solid. ES-API: [M+H].sup.+=664.3.
[0535] Step 7: the tert-butyl (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(3-isopropyl-1,5-dimethyl-1H-pyrazol-4-yl)-2-methyl-5,7-di oxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (250 mg, 0.37 mmol), 4 mL of acetone, anhydrous potassium carbonate (103.9 mg, 0.74 mmol), and iodomethane (246 mg, 1.70 mmol) were orderly added to a 15 mL sealing tube. The sealing tube was sealed, and the resulting mixture was stirred at 50° C. for 18 hours to react. The resulting reaction liquid was added with 30 mL of EtOAc, washed orderly with 12 mL of water and 15 mL of saturated salt solution, dried and concentrated to obtain product tert-butyl (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(3-isopropyl-1,5-dimethyl-1H-pyrazol-4-yl)-2,6-dimethyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (230 mg, Y: 90%), which was yellow solid. ES-API: [M+H].sup.+=678.3.
[0536] Step 8: the tert-butyl (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(3-isopropyl-1,5-dimethyl-1H-pyrazol-4-yl)-2,6-dimethyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-3-carboxylate (230 mg, 0.34 mmol) was dissolved in DCM (3 mL), and added with TFA (2 mL). After stirring at room temperature for 2 hours, the resulting reaction liquid was concentrated to obtain product (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(3-isopropyl-1,5-dimethyl-1H-pyrazol-4-yl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (230 mg, crude), which was directly used in next step. ES-API: [M+H].sup.+=578.2.
[0537] Step 9: the (2R,4aR)-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(3-isopropyl-1,5-dimethyl-1H-pyrazol-4-yl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (230 mg, crude) was dissolved in DCM (5 mL), and added with N,N-diisopropylethylamine (110 mg, 0.85 mmol). The resulting reaction liquid was cooled to 0° C., and added dropwise with acryloyl chloride (31 mg, 0.34 mmol). The resulting mixture was stirred at 0° C. for 15 minutes to react. The resulting reaction liquid was added with 25 mL of DCM, washed orderly with 15 mL of water, 15 mL of saturated solution of NaHCO.sub.3 and 15 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-100%) to obtain product (2R,4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(3-isopropyl-1,5-dimethyl-1H-pyrazol-4-yl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (190 mg, Y: 81.3%), which was faint yellow solid. ES-API: [M+H].sup.+=632.2.
[0538] Step 10: the (2R,4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-methoxyphenyl)-8-(3-isopropyl-1,5-dimethyl-1H-pyrazol-4-yl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (190 mg, 0.3 mmol) was dissolved in DCM (1.5 mL). The resulting solution was cooled to 0° C., and then added dropwise with a solution (5 mL) of 17% boron tribromide in DCM. The resulting mixture was stirred at room temperature for 4 hours to react. The resulting reaction liquid was poured into 40 mL of saturated solution of NaHCO.sub.3 and extracted with 25 mL of DCM twice. The resulting organic phase was dried and concentrated, and the resulting crude product was purified by preparative scale HPLC to obtain product (2R,4aR)-3-acryloyl-11-fluoro-10-(2-fluoro-6-hydroxyphenyl)-8-(3-isopropyl-1,5-dimethyl-1H-pyrazol-4-yl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c][1,8]naphthyridin-5,7-dione (Z47, 60 mg, Y: 31.5%), which was faint yellow solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.16 (d, J=3.7 Hz, 1H), 7.93 (t, J=9.5 Hz, 1H), 7.30 (dd, J=15.5, 8.1 Hz, 1H), 7.02 (dd, J=16.8, 10.6 Hz, 1H), 6.80-6.69 (m, 2H), 6.15 (t, J=13.1 Hz, 1H), 5.78-5.69 (m, 1H), 4.76 (s, 1H), 4.59 (dd, J=13.9, 6.2 Hz, 1H), 3.90 (d, J=18.9 Hz, 1H), 3.72 (dd, J=19.5, 4.3 Hz, 4H), 3.26 (s, 1H), 2.75 (d, J=7.7 Hz, 1H), 2.63-2.57 (m, 1H), 2.44-2.39 (m, 1H), 1.91 (d, J=70.7 Hz, 3H), 1.59-1.47 (m, 3H), 0.95 (ddd, J=34.1, 17.0, 5.2 Hz, 6H). ES-API: [M+H].sup.+=618.3.
Example 48 Preparation of Compounds Z48, Z48′-1, and Z48′-2
[0539] ##STR00568## ##STR00569## ##STR00570## ##STR00571## ##STR00572##
[0540] Step 1: 6,7-dichloro-1-(4,6-diisopropylpyrimidin-5-yl)-4-hydroxy-2-oxo-1,2-dihydro-1,8-naphthyridin-3-carbonitrile (2 g, 12 mmol) was suspended in 1,4-dioxane (12 mL), and slowly added with a mixed liquid of concentrated sulfuric acid (12 mL) and water (12 mL). The resulting mixture was stirred at 120° C. to react overnight. Cooled reaction liquid was then poured into 50 mL of ice water, mixed with aqueous solution of potassium hydroxide to adjust the pH to 6, and extracted with EtOAc (50 mL*3). The resulting organic phase was dried and concentrated to obtain target product 6,7-dichloro-1-(4,6-diisopropylpyrimidin-5-yl)-4-hydroxy-1,8-naphthyridin-2(1H)-one (1.5 g, Y: 80%), which was faint yellow solid. ES-API: [M+H].sup.+=393.1
[0541] Step 2: the 6,7-dichloro-1-(4,6-diisopropylpyrimidin-5-yl)-4-hydroxy-1,8-naphthyridin-2(1H)-one (1.50 g, 3.81 mmol) was dissolved in acetic acid (5 mL), orderly added with sodium nitrite (26 mg, 0.38 mmol) and concentrated nitric acid (721 mg, 11.44 mmol), and stirred at room temperature for 20 minutes. The resulting reaction liquid was poured into 5 mL of water. Yellow solid was precipitated and filtered. The filter cake was washed with 5 mL of water and dried in vacuum to obtain target product 6,7-dichloro-1-(4,6-diisopropylpyrimidin-5-yl)-4-hydroxy-3-nitro-1,8-naphthyridin-2(1H)-one (1.1 g, Y:66%), which was faint yellow solid. ES-API: [M+H].sup.+=438.1
[0542] Step 3: a mixed solution of 6,7-dichloro-1-(4,6-diisopropylpyrimidin-5-yl)-4-hydroxy-3-nitro-1,8-naphthyridin-2(1H)-one (1.1 g, 2.51 mmol), (2-fluoro-6-methoxyphenyl)boric acid (1.28 g, 7.53 mmol), chloro(2-dicyclohexylphosphino-2′,6′-dimethoxy-1,1′-biphenyl)(2′-amino-1,1′-biphen-2-yl)palladium(II) (166 mg, 0.23 mmol), 2-dicyclohexylphosphino-2′,6′-dimethoxybiphenyl (94 mg, 0.23 mmol), and potassium phosphate (1.45 g, 6.85 mmol) in 2 mL of water and 10 mL of dioxane was stirred at 80° C. for 1 hour under the protection of nitrogen. The resulting reaction liquid was poured into 30 mL of water, and washed with methyl tert-butyl ether (30 mL*2). The water phase was mixed with 3.0 M diluted hydrochloric acid to adjust the pH to 6.0, and extracted with EtOAc (50 mL*3). The resulting organic phase was dried and concentrated to obtain a target product, namely a mixture of 6-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-7-(2-fluoro-6-methoxyphenyl)-4-hydroxy-3-nitro-1,8-naphthyridin-2(1H)-one and 7-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-6-(2-fluoro-6-methoxyphenyl)-4-hydroxy-3-nitro-1,8-naphthyridin-2(1H)-one (1.1 g, Y: 83%, P1:P2=55:45). ES-API: [M+H].sup.+=528.1.
[0543] Step 4: phosphorus oxychloride (1.6 g, 10.42 mmol) and N,N-diisopropylethylamine (2.15 g, 16.67 mmol) were orderly added to a solution of the mixture of 6-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-7-(2-fluoro-6-methoxyphenyl)-4-hydroxy-3-nitro-1,8-naphthyridin-2(1H)-one and 7-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-6-(2-fluoro-6-methoxyphenyl)-4-hydroxy-3-nitro-1,8-naphthyridin-2(1H)-one (1.1 g, 2.08 mmol) in ACN (15 mL), and stirred at 80° C. for 1 hour. The resulting reaction liquid was concentrated, added with 50 mL of EtOAc, and washed orderly with 30 mL of saturated sodium bicarbonate twice and then with 30 mL of water and 30 mL of saturated salt solution. After the resulting organic phase was dried and concentrated, the resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-50%) to obtain a mixture of 4,6-dichloro-1-(4,6-diisopropylpyrimidin-5-yl)-7-(2-fluoro-6-methoxyphenyl)-3-nitro-1,8-naphthyridin-2(1H)-one and 4,7-dichloro-1-(4,6-diisopropylpyrimidin-5-yl)-6-(2-fluoro-6-methoxyphenyl)-3-nitro-1,8-naphthyridin-2(1H)-one (750 mg, Y: 61%), which was faint yellow solid. ES-API: [M+H].sup.+=546.1.
[0544] Step 5: the mixture of 4,6-dichloro-1-(4,6-diisopropylpyrimidin-5-yl)-7-(2-fluoro-6-methoxyphenyl)-3-nitro-1,8-naphthyridin-2(1H)-one and 4,7-dichloro-1-(4,6-diisopropylpyrimidin-5-yl)-6-(2-fluoro-6-methoxyphenyl)-3-nitro-1,8-naphthyridin-2(1H)-one (750 mg, 1.37 mmol) was dissolved in N,N-dimethylacetamide (7.5 mL), orderly added with 1-(tert-butyl)3-methyl(R)-piperazin-1,3-dicarboxylate (1.01 g, 4.12 mmol) and N,N-diisopropylethylamine (530 mg, 4.12 mmol), and stirred at 120° C. for 2 hours. After being cooled, the reaction liquid was added with 100 mL of EtOAc, washed with 50 mL of sodium bicarbonate twice, with 50 mL of dilute brine twice, with 50 mL of water once and with 50 mL of saturated salt solution once, dried and concentrated to obtain a mixture of 1-(tert-butyl)3-methyl(3R)-4-(6-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-7-(2-fluoro-6-methoxyphenyl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazin-1,3-dicarboxylate and 1-(tert-butyl)3-methyl(3R)-4-(7-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-6-(2-fluoro-6-methoxyphenyl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazin-1,3-dicarboxylate (800 mg). ES-API: [M+H].sup.+=754.2.
[0545] Step 6: the mixture of 1-(tert-butyl)3-methyl(3R)-4-(6-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-7-(2-fluoro-6-methoxyphenyl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazin-1,3-dicarboxylate and 1-(tert-butyl)3-methyl(3R)-4-(7-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-6-(2-fluoro-6-methoxyphenyl)-3-nitro-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazin-1,3-dicarboxylate (800 mg, 1.06 mmol) was dissolved in acetic acid (8 mL), added with iron powder (207 mg, 3.71 mmol), and stirred at 80° C. for 1 hour. The resulting reaction liquid was concentrated, added with 50 mL of EtOAc, and mixed with saturated sodium bicarbonate solution to adjust the pH to 8. The resulting suspension was filtered by diatomite. The filter cake was washed with EtOAc. The resulting organic phase was separated, washed orderly with 25 mL of saturated sodium bicarbonate and 25 mL of saturated salt solution, dried and concentrated, and purified by flash column chromatography on silica gel (EtOAc/PE: 0-80%) to obtain a mixture of tert-butyl(4aR)-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-10-(2-fluoro-6-methoxyphenyl)-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c] [1,8]naphthyridin-3-carboxylate and tert-butyl(4aR)-10-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-11-(2-fluoro-6-methoxyphenyl)-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c] [1,8]naphthyridin-3-carboxylate (350 mg, Y: 48%), which was yellow solid. ES-API: [M+H].sup.+=692.3.
[0546] Step 7: the mixture of tert-butyl(4aR)-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-10-(2-fluoro-6-methoxyphenyl)-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c] [1,8]naphthyridin-3-carboxylate and tert-butyl(4aR)-10-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-11-(2-fluoro-6-methoxyphenyl)-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c] [1,8]naphthyridin-3-carboxylate (350 mg, 0.51 mmol), acetone (5 mL), anhydrous potassium carbonate (209 mg, 1.52 mmol), and iodomethane (718 mg, 5.06 mmol) were orderly added to a 15 mL sealing tube. The sealing tube was sealed, and the resulting mixture was stirred at 50° C. to react overnight. The resulting reaction liquid was filtered and concentrated to obtain a mixture of tert-butyl(4aR)-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-10-(2-fluoro-6-methoxyphenyl)-6-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c] [1,8]naphthyridin-3-carboxylate and tert-butyl(4aR)-10-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-11-(2-fluoro-6-methoxyphenyl)-6-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c] [1,8]naphthyridin-3-carboxylate (350 mg), which was yellow solid. ES-API: [M+H].sup.+=706.3
[0547] Step 8: the mixture of tert-butyl(4aR)-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-10-(2-fluoro-6-methoxyphenyl)-6-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c] [1,8]naphthyridin-3-carboxylate and tert-butyl(4aR)-10-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-11-(2-fluoro-6-methoxyphenyl)-6-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c] [1,8]naphthyridin-3-carboxylate (350 mg, 0.50 mmol) was dissolved in DCM (8 mL), and added with TFA (2 mL). After stirring at room temperature for 2 hours, the resulting reaction liquid was concentrated to obtain a mixture of (4aR)-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-10-(2-fluoro-6-methoxyphenyl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c] [1,8]naphthyridin-5,7-dione and (4aR)-10-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-11-(2-fluoro-6-methoxyphenyl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c] [1,8]naphthyridin-5,7-dione (300 mg, P1:P2=53:47), which was directly used in next step. ES-API: [M+H].sup.+=606.3
[0548] Step 9: the mixture of (4aR)-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-10-(2-fluoro-6-methoxyphenyl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c] [1,8]naphthyridin-5,7-dione and (4aR)-10-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-11-(2-fluoro-6-methoxyphenyl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c] [1,8]naphthyridin-5,7-dione (300 mg, 0.50 mmol) was dissolved in DCM (3 mL), and added with N,N-diisopropylethylamine (320 mg, 2.45 mmol). The resulting reaction liquid was cooled to 0° C., and added dropwise with acryloyl chloride (90 mg, 0.99 mmol). The reaction liquid was stirred at 0° C. for 5 minutes, and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-80%) to obtain a mixture of (4aR)-3-acryloyl-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-10-(2-fluoro-6-methoxyphenyl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c] [1,8]naphthyridin-5,7-dione and (4aR)-3-acryloyl-10-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-11-(2-fluoro-6-methoxyphenyl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c] [1,8]naphthyridin-5,7-dione (250 mg, Y: 77%), which was faint yellow solid. ES-API: [M+H].sup.+=662.3.
[0549] Step 10: the mixture of (4aR)-3-acryloyl-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-10-(2-fluoro-6-methoxyphenyl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c] [1,8]naphthyridin-5,7-dione and (4aR)-3-acryloyl-10-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-11-(2-fluoro-6-methoxyphenyl)-6-methyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c] [1,8]naphthyridin-5,7-dione (250 mg, 0.38 mmol) was dissolved in DCM (3 mL). The resulting solution was cooled to 0° C., added dropwise with a solution (3 mL) of 17% boron tribromide in DCM, and stirred at room temperature overnight. The resulting reaction liquid was poured into 40 mL of saturated solution of sodium bicarbonate and extracted with 25 mL of DCM twice. The resulting organic phase was dried and concentrated, and the resulting crude product was purified by preparative scale HPLC to obtain: a compound Z48 (14.2 mg, P: 100%, retention time: 9.94 min, Y: 6%), which was faint yellow solid, ES-API: [M+H].sup.+=646.2; .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.04 (d, J=20 Hz, 1H), 9.10 (s, 1H), 8.54 (d, J=7.5 Hz, 1H), 7.28-7.17 (m, 1H), 7.14-6.77 (m, 1H), 6.73-6.68 (m, 1H), 6.68-6.63 (m, 1H), 6.21-6.11 (m, 1H), 5.79-5.71 (m, 1H), 5.15-4.66 (m, 1H), 4.51-4.02 (m, 1H), 3.99 (s, 1H), 3.66-3.53 (m, 2H), 3.32 (s, 3H), 3.27-3.18 (m, 1H), 2.84-2.74 (m, 1H), 2.74-2.61 (m, 1H), 2.52-2.46 (m, 1H), 1.10 (d, J=7 Hz, 3H), 1.04 (d, J=6.5 Hz, 3H), 0.99 (dd, J=10.5, 7 Hz, 3H), 0.86 (dd, J=15.5, 6.5 Hz, 3H); ES-API: [M+H].sup.+=646.2; a compound Z48′-1 (16.5 mg, P: 96%, retention time: 10.13 min, Y: 7%), which was faint yellow solid, ES-API: [M+H].sup.+=646.2; and a compound Z48′-2 (17.5 mg, P: 100%, retention time: 10.36 min, Y: 7%), which was faint yellow solid, ES-API: [M+H].sup.+=646.2.
Example 49 Preparation of Compounds Z49, Z49-1, and Z49-2
[0550] ##STR00573##
[0551] A compound Z49 was synthesized from 7-chloro-6-fluoro-4-hydroxy-1-(2-isopropyl-4-(trifluoromethyl)pyridin-3-yl)-3-nitro-1,8-naphthyridin-2(1H)-one with reference to the synthesis of the compound Z33. ES-API: [M+H].sup.+=669.2. The compound Z49 was resolved by preparative scale chiral HPLC (column type: IB 10 μm, 30*250 mm; mobile phase: hexane:EtOH=80:20; flow rate: 25 mL/min; and column temperature: room temperature) to obtain the following atropisomer compounds. One of the atropisomer compounds had a structure arbitrarily specified as Z49-1 (71 mg, peak 1, retention time: 7.68 min, de value: 100%). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.14 (d, J=1.4 Hz, 1H), 8.92 (d, J=4.9 Hz, 1H), 8.00 (dd, J=16.4, 8.6 Hz, 1H), 7.76 (d, J=5.0 Hz, 1H), 7.25 (dd, J=15.3, 8.3 Hz, 1H), 6.94 (ddd, J=72.2, 16.7, 10.5 Hz, 1H), 6.71 (d, J=8.3 Hz, 1H), 6.66 (t, J=8.8 Hz, 1H), 6.22-6.09 (m, 1H), 5.79-5.69 (m, 1H), 5.06-4.72 (m, 1H), 4.53 (t, J=45.4 Hz, 1H), 4.02-3.90 (m, 1H), 3.75 (dd, J=14.1, 4.1 Hz, 1H), 3.45 (d, J=11.9 Hz, 1H), 3.34 (s, 3H), 2.93 (dd, J=13.3, 6.6 Hz, 1H), 2.90-2.85 (m, 1H), 1.55 (dd, J=19.2, 6.8 Hz, 3H), 1.14 (d, J=6.7 Hz, 3H), 1.04 (d, J=6.6 Hz, 3H). ES-API:[M+H].sup.+=669.1. The other atropisomer compound had a structure arbitrarily specified as Z49-2 (80 mg, peak 2, retention time: 9.79 min, de value: 99.85%). .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.15 (s, 1H), 8.92 (d, J=5.0 Hz, 1H), 8.01 (dd, J=14.5, 8.5 Hz, 1H), 7.78 (d, J=5.0 Hz, 1H), 7.25 (dd, J=15.2, 8.2 Hz, 1H), 6.94 (ddd, J=82.1, 16.7, 10.7 Hz, 1H), 6.71 (d, J=8.3 Hz, 1H), 6.66 (t, J=8.8 Hz, 1H), 6.15 (dt, J=16.9, 3.9 Hz, 1H), 5.79-5.69 (m, 1H), 5.11-4.69 (m, 1H), 4.54 (t, J=56.0 Hz, 1H), 4.15-3.97 (m, 1H), 3.74 (dd, J=14.2, 4.3 Hz, 1H), 3.44 (d, J=11.6 Hz, 1H), 3.28 (s, 3H), 2.88 (dd, J=40.3, 9.5 Hz, 1H), 2.66-2.56 (m, 1H), 1.54 (dd, J=19.8, 6.6 Hz, 3H), 1.07 (d, J=6.6 Hz, 3H), 0.92 (d, J=6.7 Hz, 3H). ES-API: [M+H].sup.+=669.1. The isomer compounds were detected by analytical scale chiral HPLC (column type: IB 5 μm, 4.6*250 mm; mobile phase: hexane:EtOH=80:20; flow rate: 1 mL/min; and column temperature=30° C.).
Example 50 Preparation of Compounds Z50, Z50-1 and Z50-2
[0552] ##STR00574## ##STR00575## ##STR00576##
[0553] Step 1: 7-bromo-4,6-dichloro-1-(4,6-diisopropylpyrimidin-5-yl)-8-fluoro-3-nitroquinolin-2(1H)-one (450 mg, 0.87 mmol) was dissolved in N,N-dimethylacetamide (6 mL), orderly added with methyl (3R,6R)-1-N-BOC-6-methylpiperazin-3-formate (404 mg, 1.57 mmol) and N,N-diisopropylethylamine (0.45 mL, 2.61 mmol), and stirred at 120° C. for 1 hour to react. The resulting reaction liquid was added with 100 mL of EtOAc, washed with 30 mL of dilute brine for 4 times and then with 30 mL of saturated salt solution, dried and concentrated to obtain product (3R,6R)-1-(tert-butyl)3-methyl-4-(7-bromo-6-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-8-fluoro-3-nitro-2-oxo-1,2-dihydroquinolin-4-yl)-6-methylpiperazin-1,3-dicarboxylate (770 mg, crude), which was directly used in next step. ES-API: [M+H].sup.+=739.2, 741.1.
[0554] Step 2: the (3R,6R)-1-(tert-butyl)3-methyl-4-(7-bromo-6-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-8-fluoro-3-nitro-2-oxo-1,2-dihydroquinolin-4-yl)-6-methylpiperazin-1,3-dicarboxylate (777 mg, crude) was dissolved in acetic acid (8 mL), added with iron powder (170 mg, 3.05 mmol), and stirred at 80° C. for 30 minutes to react. The resulting reaction liquid was concentrated, and orderly added with 100 mL of EtOAc and 60 mL of saturated sodium bicarbonate. The resulting suspension was filtered by diatomite. The filter cake was washed with EtOAc. The resulting organic phase was separated, washed orderly with 40 mL of saturated sodium bicarbonate and 40 mL of saturated salt solution, dried and concentrated to obtain product tert-butyl (2R,4aR)-10-bromo-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-9-fluoro-2-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octa hydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]quinolin-3-carboxylate (700 mg, crude), which was light brown solid. ES-API: [M+H].sup.+=677.2, 679.2.
[0555] Step 3: the tert-butyl (2R,4aR)-10-bromo-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-9-fluoro-2-methyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octa hydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]quinolin-3-carboxylate (700 mg, crude), 20 mL of acetone, anhydrous potassium carbonate (480 mg, 3.48 mmol), and iodomethane (1.24 g, 8.70 mmol) were orderly added to a 50 mL sealing tube. The sealing tube was sealed, and the resulting mixture was stirred at 50° C. for 18 hours to react. The resulting reaction liquid was added with 80 mL of EtOAc, washed orderly with 20 mL of water and 30 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-25%) to obtain product tert-butyl (2R,4aR)-10-bromo-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-9-fluoro-2,6-dimethyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]quinolin-3-carboxylate (350 mg, Y in step 3: 58.2%), which was yellow solid. ES-API: [M+H].sup.+=691.1, 693.2.
[0556] Step 4: the tert-butyl (2R,4aR)-10-bromo-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-9-fluoro-2,6-dimethyl-5,7-dioxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]quinolin-3-carboxylate (325 mg, 0.47 mmol), (2-fluoro-6-hydroxyphenyl)boric acid (293 mg, 1.88 mmol), SPhos-Pd-G2 (34 mg, 0.047 mmol), 2-dicyclohexylphosphino-2′,6′-dimethoxybiphenyl (19 mg, 0.047 mmol), potassium phosphate (398 mg, 1.88 mmol), 3 mL of water, and 15 mL of dioxane were added to a 100 mL round-bottom flask. The resulting mixture was stirred at 90° C. for 4 hours to react under the protection of nitrogen. The resulting reaction liquid was concentrated, added with 60 mL of EtOAc, washed orderly with 10 mL of water and 15 mL of saturated salt solution, dried and concentrated. The resulting crude product was purified by flash column chromatography on silica gel (EtOAc/PE: 0-50%) to obtain product tert-butyl (2R,4aR)-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-9-fluoro-10-(2-fluoro-6-hydroxyphenyl)-2,6-dimethyl-5,7-di oxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]quinolin-3-carboxylate (220 mg, Y: 64.8%), which was yellow solid. ES-API: [M+H].sup.+=723.3.
[0557] Step 5: the tert-butyl (2R,4aR)-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-9-fluoro-10-(2-fluoro-6-hydroxyphenyl)-2,6-dimethyl-5,7-di oxo-1,2,4,4a,5,6,7,8-octahydro-3H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]quinolin-3-carboxylate (220 mg, 0.30 mmol) was dissolved in DCM (4 mL), and added with TFA (1 mL). After stirring at room temperature for 2 hours, the resulting reaction liquid was concentrated to obtain product (2R,4aR)-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-9-fluoro-10-(2-fluoro-6-hydroxyphenyl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]quinolin-5,7-dione (240 mg, crude), which was directly used in next step. ES-API: [M+H].sup.+=623.3.
[0558] Step 6: the (2R,4aR)-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-9-fluoro-10-(2-fluoro-6-hydroxyphenyl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]quinolin-5,7-dione (240 mg, crude) was dissolved in DCM (6 mL), and added with N,N-diisopropylethylamine (194 mg, 1.50 mmol). The resulting reaction liquid was cooled to 0° C., and added dropwise with acryloyl chloride (24 mg, 0.27 mmol). The resulting mixture was stirred at 0° C. for 15 minutes to react. The resulting reaction liquid was added with 50 mL of DCM, washed orderly with 15 mL of water, 15 mL of saturated solution of NaHCO.sub.3 and 15 mL of saturated salt solution, dried and concentrated to obtain product (2R,4aR)-11-chloro-8-(4,6-diisopropylpyrimidin-5-yl)-9-fluoro-10-(2-fluoro-6-hydroxyphenyl)-2,6-dimethyl-2,3,4,4a,6,8-hexahydro-1H-pyrazino[1′,2′:4,5]pyrazino[2,3-c]quinolin-5,7-dione (Z50), which was purified by preparative scale HPLC to obtain the following atropisomer compounds. One of the atropisomer compounds had a structure arbitrarily specified as Z50-1 (retention time: 10.433 min; 55 mg, Y: 26.7%), which was white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.21 (d, J=1.7 Hz, 1H), 9.13 (s, 1H), 7.89-7.88 (m, 1H), 7.27 (dd, J=15.4, 8.3 Hz, 1H), 7.02 (dd, J=16.8, 10.6 Hz, 1H), 6.75-6.69 (m, 2H), 6.19-6.11 (m, 1H), 5.78-5.70 (m, 1H), 4.84-4.75 (m, 1H), 4.60 (d, J=13.6 Hz, 1H), 4.06-3.97 (m, 1H), 3.78-3.70 (m, 1H), 3.31-3.25 (m, 4H), 2.99-2.90 (m, 1H), 2.81-2.73 (m, 1H), 2.61-2.55 (m 1H), 1.48-1.58 (m, 3H), 1.18-0.86 (m, 12H). ES-API: [M+H].sup.+=677.2. The other atropisomer compound had a structure arbitrarily specified as Z50-2 (retention time: 10.752 min; 85 mg, Y: 41.2%), which was white solid. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.22 (s, 1H), 9.13 (s, 1H), 7.89-7.88 (m, 1H), 7.27 (dd, J=15.5, 8.1 Hz, 1H), 7.02 (dd, J=16.8, 10.7 Hz, 1H), 6.76-6.69 (m, 2H), 6.20-6.09 (m, 1H), 5.80-5.70 (m, 1H), 3.83-3.73 (m, 1H), 4.60 (d, J=14.0 Hz, 1H), 4.06-3.97 (m, 1H), 3.82-3.66 (m, 1H), 3.31-3.22 (m, 4H), 3.02-2.88 (m, 1H), 2.83-2.74 (m, 1H), 2.61-2.52 (m, 1H), 1.66-1.49 (m, 3H), 1.21-1.01 (m, 9H), 0.99-0.83 (m, 3H). ES-API: [M+H].sup.+=677.2. The isomer compounds were detected by analytical scale HPLC.
[0559] Compounds of Example 51 to Example 342 were prepared with reference to the synthesis processes of the above Examples.
TABLE-US-00004 ES- Ex- API: ample [M + No. Structure and No. of Compound H].sup.+ 51
Test Example 1 Cell Proliferation Inhibition Experiment
[0560] NCI-H358 was a human NSCLC cell line with Kras G12C mutation, which was cultured in a RPMI-1640 medium with 10% fetal bovine serum (FBS). A549 was a human lung adenocarcinoma cell line with Kras G12S mutation, which was cultured in an F-12K medium with 10% FBS. Cells in logarithmic growth phase were detached with trypsin and EDTA, collected and counted, and H358 was regulated to 1.8E4 cells/ml by using an RPMI-1640 medium with 2% FBS, while A549 was regulated to 8.9E3 cells/ml with an F-12K medium containing 2% FBS. 800 (45 μl) H358 cells and 400 (45 μl) A549 cells were seeded to 384-well spheroid plates, respectively, and cultured overnight to establish 3D cell models. 1000× stock solutions of compounds at a concentration gradient of 3.16 were prepared by using DMSO, and diluted 100 times by the medium with 2% FBS into 10× stock solutions of compounds. On the day after cell seeding, 5 μl of 10× stock solution of a compound was added to each well of cell culture plate, with a final concentration being 1× and a DMSO content of 0.1%. DMSO was used as the control, and the medium with 200 FBS was used as blank control group. After 5 days of cell culture with the compound, 25 μl of CellTiter-Glo working solution was added to each well, and mixed uniformly at 400 rpm and incubation for 30 minutes. After standing at room temperature for 30 minutes, 40 μl of mixed solution was transferred to a 384-well plate with white clear bottom. A value of luminescence was then read, and a cell proliferation inhibition rate (IR) (%)=(RLU control−RLU compound)/(RLU control−RLU blank)×100%. The value of IC.sub.50 was calculated by fitting a gradiently diluted concentration of the compound and the corresponding cell proliferation inhibition rate using a Prism 6 four-parameter method. Results showed that the example compounds of the present invention had high inhibitory activity for NCI-H358 cells with Kras G12C mutation, with their IC.sub.50 values below 1000 nM, or below 500 nM, or below 100 nM, and had low inhibitory activity for A549 cells, with their IC.sub.50 values above 5000 nM. Results of the example compounds were as shown in Table 1 below.
TABLE-US-00005 TABLE 1 Inhibitory Activity of Compounds For H358 and A549 Cells H358 A549 H358 A549 Compound IC.sub.50 IC.sub.50 Compound IC.sub.50 IC.sub.50 No. (μM) (μM) No. (μM) (μM) Z1 0.031 >10 Z31 0.017 9.494 Z2 0.773 >10 Z32 0.064 >30 Z1-1 0.031 10.716 Z33-1 0.074 >30 Z1-2 0.198 16.482 Z33-2 0.034 >30 Z3a 0.017 >10 Z34-1 0.078 >30 Z3 0.016 >10 Z34-2 0.007 >30 Z6 0.035 11.312 Z35 0.006 28.208 Z9 0.032 — Z35-1 0.037 >30 Z9-1 0.218 >10 Z35-2 0.006 >30 Z9-2 0.015 >10 Z36-1 0.588 >30 Z10 0.008 12.125 Z36-2 0.018 28.025 Z10-1 0.070 >10 Z37 0.001 8.572 Z10-2 0.004 24.640 Z37-1 0.0007 >30 Z21 0.023 >30 Z37-2 0.165 3.322 Z21-1 0.027 >30 Z38-1 0.007 >30 Z21-2 0.407 >30 Z38-2 0.513 >30 Z22 0.067 >30 Z39-1 0.0006 >30 Z23 0.002 10.482 Z39-2 0.316 7.781 Z24 0.003 28.067 Z40 0.011 >30 Z24-1 0.028 >30 Z41 0.017 >30 Z24-2 0.003 11.308 Z42 0.053 >30 Z25 0.004 28.332 Z43 0.049 29.234 Z25-1 0.050 >30 Z44a 0.024 9.237 Z25-2 0.003 >30 Z44 0.011 >30 Z26 0.002 12.856 Z45 0.013 >30 Z26-1 0.042 >30 Z46 0.012 10.306 Z26-2 0.003 >30 Z47 0.031 >30 Z27 0.002 10.123 Z48 0.003 >10 Z27-1 0.012 12.705 Z49-1 0.009 9.417 Z27-2 0.001 18.995 Z49-2 0.044 >30 Z28 0.090 16.881 Z50 0.0005 28.289 Z29 0.166 21.794 Z50-1 0.0003 >30 Z30 0.051 >30 Z50-2 0.320 3.872 Z30-1 0.804 >30 Z48′-1 0.828 — Z30-2 0.047 >30 Z48′-2 >1 —
[0561] From Table 1, it could be seen that the example compounds of the present invention had high inhibitory activity against NCI-H358 cells with Kras G12C mutation and low inhibitory activity for A549 cells, and thus had obvious selective inhibitory activity.
Test Example 2 Phospho-Extracellular Signal-Regulated Kinase (p-ERK) Assay for Cells
[0562] MIA PaCa2 was a human pancreatic carcinoma cell line with Kras G12C mutation, which was cultured in a DMEM with 10% FBS and 2.5% Horse serum. Cells in logarithmic growth phase were detached with trypsin and EDTA, collected and counted, and 2.5E4 cells were seeded to a 96-well cell culture plate and cultured overnight. 1000× stock solutions of compounds at a concentration gradient of 3.16 were prepared by using DMSO, and diluted 200 times by the medium into 5× stock solutions of compounds. On the day after cell seeding, 5× stock solution of a compound was added to each well of cell culture plate, with a final concentration being 1× and a DMSO content of 0.1%. DMSO was used as the control. The residual culture was removed after culturing for 2 hours with the compound. To each well, 50 ul of cell lysis buffer was added and mixed uniformly and incubation for 30 minutes. Subsequently, 16 ul of mixed solution was transferred to a 96-well plate with white unclear bottom, and 16 ul of cell lysis buffer was added to the blank control group. After the completion of transfer, 4 ul of p-ERK HTRF antibody mixture was added to each well, and a value of fluorescence intensity was read after incubation for 4 hours. The inhibition rate (IR) of the compound was calculated by the following equation: IR (%)=(RLU control-RLU compound)/(RLU control-RLU blank)×100%. The value of IC50 was calculated by fitting a gradiently diluted concentration of the compound and the corresponding cell proliferation inhibition rate using a Prism 8 four-parameter method. Results showed that the example compounds of the present invention had good inhibitory activity for the level of phosphorylated ERK downstream of the cell passage of the Kras G12C protein mutation, with their IC50 values below 10 M, or below 1000 nM, or below 100 nM. Results of the example compounds were as shown in Table 2 below.
TABLE-US-00006 TABLE 2 Inhibitory Activity of Compounds For p-ERK p-ERK p-ERK Compound No. IC50 (μM) Compound No. IC50 (μM) Z1 0.432 Z34-1 0.578 Z1-1 0.512 Z34-2 0.051 Z3a 0.273 Z35 0.083 Z3 0.160 Z35-1 0.249 Z6 0.313 Z35-2 0.049 Z9 0.187 Z36-2 0.188 Z9-2 0.221 Z37 0.021 Z10 0.075 Z37-1 0.011 Z10-1 0.460 Z38-1 0.212 Z10-2 0.034 Z39-1 0.011 Z21 0.271 Z40 0.078 Z21-1 0.428 Z41 0.279 Z23 0.017 Z42 0.566 Z24 0.113 Z43 0.439 Z24-1 0.536 Z44a 0.170 Z24-2 0.061 Z44 0.088 Z25 0.051 Z45 0.122 Z25-2 0.029 Z46 0.224 Z26 0.039 Z48 0.052 Z26-2 0.062 Z49-1 0.114 Z27 0.029 Z50 0.007 Z27-1 0.157 Z50-1 0.006 Z27-2 0.011 Z50-2 0.316 Z30 0.676 Z72 0.106 Z30-2 0.488 Z48′-1 7.477 Z31 0.206 Z48′-2 >10 Z33-2 0.238
Test Example 3 Cell Proliferation Inhibition Experiment
[0563] MIA PaCa-2 was a human pancreatic carcinoma cell line with Kras G12C mutation, which was cultured in a DMEM with 10% FBS and 2.5% Horse serum. A549 was a human lung adenocarcinoma cell line with Kras G12S mutation, which was cultured in an F-12K medium with 10% FBS. Cells in logarithmic growth phase were detached with trypsin and EDTA, collected and counted, and 200 MIA PaCa-2 cells and 400 A549 cells were seeded to 384-well spheroid plates, respectively, and cultured overnight to establish 3D cell models. 1000× stock solutions of compounds at a concentration gradient of 3.16 were prepared by using DMSO, and diluted 100 times by the medium into 10× stock solutions of compounds. On the day after cell seeding, 10× stock solution of a compound was added to each well of cell culture plate, with a final concentration being 1× and a DMSO content of 0.1%. DMSO was used as the control, and the medium was used as the blank. After 5 days of cell culture with the compound, 30 μl of CellTiter-Glo working solution was added to each well, and mixed uniformly and incubation for 30 minutes. After standing at room temperature for 30 minutes, 40 μl of mixed solution was transferred to a 384-well plate with white unclear bottom. A value of luminescence intensity was then read, and a cell proliferation inhibition rate (IR) (%)=(RLU control−RLU compound)/(RLU control−RLU blank)×100%. The value of IC.sub.50 was calculated by fitting a gradiently diluted concentration of the compound and the corresponding cell proliferation inhibition rate using an XLFit four-parameter method. Results showed that the example compounds of the present invention had high inhibitory activity for MIA PaCa-2 cells with Kras G12C mutation, with their IC.sub.50 values below 1000 nM, or below 100 nM, or below 10 nM. Results of the example compounds were as shown in Table 3 below.
TABLE-US-00007 TABLE 3 Inhibitory Activity of Compounds For MIA-PaCa2 Compound MIA-PaCa2 Compound MIA-PaCa2 No. IC50 (μM) No. IC50 (μM) Z1 0.090 Z33-1 0.092 Z1-1 0.095 Z33-2 0.048 Z3a 0.070 Z34-1 0.135 Z3 0.018 Z34-2 0.010 Z6 0.085 Z35 0.015 Z9 0.074 Z36-2 0.133 Z9-2 0.017 Z37 0.004 Z10-2 0.008 Z37-1 0.002 Z21 0.047 Z38-1 0.027 Z21-1 0.046 Z39-1 0.001 Z22 0.180 Z40 0.012 Z23 0.002 Z41 0.044 Z24 0.009 Z42 0.128 Z24-2 0.004 Z43 0.220 Z25 0.008 Z44a 0.103 Z25-2 0.005 Z44 0.034 Z26 0.005 Z45 0.048 Z26-2 0.005 Z46 0.068 Z27 0.004 Z47 0.083 Z27-2 0.001 Z48 0.006 Z28 0.234 Z49-1 0.012 Z30 0.140 Z50 0.002 Z30-2 0.130 Z50-1 0.001 Z31 0.059 Z48′-1 >1 Z32 0.170 Z48′-2 >1
Test Example 4 Nucleotide Exchange Assay (NEA)-HTRF Assay for KRas G12C
[0564] The effects of compounds on SOS1 catalyzed displacement of GDP by GTP on KRas proteins was examined by homogeneous time-resolved fluorescence (HTRF). 30 μM 6×his labeled KRas G12C recombinant protein and 80 M fluorochrome DY647 labeled GDP were co-incubated in a labeling buffer (1 mM DTT, 7.5 mM EDTA, 25 mM Tris-HCl, 45 mM NaCl) at 20° C. away from light for 2 hours. Protein quantification was performed after purification on the NAP-5 column to determine the concentration of KRas G12C-GDP. 1000× stock solutions of compounds at a concentration gradient of 3.16 were prepared by using DMSO, and diluted 250 times by a reaction buffer (40 mM N-2-hydroxyethylpiperazine-N-2-ethane sulfonic acid (HEPES), 10 mM MgCl.sub.2, 1 mM DTT, 0.002% Triton X-100) into 4× stock solutions of compounds. KRas G12C-GDP/Tb working solution (40 nM KRas G12C-GDP, Ixanti-his Tb) and SOS1/GTP working solution (0.2 μM SOS1, 200 M GTP) were prepared by using the reaction buffer.
[0565] 5 μl of 4× stock solution of a compound and 10 μl of KRas G12C-GDP/T working solution were added to each well of 96-well plate with white unclear bottom, and 5 μl of reaction buffer was added to the well of control group in stead of the 4× stock solution of a compound. After incubation at 20° C. away from light for 15 minutes, 5 μl of SOS1/GTP working solution was added and incubated at 20° C. away from light for 2 hours, and then a value of fluorescence intensity was read (excitation wavelength: 320 nm, and emission wavelengths: 615 nm and 665 nm). Besides, group T0 was established with 10 μl of the reaction buffer and 10 μl of the KRas G12C-GDP/Tb working solution, and the value of fluorescence intensity was directly read. A relative light unit (RLU) was calculated by the following equation: RLU=(665 nm signal/615 nm signal)×104; and the inhibition rate (IR) of the compound was calculated by the following equation: IR (%)=(RLU.sub.compound−RLU.sub.control)/(RLU.sub.T0−RLU.sub.control)×100%. The value of IC.sub.50 was calculated by fitting a gradiently diluted concentration of the compound and the corresponding cell proliferation inhibition rate using a four-parameter method. Results were shown in Table 4.
TABLE-US-00008 TABLE 4 Compound No. NEA IC50 (μM) Z1 0.255 Z1-1 0.152 Z1-2 1.428 Z9-1 2.571 Z9-2 0.115 Z10 0.067 Z10-1 0.708 Z10-2 0.037 Z21 0.388 Z21-1 0.172 Z21-2 3.935 Z24 0.058 Z24-1 0.291 Z24-2 0.038 Z25 0.060 Z25-2 0.025 Z25-1 0.951 Z26 0.034 Z26-1 0.868 Z26-2 0.040 Z27 0.027 Z27-1 0.198 Z27-2 0.009 Z30 0.656 Z31 0.151 Z33-1 0.515 Z33-2 0.162 Z35 0.073 Z37 0.020 Z37-1 0.011 Z37-2 1.898 Z38-1 0.053 Z38-2 2.147 Z39-2 2.245 Z50 0.019 Z50-1 0.008 Z50-2 1.450
Test Example 5 In Vivo Pharmacodynamic Experiment
[0566] Experimental objective: to evaluate the in vivo pharmaceutical effects of tested compounds on the subcutaneous xenograft tumor model of MIA PaCa-2.
[0567] Experimental operation: 6 to 8 weeks old female BALB/c nude mice each having the body weight of 18-20 g were chosen. MIA PaCa-2 cells were cultured in DMEM containing 10% FBS, 2.5% HS and 1% penicillin-streptomycin at 37° C. in an incubator with 5% CO.sub.2. Cells were collected. The MIA PaCa-2 cells were subcutaneously inoculated to the animals at their backs on the right, 2.0×10.sup.6 cells (0.1 mL) for each animal. When tumor grew to 190-311 mm.sup.3, mice with tumor having a suitable size were chosen and treated in groups, with administration dosages shown in Table 5 below. The animals were weighed on an electronic balance each day. The tumor volume was investigated by using a vernier caliper twice a week. The tumor volume was calculated by the following equation: V=0.5 a×b.sup.2, a and b representing the long diameter and the short diameter of the tumor, respectively. The tumor volume was used to calculate a tumor growth inhibition rate (TGI), and the TGI shown in the form of a percentage was used to indicate the antineoplastic activity of a compound. The TGI was calculated by the following equation: TGI (%)=[1-avTi−0/avCi-0)]×100, with avTi-0 representing the average tumor volume of the group administrated with the compound on a specific day minus the average tumor volume of this group on the day of grouping, and avCi-0 representing the average tumor volume of the solvent control group on a specific day minus the average tumor volume of the solvent control group on the day of grouping. The tumor volume was shown in the form of an average value±standard error of mean (SEM). Experimental results were shown in Table 5 below.
TABLE-US-00009 TABLE 5 Tumor Volume on Day 14 after Group Administration (mm.sup.3) TGI(%) Solvent control group 670.99 — AMG 510 (1 mg/kg, p.o., QD) 314.28 82.8 AMG 510 (3 mg/kg, p.o., QD) 205.35 109.1 AMG 510 (10 mg/kg, p.o., QD) 81.72 138.9 Z37-1 (1 mg/kg, p.o., QD) 286.12 89.4 Z37-1 (3 mg/kg, p.o., QD) 238.82 102.8 Z37-1 (10 mg/kg, p.o., QD) 210.59 108.4 Z48 (1 mg/kg, p.o., QD) 459.33 51.2 Z48 (3 mg/kg, p.o., QD) 148.62 124.7 Z48 (10 mg/kg, p.o., QD) 90.39 138.5 Z23 (1 mg/kg, p.o., QD) 194.65 113.2 Z23 (3 mg/kg, p.o., QD) 59.52 143.5 Z23 (10 mg/kg, p.o., QD) 47.32 149.2 Z25-2 (1 mg/kg, p.o., QD) 180.14 116.2 Z25-2 (3 mg/kg, p.o., QD) 62.35 143.7 Z25-2 (10 mg/kg, p.o., QD) 18.94 154.4 Z26-2 (1 mg/kg, p.o., QD) 157.06 121.1 Z26-2 (3 mg/kg, p.o., QD) 67.66 143.8 Z26-2 (10 mg/kg, p.o., QD) 30.49 151.1 Z27-2 (1 mg/kg, p.o., QD) 140.10 126.3 Z27-2 (3 mg/kg, p.o., QD) 54.81 145.1 Z27-2 (10 mg/kg, p.o., QD) 15.16 155.8
[0568] Experimental conclusion: The compounds of the present invention exhibited excellent in vivo pharmaceutical effects for the subcutaneous xenograft tumor model of MIA PaCa-2. On day 14 after administration, the compounds of the present invention had more significant effects on tumor inhibition than the reference compound AMG 510. Besides, some compounds still exhibited significant tumor regression effect when administrated by a dosage (1 mg/kg) lower than that (3 mg/kg) of the reference compound AMG 510. This indicated that some compounds among the compounds of the present invention exhibited superior in vivo pharmaceutical effects to the reference compound AMG 510 in the subcutaneous xenograft tumor model of MIA PaCa-2, and the anti-tumor effects had dose dependency.
[0569] The structure of the reference compound AMG 510 was
##STR01337##
Test Example 6 Pharmacokinetic Evaluation Experiment on Mice
[0570] Experimental objective: male CD-1 mice were chosen as tested animals, and the LC-MS-MS method was employed to measure the concentrations of a tested compound in blood plasma of a mouse intravenously and intragastrically administrated with the tested compound at different times. The pharmacokinetic behaviors of tested compounds in mice were studied, and the pharmacokinetic characteristics thereof were evaluated.
[0571] Experimental scheme: experimental animals: 18 healthy adult male CD-1 mice (6 to 8 weeks old, and body weight about 30 g) were divided into 6 groups according to the principle of similar body weights: 3 mice in each of IV groups (3 groups), and 3 mice in each of PO groups (3 groups). The animals were purchased from Shanghai Jihui Laboratory Animal Care Co., Ltd.
[0572] Formulation: IV group: taking 10 mL of sample solution for example, 4 mg of sample was weighed, orderly added with 0.5 mL of DMSO and 10 mL of Solutol HS 15, then added with 17 g of HP-β-CD, finally added with water to a constant volume of 10 mL, and stirred and subjected to ultrasonic irradiation, thereby obtaining a 0.4 mg/mL clear solution. PO group: an appropriate amount of sample was weighed, orderly added with appropriate amounts of Labrasol and water in a volume ratio of 10:40, and stirred and subjected to ultrasonic irradiation, thereby obtaining a 1.5 mg/mL uniform solution.
[0573] Administration: after being deprived of food for one night, the IV groups were intravenously administrated with the formulated solutions, respectively, with an injection volume of 5 mL/kg and a dosage of 2 mg/kg. The PO groups were intragastrically administrated with the formulated solutions, respectively, with an injection volume of 15 mL/kg and a dosage of 15 mg/kg.
[0574] Experimental operations: after the intravenous injection groups and the intragastric administration groups of the male CD-1 mice were administrated with the tested compounds, 110 ul of blood was sampled at the following time points: 0.0833, 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours. The sampled blood was placed into an anticoagulant tube which was added with K.sub.2EDTA in advance and preserved in dry ice. The tube was centrifuged for 15 minutes to separate blood plasma, and the separated blood plasma was preserved at −70° C. The animals were allowed to be fed after 4 hours of administration. The LC-MS-MS method was employed to measure the concentration of the tested compounds in the blood plasma after the intravenous and intragastric administration to the mice. The linear range of the method was 1-3000 ng/ml. The blood plasma samples were analyzed after protein precipitation with ACN. The experimental results of the IV (2 mg/kg) groups were as shown in Table 6 below, and the experimental results of the PO (15 mg/kg) groups were as shown in Table 7 below.
TABLE-US-00010 TABLE 6 Group AMG510 Z9-2 Z10-2 Z48 Z25-2 Z27-2 Cl (L/h/kg) 4.39 0.14 0.811 1.53 0.499 0.59 V.sub.d (L/kg) 1.15 0.326 1.06 1.66 0.661 0.777 T.sub.1/2 (h) 0.318 1.74 0.941 1.02 1.00 1.08 Notes: Cl represents clearance rate; V.sub.d represents volume of distribution; and T.sub.1/2 represents half-life.
TABLE-US-00011 TABLE 7 Group AMG510 Z1 Z9-2 Z10-2 Z48 Z25-2 Z27-2 C.sub.max (ng/mL) 1221 2603 13177 3960 1250 6643 3660 T.sub.max (h) 1.00 0.25 1.00 1.00 0.5 1.93 1.74 F (%) 36.10 — 103 59.20 38.1 93.5 96.8 Notes: C.sub.max represents maximum compound concentration after oral administration; T.sub.max represents time to reach C.sub.max; and F represents bioavailability.
[0575] Experimental conclusion: in the pharmacokinetic evaluation experiment on mice, the series of compounds of the present invention exhibited lower in vivo clearance rates, higher C.sub.max and better oral bioavailability than the reference compound AMG 510.
Test Example 7 Single Crystal Cultivation
[0576] The compound Z25-2 was subjected to cultivate single crystal. The specific method was as follows: 2 mg of target compound was weighed, and dissolved in isopropanol, and single crystal was obtained by volatilization. Single-crystal X-ray diffraction analysis was conducted by using Bruker D8 Venture instrument. Results were shown in Table 8 below and
TABLE-US-00012 TABLE 8 Empirical formula C32H30C1FN6O4•C3H8O Formula Weight 677.16 Temperature 169.98 K Wavelength 1.34139 Å Crystal system Monoclinic Space group P 1 21 1 Unit cell dimensions a = 12.9184(4) Å a = 90° b = 9.0729(3) Å b = 97.305(2)° c = 14.9611(4) Å g = 90° Volume 1739.32(9) Å.sup.3 Number of formula units, 2 Z, in a unit cell Density (calculated) 1.293 Mg/m.sup.3 Absorption coefficient 0.929 mm−1 Number of electrons, F(000), 712 in a unit cell Crystal size 0.05 × 0.03 × 0.02 mm.sup.3 Theta range for data collection 3.000 to 54.999° Index ranges −15 <= h <= 15, −11 <= k <= 7, −18 <= l <= 18 Reflections collected 18587 Independent reflections 5906 [R(int) = 0.0559] Completeness to theta = 53.594° 99.7% Absorption correction Semi-empirical from equivalents Max. and min. transmissions 0.7508 and 0.6376 Refinement method Full-matrix least-squares on F.sup.2 Data/restraints/parameters 5906/1/441 Goodness-of-fit on F.sup.2 1.048 Final R indices [I > 2 sigma(I)] R1 = 0.0511, wR2 = 0.1082 R indices (all data) R1 = 0.0812, wR2 = 0.1244 Absolute structure parameter 0.064(13) Largest diff peak and hole 0.276 and −0.191 e.Å.sup.−3
Test Example 8. Single Crystal Cultivation
[0577] The compound Z27-2 was subjected to single crystal cultivation. The specific method was as follows: 20 mg of target compound was weighed, and dissolved in the mixed solvent DCM/EtOH (1:2), and single crystal was obtained by volatilization. Single-crystal X-ray diffraction analysis was conducted by using Bruker D8 Venture instrument. Results were shown in Table 9 below and
TABLE-US-00013 TABLE 9 Empirical formula C33H32C1FN6O4 Formula Weight 631.09 Temperature 172.99 K Wavelength 1.34139 Å Crystal system Monoclinic Space group P 1 21 1 Unit cell dimensions a = 12.3421(6) Å a = 90°. b = 9.5931(4) Å b = 98.435(2)°. c = 14.9580(7) Å g = 90°. Volume 1751.86(14) Å.sup.3 Number of formula units, 2 Z, in a unit cell Density (calculated) 1.196 Mg/m.sup.3 Absorption coefficient 0.887 mm.sup.−1 Number F(000) of electrons, 660 F(000), in a unit cell Crystal size 0.1 × 0.06 × 0.05 mm.sup.3 Theta range for data collection 3.149 54.861°. Index ranges −15 <= h <= 15, −11 <= k <= 11, −18 <= l <= 17 Reflections collected 21383 Independent reflections 6556 [R(int) = 0.0396] Completeness to theta = 53.594° 99.2% Absorption correction Semi-empirical from equivalents Max. and min. transmission 0.7508 and 0.5869 Refinement method) Full-matrix least-squares on F.sup.2 Data/restraints/parameters 6556/2/411 Goodness-of-fit on F.sup.2 1.043 Final R indices [I > 2sigma(I)] R1 = 0.0371, wR2 = 0.1029 R indices (all data) R1 = 0.0388, wR2 = 0.1044 Absolute structure parameter 0.039(6) Largest diff. peak and hole 0.276 and −0.311 e.Å.sup.−3
[0578] All documents mentioned in the present invention are cited in the present application for reference as if each document was cited independently for reference. Furthermore, it will be understood that various alterations or modifications can be made to the present disclosure by those skilled in the art after reading the teachings of the present invention, and these equivalent forms also fall within the scope as defined by the appended claims of the present application.