Novel tetraheterocyclic compound

Abstract

The present invention relates to a novel tetraheterocyclic compound, and more specifically, to a novel tetraheterocyclic compound effective as a KRAS protein inhibitor, an isomer, and a pharmaceutical composition for treating cancer comprising the same.

Description

Technical Field

[0001] The present invention relates to a novel tetraheterocyclic compound, and more particularly to a novel tetraheterocyclic compound effective as a KRAS protein inhibitor, an isomer thereof and a pharmaceutical composition for treating cancer comprising the same. Background Art

[0002] RAS genes are responsible for signal transduction within the mitogen activated protein kinase (MAPK) and phosphatidylinositol 3-kinase (PI3K) pathways and are known as tumor genes due to their frequent mutations. The RAS gene family is divided into KRAS, NRAS and HRAS, and these three genes encode four proteins, namely splice variants K-Ras4A and K-Ras4B and N-Ras and H-Ras. K-Ras is the most frequently mutated isoform in Ras-induced cancers (86%), followed by N-Ras (11%) and H-Ras (3%) (Non-Patent Document 1). For example, oncogenic changes in KRAS have been observed in 15.95% of cancers including pancreatic cancer, lung cancer, colon adenocarcinoma, colorectal cancer and rectal adenocarcinomas (Non-Patent Document 2).

[0003] Gain-of-function missense mutations located at most codons 12, 13, and 61 constitutively activate RAS proteins and can be detected in many types of human cancers. 98% of tumor Ras mutations are found in active site amino acid residues G12, G13, and Q61, and these mutations impair intrinsic GTP hydrolysis and GAP-mediated GTP hydrolysis, thereby abnormally activating downstream signal transduction (Non-patent Document 3). K-Ras G12 mutations (89%) dominate human cancers, followed by G13 mutations (9%) and Q61 mutations (1%). Mutation forms of codon 12 include codon 12Gly→Asp (G12D) (36%), codon 12Gly→Val (G12V) (23%), and codon 12Gly→Cys (G12C) (14%), among which G12D is the most common mutation in codon 12 variation. In addition, mutations of codon 13Gly→Asp (G13D) (7%) and codon 61Gln→His (Q61H) (0.6%) were also observed (Non-Patent Document 1).

[0004] The known role of KRAS in malignant tumors and reports of KRAS mutations in various tumor types indicate that KRAS may be an effective target for cancer therapy. The present inventors developed a novel KRAS inhibitor, thereby completing the present invention.

[0005] [Prior art literature]

[0006] [Non-patent literature]

[0007] (Non-patent document 1) Scientific Reports 6 (1): 21949

[0008] (Non-patent document 2) J Cancer Metastasis Treat 2021; 7:26

[0009] (Non-patent document 3) Cancer Biol Ther. 2006 August; 5(8): 928-932. Summary of the invention

[0010] Technical Problems to be Solved by the Invention

[0011] The present invention provides a novel tetraheterocyclic compound, an isomer, a pharmaceutically acceptable salt thereof, a pharmaceutical composition for treating cancer comprising the same, and a preparation method and an intermediate for providing the composition.

[0012] Technical Solution

[0013] To achieve the objective, one aspect of the present invention provides a compound of Chemical Formula 1, or a stereoisomer, a diastereomer, an enantiomer, a atropisomer, a solvate, an isotopic variant, a tautomer, or a pharmaceutically acceptable salt thereof.

[0014] Another aspect of the present invention provides a pharmaceutical composition comprising the compound of Chemical Formula 1, or a stereoisomer, a diastereomer, an enantiomer, an atropisomer, a solvate, an isotopic variant, a tautomer, or a pharmaceutically acceptable salt thereof.

[0015] Another aspect of the present invention provides a method for preparing a compound of Chemical Formula 1, or a stereoisomer, diastereomer, enantiomer, atropisomer, solvate, isotopic variant, tautomer or pharmaceutically acceptable salt thereof, and an intermediate used in the method.

[0016] Beneficial Effects

[0017] The present invention provides a novel tetraheterocyclic compound which is effective as a KRAS protein inhibitor, or a stereoisomer, a diastereomer, an enantiomer, a atropisomer, a solvate, an isotopic variant, a tautomer, or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition comprising the same, and can exhibit KRAS protein inhibitory activity, and thus can be effectively used for treating cancer. DETAILED DESCRIPTION

[0018] Unless otherwise defined, all technical terms used in this specification have the same meaning as those generally understood by those of ordinary skill in the art to which the present invention belongs. In addition, preferred methods or samples are described in this specification, but methods or samples similar or equivalent thereto are also included in the scope of the present invention.

[0019] Tetracyclic heterocyclic compounds

[0020] In one aspect, the present invention provides a compound of the following Chemical Formula 1, or a stereoisomer, a diastereomer, an enantiomer, an atropisomer, a solvate, an isotopic variant, a tautomer, or a pharmaceutically acceptable salt thereof:

[0021] [Chemical formula 1]

[0022]

[0023] In the chemical formula,

[0024] R 1 is independently selected from hydroxy, halogen, C 1-3 Haloalkyl, C 1-3 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, C 1-3 phenyl, pyridyl, naphthyl, indazolyl, benzothiazolyl or benzothienyl, substituted or unsubstituted by one or more substituents selected from alkoxy, amino and cyano;

[0025] R 2 is hydrogen or halogen;

[0026] R 3 For hydrogen, C 1-3 Alkyl, C 3-6 Cycloalkyl, 4- to 10-membered heterocyclic ring or -O-(L) m -A1, where A1 is respectively connected by one or more R 7 Substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, 4- to 10-membered heterocyclic ring, 6- to 10-membered aryl, 5- to 10-membered heteroaryl, or 5- to 10-membered fused heteroaryl;

[0027] L is R 7 Substituted or unsubstituted C 1-3 Alkylene or C 3-8 Cycloalkylene;

[0028] R 7 Each is independently halogen, oxo (=0), =CH2, -OCF3, -OCHF2, amino, cyano, -N(C 1-3 Alkyl)2, -NH(C 1-3 alkyl), substituted or unsubstituted C 1-3 Alkyl, substituted or unsubstituted C 1-3 Haloalkyl, substituted or unsubstituted C 1-3 Alkoxy, substituted or unsubstituted C 3-4 Cycloalkyl or substituted or unsubstituted heterocycle;

[0029] R 4 are independently hydrogen, hydroxyl, halogen, C 1-3 Haloalkyl, or C 1-3 Alkyl, C 1-3 Alkoxy;

[0030] X is O, CH2 or NR 8 ;

[0031] R 8 For hydrogen or R 9 Any C substituted 1 to 3 times 1-6 Alkyl, C 3-6 Cycloalkyl or C 3-6 Heterocycloalkyl;

[0032] R 9 is independently oxygen, hydroxyl, -C 1-4 Alkyl or -OC 1-4 alkyl;

[0033] Y is NH, O, S, SO or SO2;

[0034] Z is CR 6 or N;

[0035] R 5 For hydrogen, C 1-3 Alkyl, cyano or amino;

[0036] R 6 For hydrogen, hydroxyl, halogen, C 1-3 alkyl;

[0037] n1, n2, n3 and m are each an integer from 1 to 3;

[0038] The heterocyclic ring, heteroaryl group and fused heteroaryl group each contain one or more heteroatoms selected from N, S and O.

[0039] In one implementation example,

[0040] R 1 can be independently selected from hydroxyl, halogen, C 1-3 Haloalkyl, C 1-3 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, C 1-3 Phenyl, naphthyl, benzothiazolyl or benzothiophenyl, which may be substituted or unsubstituted by one or more substituents selected from alkoxy, amino and cyano;

[0041] X can be O, CH2 or NH;

[0042] Y can be O.

[0043] In one implementation example,

[0044] R 3 Can be -O-(L) m -A1, wherein A1 can be respectively represented by one or more R 7 A substituted or unsubstituted 4- to 10-membered heterocyclic ring or a 5- to 10-membered condensed heteroaryl group.

[0045] In one implementation example,

[0046] R 1 are independently selected from hydroxy, halogen, C 1-3 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 One or more substituents among the cycloalkyl group, amino group and cyano group may be arbitrarily substituted or unsubstituted.

[0047] The compound according to claim 1,

[0048] R 1 is independently selected from hydroxy, halogen, C 1-3 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, amino and cyano groups may be substituted or unsubstituted with one or more substituents

[0049]

[0050] R 3 O-(L) m -A1, where A1 is respectively connected by one or more R 7A substituted or unsubstituted 4- to 10-membered heterocyclic ring or a 5- to 10-membered fused heteroaryl group;

[0051] L is C 1-3 Alkylene or C 3-8 Cycloalkylene;

[0052] R 7 are independently halogen, oxo (=0), =CH2, -OCF3, -OCHF2, amino, cyano, C 1-3 Alkyl, C 1-3 Haloalkyl or C 1-3 Alkoxy;

[0053] R 4 is hydrogen;

[0054] X is O, CH2 or NH;

[0055] Z is CR 6 or N;

[0056] R 5 is hydrogen or C 1-3 alkyl;

[0057] R 6 is a halogen;

[0058] n1, n2, n3 and m are integers from 1 to 3, respectively.

[0059] In one implementation example,

[0060] When Z is N,

[0061] R 1 can be independently selected from hydroxyl, halogen, C 1-3 Haloalkyl, C 1-3 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, C 1-3 Phenyl or naphthyl which may be substituted or unsubstituted by one or more substituents selected from alkoxy, amino and cyano;

[0062] In Z is CR 6 hour,

[0063] R 1 can be independently selected from hydroxyl, halogen, C 1-3 Haloalkyl, C 1-3 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, C 1-3 One or more substituents selected from the group consisting of alkoxy, amino and cyano may be substituted or unsubstituted benzothiazolyl or benzothienyl.

[0064] In one implementation example,

[0065] R 3 Can be -O-(L) m -A2, where A2 can be They are each independently selected from halogen, C 1-3 Alkyl, C 1-3 Alkoxy and =CH2 are substituted or unsubstituted with one or more substituents;

[0066] -(L) m - can be methylene or

[0067] In one implementation example,

[0068] R 1 can be independently selected from hydroxyl, halogen, C 1-3 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, amino and cyano groups may be substituted or unsubstituted with one or more substituents

[0069]

[0070] R 2 It can be halogen;

[0071] R 3 Can be -O-(L) m -A1, wherein A1 is independently selected from halogen, C 1-3 Alkyl, C 1-3 Alkoxy and =CH2 substituted or unsubstituted with one or more substituents

[0072] Among them, -(L) m - can be methylene or

[0073] R 4 It can be hydrogen;

[0074] X can be O, CH2 or NH;

[0075] Y can be O;

[0076] Z can be CH, C(C 1-3 Alkyl) or N;

[0077] R 5 It can be hydrogen or C 1-3 alkyl.

[0078] In one implementation example, the compound of Chemical Formula 1 may be selected from any one of the following compounds, or its stereoisomers, diastereomers, enantiomers, atropisomers, solvates, isotopic variants, tautomers, or pharmaceutically acceptable salts:

[0079]

[0080]

[0081] On the other hand, the present invention provides a pharmaceutical composition for treating cancer, which comprises as an active ingredient any of the above-mentioned compounds, or its stereoisomers, diastereomers, enantiomers, atropisomers, solvates thereof, isotopic variants thereof, tautomers thereof, or pharmaceutically acceptable salts thereof.

[0082] In one embodiment, the composition can exhibit KRAS protein inhibitory activity.

[0083] definition

[0084] Unless otherwise mentioned, the term "halogen" used in the present specification means fluorine, chlorine, bromine or iodine.

[0085] Unless otherwise mentioned, the term "alkyl group" used in the present specification refers to a linear or branched saturated monovalent hydrocarbon group.

[0086] The term "alkylene" used in this specification refers to a group having (-CH2-) n The divalent straight or branched hydrocarbon group includes, but is not limited to, methylene, ethylene, propylene, butylene and isobutylene.

[0087] Unless otherwise mentioned, the term "alkenyl" used in the present invention refers to a monovalent hydrocarbon group containing at least one carbon-carbon double bond, and each double bond may have an E- or Z-shaped stereoconfiguration.

[0088] As used herein, unless otherwise noted, the term "alkynyl" refers to a monovalent hydrocarbon radical containing at least one carbon-carbon triple bond.

[0089] Unless otherwise mentioned, the term "alkoxy" as used in this specification refers to a straight or branched chain hydrocarbon residue attached through an oxygen.

[0090] Unless otherwise mentioned, the term "aryl" used in this specification refers to a substituted or unsubstituted aromatic group, including a monocyclic or bicyclic, or more than bicyclic substituted or unsubstituted aromatic group, and includes unsaturated or partially saturated aromatic groups, for example, may include C 6-15The aryl group may include, for example, but is not limited to, phenyl, biphenyl, naphthyl, toluoyl or naphthyl.

[0091] Unless otherwise mentioned, the term "heteroaryl" used in this specification refers to a monocyclic or bicyclic, or bicyclic or more substituted or unsubstituted aromatic group containing one or more heteroatoms selected from N, O and S, and includes unsaturated or partially saturated heteroaryl groups, for example, C 4-15 Heteroaryl groups include, but are not limited to, morpholinyl, piperidinyl, pyrrolidinyl, or pyrrolazinyl.

[0092] The term "fused heteroaryl" used in this specification refers to an unsaturated or partially saturated substituted or unsubstituted ring system in which the heteroaryl is connected to another aryl, heteroaryl or heterocycloalkyl in a fused manner. For example, it may include C 8-20 Heteroaryl, for example, can be a 9-, 10-, 11-, 12-, 13-, 14- or 15-membered benzo-fused heteroaryl. For example, the fused heteroaryl can form a 5+5-, 5+6-, 5+7-, 6+6- or 6+7-membered fused ring system. In addition, for example, the fused heteroaryl includes but is not limited to pyrrolizine, benzothiazole, benzothiazolinyl, benzothienyl, benzofuranyl, isobenzofuranyl, benzothionyl, indolyl, isoindolyl, indazolinyl, benzimidazolinyl, benzoxazolinyl, benzisoxazolinyl, benzothiadiazolyl, benzoxadiazolinyl, benzotriazolinyl, quinolyl, isoquinolyl or quinazolinyl, etc.

[0093] The term "partially saturated" as used in this specification refers to the above-defined aryl, heteroaryl or fused heteroaryl rings including at least one saturated site, i.e., at least one single bond. In this specification, the term "unsaturated" refers to the above-defined aryl, heteroaryl or fused heteroaryl rings not containing saturated sites, i.e., single bonds.

[0094] Unless otherwise mentioned, the term "cycloalkyl" used in this specification refers to a substituted or unsubstituted, saturated or partially unsaturated monocyclic, bicyclic or polycyclic hydrocarbon ring, including bridged cycloalkyl, fused cycloalkyl and spirocycloalkyl. For example, it may include C 3-10 Cycloalkyl or C 3-6 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or cycloheptyl.

[0095] The term "cycloalkylene" used in the present specification refers to a divalent radical derived from a cycloalkene.

[0096] Unless otherwise mentioned, the term "heterocycle" or "heterocycloalkyl" used in this specification refers to a substituted or unsubstituted, saturated or partially unsaturated, monocyclic or bicyclic, or polycyclic non-aromatic ring containing one or more heteroatoms selected from N, O and S, including bridged heterocycles, fused heterocycles and spiro heterocycles. The term may include monocyclic or polycyclic cyclic alkyl groups, for example, C 4-15 Heterocycloalkyl, for example, includes but is not limited to piperidinyl, piperazinyl, pyrrolidinyl, morpholinyl, thiomorpholinyl, tetrahydrofuranyl (tetrahydrofuranyl), tetrahydropyranyl (tetrahydro-2H-pyranyl), imidazolidinyl, pyrrolidin-2-one (pyrrolidin-2-one), pyrrolidinyl or pyrrolyl etc.Heterocycle can be a carbon linking group or a heteroatom linking group.For example, the heterocycle connected to the basic molecule by the nitrogen in the ring atom of the heterocycle can be N-morpholinyl, N-piperidinyl, N-pyrrolidinyl or N-pyrrolyl, but is not limited thereto.

[0097] Unless otherwise mentioned, the term "fused heterocycle" or "fused cycloalkyl" used in this specification refers to a substituted or unsubstituted ring system. Depending on the number of rings, bicyclic, tricyclic, tetracyclic or more polycyclic fused cycloalkyl groups can be distinguished. A fused cycloalkyl group refers to a polycyclic cycloalkyl group in which each ring shares adjacent pairs of carbon atoms with another ring, more than one ring may share more than one double bond, but none of these rings have a completely conjugated π-electron system, for example, it may include C 3-20 Fused cycloalkyl. For example, a bicyclic fused cycloalkyl is also referred to as a "bicycloalkyl", and may be a 5+6 or 6+6 bicycloalkyl, depending on the number of atoms forming the two fused rings.

[0098] Unless otherwise mentioned, the term "fused heterocycloalkyl" used in this specification refers to a substituted or unsubstituted fused cycloalkyl group containing one or more heteroatoms selected from N, O and S, for example, C 8-20 Fused heterocycloalkyl. For example, a bicyclic fused heterocycloalkyl is also referred to as a "heterobicycloalkyl", and may be a 5+6 fused heterobicycloalkyl or a 6+6 fused heterobicycloalkyl, depending on the number of atoms forming the two fused rings. For example, it may be hexahydro-1H-pyrrolizine, but is not limited thereto.

[0099] The term "stereoisomer" used in this specification may refer to compounds of the present invention or their salts that have the same chemical formula or molecular formula but are optically or stereologically different. In this specification, the term "enantiomer" refers to various stereoisomers and geometric isomers that may exist for the compounds of the present invention.

[0100] The compounds described in the present invention may have asymmetric centers, geometric centers (e.g., double bonds), or both. Unless a specific stereochemistry or isomeric form is specifically indicated, all chiral, diastereoisomer, racemic forms, and all geometric isomeric forms of the structure are intended.

[0101] The compound of Chemical Formula 1 of one aspect of the present invention may have an asymmetric carbon center (asymmetric carbon) and thus can exist in the form of enantiomers (R or S isomers), racemates, diastereomers or any mixture thereof, and all these isomers and mixtures are included in the scope of the present invention.

[0102] The compounds described in the present invention may have chiral centers, and unless otherwise mentioned, it is understood that each chiral center is independently R- or S-configuration, or the compounds of the present invention may be a mixture of two configurations. Therefore, the compounds described in this specification are included in any or all asymmetric atoms enriched (enriched) or separated (resolved) optical isomers. Not only the racemic mixture of R- and S-enantiomers, but also R- and S-enantiomers, separated or synthesized enantiomer-stereoisomer enriched mixtures containing individual optical isomers (substantially free of their enantiomers or diastereoisomer partners) are included in the scope of the present technology.

[0103] The compounds of the present invention containing asymmetrically substituted atoms can be isolated in optically active or racemic forms. Methods for preparing optically active forms (such as resolution of racemic forms, synthesis from optically active starting materials or use of chiral auxiliaries) are known in the art.

[0104] The term "atropisomer" used in this specification refers to all stereoisomers that can be separated from each other. It is an isomer that occurs because a single bond formed between carbon atoms and carbon atoms cannot rotate freely due to bulky substituents in the bonding constituting the compound. It refers to stereoisomers generated by an asymmetric axis, including completely separated stable diastereomers or enantiomeric species that do not convert to each other, which can be generated by restricted rotation around a single bond with a rotation barrier that is high enough to allow differentiation of isomeric species.

[0105] The compound of one embodiment has a high probability of producing atropisomers.

[0106] Compounds of one embodiment may also include "tautomeric" forms. Tautomeric forms result from the exchange of a single bond with an adjacent double bond and the concomitant transfer of a proton. Tautomeric forms include proton tautomers, which are isomeric protonation states having the same empirical formula and total charge. Tautomeric forms may be in equilibrium or may be stereofixed into one form by appropriate substitution.

[0107] Compounds of one embodiment may contain "isotopic variants". The present invention also includes isotopically labeled compounds identical to the compounds described herein, except that one or more atoms are replaced by atoms having an atomic mass or mass number different from the atomic mass or mass number normally found in nature ("isotopes"). Compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more atoms constituting such compounds. Examples of isotopes that may be incorporated into the compounds described herein include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, such as 2 H("D"), 3 H. 13 C. 14 C. 15 N. 18 O. 17 O. 31 P. 32 P. 35 S. 18 F and 36 Cl. For example, the compounds described in the present invention may have one or more H atoms replaced by deuterium. Generally, reference or description to a particular element (hydrogen or H) means all isotopes of the element. For example, when the definition R group comprises hydrogen or H, deuterium and tritium are included. Deuterated starting materials are easily available and, in order to provide the synthesis of deuterated compounds, can be applied to the synthesis methods described in the present invention. A large number of deuterated reagents and molecular building blocks can be purchased commercially from chemical suppliers (such as Aldrich Chemical Co.).

[0108] The term "solvate" used in this specification may include the compound of Chemical Formula 1 and one or more pharmaceutically acceptable solvent molecules (eg, a molecular complex containing ethanol or water). The complex in which the solvent molecule is water is also called a "hydrate".

[0109] The term "pharmaceutically acceptable salt" used in this specification includes any salt that has low toxicity to humans and does not adversely affect the biological activity and physicochemical properties of the parent compound.

[0110] KRAS protein inhibitors

[0111] Kirsten Rat Sarcoma Virus Oncogene Homolog (KRAS) is a GTPase that integrates extracellular signals into intracellular proliferation and survival signals. It is a member of the small GTPase family of Ras, Rho, Rab, Arf, and Ran. It receives signals from multiple receptor tyrosine kinases, especially the upstream EGFR, and transmits the signals mainly to the downstream Raf and PI3K through the KRAS GTPase cycle, thereby regulating various processes including cell proliferation.

[0112] In the GTPase cycle, it is able to bind to effector proteins in the GTP-bound activated state through the action of guanine nucleotide exchange factors (GEF), and to assume the GDP-bound inactivated state through the interaction with GTPase activating protein (GAP).

[0113] In addition to the KRAS GTP / GDP binding site, Swich I / II protein structural changes associated with GTP / GDP binding were also reported, demonstrating inhibition of the GTPase cycle.

[0114] KRAS has been reported to be involved in cancer, and various attempts have been made to inhibit KRAS activity, but its potential as a direct drug target seems low. However, with the approval of Sotorasib and Adagrasib for the treatment of non-small cell lung cancer, its potential for drug development has been confirmed.

[0115] The compound of the present invention is a compound that inhibits KRAS, and relates to a novel KRAS inhibitor compound, which includes but is not limited to any mutation in KRAS codons 12, 13 and 61, such as at least one of KRAS G12D, KRASG12V, KRAS G12C, KRAS G13D and KRAS Q61H mutations.

[0116] Pharmaceutical composition

[0117] On the other hand, the present invention provides a pharmaceutical composition for treating cancer, which comprises, as an active ingredient, a compound of Chemical Formula 1, or a stereoisomer, a diastereomer, an enantiomer, a transisomer, a solvate, an isotopic variant, a tautomer, or a pharmaceutically acceptable salt thereof.

[0118] In one implementation, the composition may include a therapeutically effective amount of the compound of Formula 1, or a stereoisomer, diastereomer, enantiomer, atropisomer, solvate, isotopic variant, tautomer, or pharmaceutically acceptable salt thereof.

[0119] In one implementation, in addition to the compound of Formula 1, or its stereoisomers, diastereomers, enantiomers, atropisomers, solvates, isotopic variants, tautomers, or pharmaceutically acceptable salts, the composition may further contain other therapeutic drugs.

[0120] In one embodiment, the composition may further include a pharmaceutically acceptable carrier or excipient.

[0121] In one implementation example, the composition may contain the compound of Chemical Formula 1, or any one of its stereoisomers, diastereomers, enantiomers, atropisomers, solvates thereof, isotopic variants thereof, tautomers thereof, or pharmaceutically acceptable salts thereof, or other therapeutic drugs in a content range of 0.005% to 100%, and a pharmaceutically acceptable carrier or excipient is included as the remaining range.

[0122] In one implementation example, the compound of Chemical Formula 1, or its stereoisomer, diastereomer, enantiomer, atropisomer, solvate, isotopic variant, tautomer, or pharmaceutically acceptable salt, or a pharmaceutical composition comprising the same can be used for the treatment of cancer.

[0123] In one implementation example, the compound of Chemical Formula 1, or its stereoisomer, diastereomer, enantiomer, atropisomer, solvate, isotopic variant, tautomer, or pharmaceutically acceptable salt, or a pharmaceutical composition comprising the same may exhibit KRAS protein inhibitory activity.

[0124] Route of administration and dosage form

[0125] The compounds and pharmaceutical compositions of the present invention can be provided by any suitable administration route and dosage form acceptable in the pharmaceutical field.

[0126] Preparation method

[0127] The compounds of the present invention can be synthesized using organic synthesis techniques known to those of ordinary skill in the relevant technical field. Some compounds and / or intermediates of the present invention are commercially available, or are known based on literature, or can be prepared by those skilled in the art by selecting appropriate synthesis methods in known organic synthesis techniques. Some compounds of the present invention can be synthesized by using the reaction formulas, embodiments or intermediates recorded in this specification. Those of ordinary skill in the art can recognize from the synthesis methods recorded in this specification that the reaction time, reagent equivalents and / or temperature can be changed, and can recognize that different post-treatment and / or purification techniques can be used.

[0128] The structures of the synthesized compounds can be verified by methods known to those of ordinary skill in the art, such as by nuclear magnetic resonance (NMR) spectroscopy and / or mass spectrometry.

[0129] General information

[0130] Unless otherwise mentioned, reagents and solvents were obtained from commercial suppliers and used directly without purification or drying. 1 HNMR was recorded using a Broker 400 MHz and 500 MHz, TMS was used as an internal standard. LCMS analysis was performed in a Waters UPLC equipped with a SQD-2 mass detector (Single quadruple), and HPLC analysis was performed in an ACQUIT UPLC H CLASS (Waters).

[0131] Even if not explicitly stated, the numerical values ​​described in this specification are deemed to include the meaning of “about.” The term “about” used in this specification means within 5%, preferably within 1% to 2% of a specified value or range.

[0132] In the present specification, the numerical range expressed using the term "to" is a range including the numerical values ​​described before and after the term "to" as the lower limit and the upper limit, respectively.

[0133] All references, publications, published patents, and patent applications cited herein in this specification are hereby incorporated by reference in their entirety for all purposes.

[0134] Hereinafter, the present invention will be described in more detail by the following synthesis reaction formulas, embodiments or test examples. However, these are only used to help understand the present invention, and the scope of the present invention is not limited thereto.

[0135] The synthetic reaction formula of Example 1

[0136]

[0137] Example 1: Preparation of 5-ethyl-6-fluoro-4-((S)-1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-6,6a,7,8,9,10-hexahydro-5H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctano[1,2,3-de]naphthalen-2-yl)naphthalen-2-ol

[0138] Step 1) Preparation of Intermediate 2 of Example 1: 2,6-dichloro-3-fluoropyridin-4-amine

[0139] At room temperature, a selective fluorine reagent (60.1 g, 169 mmol) was added to a stirred solution of a mixture of solvent MeOH and water (1:1, 500 mL, 20 vol) containing 2,6-dichloropyridin-4-amine (Intermediate 1 of Example 1 (25.0 g, 154 mmol), and the reaction mixture was heated to 45°C and stirred for 16 hours. The progress of the reaction was monitored by TLC. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to obtain a residue, which was diluted with water and extracted with ethyl acetate. The separated organic layer was dried over anhydrous Na2SO4, filtered and evaporated under reduced pressure to obtain a crude compound. The crude compound was purified by silica gel (100-200 mesh) column chromatography and eluted with petroleum ether (pet ether) containing 2% ethyl acetate to obtain 2,6-dichloro-3-fluoropyridin-4-amine (Intermediate 2 of Example 1) (20.0 g, yield was 72%) as a white solid. 1 H NMR (400MHz, CDCl3) δ4.52 (brs, 2H), 6.62 (d, J = 5.2Hz, 1H). MS (LCMS): 180.96m / z [M+H].

[0140] Step 2) Intermediate 3 of Example 1: tert-(tert-butyloxycarbonyl)(2,6-dichloro-3-fluoropyridin-4-yl)carbamic acid Preparation of Butyl Ester

[0141] At room temperature, DMAP (4-dimethylaminopyridine) (0.06 g, 0.26 mmol) and Boc anhydride (1.10 mL, 5.26 mmol) were added to a stirred solution of THF (10 mL, 10 vol) containing 2,6-dichloro-3-fluoropyridin-4-amine (Intermediate 2 of Example 1) (1.00 g, 2.63 mmol), and the reaction mixture was heated to 60°C and maintained for 6 hours. The progress of the reaction was monitored by TLC. After the reaction was completed, the reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate, and the separated organic layer was dried over anhydrous Na2SO4, filtered and evaporated under reduced pressure to obtain a crude compound. The crude compound was purified by silica gel (100-200 mesh) column chromatography and eluted with petroleum ether containing 1% ethyl acetate to obtain tert-butyl (tert-butyloxycarbonyl) (2,6-dichloro-3-fluoropyridin-4-yl) carbamate (Intermediate 3 of Example 1) (0.50 g, yield 50%). 1 H NMR (400MHz, CDCl3) δ1.47 (s, 18H), 7.15 (d, J = 4.40Hz, 1H).

[0142] Step 3) Intermediate 4 of Example 1: tert-butyl 4-((tert-butoxycarbonyl)amino)-2,6-dichloro-5-fluoronicotinate preparation

[0143] Under the temperature condition of -78 ℃, LDA (1M in THF) (147mL, 147mmol) was added to the stirred solution of THF (tetrahydrofuran) (280mL, 10vol) containing (tert-butyloxycarbonyl) (2,6-dichloro-3-fluoropyridin-4-yl) carbamate (intermediate 3 of Example 1) (28.0g, 73.7mmol) and stirred for 30 minutes, and then, Boc anhydride (14.4g, 66.3mmol) was added. The reaction mixture was stirred for 2 hours under the temperature condition of -78 ℃. The reaction progress was monitored by TLC. After the reaction was completed, the reaction mixture was diluted with water and extracted with ethyl acetate. The separated organic layer was dried over anhydrous Na2SO4, filtered and evaporated under reduced pressure to obtain a crude compound. The crude compound was purified by silica gel (100-200 mesh) column chromatography and eluted with petroleum ether containing 5% ethyl acetate to give tert-butyl 4-((tert-butoxycarbonyl)amino)-2,6-dichloro-5-fluoronicotinate (Intermediate 4 of Example 1) (26.0 g, yield 92%) as an off-white solid. 1 H NMR (400MHz, CDCl3) δ1.50 (s, 9H), 1.62 (s, 9H), 7.21 (brs, 1H). MS (LC-MS): 381.17m / z [M+H].

[0144] Step 4) Preparation of Intermediate 5 of Example 1: 4-amino-2,6-dichloro-5-fluoronicotinic acid

[0145] At a temperature of 0°C, concentrated HCl (130 mL, 5 vol) was added to a stirred solution of 1,4-dioxane (130 mL, 5 vol) containing 4-((tert-butyloxycarbonyl)amino)-2,6-dichloro-5-fluoronicotinic acid tert-butyl ester (Intermediate 4 of Example 1) (26.0 g, 68.4 mmol), and the reaction mixture was heated to room temperature and stirred for 16 hours. The reaction progress was monitored by TLC. After the reaction was completed, the reaction mixture was concentrated under reduced pressure, then washed with DCM and filtered to obtain a crude compound 4-amino-2,6-dichloro-5-fluoronicotinic acid (Intermediate 5 of Example 1) (18.0 g, crude compound) as a white solid. The crude compound was used directly in the next step without further purification. 1 H NMR (400MHz, CDCl3) δ5.71 (brs, 2H), 7.31 (brs, 1H). MS (LC-MS): 224.82m / z [M+H].

[0146] Step 5) Intermediate 6 of Example 1: 5,7-dichloro-8-fluoro-2-mercaptopyrido[4,3-d]pyrimidin-4(3H)-one Preparation A solution of SOCl2 (540 mL, 30 vol) containing 4-amino-2,6-dichloro-5-fluoronicotinic acid (intermediate 5 of Example 1) (18.0 g, 80.3 mmol) was heated to 50 ° C and stirred for 8 hours. The reaction progress was monitored by TLC. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to obtain a residue, which was dissolved in acetone (20 mL), and NH4SCN (1.0 g in 20 mL of acetone) was added dropwise at room temperature and stirred for 1 hour. After completion, the reaction mixture was diluted with water, filtered and washed with water, and dried under high vacuum to obtain a crude compound 5,7-dichloro-8-fluoro-2-thiolpyrido [4,3-d] pyrimidine-4 (3H) -one (intermediate 6 of Example 1) (10.0 g, yield 49%), which was an off-white solid. The compound was used directly in the next step without further purification. 1 H NMR (400MHz, CDCl3) δ13.31 (brs, 1H), 12.91 (s, 1H). MS (LC-MS): 265.94m / z [M+H].

[0147] Step 6) Intermediate 7 of Example 1: 5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidine-4 Preparation of (3H)-ketoneAt room temperature, 0.1 M NaOH (1500 mL, 150 vol) and MeI (iodomethane) (2.50 mL, 41.5 mmol) were added to a stirred solution of 5,7-dichloro-8-fluoro-2-thiolpyrido[4,3-d]pyrimidin-4(3H)-one (Intermediate 6 of Example 1) (10.0 g, 37.7 mmol) in MeOH (1500 mL, 150 vol), and the reaction mixture was stirred at room temperature for 2 hours. The progress of the reaction was monitored by TLC. After the reaction was completed, the reaction mixture was diluted with water (500 mL), acidified to about pH 6 with concentrated HCl and a solid was obtained, which was filtered, washed with water, and then dried under high vacuum to obtain a crude compound 5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (Intermediate 7 of Example 1) (8.0 g, crude compound) as a light brown solid. The crude compound was used directly in the next step without further purification. 1 H NMR (400MHz, CDCl3) δ2.60 (s, 3H), 13.33 (s, 1H). MS (LC-MS): 279.95m / z [M+H].

[0148] Step 7) Intermediate 9 of Example 1: (S)-3-(2-((7-chloro-8-fluoro-2-(methylthio)-4-oxo-3,4-diol Preparation of tert-butyl (4,3-d)pyrido[4,3-d]pyrimidin-5-yl)oxy)ethyl)piperazine-1-carboxylate

[0149] At 0°C, NaH (4.90 g, 129 mmol) was added to a stirred solution of THF (80 mL, 10 vol) containing 5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (intermediate 7 of Example 1) (8.00 g, 28.7 mmol) and stirred for 30 minutes. Then, (S)-3-(2-hydroxyethyl)piperazine-1-carboxylic acid tert-butyl ester (7.90 g, 34.4 mmol) was added at room temperature and the reaction mixture was stirred at room temperature for 2 hours. The progress of the reaction was monitored by TLC. After the reaction was completed, the reaction mixture was diluted with cold ice water and extracted with ethyl acetate, and the separated organic layer was washed with a saline solution, dried over anhydrous Na2SO4, filtered and evaporated under reduced pressure to obtain a crude compound. The crude compound was purified by silica gel (100-200 mesh) column chromatography and eluted with 5% methanol in DCM to give (S)-tert-butyl 3-(2-((7-chloro-8-fluoro-2-(methylthio)-4-oxo-3,4-dihydropyrido[4,3-d]pyrimidin-5-yl)oxy)ethyl)piperazine-1-carboxylate (Intermediate 9 of Example 1) (6.0 g, 44% yield) as a brown solid. 1H NMR (400MHz, DMSO-d6): δ1.42(s,9H),2.06(brs,2H),2.42(s,3H),3.04(d,J=10.1Hz,3H),3.46(brs, 2H), 4.02 (brs, 2H), 4.27 (brs, 1H), 4.52 (d, J = 8.8, 1H), 9.45 (brs, 1H). MS (LC-MS): 474.37m / z [M+H].

[0150] Step 8) Intermediate 10 of Example 1: (S)-2-chloro-1-fluoro-12-(methylthio)-5,6,6a,7,9,10-hexahydro- Preparation of tert-butyl 8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctano[1,2,3-de]naphthalene-8-carboxylate

[0151] DIPEA (N,N-diisopropylethylamine) (32.9 mL, 190 mmol) was added to a stirred solution of (S)-tert-butyl 3-(2-((7-chloro-8-fluoro-2-(methylthio)-4-oxo-3,4-dihydropyrido[4,3-d]pyrimidin-5-yl)oxy)ethyl)piperazine-1-carboxylate (Intermediate 9 of Example 1) (6.00 g, 12.7 mmol) in DCM (60 mL, 10 vol) at 0°C and stirred for 10 minutes, followed by the addition of BOP-Cl (11.6 g, 45.6 mmol) at 0°C. The reaction mixture was heated to room temperature and stirred for 5 hours. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was diluted with water and extracted with DCM, and the separated organic layer was washed with brine solution, dried over anhydrous Na2SO4, and concentrated under reduced pressure to give a crude compound. The crude compound was purified by preparative HPLC (prep-HPLC) to give (S)-2-chloro-1-fluoro-12-(methylthio)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctane[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester (Intermediate 10 of Example 1) (0.80 g, yield 14%) as a brown solid. 1 H NMR (400MHz, DMSO-d6): δ1.42(s,9H),1.97-2.10(m,2H),2.54(s,3H),3.07-3.22(m,2H),3.55(d,J=5.60Hz,1H),3.78(d,J=12.8Hz,1H),3.8 6-3.91 (m, 1H), 3.97 (d, J = 12.8Hz, 1H), 4.22 (t, J = 10.8Hz, 1H), 4.46 (dd, J = 12.8, 4.8Hz, 1H), 4.66-4.76 (m, 1H). MS (LC-MS): 456.37m / z [M+H].

[0152] Step 9) Intermediate 11 of Example 1: (S)-2-chloro-1-fluoro-12-(methylsulfonyl)-5,6,6a,7,9,10- Hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctano[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester preparation

[0153] Potassium peroxymonosulfonate (Oxone) (0.65 g, 1.05 mmol) was added to a stirred solution of a mixture of ACN and water (2:1, 48 mL) containing (S)-2-chloro-1-fluoro-12-(methylthio)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctane[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester (Intermediate 10 of Example 1) (0.40 g, 0.88 mmol) at 0° C., and the reaction mixture was stirred at room temperature for 20 minutes. The progress of the reaction was monitored by TLC. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to obtain a crude compound (S)-2-chloro-1-fluoro-12-(methylsulfonyl)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctane[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester (Intermediate 11 of Example 1) (0.34 g, crude compound) in the form of an off-white solid. MS (LCMS): 488.32 m / z [M+H].

[0154] Step 10) Intermediate 13 of Example 1: (S)-2-chloro-1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrole oxazine-7a(5H)-yl)methoxy)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4, 5] Preparation of tert-butyl cyclooctano[1,2,3-de]naphthalene-8-carboxylate

[0155] At 0°C, NaH (0.031 g, 0.82 mmol) was added to a stirred solution of ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methanol (0.01 g, 0.62 mmol) in THF (2.0 mL, 10 vol) and stirred for 30 minutes. Then, (S)-2-chloro-1-fluoro-12-(methylsulfonyl)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctane[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester (Intermediate 11 of Example 1) (0.20 g, 0.41 mmol) was added at 0°C. The reaction mixture was heated to room temperature and stirred for 4 hours. The reaction progress was monitored by TLC. After the reaction was completed, the reaction mixture was diluted with water and extracted with ethyl acetate. The separated organic layer was washed with brine solution, dried over Na2SO4, filtered and evaporated under reduced pressure to give a crude compound. The crude compound was purified by silica gel (100-200 mesh) column chromatography and eluted with 2% methanol in DCM to give (S)-2-chloro-1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctano[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester (Intermediate 13 of Example 1) (0.02 g, yield 9%) as an off-white solid. MS (LC-MS): 567.44 m / z [M+H].

[0156] Step 11) Intermediate 15 of Example 1: (S)-2-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)- 1-Fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-5,6,6a,7,9,10-hexahydro- Preparation of tert-butyl 8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctano[1,2,3-de]naphthalene-8-carboxylate

[0157] At room temperature, in the presence of (S)-2-chloro-1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctano[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester (Intermediate 13 of Example 1) (0.20 g, 0.35 mmol ) and 2-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (Intermediate 14 of Example 1) (0.19 g, 0.53 mmol) in a solvent mixture of 1,4-dioxane and water (2.0 mL, 10 vol) were added Cs2CO3 (0.34 g, 1.06 mmol) and degassed under N2 gas for 10 minutes. Pd(dppf)Cl2·DCM (0.028 g, 0.03 mmol) was added at room temperature, and the reaction mixture was heated to 80°C and stirred for 16 hours. The progress of the reaction was monitored by TLC. After the reaction was completed, the reaction mixture was diluted with water and extracted with ethyl acetate. The separated organic layer was washed with a saline solution, dried over Na2SO4, filtered and evaporated under reduced pressure to obtain a crude compound. The crude compound was purified by preparative HPLC to give (S)-2-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cycloocta[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester (Intermediate 15 of Example 1) (0.04 g, yield 15%) as a light yellow solid. MS (LC-MS): 765.71 m / z [M+H].

[0158] Step 12) Example 1: 5-ethyl-6-fluoro-4-((S)-1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrole oxazine-7a(5H)-yl)methoxy)-6,6a,7,8,9,10-hexahydro-5H-4-oxa-3,8,10a,11,13-pentaazabenzo[4, Preparation of cyclooctano[1,2,3-de]naphthalene-2-yl)naphthalene-2-ol

[0159] At 0°C, 1,4-dioxane (0.80 mL) containing 4M HCl was added to a stirred solution of (S)-2-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctane[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester (Intermediate 15 of Example 1) (0.04 g, 0.05 mmol) in DCM (1.2 mL, 30 vol), and the reaction mixture was heated to room temperature and stirred for 4 hours. The reaction progress was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain a crude compound. The crude compound was purified by preparative HPLC to give 5-ethyl-6-fluoro-4-((S)-1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-6,6a,7,8,9,10-hexahydro-5H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cycloocta[1,2,3-de]naphthalen-2-yl)naphthalen-2-ol (Example 1) (0.006 g, 19% yield) as an off-white solid.

[0160] 1 H NMR(400MHz, DMSO-d6)δ0.76(t,J=7.2Hz,2H),0.85(t,J=7.2Hz,1H),1.75-1.79(m,2H),1.79-1.81(m,2H),1.99(brs,2H),2 .05(brs,1H),2.12-2.28(m,2H),2.61(d,J=12.8Hz,1H),2.82-2.86(m,2H),2.93(brs,1H),3.02(s,1H),3.09(d,J=4.12Hz,4 H), 3.74-3.86 (m, 1H), 3.98 (d, J = 10.26 Hz, 1H), 4.02-4.20 (m, 2H), 4.43-4.48 (m, 1H), 5.03 (t, J = 12.8 Hz, 1H), 5.21 & 5.35 (brs, 1H), 6.91 & 7.11 (d, J = 2.50 Hz, 1H), 7.28-7.30 (m, 1H), 7.31 (d, J = 8.06 Hz, 1H), 7.74 (dd, J = 8.88, 5.88 Hz, 1H), 9.92 (brs, 1H). NH and -OH protons are not visible in 1H-NMR. MS (LC-MS): 621.40 m / z [M+H].

[0161] The synthetic reaction formula of Example 2

[0162]

[0163] Example 2: Preparation of 5-ethynyl-6-fluoro-4-((S)-1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-6,6a,7,8,9,10-hexahydro-5H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctano[1,2,3-de]naphthalen-2-yl)naphthalen-2-ol

[0164] Step 1) Intermediate 2 of Example 2: (S)-3-(2-((2-(ethylthio)-8-fluoro-7-(7-fluoro-3-(methoxymethyl) Oxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-4-oxo-3,4-dihydropyrido[4,3-d]pyrimidine- Preparation of tert-butyl 5-(5-yl)oxy)ethyl)piperazine-1-carboxylate

[0165] To a stirred solution of a mixture of solvent 1,4-dioxane and water (4:1, 2.5 mL) containing tert-butyl (S)-3-(2-((7-chloro-2-(ethylthio)-8-fluoro-4-oxo-3,4-dihydropyrido[4,3-d]pyrimidin-5-yl)oxy)ethyl)piperazine-1-carboxylate (Intermediate 9 of Example 1) (0.10 g, 0.20 mmol) and ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-1-yl)ethynyl)triisopropylsilane (Intermediate 1 of Example 2) (0.13 g, 0.25 mmol) was added K3PO4 (0.13 g, 0.62 mmol) at room temperature and degassed under N2 gas for 10 min. Wherein, Ruphos-Pd-G3 (0.02 g, 0.02 mmol) was added at room temperature, and the reaction mixture was heated to 90 ° C and stirred for 3 hours. The reaction progress was monitored by TLC. After the reaction was completed, the reaction mixture was diluted with water and extracted with ethyl acetate (2 times), and the combined organic layer was washed with saline solution and dried over anhydrous Na2SO4, filtered and evaporated under reduced pressure to obtain a crude compound. The crude compound was purified by silica gel (100-200 mesh) column chromatography and eluted with 5% methanol in DCM to give (S)-tert-butyl 3-(2-((2-(ethylthio)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-4-oxo-3,4-dihydropyrido[4,3-d]pyrimidin-5-yl)oxy)ethyl)piperazine-1-carboxylate (Intermediate 2 of Example 2) (0.35 g, 4 batches (4×100 mg) yield, crude compound) as a light yellow solid. MS (LC-MS): 838.54 m / z [M+H].

[0166] Step 2) Intermediate 3 of Example 2: (S)-12-(ethylthio)-1-fluoro-2-(7-fluoro-3-(methoxymethoxy)- 8-((Triisopropylsilyl)ethynyl)naphthalen-1-yl)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11, Preparation of tert-butyl 13-pentaazabenzo[4,5]cyclooctano[1,2,3-de]naphthalene-8-carboxylate

[0167] To a stirred solution of (S)-tert-butyl 3-(2-((2-(ethylthio)-8-fluoro-7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-4-oxo-3,4-dihydropyrido[4,3-d]pyrimidin-5-yl)oxy)ethyl)piperazine-1-carboxylate (Intermediate 2 of Example 2) (0.15 g, 0.28 mmol) in DCM (3.0 mL, 20 vol) was added BOP-Cl (0.254 g, 1.00 mmol) and DIPEA (0.70 mL, 4.04 mmol) (3.0 mL, 20 vol) at room temperature, the reaction mixture was heated to 50°C and stirred for 5 hours. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to give the crude compound. The crude compound was purified by silica gel (100-200 mesh) column chromatography and eluted with 5% MeOH in DCM to give (S)-12-(ethylthio)-1-fluoro-2-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctane[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester (Intermediate 3 of Example 2) (0.13 g, 2 batches in total, yield 57%) as a brown solid. MS (LCMS): 820.49 m / z [M+H].

[0168] Step 3) Intermediate 4 of Example 2: (S)-12-(ethylsulfonyl)-1-fluoro-2-(7-fluoro-3-(methoxymethoxy) 1-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8, Preparation of tert-butyl 10a,11,13-pentaazabenzo[4,5]cyclooctano[1,2,3-de]naphthalene-8-carboxylate

[0169] Potassium peroxymonosulfonate (Oxone) (0.24 g, 0.79 mmol) was added to a stirred solution of a mixture of (S)-12-(ethylthio)-1-fluoro-2-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cycloocta[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester (Intermediate 3 of Example 2) (0.13 g, 0.16 mmol) in a solvent of ACN and water (2:1, 15.6 mL) at room temperature, and the reaction mixture was stirred for 1 hour at room temperature. The progress of the reaction was monitored by TLC for 2 hours. After the reaction was completed, the reaction mixture was diluted with water (10 mL) and ethyl acetate (2×20 mL), and the combined organic layer was washed with brine solution, dried over anhydrous Na2SO4 and concentrated under reduced pressure to obtain a crude compound (S)-12-(ethylsulfonyl)-1-fluoro-2-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctane[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester (Intermediate 4 of Example 2) (0.135 g, crude compound) as a light yellow solid. The crude compound was used directly in the next step without further purification. MS (LC-MS): 852.61 m / z [M+H].

[0170] Step 4) Intermediate 6 of Example 2: (S)-1-fluoro-2-(7-fluoro-3-(methoxymethoxy)-8-(triisopropyl 1-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)- 5,6,6a,7,9,10-Hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctano[1,2,3-de]naphthalene Preparation of tert-butyl 8-carboxylate

[0171] At 0°C, NaO was added to a stirred solution of ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizine-7a(5H)-yl)methanol (Intermediate 5 of Example 2) (0.031 g, 0.20 mmol) in THF (2.0 mL, 15 vol). tBu(sodium tert-butoxide) (0.025 g, 0.26 mmol) was added and stirred for 10 minutes, followed by the addition of (S)-12-(ethylsulfonyl)-1-fluoro-2-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctane[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester (Intermediate 4 of Example 2) (0.135 g, 0.16 mmol) at 0°C. The reaction mixture was stirred for 20 minutes at 0°C. The progress of the reaction was monitored by TLC. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to obtain a residue, which was diluted with water and extracted with ethyl acetate (2 times), and the combined organic layer was washed with saline solution, dried over anhydrous Na2SO4, filtered and evaporated. Under reduced pressure, a crude compound (S)-1-fluoro-2-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalene-1-yl)-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazine-7a(5H)-yl)methoxy)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctane[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester (intermediate 6 of Example 2) (0.135 g, crude compound) was obtained as a light yellow solid. The crude compound was used in the next step without further purification. MS (LC-MS): 917.64 m / z [M+H].

[0172] Step 5) Intermediate 7 of Example 2: (S)-2-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)- 1-Fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-5,6,6a,7,9,10-hexahydro- Preparation of tert-butyl 8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctano[1,2,3-de]naphthalene-8-carboxylate

[0173] TBAF (tetrabutylammonium fluoride) (2.0 mL, 15 vol) was added to a stirred solution of (S)-1-fluoro-2-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cycloocta[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester (Intermediate 6 of Example 2) (0.135 g, 0.15 mmol) in THF (3.4 mL, 25 vol) at 0°C and the reaction mixture was stirred for 1 hour at 0°C. The progress of the reaction was monitored by TLC. After the reaction is completed, the reaction mixture is quenched with NaHCO3 aqueous solution (5mL) and extracted with ethyl acetate (2 times), the combined organic layer is washed with saline solution, and dried with anhydrous Na2SO4, filtered and evaporated under reduced pressure to obtain a crude compound. The crude compound is purified by preparative HPLC to obtain (S)-2-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphthalene-1-yl)-1-fluoro-12-(((2R, 7aS)-2-fluorotetrahydro-1H-pyrrolazine-7a(5H)-yl)methoxy)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctane[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester (intermediate 7 of embodiment 2) (0.03g, yield 26%), as an off-white solid. MS (LC-MS): 761.55 m / z [M+H].

[0174] Step 6) Example 2: 5-ethynyl-6-fluoro-4-((S)-1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidone Alloxazine-7a(5H)-yl)methoxy)-6,6a,7,8,9,10-hexahydro-5H-4-oxa-3,8,10a,11,13-pentaazabenzo [4,5]cyclooctano[1,2,3-de]naphthalen-2-yl)

[0175] Preparation of naphthalene-2-ol

[0176] At 0°C, 4M HCl was added to a stirred solution of 1,4-dioxane (1.2 mL) containing (S)-2-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-1-fluoro-12-(((2R),7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctane[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester (Intermediate 7 of Example 2) (0.12 g, 0.16 mmol). The reaction mixture was stirred for 30 minutes at 0°C. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to give a crude compound. The crude compound was purified by preparative HPLC to give 5-ethynyl-6-fluoro-4-((S)-1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-6,6a,7,8,9,10-hexahydro-5H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cycloocta[1,2,3-de]naphthalen-2-yl)naphthalen-2-ol (Example 2) (0.0061 g, 6% yield) as an off-white solid.

[0177] 1 H NMR (401MHz, DMSO-d6) δ1.72-2.23(m,8H),2.58-2.63(m,1H),2.82-2.87(m,2H),2.98 -3.16(m,6H),3.73(dd,J=10.88,1.38Hz,1H),3.94-3.98(m,1H),4.06-4.17(m,3H),4 .42 (dd, J = 12.51, 5.50 Hz, 1H), 5.03 (d, J = 12.51 Hz, 1H), 5.21-5.34 (m, 1H), 7.04-7.24 (m, 1H), 7.35-7.36 (m, 1H), 7.42-7.48 (m, 1H), 7.95 (dd, J = 9.13, 5.88 Hz, 1H), exchangeable protons 1 Not observed in H NMR. MS (LC-MS): 617.31 m / z [M+H].

[0178] The synthetic reaction formula of Example 3

[0179]

[0180] Example 3: Preparation of 4-((14aS)-11-chloro-9-fluoro-7-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-1,3,4,13,14,14a-hexahydro-2H-pyrazino[1',2':5,6][1,5]oxazocin[4,3,2-de]quinazolin-10-yl)-7-fluorobenzo[d]thiazole-2-amine

[0181] Step 1) Preparation of Intermediate 2 of Example 3: 3-bromo-2,5-difluoroaniline

[0182] At room temperature, Fe (7.03 g, 126 mmol) and NH4Cl (13.5 g, 252 mmol) were added to a stirred solution (4:1, 250 mL) of a mixture of solvent EtOH and H2O containing 1-bromo-2,5-difluoro-3-nitrobenzene (intermediate 1 of Example 3) (10.0 g, 42.0 mmol), and the reaction mixture was heated to 80 ° C and stirred for 6 hours. The reaction progress was monitored by TLC. After the reaction was completed, ethyl acetate (100 mL) was used as a washing solvent for the reaction mixture, and filtered through a diatomaceous earth pad to obtain a filtrate, which was then washed with water (50 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a crude compound. The crude compound was purified by silica gel (100-200 mesh) column chromatography and eluted with petroleum ether containing 10% ethyl acetate to obtain 3-bromo-2,5-difluoroaniline (Intermediate 2 of Example 3) (10.0 g, crude compound) as a light yellow viscous liquid. 1 H NMR (400MHz, CDCl3) δ3.93 (brs, 2H), 6.40-6.45 (m, 1H), 6.59-6.64 (m, 1H). MS (LC-MS): 208.02m / z[M+H].

[0183] Step 2) Intermediate 3 of Example 3: (Z)-N-(3-bromo-2,5-difluorophenyl)-2-(hydroxyimino)acetamide Preparation

[0184] To a stirred solution of 3-bromo-2,5-difluoroaniline (Intermediate 2 of Example 3) (5.00 g, 24.1 mmol) in H2O (100 mL, 20 vol) was added NH2OH.HCl (5.02 g, 72.5 mmol), Na2SO4 (27.4 g, 217 mmol) and 1,2,2-trichloroethane-1,1-diol (5.99 g, 36.2 mmol) in EtOH (ethanol) (14.0 mL), and concentrated HCl (3.5 mL) was added at 0°C, and the reaction mixture was heated to 60°C and stirred for 16 hours. The reaction progress was monitored by TLC. After the reaction was completed, the reaction mixture was cooled to room temperature, thereby forming a solid. The solid formed was filtered using water (50 mL) as a washing solvent and dried under high vacuum to obtain a crude compound (Z)-N-(3-bromo-2,5-difluorophenyl)-2-(hydroxyimino)acetamide (Intermediate 3 of Example 3) (10.0 g, a total of 2 batches (2×5 g) yield, crude compound) as a brown solid. The crude compound was used directly in the next step without further purification. 1 H NMR (401MHz, DMSO-d6) δ7.50-7.54(m,1H),7.78(s,1H),7.82-7.87(m,1H),10.11(s,1H),12.45(s,1H).MS(LC-MS): 279.02m / z[M+H].

[0185] Step 3) Preparation of Intermediate 4 of Example 3: 6-bromo-4,7-difluoroindoline-2,3-dione

[0186] At 60°C, (Z)-N-(3-bromo-2,5-difluorophenyl)-2-(hydroxyimino)acetamide (Intermediate 3 of Example 3) (5.00 g, 17.9 mmol) was added to H2SO4 (50 mL, 10 vol), and the reaction mixture was stirred for 2 hours at 90°C. The progress of the reaction was monitored by TLC. After the reaction was completed, the reaction mixture was diluted with ice-cold water (50 mL) to obtain a solid, which was filtered and dried under vacuum to obtain a crude compound 6-bromo-4,7-difluoroindoline-2,3-dione (Intermediate 4 of Example 3) (4.0 g, crude compound) as a brown solid. The crude compound was used directly in the next step without further purification. MS (LC-MS): 259.88 m / z [MH].

[0187] Step 4) Preparation of Intermediate 5 of Example 3: 2-amino-4-bromo-3,6-difluorobenzoic acid

[0188] At 0°C, 30% H2O2 (9.53 mL) was added to a stirred solution of 2M NaOH solution (84 mL, 11 vol) containing 6-bromo-4,7-difluoroindoline-2,3-dione (intermediate 4 of Example 3) (4.00 g, 15.3 mmol), and the reaction mixture was heated to room temperature and stirred for 16 hours. The progress of the reaction was monitored by TLC. After the reaction was completed, the reaction mixture was acidified to about pH 3 with concentrated HCl at 0°C to obtain a solid, which was filtered and dried under high vacuum to obtain a crude compound 2-amino-4-bromo-3,6-difluorobenzoic acid (intermediate 5 of Example 3) (4.0 g, a total of 2 batches (2×4 g) yield, crude compound) as a brown solid. The crude compound was used directly in the next step without further purification. 1 HNMR (400MHz, DMSO-d6) δ6.72 (dd, J = 10.76, 5.25Hz, 1H), 6.77-7.00 (m, 2H), 13.42 (brs, 1H). MS (LCMS): 252.06m / z [M+H].

[0189] Step 5) Preparation of Intermediate 6 of Example 3: 2-amino-4-bromo-5-chloro-3,6-difluorobenzoic acid

[0190] At room temperature, NCS (N-chlorosuccinimide) (2.48 g, 14.3 mmol) was added to a stirred solution of DMF (40 mL, 13 vol) containing 2-amino-4-bromo-3,6-difluorobenzoic acid (intermediate 5 of Example 3) (3.00 g, 11.9 mmol), and the reaction mixture was heated to 90 ° C and stirred for 2 hours. The progress of the reaction was monitored by TLC. After the reaction was completed, the reaction mixture was diluted with water (100 mL) to obtain a solid, and the solid was filtered and dried under high vacuum to obtain a crude compound 2-amino-4-bromo-5-chloro-3,6-difluorobenzoic acid (intermediate 6 of Example 3) (3.0 g, crude compound) as a brown solid. The crude compound was used directly in the next step without further purification. MS (LCMS): 286.09 m / z [M+H].

[0191] Step 6) Preparation of Intermediate 7 of Example 3: 2-amino-4-bromo-5-chloro-3,6-difluorobenzamide

[0192] At 0°C, NH4Cl (0.56 g, 10.5 mmol), HATU (2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate) (2.20 g, 5.79 mmol) and DIPEA (2.03 g, 15.8 mmol) were added to a stirred solution of DMF (15 mL, 10 vol) containing 2-amino-4-bromo-5-chloro-3,6-difluorobenzoic acid (Intermediate 6 of Example 3) (1.50 g, 5.26 mmol) and the reaction mixture was allowed to stand. It was heated to room temperature and stirred for 2 hours. The progress of the reaction was monitored by TLC. After the reaction was completed, the reaction mixture was diluted with water (20 mL) to obtain a solid, which was filtered and dried under high vacuum to obtain a crude compound 2-amino-4-bromo-5-chloro-3,6-difluorobenzamide (Intermediate 7 of Example 3) (0.80 g, crude compound) as a brown solid. The crude compound was used in the next step without further purification. MS (LCMS): 285.08 m / z [M+H].

[0193] Step 7) Preparation of Intermediate 8 of Example 3: 7-bromo-2,6-dichloro-5,8-difluoroquinazolin-4(3H)-one

[0194] At a temperature of 0°C, thiophosgene (0.96 g, 8.45 mmol) was added to a stirred solution of 1,4-dioxane (8.0 mL, 10 vol) containing 2-amino-4-bromo-5-chloro-3,6-difluorobenzamide (Intermediate 7 of Example 3) (0.80 g, 2.82 mmol) and stirred at room temperature for 1 hour. The reaction mixture was heated to 105°C and stirred for 2 hours. The reaction progress was monitored by TLC. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to obtain a residue, and the residue was washed with petroleum ether containing 20% ​​ethyl acetate to obtain a solid, which was filtered and dried under high vacuum to obtain a crude compound 7-bromo-2,6-dichloro-5,8-difluoroquinazoline-4 (3H) -one (Intermediate 8 of Example 3) (0.70 g, crude compound) as a light yellow solid. The crude compound was used directly in the next step without further purification. MS (LC-MS): 329.02m / z[M+H].

[0195] Step 8) Intermediate 9 of Example 3: (S)-3-(2-((7-bromo-2,6-dichloro-8-fluoro-4-oxo-3,4-dihydro Preparation of tert-butyl quinazolin-5-yl)oxy)ethyl)piperazine-1-carboxylate

[0196] At 0°C, NaH (0.58 g, 24.3 mmol) was added to a stirred solution of (S)-3-(2-hydroxyethyl)piperazine-1-carboxylic acid tert-butyl ester (1.19 g, 3.65 mmol) in THF (10 mL, 14 vol), and stirred for 30 minutes at 0°C. A solution of 7-bromo-2,6-dichloro-5,8-difluoroquinazoline-4(3H)-one (Intermediate 8 of Example 3) (0.70 g, 3.04 mmol) in THF (7.0 mL, 10 vol) was added at 0°C, and the reaction mixture was heated to room temperature and stirred for 3 hours. The reaction progress was monitored by TLC. After the reaction was completed, the reaction mixture was diluted with supersaturated NH4Cl solution (20 mL) and extracted with ethyl acetate (2×20 mL), and the combined organic layer was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a crude compound (S)-3-(2-((7-bromo-2,6-dichloro-8-fluoro-4-oxo-3,4-dihydroquinazolin-5-yl)oxy)ethyl)piperazine-1-carboxylic acid tert-butyl ester (Intermediate 9 of Example 3) (0.45 g, crude compound) as a brown solid. The crude compound was used directly in the next step without further purification. 1 H NMR (400 MHz, DMSO-d6) δ 1.16-1.23 (m, 2H), 1.37-1.43 (m, 9H), 1.99-2.10 (m, 2H), 2.81-3.13 (m, 3H), 3.96-4.13 (m, 4H), exchangeable protons 1 Not observed in HNMR. MS (LC-MS): 539.16 m / z [M+H].

[0197] Step 9) Intermediate 10 of Example 3: (S)-10-bromo-7,11-dichloro-9-fluoro-1,3,4,13,14,14a-hexa Preparation of tert-butyl 2H-pyrazino[1',2':5,6][1,5]oxazocin[4,3,2-de]quinazoline-2-carboxylate

[0198] BOP-Cl (0.68 g, 2.69 mmol) and DIPEA (1.48 g, 11.5 mmol) were added to a stirred solution of (S)-tert-butyl 3-(2-((7-bromo-2,6-dichloro-8-fluoro-4-oxo-3,4-dihydroquinazolin-5-yl)oxy)ethyl)piperazine-1-carboxylate (Intermediate 9 of Example 3) (0.45 g, 0.77 mmol) in DCM (5 mL, 11 vol) at 0°C, and the reaction mixture was heated to room temperature and stirred for 16 hours. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was diluted with DCM (15 mL) and washed with water (10 mL), and the separated organic layer was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a crude compound. The crude compound was purified by silica gel (100-200 mesh) and washed with petroleum ether containing 40% ethyl acetate to obtain (S)-10-bromo-7,11-dichloro-9-fluoro-1,3,4,13,14,14a-hexahydro-2H-pyrazino[1',2':5,6][1,5]oxazocin[4,3,2-de]quinazoline-2-carboxylic acid tert-butyl ester (Intermediate 10 of Example 3) (0.15 g, crude compound) as a brown solid. MS (LC-MS): 521.23 m / z [M+H].

[0199] Step 10) Intermediate 11 of Example 3: (14aS)-10-(2-((tert-butyloxycarbonyl)amino)-7-fluorobenzo[d] Thiazol-4-yl)-7,11-dichloro-9-fluoro-1,3,4,13,14,14a-hexahydro-2H-pyrazino[1',2':5,6][1,5]oxazine Preparation of tert-butyl octyl[4,3,2-de]quinazoline-2-carboxylate

[0200] To a stirred solution of a mixture of solvent 1,4-dioxane and water (2:1, 3.0 mL) containing tert-butyl (S)-10-bromo-7,11-dichloro-9-fluoro-1,3,4,13,14,14a-hexahydro-2H-pyrazino[1',2':5,6][1,5]oxazocine[4,3,2-de]quinazoline-2-carboxylate (Intermediate 10 of Example 3) (0.15 g, 0.28 mmol) and tert-butyl (7-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[d]thiazol-2-yl)carbamate (0.095 g, 0.31 mmol) was added Cs2CO3 (0.27 g, 0.84 mmol) at room temperature and degassed for 5 minutes. Wherein, dichloro[bis(diphenylphosphinophenyl)ether]palladium(II) (0.02 g, 0.03 mmol) was added at room temperature, and the reaction mixture was heated to 80°C and stirred for 4 hours. The reaction progress was monitored by TLC. After the reaction was completed, the reaction mixture was diluted with ice-cold water (10 mL) and extracted with ethyl acetate (2×15 mL), and the combined organic layer was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a crude compound. The crude compound was purified by silica gel (100-200 mesh) and eluted with 40% ethyl acetate in petroleum ether to give (14aS)-10-(2-((tert-butyloxycarbonyl)amino)-7-fluorobenzo[d]thiazol-4-yl)-7,11-dichloro-9-fluoro-1,3,4,13,14,14a-hexahydro-2H-pyrazino[1',2':5,6][1,5]oxazocino[4,3,2-de]quinazoline-2-carboxylic acid tert-butyl ester (Intermediate 11 of Example 3) (0.07 g, 35% yield) as a light yellow sticky solid. MS (LCMS): 709.35 m / z [M+H].

[0201] Step 11) Intermediate 12 of Example 3: (14aS)-10-(2-((tert-butyloxycarbonyl)amino)-7-fluorobenzo[d] Thiazol-4-yl)-11-chloro-9-fluoro-7-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-1,3, 4,13,14,14a-Hexahydro-2H-pyrazino[1',2':5,6][1,5]oxazocino[4,3,2-de]quinazoline-2-carboxylic acid Preparation of tert-butyl ester

[0202] To a stirred solution of ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methanol (0.04 g, 0.25 mmol) in DMF (1.0 mL, 6 vol) was added NaH (0.008 g, 0.32 mmol) at 0°C and stirred for 1 hour. Wherein, (14aS)-10-(2-((tert-butyloxycarbonyl)amino)-7-fluorobenzo[d]thiazol-4-yl)-7,11-dichloro-9-fluoro-1,3,4,13,14,14a-hexahydro-2H-pyrazino[1',2':5,6][1,5]oxazocine[4,3,2-de]quinazoline-2-carboxylic acid tert-butyl ester (Intermediate 11 of Example 3) (0.15 g, 0.21 mmol) was added at 0°C, and the reaction mixture was heated to room temperature and stirred for 3 hours. The progress of the reaction was monitored by TLC. After the reaction was completed, the reaction mixture was diluted with saturated NH4Cl solution (10 mL) and extracted with ethyl acetate (2×15 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain the crude compound. The crude compound was purified on silica gel (100-200 mesh) and eluted with 5% methanol in DCM to give (14aS)-10-(2-((tert-butyloxycarbonyl)amino)-7-fluorobenzo[d]thiazol-4-yl)-11-chloro-9-fluoro-7-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-1,3,4,13,14,14a-hexahydro-2H-pyrazino[1',2':5,6][1,5]oxazocino[4,3,2-de]quinazoline-2-carboxylic acid tert-butyl ester (Intermediate 12 of Example 3) (0.05 g, 28% yield) as a brown solid. MS (LCMS): 832.41 m / z [M+H].

[0203] Step 12) Example 3: 4-((14aS)-11-chloro-9-fluoro-7-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizine- 7a(5H)-yl)methoxy)-1,3,4,13,14,14a-hexahydro-2H-pyrazino[1',2':5,6][1,5]oxazocine Preparation of [4,3,2-de]quinazolin-10-yl)-7-fluorobenzo[d]thiazol-2-amine

[0204] A solution of (14aS)-10-(2-((tert-butyloxycarbonyl)amino)-7-fluorobenzo[d]thiazol-4-yl)-11-chloro-9-fluoro-7-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-1,3,4,13,14,14a-hexahydro-2H-pyrazino[1',2':5,6][1,5]oxazocino[4,3,2-de]quinazoline-2-carboxylic acid tert-butyl ester (Intermediate 12 of Example 3) (0.05 g, 0.06 mmol) in dioxane (1.0 mL, 20 vol) (containing 4 M HCl) was stirred at room temperature for 5 hours. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to give the crude compound. The crude compound was purified by preparative HPLC to give 4-((14aS)-11-chloro-9-fluoro-7-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-1,3,4,13,14,14a-hexahydro-2H-pyrazino[1',2':5,6][1,5]oxazocino[4,3,2-de]quinazolin-10-yl)-7-fluorobenzo[d]thiazol-2-amine (Example 3) (0.0075 g, 20% yield) as an off-white solid.

[0205] 1 H NMR(400MHz,DMSO-d6)δ1.70-1.91(m,4H),1.93-2.16(m,3H),2.54-2.56(m,1H),2.64-2.70(m,1 H),2.77-2.82(m,3H),3.00-3.10(m,4H),3.19-3.24(m,1H),3.64-3.74(m,1H),3.94(dd,J=10.51 ,2.25Hz,1H),4.06(dd,J=10.51,2.75Hz,1H),4.24(q,J=11.6Hz,1H),4.43(brs,1H),4.55-4.93(m,1H),5.20-5.34(m,1H),7.01-7.06(m,1H),7.14-7.23(m,1H),7.89(d,J=16.76Hz,2H),-NH proton in 1 Not observed in H NMR. MS (LCMS): 632.20 m / z [M+H].

[0206] The synthetic reaction formula of Example 4

[0207]

[0208] Example 4: Preparation of 3-chloro-4-cyclopropyl-5-((S)-1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-6,6a,7,8,9,10-hexahydro-5H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctano[1,2,3-de]naphth-2-yl)phenol

[0209] Step 1) Preparation of Intermediate 2 of Example 4: 1-bromo-3-chloro-2-cyclopropylbenzene

[0210] A mixture of dioxane: water (100 mL: 30 mL) containing 1-bromo-3-chloro-2-iodobenzene (22 g, 69.65 mmol), cyclopropylboronic acid (15.21 g, 90.54 mmol), Pd(dppf)Cl2 (2.3 g, 3.16 mmol) and K3PO4 (24.2 g, 113.96 mmol) was stirred under N2 gas at 100°C for 7 hours. TLC (PE / EA=1 / 0) showed consumption of SM1 (Rf=0.6) and formation of a new spot (Rf=0.7). The reaction mixture was diluted with water and extracted with ethyl acetate (200 mL×2). The combined organic layers were washed with 70 mL of brine, dried over Na2SO4, filtered and concentrated in vacuo to obtain a residue. The residue was purified by column chromatography (PE / EA=1 / 0) to give 1-bromo-3-chloro-2-cyclopropyl-benzene (13 g, yield 81.25%) as a colorless liquid.

[0211] 1 H NMR (400MHz, CDCl3) δ7.46 (dd, J=8.0, 0.9Hz, 1H), 7.30 (dd, J=8.0, 0.9Hz, 1H), 6.99(t,J=8.0Hz,1H),1.81-1.71(m,1H),1.22-1.14(m,2H),0.81-0.73(m,2H).

[0212] Step 2) Intermediate 3 of Example 4: 2-(3-bromo-5-chloro-4-cyclopropylphenyl)-4,4,5,5-tetramethyl-1,3, Preparation of 2-dioxaborolane

[0213] A mixture of 1-bromo-3-chloro-2-cyclopropylbenzene (13 g, 56.52 mmol), [Ir(OMe)(COD))2 (3.74 g, 5.65 mmol), 4,4'-di-tert-butyl-2,2'-bipyridine (1.82 g, 6.78 mmol) and 4,4,5,5-tetramethyl-1,3,2-dioxaborolane (21.70 g, 169.56 mmol) in hexane (130 mL) was stirred at 60°C under N2 gas for 3 hours. Consumption of SM1 (Rf=0.3) and formation of a new spot (Rf=0.5) were shown in TLC (PE / EA=50 / 1). The solvent was removed by concentrating the reaction mixture under reduced pressure. The residue was purified by column chromatography (PE:EA=50:1) to give 2-(3-bromo-5-chloro-4-cyclopropyl-phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (10.7 g, yield 53.18%) as a colorless oil.

[0214] 1 H NMR (400MHz, CDCl3) δ7.87 (s, 1H), 7.70 (s, 1H), 1.78 (tt, J = 8.5, 5.8Hz, 1H), 1.33 (s, 12H), 1.22-1.14 (m, 2H), 0.77 (q, J = 5.8Hz, 2H).

[0215] Step 3) Intermediate 4 of Example 4: 2-(3-bromo-5-chloro-4-cyclopropylphenyl)-4,4,5,5-tetramethyl-1,3, Preparation of 2-dioxaborolane

[0216] AcOH (113.4 g, 1888.42 mmol, 107.5 mL) and H2O2 (2.05 g, 4.79 mmol, 15.96 mL, purity 30%) were added to a solution of 2-(3-bromo-5-chloro-4-cyclopropyl-phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (10.5 g, 29.49 mmol) in THF / H2O (100 / 50 mL) at 0°C, and the mixture was stirred at 0°C for 1 hour. TLC (PE / EA=15 / 1) showed that SM1 (Rf=0.7) was consumed and a new spot (Rf=0.4) was formed. The reaction mixture was quenched with Na2S2O3 (10% in aq, 80 mL) at 0°C. The mixture was extracted with EA (100 mL×2). The combined organic layer was washed with 100 mL of brine and dried over Na2SO4. The mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (PE:EA=10:1) to give 3-bromo-5-chloro-4-cyclopropyl-phenol (5.7 g, yield 78.62%) as a colorless oil.

[0217] 1 H NMR (400MHz, CDCl3) δ7.00(d,J=2.6Hz,1H),6.83(d,J=2.6Hz,1H),3.73(d,J=47.7H z, 1H), 1.66 (ddd, J = 14.0, 7.0, 4.2Hz, 1H), 1.15-1.08 (m, 2H), 0.71 (q, J = 5.7Hz, 2H).

[0218] Step 4) Preparation of Intermediate 5 of Example 4: 1-bromo-3-chloro-2-cyclopropyl-5-(methoxymethoxy)benzene

[0219] At 0°C, DIEA (4.49 g, 34.76 mmol) and methoxymethyl hypobromite (3.19 g, 25.49 mmol) were added to a solution of 3-bromo-5-chloro-4-cyclopropylphenol (5.7 g, 23.17 mmol) in DCM (57 mL). Then, the reaction mixture was stirred for 2 hours at 25°C. The reaction mixture was diluted with water and extracted with DCM (50 mL×2). The combined organic layers were washed with 70 mL of brine and dried with Na2SO4, filtered and concentrated in vacuo to obtain a residue. The residue was purified by column chromatography (PE / EA=20 / 1) to obtain 1-bromo-3-chloro-2-cyclopropyl-5-(methoxymethoxy)benzene (4 g, yield 59.52%) as a colorless oil.

[0220] 1 H NMR (400MHz, CDCl3) δ7.12(d,J=2.5Hz,1H),6.96(d,J=2.5Hz,1H),5.04(s,2 H), 3.38 (s, 3H), 1.61 (s, 1H), 1.08-1.02 (m, 2H), 0.65 (dd, J = 5.6, 1.1Hz, 2H).

[0221] Step 5) Intermediate 6 of Example 4: 2-(3-chloro-2-cyclopropyl-5-(methoxymethoxy)phenyl)-4,4,5, Preparation of 5-Tetramethyl-1,3,2-dioxaborolane

[0222] Pd(dppf)Cl2 (1.0 g, 1.38 mmol) was added to a solution of 1-bromo-3-chloro-2-cyclopropyl-5(methoxymethoxy)benzene (4 g, 13.79 mmol), KOAc (4.06 g, 41.38 mmol) and 4,4,5,5-tetramethyl-2-(4,5,5-trimethyl-4-methyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolan (7.0 g, 27.59 mmol) in dioxane (80 mL). The mixture was stirred at 100°C for 16 hours. The reaction mixture was diluted with water and extracted with ethyl acetate (50 mL×3). The combined organic layers were washed with 70 mL of brine and dried over Na2SO4, filtered and concentrated in vacuo to obtain a residue. The residue was purified by column chromatography (PE / EA=10 / 1) to give 2-(3-chloro-2-cyclopropyl-5-(methoxymethoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.4 g, yield 30.04%) as a colorless oil. MS: m / z=339.1 (M+H + ,ESI+).

[0223] Step 6) Intermediate 7 of Example 4: (3S)-3-(2-((7-(3-chloro-2-cyclopropyl-5-(methoxymethoxy) phenyl)-8-fluoro-2-(methylthio)-4-oxo-4,4a-dihydropyrido[4,3-d]pyrimidin-5-yl)oxy)ethyl)piperazine- Preparation of tert-butyl 1-carboxylate

[0224] A solution of (3S)-tert-butyl 3-(2-((7-chloro-8-fluoro-2-(methylthio)-4-oxo-4,4a-dihydropyrido[4,3-d]pyrimidin-5-yl)oxy)ethyl)piperazine-1-carboxylate (Intermediate 9 of Example 1, 1.4 g, 2.96 mmol), 2-(3-chloro-2-cyclopropyl-5-(methoxymethoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (2.0 g, 5.92 mmol), Ruphos-Pd-G3 (742 mg, 0.89 mmol) and K3PO4 (1.88 g, 8.88 mmol) in dioxane / H2O (16 mL / 4 mL) was stirred under N2 gas at 90°C for 3 hours. LCMS indicated consumption of starting material and formation of the desired mass. The reaction mixture was quenched with water (60 mL) and extracted with ethyl acetate (60 mL × 3). The combined organic layer was washed with 60 mL of brine and dried with Na2SO4, filtered and concentrated in vacuo to give a residue. The residue was purified by column chromatography (PE / EA = 0 / 1) to give (3S) -3- (2- ((7- (3- chloro-2-cyclopropyl -5- (methoxymethyl) phenyl) -8- fluoro-2- (methylthio) -4- oxo -4,4a- dihydropyrido [4,3-d] pyrimidin-5-yl) oxy) ethyl) piperazine -1- carboxylic acid tert-butyl ester (700 mg, yield 36.46%), as a yellow solid. MS: m / z = 650.3 (M + H + ,ESI+).

[0225] Step 7) Intermediate 8 of Example 4: (S)-2-(3-chloro-2-cyclopropyl-5-(methoxymethoxy)phenyl)-1- Fluoro-12-(methylthio)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctane Preparation of tert-butyl 1,2,3-de-naphthalene-8-carboxylate

[0226] To a solution of tert-butyl (3S)-3-(2-((7-(3-chloro-2-cyclopropyl-5-(methoxymethyl)phenyl)-8-fluoro-2-(methylthio)-4-oxo-4,4a-dihydropyrido[4,3-d]pyrimidin-5-yl)oxy)ethyl)piperazine-1-carboxylate (700 mg, 1.08 mmol) in DCM (47 mL) was added BoPCl (822 mg, 3.24 mmol) and DIEA (1.25 g, 9.71 mmol). The mixture was then stirred at 25°C for 16 hours. LCMS indicated that the starting material was consumed and the desired mass was formed. The reaction mixture was diluted with water (20 mL) and extracted with DCM (40 mL x 2). The combined organic layers were washed with 60 mL of brine and dried over Na2SO4, filtered and concentrated in vacuo to give a residue. The residue was purified by column chromatography (PE / EA=1 / 1) to give (S)-2-(3-chloro-2-cyclopropyl-5-(methoxymethoxy)phenyl)-1-fluoro-12-(methylthio)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctane[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester (140 mg, yield: 68.06%) as a yellow solid. MS: m / z=632.4 (M+H + ,ESI+).

[0227] Step 8) Intermediate 9 of Example 4: (6aS)-2-(3-chloro-2-cyclopropyl-5-(methoxymethoxy)phenyl)- 1-Fluoro-12-(methylsulfinyl)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo Preparation of tert-butyl [4,5]cyclooctano[1,2,3-de]naphthalene-8-carboxylate

[0228] Potassium peroxymonosulfonate (536 mg, 0.87 mmol) was added to a solution of (S)-2-(3-chloro-2-cyclopropyl-5-(methoxymethoxy)phenyl)-1-fluoro-12-(methylthio)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctane[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester (110 mg, 0.17 mmol) in ACN / H2O (10 mL / 10 mL) and stirred at 25°C for 1 hour. LCMS indicated that the starting material was consumed and the desired mass was detected. The reaction mixture was diluted with water (20 mL) and extracted with DCM (30 mL×2). The combined organic layers were washed with 20 mL of brine and dried over Na2SO4, filtered and concentrated in vacuo to give (6aS)-2-(3-chloro-2-cyclopropyl-5-(methoxymethoxy)phenyl)-1-fluoro-12-(methylsulfinyl)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctano[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester (120 mg, crude compound) as a yellow solid. MS: m / z=648.2 (M+H + ,ESI+).

[0229] Step 9) Intermediate 10 of Example 4: (S)-2-(3-chloro-2-cyclopropyl-5-(methoxymethoxy)phenyl)-1- Fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-5,6,6a,7,9,10-hexahydro-8H- Preparation of tert-butyl 4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctano[1,2,3-de]naphthalene-8-carboxylate

[0230] To a solution of (6aS)-2-(3-chloro-2-cyclopropyl-5-(methoxymethoxy)phenyl)-1-fluoro-12-(methylsulfinyl)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cycloocta[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester (120 mg, 0.18 mmol) and ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol (44 mg, 0.28 mmol) in THF (20 mL) was added t-BuONa (36 mg, 0.37 mmol) and the mixture was stirred at 0°C for 1 hour. LCMS indicated consumption of starting material and the desired mass was detected. The reaction mixture was diluted with water (30 mL) and extracted with DCM (30 mL × 2). The combined organic layer was washed with 20 mL of brine and dried over Na2SO4, filtered and concentrated in vacuo to give a residue. The residue was purified by preparative (pre)-TLC (DCM / MeOH=10 / 1) to give (S)-2-(3-chloro-2-cyclopropyl-5-(methoxymethoxy)phenyl)-1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazine-7a(5H)-yl)methoxy)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctane[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester (70 mg, 52.24% yield) as a yellow solid. MS: m / z = 743.3 (M + H + ,ESI+).

[0231] Step 10) Example 4: 3-chloro-4-cyclopropyl-5-((S)-1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidone Alloxazine-7a(5H)-yl)methoxy)-6,6a,7,8,9,10-hexahydro-5H-4-oxa-3,8,10a,11,13-pentaazabenzo [4,5]cyclooctano[1,2,3-de]naphthalen-2-yl)

[0232] Preparation of phenol

[0233] To a solution of (S)-tert-butyl 2-(3-chloro-2-cyclopropyl-5-(methoxymethoxy)phenyl)-1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cycloocta[1,2,3-de]naphthalene-8-carboxylate (70 mg, 0.09 mmol) in EA (2 mL) was added 3 M HCl (7 mL) and the mixture was stirred at 0° C. for 1 hour and LCMS indicated that the starting material was consumed and the desired mass was formed. The reaction mixture was concentrated in vacuo to give a residue. The residue was purified by preparative (pre)-HPLC (FA conditions) to give 3-chloro-4-cyclopropyl-5-((S)-1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-6,6a,7,8,9,10-hexahydro-5H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cycloocta[1,2,3-de]naphth-2-yl)phenol (Example 4) (2.76 mg, 4.9% yield) as a white solid.

[0234] 1 H NMR(400MHz,MeOD)δ8.35(brs,2H,FA),6.95(d,J=2.4Hz,1H),6.77(s,1H),5.52(d,J=52.6 Hz,1H),5.35-5.17(m,1H),4.69-4.45(m,3H),4.40-4.25(m,1H),4.20-4.10(m,1H),4.01-3 .57(m,3H),3.48-3.32(m,3H),3.25-3.13(m,2H),3.06-2.93(m,1H),2.73-2.44(m,3H),2.4 0-2.22(m,3H),2.20-2.02(m,2H),1.90-1.77(m,1H),0.75-0.57(m,2H),0.23-0.03(m,2H). MS: m / z=599.6(M+H + ,ESI+).

[0235] The synthetic reaction formula of Example 5

[0236]

[0237] Example 5: Preparation of 5-ethyl-6-fluoro-4-((6aS)-1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methyl-6,6a,7,8,9,10-hexahydro-5H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctano[1,2,3-de]naphthalen-2-yl)naphthalen-2-ol

[0238] Step 1) Intermediate 2 of Example 5: (S)-3-(2-((tert-butyldiphenylsilyl)oxy)ethyl)piperazine-1- Preparation of tert-butyl carboxylate

[0239] To a mixture of (S)-tert-butyl 3-(2-hydroxyethyl)piperazine-1-carboxylate (8 g, 34.78 mmol), DMAP (848 mg, 6.95 mmol) and imidazole (4.73 g, 69.65 mol) in DCM (80 mL) was added tert-butyldiphenylsilyl chloride (TBDPSCl) (19.11 g, 69.65 mmol) and the reaction mixture was stirred at 25°C for 16 hours. TLC (PE / EA=2 / 1) showed complete consumption of SM1 (Rf=0.2) and formation of a new spot (Rf=0.4). The reaction mixture was diluted with H2O (100 mL) and extracted with DCM (150 mL×3). The combined organic layers were washed with brine (150 mL) and dried over Na2SO4. The residue was obtained by filtration and vacuum concentration. The residue was purified by column chromatography (PE:EA=3:1) to give (S)-tert-butyl 3-(2-((tert-butyldiphenylsilyl)oxy)ethyl)piperazine-1-carboxylate (14.55 g, yield 91%) as a colorless oil. MS: m / z=469.4 (M+H + ,ESI+).

[0240] Step 2) Intermediate 3 of Example 5: 1-benzyl 4-(tert-butyl)(S)-2-(2-((tert-butyldiphenylsilyl)oxy) Preparation of 1,4-dicarboxylic acid ester

[0241] Benzyloxycarbonyl chloride (CBZCl) (13.21 g, 77.7 mmol) was added to a mixture of (S)-tert-butyl 3-(2-((tert-butyldiphenylsilyl)oxy)ethyl)piperazine-1-carboxylate (14.55 g, 31.08 mmol) and DIEA (6.01 g, 46.62 mmol) in DCM (150 mL) at 0°C, and the reaction mixture was stirred for 1 hour under N2 gas at 0°C. The reaction mixture was diluted with H2O (100 mL) and extracted with EA (150 mL×3). The combined organic layers were washed with brine (150 mL) and dried over Na2SO4. Filtered and concentrated in vacuo to obtain a residue. The residue was purified by column chromatography (PE:EA=50:1) to give 1-benzyl 4-(tert-butyl)(S)-2-(2-((tert-butyldiphenylsilyl)oxy)ethyl)piperazine-1,4-dicarboxylate (13.806 g, yield 73%) as a colorless oil. MS: m / z=688.2 (M+H + ,ESI+).

[0242] 1 H NMR(400MHz, CDCl3) δ7.64(d,J=6.7Hz,4H),7.35(ddd,J=40.3,20.2,11.1Hz,11H),5.11(d,J=17.9Hz,2H), 4.44(s,1H),4.02-3.56(m,5H),2.92(td,J=12.8,2.9Hz,3H),1.89-1.70(m,2H),1.40(s,9H),1.03(s,9H).

[0243] Step 3) Intermediate 4 of Example 5: 1-benzyl 4-(tert-butyl)(S)-2-(2-hydroxyethyl)piperazine-1,4-dicarboxylic acid Preparation of acid esters After adding tetrabutylammonium fluoride (TBAF) (91.7 mL, including 1M in THF) to a solution of 1-benzyl 4-(tert-butyl)(S)-2-(2-((tert-butyldiphenylsilyl)oxy)ethyl)piperazine-1,4-dicarboxylate (13.806 g, 22.93 mmol) in THF (150 mL), the mixture was stirred for 2 hours under N2 gas and room temperature. LCMS showed that the starting material was consumed and the target compound was detected. The reaction mixture was diluted with H2O (100 mL) and extracted with EA (150 mL×3). The combined organic layer was washed with brine (150 mL) and dried over Na2SO4. The residue was obtained by filtration and vacuum concentration. The residue was purified by column chromatography (PE:EA=5:1) to give 1-benzyl 4-(tert-butyl)(S)-2-(2-hydroxyethyl)piperazine-1,4-dicarboxylate (7.4 g, yield 92%) as a colorless oil. MS: m / z=387.2 (M+H +,ESI+).

[0244] Step 4) Intermediate 5 of Example 5: 1-benzyl 4-(tert-butyl) (S)-2-(2-hydroxyethyl)piperazine-1,4-dicarboxylic acid Preparation of acid esters

[0245] At -78°C, a solution of dimethyl sulfoxide (5.51 mg, 70.7 mmol) in dichloromethane (50 mL) was added to a solution of oxalyl chloride (6.41 g, 50.5 mmol) in DCM (60 mL). After stirring at -78°C for 0.5 h, a solution of (S)-1-benzyl 4-tert-butyl 2-(2-hydroxyethyl)piperazine-1,4-dicarboxylate (7.4 g, 20.2 mmol) in DCM (20 mL) was added to the mixture. After stirring at -78°C for 3 h, TEA (15.3 g, 151.5 mmol) was added dropwise. The reaction mixture was further heated at 25°C for 0.5 h. LCMS indicated that the starting material was consumed and the target compound was detected. The reaction mixture was diluted with H2O (60 mL) and extracted with DCM (60 mL×3). The combined organic layers were washed with brine (30 mL) and dried over Na2SO4. The residue was obtained by filtration and vacuum concentration. The residue was purified by column chromatography (PE:EA=5:1) to give 1-benzyl 4-(tert-butyl)(S)-2-(2-hydroxyethyl)piperazine-1,4-dicarboxylate (5.4 g, yield 74.5%) as a colorless oil. MS: m / z=363.2 (M+H + ,ESI+).

[0246] Step 5) Intermediate 6 of Example 5: 1-benzyl 4-(tert-butyl)(2S)-2-(2-hydroxypropyl)piperazine-1,4-dicarboxylic acid Preparation of acid esters

[0247] At 0°C, 1M CH3MgBr (1M in THF, 14.9mL) was added to a stirred solution of 1-benzyl 4-(tert-butyl) (S)-2-(2-hydroxyethyl) piperazine-1,4-dicarboxylate (5.4g, 14.9mmol) in dry THF (25mL). The reactants were stirred for 2 hours at 25°C, and LCMS indicated that the starting material was consumed and the target compound was detected. The reaction mixture was quenched with saturated NH4Cl (40mL) and extracted with DCM (60mL×3). The combined organic layers were washed with brine (30mL) and dried with Na2SO4. The residue was obtained by filtration and vacuum concentration. The residue was purified by column chromatography (PE:EA=5:1) to obtain 1-benzyl 4-(tert-butyl) (2S)-2-(2-hydroxypropyl) piperazine-1,4-dicarboxylate (2g, yield 35.5%) as a colorless oil. MS: m / z=401.2 (M+Na+, ESI+).

[0248] Step 6) Preparation of Intermediate 7 of Example 5: (3S)-3-(2-hydroxypropyl)piperazine-1-carboxylic acid tert-butyl ester

[0249] Pd(OH)2 (200 mg) was added to a suspension of 1-benzyl 4-(tert-butyl) (2S)-2-(2-hydroxypropyl) piperazine-1,4-dicarboxylate (2 g, 5.29 mmol) in MeOH (20 mL), and the mixture was stirred for 2 hours under a hydrogen atmosphere at 25°C. The mixture was filtered through a celite pad and the filtrate was evaporated to give (3S)-3-(2-hydroxypropyl) piperazine-1-carboxylic acid tert-butyl ester (1.01 g, yield 83%) as a colorless oil. MS: m / z = 245.2 (M+H + ,ESI+).

[0250] Step 7) Intermediate 8 of Example 5: (3S)-3-(2-((7-chloro-8-fluoro-2-(methylthio)-4-oxo-3,4-di Preparation of tert-butyl (4,3-d)pyrido[4,3-d]pyrimidin-5-yl)oxy)propyl)piperazine-1-carboxylate

[0251] NaH (553 mg, 13.8 mmol, 60% suspension in mineral oil) was added to a mixture of (3S)-3-(2-hydroxypropyl)piperazine-1-carboxylic acid tert-butyl ester (1.01 g, 4.14 mmol) in THF (15 mL), and the mixture was stirred at 0°C for 0.5 h. 5,7-Dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4(3H)-one (1.29 g, 4.61 mmol) was added to the mixture, and the mixture was further stirred at 60°C for 1 h. TLC (EA / MeOH=15 / 1) indicated consumption of SM1 (Rf=0.2) and formation of a new spot (Rf=0.4). The reaction mixture was quenched with water (40 mL) and extracted with ethyl acetate (50 mL×3). The combined organic layers were washed with 60 mL of brine and dried over Na2SO4, filtered and concentrated in vacuo to give a residue. The residue was purified by column chromatography (PE / EA=0 / 1) to give (3S)-tert-butyl 3-(2-((7-chloro-8-fluoro-2-(methylthio)-4-oxo-3,4-dihydropyrido[4,3-d]pyrimidin-5-yl)oxy)propyl)piperazine-1-carboxylate (934 mg, yield 46.3%) as a white solid. MS: m / z=488.1 (M+H + ,ESI+).

[0252] Step 8) Intermediate 9 of Example 5: (3S)-3-(2-((7-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalene- 1-yl)-8-fluoro-2-(methylthio)-4-oxo-3,4-dihydropyrido[4,3-d]pyrimidin-5-yl)oxy)propyl)piperazine-1- Preparation of tert-butyl carboxylate

[0253] A mixture of (3S)-tert-butyl 3-(2-((7-chloro-8-fluoro-2-(methylthio)-4-oxo-3,4-dihydropyrido[4,3-d]pyrimidin-5-yl)oxy)propyl)piperazine-1-carboxylate (934 mg, 1.92 mmol), 2-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (691.2 mg, 1.92 mmol), Ruphos-Pd-G3 (481.5 mg, 0.576 mmol) and K3PO4 (1.22 g, 5.76 mmol) in dioxane / H2O (8 mL / 2 mL) was stirred under N2 gas at 90°C for 3 hours. LCMS indicated that the starting material was consumed and the target compound was detected. The reaction mixture was diluted with water (30 mL) and extracted with DCM (40 mL × 2). The combined organic layer was washed with 60 mL of brine and dried with Na2SO4, filtered and concentrated in vacuo to give a residue. The residue was purified by column chromatography (PE / EA=0 / 1) to give (3S)-3-(2-((7-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalene-1-yl)-8-fluoro-2-(methylthio)-4-oxo-3,4-dihydropyrido[4,3-d]pyrimidin-5-yl)oxy)propyl)piperazine-1-carboxylic acid tert-butyl ester (235 mg, yield 20%) as a light yellow solid. MS: m / z=686.3 (M+H + ,ESI+).

[0254] Step 9) Intermediate 10 of Example 5: (6aS)-2-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalene-1- 1-fluoro-5-methyl-12-(methylthio)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaaza Preparation of tert-butyl benzo[4,5]cyclooctano[1,2,3-de]naphthalene-8-carboxylate

[0255] To a solution of tert-butyl (3S)-3-(2-((7-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-8-fluoro-2-(methylthio)-4-oxo-3,4-dihydropyrido[4,3-d]pyrimidin-5-yl)oxy)propyl)piperazine-1-carboxylate (100 mg, 0.15 mmol) in DCM (4 mL) was added DIEA (169 mg, 1.31 mmol) and BOPCl (114 mg, 0.45 mmol). The mixture was then stirred at 25° C. for 16 hours. LCMS indicated that the starting material was consumed and the target compound was detected. The reaction mixture was diluted with water (20 mL) and extracted with DCM (30 mL×2). The combined organic layers were washed with 60 mL of brine and dried over Na2SO4, filtered and concentrated in vacuo to give a residue. The residue was purified by column chromatography (PE / EA=1 / 1) to give (6aS)-2-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-1-fluoro-5-methyl-12-(methylthio)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctane[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester (40 mg, yield 41.2%) as a white solid. MS: m / z=668.3 (M+H + ,ESI+).

[0256] Step 10) Intermediate 11 of Example 5: (6aS)-2-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalene-1- 1-fluoro-5-methyl-12-(methylsulfonyl)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentahydro- Preparation of tert-butyl azabenzo[4,5]cyclooctano[1,2,3-de]naphthalene-8-carboxylate

[0257] Potassium peroxymonosulfonate (Oxone) (190.3 mg, 0.55 mmol) was added to a solution of (6aS)-2-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-1-fluoro-5-methyl-12-(methylthio)-56,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cycloocta[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester (40 mg, 0.11 mmol) in MeCN / H2O (1 mL / 1 mL) and stirred at 25°C for 1 hour. LCMS indicated that the starting material was consumed and the desired mass was detected. The reaction mixture was diluted with water (20 mL) and extracted with DCM (30 mL×2). The combined organic layers were washed with 20 mL of brine and dried over Na2SO4, filtered and concentrated in vacuo to give (6aS)-2-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl-1-fluoro-5-methyl-12-(methylsulfonyl)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctano[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester (50 mg, 65% yield of crude compound) as a white solid. MS: m / z=700.2 (M+H + ,ESI+).

[0258] Step 10) Intermediate 12 of Example 5: (6aS)-2-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalene-1- 1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methyl-5,6,6a,7, 9,10-Hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctano[1,2,3-de]naphthalene-8-carboxylic acid tert- Preparation of Butyl Ester

[0259] To a solution of (6aS)-2-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-1-fluoro-5-methyl-12-(methylsulfonyl)-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cycloocta[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester (50 mg, 0.071 mmol) and ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol (16.9 mg, 0.106 mmol) in dry THF (2 mL) was added t-BuONa (11 mg, 0.14 mmol) and the mixture was stirred at -60°C under N2 atmosphere for 1 hour. LCMS indicated that the starting material was consumed and the desired mass was formed. The reaction mixture was quenched with water (60 mL) and extracted with ethyl acetate (60 mL×3). The combined organic layer was washed with 60 mL of brine and dried over Na2SO4, filtered and concentrated in vacuo to give a residue. The residue was purified by preparative (pre)-TLC (PE / EA=0 / 1) to give (6aS)-2-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methyl-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cyclooctane[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester (20 mg, 36% yield). LC-MS MS: m / z = 779.3 (M+H + ,ESI+).

[0260] Step 11) Example 5: 5-ethyl-6-fluoro-4-((6aS)-1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidone Alloxazin-7a(5H)-yl)methoxy)-5-methyl-6,6a,7,8,9,10-hexahydro-5H-4-oxa-3,8,10a,11,13-pentahydro- Preparation of heterobenzo[4,5]cyclooctano[1,2,3-de]naphthalen-2-yl)naphthalen-2-ol

[0261] To a solution of (6aS)-2-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methyl-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cycloocta[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester (20 mg, 0.25 mmol) in EA (1 mL) was added 3 M HCl (2 mL) and the mixture was stirred at 0° C. for 1 min. After 1 h, LCMS indicated consumption of starting material and formation of the desired mass. The mixture was concentrated in vacuo to give a residue. The residue was purified by preparative (pre)-HPLC to give (6aS)-2-(8-ethyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-1-fluoro-12-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5-methyl-5,6,6a,7,9,10-hexahydro-8H-4-oxa-3,8,10a,11,13-pentaazabenzo[4,5]cycloocta[1,2,3-de]naphthalene-8-carboxylic acid tert-butyl ester (2.63 mg, yield 16.1%).

[0262] 1 H NMR(400MHz,MeOD)δ8.49(brs,1H,FA),7.71-7.57(m,1H),7.32-6.90(m,3H),5.55-5.20 (m,2H),4.64-4.52(m,1H),4.41(dd,J=59.4,11.3Hz,2H),4.02(dd,J=30.6,12.5Hz,1H) ,3.66-3.40(m,3H),3.26-2.92(m,5H),2.91-2.65(m,2H),2.60-2.34(m,3H),2.28-2.08 (m,4H),2.05-1.92(m,1H),1.88-1.74(m,1H),1.47(d,J=5.9Hz,3H),0.98-0.82(m,3H). MS: m / z=635.4(M+H + ,ESI+).

[0263] The synthetic reaction formula of Example 6

[0264]

[0265] Example 6: Preparation of 4-(3-chloro-1-fluoro-13-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-5a,6,7,8,9,10-hexahydro-5H-azepin[2',1':3,4][1,4]oxazepin[5,6,7-de]quinazolin-2-yl)-7-fluorobenzo[d]thiazole-2-amine

[0266] Step 1) Preparation of Intermediate 2 of Example 6: 2-amino-4-bromo-5-chloro-3,6-difluorobenzoyl chloride

[0267] At 50°C, 2-amino-4-bromo-5-chloro-3,6-difluorobenzoic acid (2 g, 6.99 mmol) was added to a solution of SOCl 2( The mixture was stirred for 3 hours with 20 mL of 4-bromo-5-chloro-3,6-difluorobenzoyl chloride (2.13 g, 100% yield) as a brown oil. The crude product was used directly in the next step without further purification.

[0268] Step 2) Preparation of Intermediate 3 of Example 6: 7-bromo-6-chloro-5,8-difluoro-2-mercaptoquinazoline-4-ol

[0269] A solution of 2-amino-4-bromo-5-chloro-3,6-difluorobenzoyl chloride (2.13 g, 6.98 mmol) dissolved in acetone (10 mL) was added to a mixture of NH4SCN (1.6 g, 21.02 mmol) dissolved in acetone (20 mL). The reaction mixture was stirred for 1 hour at room temperature. The reaction mixture was quenched with water (200 mL) and the solution was extracted with EtOAc (100 mL×3). The combined organic phases were washed with brine (50 mL) and dried with Na2SO4(s), filtered and concentrated to give the desired product, 7-bromo-6-chloro-5,8-difluoro-2-mercaptoquinazoline-4-ol (2.78 g, crude compound), as a yellow solid. MS: m / z=326.9 (M+H + , ESI+). The crude product was used directly in the next step without further purification.

[0270] Step 3) Preparation of Intermediate 4 of Example 6: 7-bromo-6-chloro-5,8-difluoro-2-(methylthio)quinazoline-4-olAt room temperature, NaOH (1M, 17mL, 17mmol) and MeI (1.06mL, 17.05mmol) were added to a mixture of MeOH (30mL) containing 7-bromo-6-chloro-5,8-difluoro-2-mercaptoquinazolin-4-ol (2.78g, 8.53mmol). The reaction mixture was stirred for 1.5 hours at room temperature. The reaction mixture was adjusted to pH=7 with 1M HCl, and then the mixture was diluted with water (150mL) and extracted with EtOAc (100mL×3). The combined organic phases were washed with brine (50mL) and dried over Na2SO4(s), filtered and concentrated to obtain a residue, which was purified by silica gel column chromatography and purified with PE / EA=5:1 to obtain the desired product 7-bromo-6-chloro-5,8-difluoro-2-(methylthio)quinazolin-4-ol (1.85g, yield 64%) as a yellow solid. MS: m / z = 340.9 (M + H + ,ESI+).

[0271] Step 4) Intermediate 5 of Example 6: 5-(azepan-2-ylmethoxy)-7-bromo-6-chloro-8-fluoro-2-(methyl Preparation of (4-(2-thio)quinazoline-4-ol

[0272] At 0°C, NaH (60% in oil, 280 mg, 11.67 mmol) was added to a mixture of THF (5 mL) containing azepan-2-ylmethanol hydrochloride (387 mg, 2.34 mmol). The reaction mixture was stirred for 10 minutes under N2 gas and 0°C, and a THF (15 mL) solution containing 7-bromo-6-chloro-5,8-difluoro-2-(methylthio)quinazolin-4-ol (794 mg, 2.33 mmol) was added to the mixture. The reaction mixture was stirred for 16 hours under N2 gas and 25°C. The reaction mixture was quenched with concentrated NH4Cl(aq) (150 mL) and the mixture was extracted with EA (80 mL×3). The organic phase was washed with brine (50 mL), dried over Na2SO4(s), filtered and concentrated in vacuo to give a yellow solid. The yellow solid was purified by silica gel column chromatography and eluted with DCM / MeOH=5:1 to give the desired product 5-(azepan-2-ylmethoxy)-7-bromo-6-chloro-8-fluoro-2-(methylthio)quinazolin-4-ol (706 mg, yield 67.4%) as a yellow solid. MS: m / z=451.9 (M+H + ,ESI+).

[0273] Step 5) Intermediate 6 of Example 6: 2-bromo-3-chloro-1-fluoro-13-(methylthio)-5a,6,7,8,9,10-hexahydro- Preparation of 5H-aza[2',1':3,4][1,4]oxaza[5,6,7-de]quinazoline

[0274] At 0°C under N2, BOPCl (1.2 g, 4.71 mmol) was added to a mixture of 5-(azepan-2-ylmethoxy)-7-bromo-6-chloro-8-fluoro-2-(methylthio)quinazolin-4-ol (706 mg, 1.57 mmol) and DIEA (2.4 mL, 14.08 mmol) in DCM (20 mL). The reaction mixture was stirred for 2 hours at room temperature under N2. The reaction was diluted with water (200 mL) and the solution was extracted with EtOAc (100 mL×3). The combined organic phases were washed with brine (80 mL) and dried over Na2SO4(s), filtered and concentrated in vacuo to give a yellow solid. The yellow solid was purified by silica gel column chromatography using PE / EA=10:1 as eluent to give the desired product, 2-bromo-3-chloro-1-fluoro-13-(methylthio)-5a,6,7,8,9,10-hexahydro-5H-azepin[2',1':3,4][1,4]oxazepin[5,6,7-de]quinazoline (640 mg, yield 74.5%), as a yellow solid. MS: m / z=433.9 (M+H + ,ESI+).

[0275] Step 6) Intermediate 7 of Example 6: (4-(3-chloro-1-fluoro-13-(methylthio)-5a,6,7,8,9,10-hexahydro- 5H-Aza[2',1':3,4][1,4]oxaza[5,6,7-de]quinazolin-2-yl)-7-fluorobenzo[d]thiazole- Preparation of tert-butyl 2-aminocarbamate

[0276] Ruphos-Pd-G3 (174 mg, 0.21 mmol) was added to a mixture of dioxane / H2O (9:1, 5 mL) containing 2-bromo-3-chloro-1-fluoro-13-(methylthio)-5a,6,7,8,9,10-hexahydro-5H-azepino[2',1':3,4][1,4]oxazepino[5,6,7-de]quinazoline (300 mg, 0.7 mol), K3PO4 (443 mg, 2.1 mmol) and tert-butyl (7-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[d]thiazol-2-yl)carbamate (548 mg, 1.39 mmol). The solution was purged with N2 three times and stirred at 100°C under N2 gas for 2 hours. The reactant was diluted with water (100mL), and the solution was extracted with EtOAc (50mL×3). The combined organic phase was washed with brine (50mL), dried with Na2SO4(s), filtered and concentrated in vacuo to give a yellow solid. The yellow solid was purified by silica gel column chromatography eluted with PE / EA=10:1 to give the desired product (4-(3-chloro-1-fluoro-13-(methylthio)-5a,6,7,8,9,10-hexahydro-5H-azepine [2',1':3,4][1,4]oxazepine [5,6,7-de]quinazoline-2-yl)-7-fluorobenzo [d] thiazole-2-yl) tert-butyl carbamate (454mg, crude compound) as a yellow solid. MS: m / z=620.3 (M+H + ,ESI+).

[0277] Step 7) Intermediate 8 of Example 6: (4-(3-chloro-1-fluoro-13-(methylsulfonyl)-5a,6,7,8,9,10-hexa Hydrogen-5H-azepino[2',1':3,4][1,4]oxazepino[5,6,7-de]quinazolin-2-yl)-7-fluorobenzo[d]thiol Preparation of tert-butyl oxazol-2-yl)carbamate

[0278] Potassium peroxymonosulfonate (Oxone) (2.2 g, 3.58 mmol) was added to a mixture of ACN:H2O (3:1, 6 mL) containing tert-butyl (4-(3-chloro-1-fluoro-13-(methylthio)-5a,6,7,8,9,10-hexahydro-5H-azepine [2',1':3) at 0°C. The reaction mixture was stirred at room temperature for 5 hours. The reaction was quenched with aq. Na2SO3 (60 mL) and extracted with EtOAc (30 mL×3). The combined organic phases were washed with aq. Na2SO 3 (20 mL × 2) and brine (20 mL) and dried over Na2SO4 (s), filtered and concentrated in vacuo to give the desired product (4-(3-chloro-1-fluoro-13-(methylsulfonyl)-5a,6,7,8,9,10-hexahydro-5H-azepino[2',1':3,4][1,4]oxazepino[5,6,7-de]quinazolin-2-yl)-7-fluorobenzo[d]thiazol-2-yl)carbamic acid tert-butyl ester (434 mg, crude compound) as a yellow solid. MS: m / z = 652.2 (M+H + ,ESI+).

[0279] Step 8) Intermediate 9 of Example 6: (4-(3-chloro-1-fluoro-13-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrole Oxazine-7a(5H)-yl)methoxy)-5a,6,7,8,9,10-hexahydro-5H-azepin[2',1':3,4][1,4]oxazepin Preparation of tert-butyl (5,6,7-de)quinazolin-2-yl)-7-fluorobenzo[d]thiazol-2-yl)carbamate

[0280] NaH (60% in oil, 39 mg, 1.62 mmol) was added to a mixture of ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol (51 mg, 0.32 mmol) in THF (2 mL) at 0° C. The reaction mixture was stirred for 10 min under N2 gas at 0° C. and tert-butyl (4-(3-chloro-1-fluoro-13-(methylsulfonyl)-5a,6,7,8,9,10-hexahydro-5H-azepino[2',1':3,4][1,4]oxazepino[5,6,7-de]quinazolin-2-yl)-7-fluorobenzo[d]thiazol-2-yl)carbamate (210 mg, 0.32 mmol) in THF (1 mL) was added to the solution. Under N2 gas and 25 ℃, the reaction mixture was stirred for 1 hour. The reaction mixture was quenched with concentrated NH4Cl(aq) (40 mL), and the mixture was extracted with EA (20 mL×3). The organic phase was washed with brine (20 mL) and dried over Na2SO4(s), filtered and concentrated in vacuo to give a brown solid. The brown solid was purified by silica gel column chromatography and eluted with PE / EA=1:2 to give the desired product (tert-butyl 4-(3-chloro-1-fluoro-13-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5a,6,7,8,9,10-hexahydro-5H-azepin[2',1':3,4][1,4]oxazepin[5,6,7-de]quinazolin-2-yl)-7-fluorobenzo[d]thiazol-2-yl)carbamate (65 mg, yield 27.6%) as a white solid. m / z=731.3 (M+H + ,ESI+).

[0281] Step 9) Example 6: 4-(3-chloro-1-fluoro-13-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazine-7a(5H)- 1-(2-(2-(2-nitrophenyl)methoxy)-5a,6,7,8,9,10-hexahydro-5H-azepin[2',1':3,4][1,4]oxazepin[5,6,7-de] Preparation of quinazolin-2-yl)-7-fluorobenzo[d]thiazol-2-amine

[0282] To a mixture of tert-butyl (4-(3-chloro-1-fluoro-13-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-5a,6,7,8,9,10-hexahydro-5H-azepino[2',1':3,4][1,4]oxazepino[5,6,7-de]quinazolin-2-yl)-7-fluorobenzo[d]thiazol-2-yl)carbamate (65 mg, 0.089 mmol) in DCM (1.5 mL) was added HCl / dioxane (1.5 mL) at room temperature. The reaction mixture was stirred at room temperature for 23 hours. The reaction was concentrated to dryness in vacuo to give a yellow solid. The yellow solid was purified by preparative HPLC (Waters 2767 / Qda, column: SunFireSunfire C18, 19*250 mm, 10 um; mobile phase A: 0.1% FA / H2O, B: ACN, flow rate: 20 mL / min, gradient: 20% to 30%) to give the desired product Example 6 (22.82 mg, yield 40.7%) as a white solid.

[0283] 1 H NMR(400MHz, DMSO-d6)δ8.33(s,0.31H,FA),7.96-7.85(m,2H),7.20(t,J=6.6Hz,1H),7.04(q,J=8.0Hz,1H),5.27(d,J=55.1Hz,1H),4 .87-4.59(m,2H),4.48-4.37(m,1H),4.11-3.95(m,3H),3.14-2.96(m,4H),2.87-2.76(m,1H),2.14-1.55(m,13H),1.30-1.16(m,1H). m / z=631.4(M+H + ,ESI+).

[0284] The synthetic reaction formula of Example 7

[0285]

[0286] Example 7: Preparation of 4-((14aS)-12-chloro-10-fluoro-8-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepin[3',4':3,4][1,4]oxazepin[5,6,7-de]quinazolin-11-yl)-7-fluorobenzo[d]thiazole-2-amine

[0287] Step 1) Intermediate 2 of Example 7: (S)-3-(3-(benzyloxy)-2-((tert-butyloxycarbonyl)amino)propoxy) Preparation of tert-butyl propionate

[0288] Tert-butyl acrylate (72 mL) was added to a t-BuOH (72 mL) solution containing (S)-(1-(benzyloxy)-3-hydroxypropane-2-yl)carbamate (9 g, 31.99 mmol) and Cs2CO3 (10.6 g, 32.53 mmol). The reaction mixture was stirred for 16 hours at room temperature. The reaction mixture was diluted with water (250 mL), and the solution was extracted with ethyl acetate (150 mL×3). The combined organic phase was washed with brine (50 mL) and dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by silica gel column chromatography and purified with petroleum ether (pet ether) / ethyl acetate = 10:1 to obtain the desired product (S)-tert-butyl 3-(3-(benzyloxy)-2-((tert-butoxycarbonyl)amino)propoxy)propanoate (10.05 g, yield 76.8%) as a colorless oil. MS: m / z = 410.2 (M+H + ,ESI+).

[0289] Step 2) Preparation of Intermediate 3 of Example 7: (S)-3-(2-amino-3-(benzyloxy)propoxy)propanoic acid

[0290] A mixture of HCl / dioxane (20 mL) containing tert-butyl (S)-3-(3-(benzyloxy)-2-((tert-butoxycarbonyl)amino)propoxy)propanoate (2 g, 4.89 mmol) was stirred at room temperature for 2 hours. The reaction was concentrated in vacuo to give the crude product (S)-3-(2-amino-3-(benzyloxy)propoxy)propanoic acid (1.81 g, HCl salt) as a yellow oil. MS: m / z = 254.0 (M+H + , ESI+). The crude product was used directly in the next step without further purification.

[0291] Step 3) Preparation of Intermediate 4 of Example 7: (S)-3-((Benzyloxy)methyl)-1,4-oxazepan-5-one

[0292] HATU (3.08 g, 8.1 mmol) was added to a DCM (18 mL) solution containing (S)-3-(2-amino-3-(benzyloxy)propoxy)propanoic acid (1.71 g, 6.72 mmol) and TEA (7.5 mL, 53.96 mmol). The reaction mixture was stirred for 1 hour at room temperature. The reaction mixture was diluted with water (150 mL), and the solution was extracted with ethyl acetate (50 mL×3). The combined organic phases were washed with brine (50 mL) and dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by silica gel column chromatography and purified with petroleum ether / ethyl acetate = 1:1 to give the desired product (S)-3-((benzyloxy)methyl)-1,4-oxazepan-5-one (1.43 g, yield 90.5%) as a yellow solid. MS: m / z = 236.1 (M+H+ ,ESI+).

[0293] Step 4) Preparation of Intermediate 5 of Example 7: (S)-3-((Benzyloxy)methyl)-1,4-oxazepane

[0294] At 0°C and N2 gas conditions, THF (5mL) containing (S)-3-((benzyloxy)methyl)-1,4-oxazepan-5-one (1.33g, 5.66mmol) was added to a mixture of THF (5mL) containing LAH (1M in THF, 11.5mL, 11.33mmol). The reaction mixture was stirred for 5 hours under N2 gas and 25°C. At 0°C, the reaction mixture was quenched with H2O (1mL), and Na2SO4 was added to the solution. The mixture was stirred for 10 minutes at room temperature. The mixture was filtered and the filtrate was collected and concentrated in vacuo. The residue was purified by silica gel column chromatography and purified with petroleum ether / ethyl acetate = 1:2 to obtain the desired product (S)-3-((benzyloxy)methyl)-1,4-oxazepane (1g, yield 80%) as a yellow oil. MS: m / z = 222.2 (M+H + ,ESI+).

[0295] Step 5) Preparation of Intermediate 6 of Example 7: (R)-(1,4-oxazepan-3-yl)methanol

[0296] Pd / C (0.5 g, 60%) was added to a mixture of MeOH (10 mL) containing (S)-3-((benzyloxy)methyl)-1,4-oxazepane (0.95 g, 4.30 mmol) at room temperature. The reaction mixture was stirred for 40 hours under H2 (15 psi) and 25°C. The reactants were filtered through celite and the filtrate was collected and concentrated in vacuo to give a crude product of (R)-(1,4-oxazepane-3-yl)methanol (637 mg, crude compound) as a white solid. MS: m / z = 132.1 (M+H + , ESI+). The crude product was used directly in the next step without further purification.

[0297] Step 6) Intermediate 7 of Example 7: (S)-5-((1,4-oxazepan-3-yl)methoxy)-7-bromo-6- Preparation of chloro-8-fluoro-2-(methylthio)quinazolin-4-ol

[0298] NaH (included in oil at 60%, 275 mg, 11.46 mmol) was added to a mixture of (R)-(1,4-oxazepan-3-yl)methanol (300 mg, 2.29 mmol) in THF (4 mL) at 0°C. The reaction mixture was stirred for 10 minutes under N2 gas and 0°C, and the solution was added to a solution of 7-bromo-6-chloro-5,8-difluoro-2-(methylthio)quinazolin-4-ol (779 mg, 2.29 mmol) in THF (4 mL). The reaction mixture was stirred for 16 hours under N2 gas and 25°C. The reaction mixture was quenched with concentrated NH4Cl(aq) (150 mL), and the mixture was extracted with ethyl acetate (50 mL×3). The organic phase was washed with brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by silica gel column chromatography and eluted with DCM / MeOH=5:1 to give the desired product (S)-5-((1,4-oxazepan-3-yl)methoxy)-7-bromo-6-chloro-8-fluoro-2-(methylthio)quinazolin-4-ol (319 mg, yield 30.9%) as a yellow solid. MS: m / z=454.0 (M+H + ,ESI+).

[0299] Step 7) Intermediate 8 of Example 7: (S)-11-bromo-12-chloro-10-fluoro-8-(methylthio)-4,5,14,14a-tetra Preparation of Hydrogen-1H,3H-[1,4]Oxazepin[3',4':3,4][1,4]Oxazepin[5,6,7-de]quinazoline

[0300] Under N2 gas and 0°C, BOPCl (540 mg, 2.12 mmol) was added to a mixture of (S)-5-((1,4-oxazepan-3-yl)methoxy)-7-bromo-6-chloro-8-fluoro-2-(methylthio)quinazolin-4-ol (319 mg, 0.71 mmol) and DIEA (1 mL, 6.05 mmol) in DCM (5 mL). The reaction mixture was stirred for 16 hours under N2 gas and room temperature. The reaction was quenched with water (150 mL) and the solution was extracted with ethyl acetate (70 mL×3). The combined organic phases were washed with brine (50 mL) and dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by silica gel column chromatography and eluted with petroleum ether / ethyl acetate = 4:1 to obtain the desired product (S)-11-bromo-12-chloro-10-fluoro-8-(methylthio)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepin[3',4':3,4][1,4]oxazepin[5,6,7-de]quinazoline (142 mg, yield 46.4%) as a yellow solid. MS: m / z = 435.9 (M+H + ,ESI+).

[0301] Step 8) Intermediate 9 of Example 7: (4-((14aS)-12-chloro-10-fluoro-8-(methylthio)-4,5,14,14a- Tetrahydro-1H,3H-[1,4]oxazepino[3',4':3,4][1,4]oxazepino[5,6,7-de]quinazolin-11-yl)- Preparation of tert-butyl 7-fluorobenzo[d]thiazol-2-yl)carbamate

[0302] Ruphos-Pd-G3 (82 mg, 0.10 mmol) was added to a mixture of (S)-11-bromo-12-chloro-10-fluoro-8-(methylthio)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepin[3',4':3,4][1,4]oxazepin[5,6,7-de]quinazoline (142 mg, 0.33 mol), K3PO4 (209 mg, 0.98 mmol) and tert-butyl (7-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[d]thiazol-2-yl)carbamate (259 mg, 0.66 mmol) in dioxane / H2O (9:1, 5 mL). The solution was purged with N2 three times and stirred for 2 hours under N2 gas and 100°C. The reactant was diluted with water (100 mL) and the solution was extracted with ethyl acetate (50 mL×3). The combined organic phase was washed with brine (50 mL) and dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by silica gel column chromatography and eluted with petroleum ether / ethyl acetate = 1:1 to obtain the desired product (4-((14aS)-12-chloro-10-fluoro-8-(methylthio)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepine [3',4':3,4][1,4]oxazepine [5,6,7-de]quinazolin-11-yl)-7-fluorobenzo [d] thiazol-2-yl) carbamic acid tert-butyl ester (143 mg, yield 70%) as a yellow solid. MS: m / z = 622.2 (M + H + ,ESI+).

[0303] Step 9) Intermediate 10 of Example 7: (4-((14aS)-12-chloro-10-fluoro-8-(methylsulfonyl)-4,5,14, 14a-Tetrahydro-1H,3H-[1,4]oxazepino[3',4':3,4][1,4]oxazepino[5,6,7-de]quinazoline-11- Preparation of tert-butyl (7-fluorobenzo[d]thiazol-2-yl)carbamate

[0304] Potassium peroxymonosulfonate (Oxone) (693 mg, 1.13 mmol) was added to a mixture of THF:H2O (1:1, 4 mL) containing tert-butyl (4-((14aS)-12-chloro-10-fluoro-8-(methylthio)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepin[3',4':3,4][1,4]oxazepin[5,6,7-de]quinazolin-11-yl)-7-fluorobenzo[d]thiazol-2-yl)carbamate (140 mg, 0.23 mmol) at 0°C. The reaction mixture was stirred at room temperature for 1 hour. The reaction was quenched with aq.Na2SO3 (80 mL) and the solution was extracted with ethyl acetate (30 mL×3). The combined organic phases were washed with aq. Na2SO3 (20 mL×2) and brine (20 mL) and dried over Na2SO4, filtered and concentrated in vacuo to give the crude product (tert-butyl (4-((14aS)-12-chloro)-10-fluoro-8-(methylsulfonyl)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepin[3',4':3,4][1,4]oxazepin[5,6,7-de]quinazolin-11-yl)-7-fluorobenzo[d]thiazol-2-yl)carbamate (140 mg, crude compound) as a yellow solid. MS: m / z=654.2 (M+H + ,ESI+).

[0305] Step 10) Intermediate 11 of Example 7: (4-((14aS)-12-chloro-10-fluoro-8-(((2R,7aS)-2-fluorotetra Hydrogen-1H-pyrrolizin-7a(5H)-yl)methoxy)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepine[3',4':3, tert-Butyl 4-[1,4]oxazepino[5,6,7-de]quinazolin-11-yl)-7-fluorobenzo[d]thiazol-2-yl)carbamate Preparation

[0306] To a mixture of ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methanol (34 mg, 0.21 mmol) in THF (2 mL) was added NaH (60% in oil, 26 mg, 1.08 mmol) at 0°C. The reaction mixture was stirred for 10 minutes under N2 gas at 0°C, and a solution of tert-butyl (4-((14aS)-12-chloro-10-fluoro-8-(methylsulfonyl)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepin[3',4':3,4][1,4]oxazepin[5,6,7-de]quinazolin-11-yl)-7-fluorobenzo[d]thiazol-2-yl)carbamate (140 mg, 0.21 mmol) in THF (3 mL) was added to the solution. The reaction mixture was stirred for 16 hours under N2 gas at 25°C. The reaction mixture was quenched with concentrated NH4Cl(aq) (40 mL), and the mixture was extracted with ethyl acetate (20 mL×3). The organic phase was washed with brine (20 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by preparative (Pre)-TLC (ethyl acetate, Rf = 0.4) to give the desired product (tert-butyl 4-((14aS)-12-chloro-10-fluoro-8-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepin[3',4':3,4][1,4]oxazepin[5,6,7-de]quinazolin-11-yl)-7-fluorobenzo[d]thiazol-2-yl)carbamate (30 mg, yield 19%) as a white solid. MS: m / z = 733.3 (M+H + ,ESI+).

[0307] Step 11) Example 7: 4-((14aS)-12-chloro-10-fluoro-8-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizine- 7a(5H)-yl)methoxy)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepina[3',4':3,4][1,4]oxazepin Preparation of 7-fluorobenzo[5,6,7-de]quinazolin-11-yl)-7-fluorobenzo[d]thiazol-2-amine

[0308] A mixture of tert-butyl (4-((14aS)-12-chloro-10-fluoro-8-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepin[3',4':3,4][1,4]oxazepin[5,6,7-de]quinazolin-11-yl)-7-fluorobenzo[d]thiazol-2-yl)carbamate (30 mg, 0.04 mmol) in HCl / dioxane (2 mL) was stirred at room temperature for 16 hours. The reaction was concentrated in vacuo. The residue was purified by preparative HPLC (Waters 2767 / Qd, column: SunFireSunfire C18, 19*250mm, 10um; mobile phase A: 0.1% FA / H2O, B: ACN, flow rate: 20mL / min; gradient: 20% to 30%) to give the desired product Example 7 (2.47 mg, yield: 9.5%) as a white solid.

[0309] 1 H NMR (400MHz, CD3OD) δ8.56 (s, 0.72H-FA), 7.24-7.14 (m, 1H), 6.97 (t, J = 8.8Hz, 1H), 5.39-5.25 (m, 1H), 5.11-5.00 (m, 1H), 4.83-4. 70(m,3H),4.62-4.54(m,1H),4.47-4.42(m,1H),4.39-4.12(m,4H),4.08-3.97(m,1H),3.85-3.76(m,1H),3.60-3.46(m,2H),3.14 -3.05(m,1H),2.36-2.17(m,4H),2.06-1.87(m,4H).MS:m / z=633.3(M+H + ,ESI+).

[0310] The synthetic reaction formula of Example 8

[0311]

[0312] Example 8: Preparation of 2-amino-4-((S)-12-chloro-10-fluoro-8-((1-(pyrrolidin-1-ylmethyl)cyclopropyl)methoxy)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepin[3',4':3,4][1,4]oxazepin[5,6,7-de]quinazolin-11-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile

[0313] Step 1) Intermediate 1 of Example 8: (S)-11-bromo-12-chloro-10-fluoro-8-(methylsulfonyl)-4,5,14, Preparation of 14a-tetrahydro-1H,3H-[1,4]oxazepino[3',4':3,4][1,4]oxazepino[5,6,7-de]quinazoline Preparation

[0314] Potassium peroxymonosulfonate (Oxone) (1.4 g, 2.28 mmol) was added to a mixture of THF / H2O (1:1, 8 mL) containing (S)-11-bromo-12-chloro-10-fluoro-8-(methylthio)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepino[3',4':3,4][1,4]oxazepino[5,6,7-de]quinazoline (197 mg, 0.45 mmol) at 0°C. The reaction mixture was stirred at room temperature for 1 hour. The reaction was quenched with aq.Na2SO3 (80 mL) and the solution was extracted with ethyl acetate (30 mL×3). The combined organic phases were washed with aq. Na2SO3 (20 mL×2) and brine (20 mL) and dried over anhydrous Na2SO4, filtered and concentrated in vacuo to give the crude product (S)-11-bromo-12-chloro-10-fluoro-8-(methylsulfonyl)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepin[3',4':3,4][1,4]oxazepin[5,6,7-de]quinazoline (227 mg, crude compound) as a yellow solid. MS: m / z=467.9 (M+H + , ESI+). The crude product was used directly in the next step without further purification.

[0315] Step 2) Intermediate 2 of Example 8: (S)-11-bromo-12-chloro-10-fluoro-8-((1-(pyrrolidin-1-ylmethyl) Cyclopropyl)methoxy)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepine[3',4':3,4][1,4]oxazepine 5,6,7-de]quinazoline

[0316] Under nitrogen and 0°C, NaH (60% in mineral oil, 20 mg, 0.51 mmol) was added to a stirred solution of (1-(pyrrolidin-1-ylmethyl)cyclopropyl)methanol (79 mg, 0.51 mmol) in THF (2 mL) and stirred for 20 minutes. Then, (S)-11-bromo-12-chloro-10-fluoro-8-(methylsulfonyl)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepin[3',4':3,4][1,4]oxazepin[5,6,7-de]quinazoline (160 mg, 0.34 mmol) was added and stirred for further 16 hours at room temperature. After the reaction was completed, aqueous ammonium chloride solution (20 mL) was added to terminate the reaction, and the organic layer was extracted by adding ethyl acetate (3×25 mL). The obtained organic layer was washed with brine (60 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The obtained residue was purified by silica gel column chromatography with an elution condition of methanol / DCM=1:6 to obtain the desired compound (S)-11-bromo-12-chloro-10-fluoro-8-((1-(pyrrolidin-1-ylmethyl)cyclopropyl)methoxy)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepin[3',4':3,4][1,4]oxazepin[5,6,7-de]quinazoline (50 mg, yield 26.6%).

[0317] Step 3) Intermediate 3 of Example 8: (4-((S)-12-chloro-10-fluoro-8-((1-(pyrrolidin-1-ylmethyl)cyclohexane propyl)methoxy)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepin[3',4':3,4][1,4]oxazepin tert-Butyl (5,6,7-de)quinazolin-11-yl)-3-cyano-7-fluorobenzo[b]thiophen-2-yl)carbamate

[0318] To a stirred solution of (S)-11-bromo-12-chloro-10-fluoro-8-((1-(pyrrolidin-1-ylmethyl)cyclopropyl)methoxy)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepin[3',4':3,4][1,4]oxazepin[5,6,7-de]quinazoline (50 mg, 0.09 mmol) in 1,4-dioxane (3 mL) was added (3-cyano-4 -(5,5-dimethyl-1,3,2-dioxaborinane-2-yl)-7-fluorobenzo[b]thiophene-2-yl)carbamic acid tert-butyl ester (73 mg, 0.18 mmol), K3PO4 (57 mg, 0.27 mmol), KF (10 mg, 0.18 mmol) and DPEphos-PdCl2 (14 mg, 0.02 mmol) were added, and stirred under nitrogen at 105° C. for 4 hours. After the reaction was completed, the reaction mixture was cooled to room temperature and concentrated under reduced pressure to obtain a residue. The residue was purified by reverse column chromatography (column: SunFire Sunfire C18, 19*250mm, 10um; mobile phase A: 0.1% HCOOH / H2O, B: acetonitrile; flow rate: 20mL / min; gradient: 35%-50%) to give the desired compound (tert-butyl 4-((S)-12-chloro-10-fluoro-8-((1-(pyrrolidin-1-ylmethyl)cyclopropyl)methoxy)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepin[3',4':3,4][1,4]oxazepin[5,6,7-de]quinazolin-11-yl)-3-cyano-7-fluorobenzo[b]thiophen-2-yl)carbamate (12 mg, yield 17.3%) as a white solid. MS: m / z = 753.4 (M + H + ,ESI+).

[0319] Step 4) Example 8: 2-amino-4-((S)-12-chloro-10-fluoro-8-((1-(pyrrolidin-1-ylmethyl)cyclopropane 4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepino[3',4':3,4][1,4]oxazepino[5, 6,7-de]quinazolin-11-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile

[0320] A stirred solution of tert-butyl (4-((S)-12-chloro-10-fluoro-8-((1-(pyrrolidin-1-ylmethyl)cyclopropyl)methoxy)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepin[3',4':3,4][1,4]oxazepin[5,6,7-de]quinazolin-11-yl)-3-cyano-7-fluorobenzo[b]thiophen-2-yl)carbamate (12 mg, 0.16 mmol) in a 1,4-dioxane / hydrochloric acid mixed solution (4M, 2.5 mL) was placed at room temperature for 6 hours. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to obtain a residue. The obtained residue was purified by preparative (Prep)-HPLC (column: XBridge C1819*250mm, 10um; mobile phase A: 0.1% NH4HCO3 / H2O, B: acetonitrile; flow rate: 20mL / min; gradient: 45% to 50%; retention time: 6.4 to 9 minutes in 16 minutes) to obtain the desired compound 2-amino-4-((S)-12-chloro-10-fluoro-8-((1-(pyrrolidin-1-ylmethyl)cyclopropyl)methoxy)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepin[3',4':3,4][1,4]oxazepin[5,6,7-de]quinazolin-11-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile (0.70 mg, yield was 6.7%).

[0321] 1 H NMR(400MHz,MeOD-d4)δ8.55(brs,2.25H,FA),7.22(dd,J=8.4,5.1Hz,1H),7.10-7.02(m,1H),4.53-4.44(m,4H),4.40-3.33(m,1H),4.30-4.1 6(m,3H),4.10-3.94(m,2H),3.91-3.74(m,2H),3.62-3.45(m,5H),2.28 -2.16(m,1H),2.08-1.92(m,5H),0.94-0.74(m,4H).MS:m / z=653.0(M+H + ,ESI+).

[0322] The synthetic reaction formula of Example 11

[0323]

[0324] Example 11: Preparation of 2-amino-4-(3-chloro-1-fluoro-13-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-5a,6,7,8,9,10-hexahydro-5H-azepina[2',1':3,4][1,4]oxazepina[5,6,7-de]quinazolin-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile

[0325] Step 1) Intermediate 1 of Example 11: 2-bromo-3-chloro-1-fluoro-13-(methylsulfonyl)-5a,6,7,8,9,10- Preparation of Hexahydro-5H-Aza[2',1':3,4][1,4]Oxaza[5,6,7-de]Quinazoline

[0326] Potassium peroxymonosulfonate (Oxone) (2.14 g, 3.48 mmol) was added to a mixture of THF / H2O (1:1, 4 mL) containing 2-bromo-3-chloro-1-fluoro-13-(methylthio)-5a,6,7,8,9,10-hexahydro-5H-azepino[2',1':3,4][1,4]oxazepino[5,6,7-de]quinazoline (300 mg, 0.70 mmol) at 0°C, and the reaction mixture was stirred at room temperature for 2 hours. The reaction was quenched with aq. Na2SO3 (80 mL) and the solution was extracted with ethyl acetate (30 mL×3). The combined organic phases were washed with aq. Na2SO3 (20 mL×2) and brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated in vacuo to give the crude product 2-bromo-3-chloro-1-fluoro-13-(methylsulfonyl)-5a,6,7,8,9,10-hexahydro-5H-azepin[2',1':3,4][1,4]oxazepin[5,6,7-de]quinazoline (365 mg, crude compound) as a yellow solid. MS: m / z=466.0 (M+H + , ESI+). The crude product was used directly in the next step without further purification.

[0327] Step 2) Intermediate 2 of Example 11: 2-bromo-3-chloro-1-fluoro-13-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrole Oxazine-7a(5H)-yl)methoxy)-5a,6,7,8,9,10-hexahydro-5H-azepin[2',1':3,4][1,4]oxazepin Preparation of [5,6,7-de]quinazoline

[0328] NaH (60% in oil, 95 mg, 3.96 mmol) was added to a mixture of ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol (125 mg, 0.79 mmol) in THF (2 mL) at 0° C. The reaction mixture was stirred for 10 minutes under N2 gas at 0° C., and a solution of 2-bromo-3-chloro-1-fluoro-13-(methylsulfonyl)-5a,6,7,8,9,10-hexahydro-13-(methylsulfonyl)-5a,6,7,8,9,10-hexahydro-5H-azepino[2',1':3,4][1,4]oxazepino[5,6,7-de]quinazoline (365 mg, 0.79 mmol) in THF (10 mL) was added to the solution. The reaction mixture was stirred for 16 hours under N2 gas and 25°C. The reaction mixture was quenched with concentrated NH4Cl(aq) (100 mL), and the mixture was extracted with ethyl acetate (50 mL×3). The organic phase was washed with brine (30 mL), dried over anhydrous Na2SO4, filtered and concentrated in vacuo to dryness. The residue was purified by silica gel column chromatography and eluted with petroleum ether / ethyl acetate = 1:2 to give the desired product 2-bromo-3-chloro-1-fluoro-13-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazine-7a(5H)-yl)methoxy)-5a,6,7,8,9,10-hexahydro-5H-azepin [2',1':3,4][1,4]oxazepin [5,6,7-de]quinazoline (319 mg, yield 74.7%) as a yellow solid. MS: m / z = 545.0 (M+H + ,ESI+).

[0329] Step 3) Intermediate 3 of Example 11: (4-(3-chloro-1-fluoro-13-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrole Oxazine-7a(5H)-yl)methoxy)-5a,6,7,8,9,10-hexahydro-5H-azepin[2',1':3,4][1,4]oxazepin Preparation of tert-butyl (5,6,7-de)quinazolin-2-yl)-3-cyano-7-fluorobenzo[b]thiophen-2-yl)carbamateA 5 mL toluene mixture containing 2-bromo-3-chloro-1-fluoro-13-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-5a,6,7,8,9,10-hexahydro-5H-azepino[2',1':3,4][1,4]oxazepino[5,6,7-de]quinazoline (220 mg, 0.41 mol), Cs2CO3 (396 mg, 1.22 mmol) and DPEphos-PdCl2 (73 mg, 0.10 mmol) was purged with N2 three times. The solution was stirred at 80° C. under N2 gas for 10 minutes. Next, tert-butyl (3-cyano-4-(5,5-dimethyl-1,3,2-dioxaborinane-2-yl)-7-fluorobenzo[b]thiophene-2-yl)carbamate (328 mg, 0.81 mmol) was added to the mixture. The mixture was purged with N2 for 3 times and stirred at 110°C for 2 hours. The reactant was diluted with water (200 mL) and the solution was extracted with ethyl acetate (100 mL×3). The combined organic phase was washed with brine (80 mL), dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by silica gel column chromatography and purified with petroleum ether / ethyl acetate = 1:2 to obtain the desired product (tert-butyl 4-(3-chloro-1-fluoro-13-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizine-7a(5H)-yl)methoxy)-5a,6,7,8,9,10-hexahydro-5H-azepino[2',1':3,4][1,4]oxazepino[5,6,7-de]quinazolin-2-yl)-3-cyano-7-fluorobenzo[b]thiophen-2-yl)carbamate (176 mg, crude compound) as a yellow oil. MS: m / z = 755.4 (M+H + ,ESI+).

[0330] Step 4) Example 11: 2-amino-4-(3-chloro-1-fluoro-13-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizine- 7a(5H)-yl)methoxy)-5a,6,7,8,9,10-hexahydro-5H-azepin[2',1':3,4][1,4]oxazepin[5, Preparation of 6,7-de]quinazolin-2-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile

[0331] A mixture of tert-butyl (4-(3-chloro-1-fluoro-13-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-5a,6,7,8,9,10-hexahydro-5H-azepino[2',1':3,4][1,4]oxazepino[5,6,7-de]quinazolin-2-yl)-3-cyano-7-fluorobenzo[b]thiophen-2-yl)carbamate (166 mg, 0.22 mmol) in HCl / dioxane (5 mL) was stirred at room temperature for 16 hours. The reaction was concentrated to dryness in vacuo. The residue was purified by preparative HPLC (Waters 2767 / Qda, column: SunFireSunfire C18, 19*250 mm, 10 um; mobile phase A: 0.1% FA / H2O, B: ACN, flow rate: 20 mL / min; gradient: 20%-30%) to obtain the desired product as a white solid. Example 11a (40 mg) and white solid Example 11b (17.08 mg). HPLC results showed that Example 11a was not clean and was purified by preparative HPLC (Waters 2767 / Qda, column: XBridge C18, 19*250 mm, 10 um; mobile phase A: 10 mmol of NH4HCO3 / H2O, B: ACN, flow rate: 20 mL / min, gradient: 30%-40%, retention time: 7-8 min, 16 min) to obtain pure Example 11a (15.29 mg) as a white solid. MS: m / z=655.4 (M+H + ,ESI+).

[0332] HNMR Example 11a ): 1 H NMR (400MHz, DMSO-d6) δ8.04 (s, 2H), 7.30-7.21 (m, 1H), 7.18-7.10 (m, 1H), 5.27 (d, J = 54.7Hz, 1H), 4.86-4.71 (m, 1H), 4.70-4. 58(m,1H),4.52-4.41(m,1H),4.12-4.02(m,2H),4.01-3.95(m,1H),3.15-2.94(m,4H),2.88-2.77(m,1H),2.14-1.45(m,14H).

[0333] HNMR Example 11b ): 1H NMR (400MHz, DMSO-d6) δ8.14(s,0.40H-FA),8.08(s,2H),7.29-7.21(m,1H),7.19-7.04(m,1H),5.29(d,J=53.8Hz,1H ),4.75-4.56(m,2H),4.39-4.25(m,1H),4.17-3.98(m,3H),3.18-2.99(m,4H),2.91-2.81(m,1H),2.20-1.50(m,14H).

[0334] The synthetic reaction formula of Example 12

[0335]

[0336] Example 12: Preparation of 2-amino-4-((14aS)-12-chloro-10-fluoro-8-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepin[3',4':3,4][1,4]oxazepin[5,6,7-de]quinazolin-11-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile

[0337] Step 1) Intermediate 1 of Example 12: (S)-11-bromo-12-chloro-10-fluoro-8-(((2R,7aS)-2-fluorotetrahydro- 1H-pyrrolizin-7a(5H)-yl)methoxy)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepine[3',4':3,4] Preparation of [1,4]oxazepine[5,6,7-de]quinazoline

[0338] To a mixture of ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methanol (78 mg, 0.49 mmol) in THF (5 mL) was added NaH (60% in oil, 60 mg, 2.5 mmol) at 0°C. The reaction mixture was stirred for 10 minutes under N2 gas at 0°C, and a solution of (S)-11-bromo-12-chloro-10-fluoro-8-(methylsulfonyl)-4,5,14,14a-tetrahydro-14-tetrahydro-12-chloro-10-fluoro-8-(methylsulfonyl)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepino[3',4':3,4][1,4]oxazepino[5,6,7-de]quinazoline (227 mg, 0.49 mmol) in THF (5 mL) was added to the solution. The reaction mixture was stirred for 16 hours under N2 gas at 25°C. The reaction mixture was quenched with concentrated NH4Cl(aq) (100 mL), and the mixture was extracted with ethyl acetate (50 mL×3). The organic phase was washed with brine (30 mL), dried over anhydrous Na2SO4, filtered and concentrated to dryness in vacuo. The residue was purified by silica gel column chromatography and eluted with ethyl acetate to give the desired product (S)-11-bromo-12-chloro-10-fluoro-8-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepin[3',4':3,4][1,4]oxazepin[5,6,7-de]quinazoline (122 mg, yield 46%) as a yellow oil. MS: m / z=547.1 (M+H + ,ESI+).

[0339] Step 2) Intermediate 2 of Example 12: (4-((14aS)-12-chloro-10-fluoro-8-(((2R,7aS)-2-fluorotetrahydro- 1H-pyrrolizin-7a(5H)-yl)methoxy)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepine[3',4':3,4] tert-[1,4]oxazepino[5,6,7-de]quinazolin-11-yl)-3-cyano-7-fluorobenzo[b]thiophen-2-yl)carbamic acid Preparation of Butyl Ester

[0340] The mixture contained (S)-11-bromo-12-chloro-10-fluoro-8-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepin[3',4':3,4][1,4]oxazepin[5,6,7-de]quinazoline (91 mg, 0.17 mmol), (3-cyano-4-(5, A mixture of tert-butyl 5-dimethyl-1,3,2-dioxaborinane-2-yl)-7-fluorobenzo[b]thiophene-2-yl)carbamate (135 mg, 0.33 mmol), K3PO4 (107 mg, 0.5 mmol), KF (19 mg, 0.33 mmol) and DPEphos-PdCl2 (24 mg, 0.034 mmol) in 4 mL of dioxane was purged with N2 three times. Then, the solution was stirred at 105°C for 5 hours. The reaction mixture was cooled to room temperature and diluted with water (80 mL). The solution was extracted with ethyl acetate (40 mL×3). The organic phase was washed with brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated to dryness in vacuo. The residue was purified by silica gel column chromatography and eluted with ethyl acetate to give the desired product (tert-butyl 4-((14aS)-12-chloro-10-fluoro-8-(((2R,7aS)-2-fluorotetrahydro-1H)-pyrrolazin-7a(5H)-yl)methoxy)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepin[3',4':3,4][1,4]oxazepin[5,6,7-de]quinazolin-11-yl)-3-cyano-7-fluorobenzo[b]thiophen-2-yl)carbamate (40 mg, yield 31.7%) as a yellow oil. MS: m / z=757.3 (M+H + ,ESI+).

[0341] Step 3) Example 12: 2-amino-4-((14aS)-12-chloro-10-fluoro-8-(((2R,7aS)-2-fluorotetrahydro-1H- Pyrrolizin-7a(5H)-yl)methoxy)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepine[3',4':3,4][1, Preparation of 4-oxazepine (5,6,7-de)quinazolin-11-yl)-7-fluorobenzo[b]thiophene-3-carbonitrile

[0342] A mixture of tert-butyl (4-((14aS)-12-chloro-10-fluoro-8-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methoxy)-4,5,14,14a-tetrahydro-1H,3H-[1,4]oxazepin[3',4':3,4][1,4]oxazepin[5,6,7-de]quinazolin-11-yl)-3-cyano-7-fluorobenzo[b]thiophen-2-yl)carbamate (40 mg, 0.05 mmol) in HCl / dioxane (2 mL) was stirred at room temperature for 16 hours. The solvent was removed under reduced pressure. The residue was purified by preparative HPLC (Waters 2767 / Qda, column: SunFireSunfire C18, 19*250mm, 10um; mobile phase A: 0.1% FA / H2O, B: ACN, flow rate: 20mL / min; gradient: 20%-30%) to obtain the desired product. Example 12a (1.94 mg, yield 5.6%) and Example 12b (1.32 mg, yield 3.8%), in the form of a white solid. MS: m / z = 657.3 (M+H + ,ESI+).

[0343] HNMR Example 12a ): 1 H NMR (400MHz, CD3OD) δ7.15 (dd, J=8.4, 5.1Hz, 1H), 7.02-6.92

[0344] (m,1H),5.59 -5.33(m,1H),5.06-4.96(m,1H),4.69-4.36(m,4H),4.27-4.10(m,2H),4.01-3.69( m,5H),3.55-3.34(m,3H),2.70-2.42(m,2H),2.32-2.23(m,2H),2.17-1.88(m,4H).

[0345] HNMR Example 12b ): 1H NMR(400MHz,CD3OD)δ8.43(brs,0.61H-FA),7.18-7.09(m,1H),7.03-6.90(m,1H), 5.52-5.31(m,1H),5.04-4.92(m,1H),4.73-4.63(m,2H),4.59-4.37(m,4H),4.26- 4.18(m,1H),4.17-4.08(m,1H),3.99-3.89(m,1H),3.85-3.58(m,4H),3.57-3.43( m,2H),2.57-2.37(m,2H),2.35-2.24(m,1H),2.22-2.11(m,3H),1.99-1.89(m,2H).

[0346] The compounds of Examples 1 to 15 in Table 1 below were prepared by methods similar to those described above using samples suitable for the preparation of the compounds described in each Example.

[0347] Table 1

[0348]

[0349]

[0350]

[0351]

[0352]

[0353]

[0354] <Test Example 1: KRAS Nucleotide Exchange Assay> Purpose of the test

[0355] Evaluation of compounds targeting KRAS WT 、KRAS G12D and KRAS G12V Mutated SOS1 mediates the inhibitory effect of nucleotide exchange activity.

[0356] Test Principle

[0357] The method is a method for monitoring the SOS1-mediated exchange of unlabeled KRAS bound to GDP and fluorescently labeled GTP, based on the detection of energy transfer between two fluorescent bodies when a GST antibody labeled with a donor Tb cryptate and an acceptor DY-647P1GTP are brought into proximity.

[0358] Test conditions and procedures

[0359] Material

[0360] GST-tagged KRAS WT or mutant protein (amino acids 2-169);

[0361] SOS1 (amino acids 564-1049);

[0362] Labeled GTP (GTP-DY-647P1);

[0363] Assay Buffer (20 mM HEPES pH 7.4, 150 mM NaCl, 5 mM MgCl2, 1 mM DTT, 0.05% BSA and 0.0025% NP40).

[0364] Test Procedure

[0365] 1. Dilute KRAS protein with assay buffer to 1.5 times the final concentration, then mix Tb cryptate anti-GST antibody and add 10 μL (KRAS WT 、KRAS G12D and KRAS G12V The final concentration was 20 nM).

[0366] 2. Dissolve the compound in DMSO and then dilute to 100 times the final concentration.

[0367] 3. Using an ECHO acoustic dispenser (Beckman), compounds were added to the assay wells and gently mixed with the KRAS / Ab mixture and incubated for 60 minutes.

[0368] 4. Mix SOS1 and labeled GTP, dilute with assay buffer to 3 times the final concentration, and add 5 μL of the solution to the assay well to start the reaction (the final concentration of labeled GTP is 0.15 μM, the final concentration of SOS1 is 7.5 nM in WT, 12.5 nM in G12D, and 50 nM in G12V). Only assay buffer and labeled GTP are placed in the blank well.

[0369] 5. The reaction was monitored using a PHERASTAR Plate Reader (BMG) at Ex / Em = (337 / 665; 337 / 620).

[0370] 6. HTRF signal was analyzed about 25 minutes after the start of the reaction (60 minutes for G12V reaction).

[0371] 7. Nucleotide exchange activity is expressed as a percentage of the difference compared to the DMSO reaction value, and IC 50 Values ​​were calculated based on the four-parameter logistic equation in GraphPad 4.0 software.

[0372] HTRF signal = ems 665 / ems 620*10000.

[0373] Table 2 shows the test results.

[0374] Table 2

[0375] Test results

[0376]

[0377] <Test Example 2: KRAS NanoBRET Assay>

[0378] Purpose of the test

[0379] NanoBRET target binding evaluation of compounds against KRAS (WT, G12D or G12V), HEK293 cells.

[0380] Test conditions and procedures

[0381] Preparation of compounds

[0382] The test compound was dissolved as a 10 mM stock solution, and BI-2582 and MRTX1133 (MedChemExpress) as standard compounds were dissolved in DMSO as 10 mM and 1 mM stock solutions, respectively.

[0383] Cell culture

[0384] NanoBRET KRAS (WT, G12D or G12V)-NanoLuc fusion vector and tracer K-2 are products of PROMEGA, and HEK293 cell line was purchased from ATCC. HEK293 cells were sprayed with EMEM medium supplemented with 10% FBS and 100 μg / mL penicillin-streptomycin, and cultured under the conditions of 5% CO2, 95% air humidity and 37°C.

[0385] Test Procedure

[0386] 1. Transfect (transfection) NanoBRET KRAS (WT, G12D or G12V)-NanoLuc fusion vector (Fusion vector) into HEK293 cells.

[0387] 2. In Opti-MEM without phenol red, adjust the density of transfected cells to 2×10 5 cells / mL, and mixed 20×K-2 tracer with the cells.

[0388] 3. A mixture of cells and tracer was inoculated into 384 wells and placed in a culture medium at 37°C and 5% CO2 for one hour, and then placed at room temperature for 15 minutes.

[0389] 4. Treat the 384 wells inoculated with cells and tracers with substrate and test compound solutions and react at room temperature for 15 minutes.

[0390] 5. Use Envision 2104 Plate Reader to measure at the donor emission wavelength (460nm) and the acceptor emission wavelength (600nm).

[0391] 6. Calculate the BRET ratio by dividing the acceptor emission (600 nm) by the donor emission (460 nm) and correct for background by removing the BRET ratio without tracer.

[0392] 7. Calculate the BRET response by calculating the BRET ratio when treated with the compound relative to the BRET ratio when treated with DMSO*100.

[0393] 8. Calculate IC based on the sigmoidal dose-response equation in the GraphPad Prism 4 program 50 value.

[0394] <Test Example 3: Cell proliferation assay>

[0395] Purpose of the test

[0396] Test compounds were tested for cell viability over 72 hours in AsPC-1 pancreatic cancer (KRAS G12D mutation) or SW480 colon cancer (KRAS G12V mutation) cells.

[0397] Test conditions and procedures

[0398] Material

[0399] The standard compound Staurosporine was purchased from Sigma-Aldrich (Saint Louis, MI). 2.0 Luminescent cell viability assay reagent (cat#G9243) was purchased from PROMEGA (Madison, WI). AsPC-1 and SW480 cell lines were purchased from the American Type Culture Collection (Manassas, VA). AsPC-1 cells were cultured in RPMI-1640 (ATCC, cat#30-2001), and SW480 cells were cultured in DMEM (ATCC cat#30-2002), and the culture medium was supplemented with 10% FBS (Sigma-Aldrich, cat#F2442) and 100 μg / mL penicillin-streptomycin (Sigma-Aldrich, cat#P4333). Cells were cultured under the conditions of 5% CO2, 95% air humidity and 37°C.

[0400] Test Procedure

[0401] 1. The test compound and the standard compound Staurosporine were dissolved in DMSO solution, 20 mM (test compound) and 10 mM (control compound Staurosporine) were prepared in the source plate, and 10 doses were diluted 3-fold with DMSO.

[0402] 2. In the Source Plate, use Echo 655 to inoculate 10 doses of 125 nL of test compound or 25 nL of standard compound into the wells of a 384-well plate (VWR, cat#82050-076).

[0403] 3. In a 384-well cell culture plate, inoculate 25 μL of culture medium containing 2000 AsPC-1 or SW480 cells.

[0404] 4. The cells were cultured with the compounds at 37°C and 5% CO2 for 72 hours.

[0405] 5. Add 25 μL of CellTiter-Glo 2.0 reagent to each well of the plate.

[0406] 6. After mixing the contents using an orbital shaker for 2 minutes, stabilize the luminescence signal at room temperature for 15 minutes.

[0407] 7. Luminescence signals were measured using Envision 2104 Plate Reader (PerkinElmer, Santa Clara, CA), and the number of viable cells was determined by quantifying the ATP present in each culture medium.

[0408] 8. Calculate IC based on the sigmoidal dose-response equation in the GraphPad Prism 4 program 50 value.

[0409] Table 3 shows the test results.

[0410] Table 3

[0411] Test results

[0412] Example AsPC-1 SW480 1 C - 2 C C 3 B - 6 A - 7 A - 11a B - 11b A - 12a B - 12b A -

[0413] (IC 50 <2μM=A;IC 50 ≥2μM, <20μM=B; IC 50 >20 μM = C).

[0414] The present invention has been studied above by specific examples. The present invention is specifically shown and described with reference to the preferred embodiment and various optional embodiments, but it should be understood by those skilled in the art that various changes in form and details can be made without departing from the spirit and scope of the present invention.

Claims

1. A compound, wherein The compound is a compound of the following Chemical Formula 1, or a stereoisomer, a diastereomer, an enantiomer, an atropisomer, a solvate, an isotopic variant, a tautomer, or a pharmaceutically acceptable salt thereof: [Chemical formula 1] In the chemical formula, R 1 is independently selected from hydroxy, halogen, C 1-3 Haloalkyl, C 1-3 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, C 1-3 phenyl, pyridyl, naphthyl, indazolyl, benzothiazolyl or benzothienyl, substituted or unsubstituted by one or more substituents selected from alkoxy, amino and cyano; R 2 is hydrogen or halogen; R 3 For hydrogen, C 1-3 Alkyl, C 3-6 Cycloalkyl, 4- to 10-membered heterocyclic ring or -O-(L) m -A1, where A1 is respectively connected by one or more R 7 Substituted or unsubstituted C 1-3 Alkyl, C 3-6 Cycloalkyl, 4- to 10-membered heterocyclic ring, 6- to 10-membered aryl, 5- to 10-membered heteroaryl, or 5- to 10-membered fused heteroaryl; L is R 7 Substituted or unsubstituted C 1-3 Alkylene or C 3-8 Cycloalkylene; R 7 Each is independently halogen, oxo (=0), =CH2, -OCF3, -OCHF2, amino, cyano, -N(C 1-3 Alkyl)2, -NH(C 1-3 alkyl), substituted or unsubstituted C 1-3 Alkyl, substituted or unsubstituted C 1-3 Haloalkyl, substituted or unsubstituted C 1-3 Alkoxy, substituted or unsubstituted C 3-4 Cycloalkyl or substituted or unsubstituted heterocycle; R 4 are independently hydrogen, hydroxyl, halogen, C 1-3 Haloalkyl, or C 1-3 Alkyl, C 1-3 Alkoxy; X is O, CH2 or NR 8 ; R 8 For hydrogen or R 9 Any C substituted 1 to 3 times 1-6 Alkyl, C 3-6 Cycloalkyl or C 3-6 Heterocycloalkyl; R 9 is independently oxygen, hydroxyl, -C 1-4 Alkyl or -OC 1-4 alkyl; Y is NH, O, S, SO or SO2; Z is CR 6 or N; R 5 For hydrogen, C 1-3 Alkyl, cyano or amino; R 6 For hydrogen, hydroxyl, halogen, C 1-3 alkyl; n1, n2, n3 and m are each an integer from 1 to 3; The heterocyclic ring, heteroaryl group and fused heteroaryl group each contain one or more heteroatoms selected from N, S and O.

2. The compound according to claim 1, wherein R 1 is independently selected from hydroxy, halogen, C 1-3 Haloalkyl, C 1-3 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, C 1-3 Phenyl, naphthyl, benzothiazolyl or benzothiophenyl, which may be substituted or unsubstituted by one or more substituents selected from alkoxy, amino and cyano; X is O, CH2 or NH; Y is O.

3. The compound according to claim 1, wherein R 3 O-(L) m -A1, where A1 is respectively connected by one or more R 7 A substituted or unsubstituted 4- to 10-membered heterocyclic ring or a 5- to 10-membered condensed heteroaryl group.

4. The compound according to claim 1, wherein R 1 is independently selected from hydroxy, halogen, C 1-3 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, amino and cyano groups may be substituted or unsubstituted with one or more substituents R 3 O-(L) m -A1, where A1 is respectively connected by one or more R 7 A substituted or unsubstituted 4- to 10-membered heterocyclic ring or a 5- to 10-membered fused heteroaryl group; L is C 1-3 Alkylene or C 3-8 Cycloalkylene; R 7 are independently halogen, oxo (=0), =CH2, -OCF3, -OCHF2, amino, cyano, C 1-3 Alkyl, C 1-3 Haloalkyl or C 1-3 Alkoxy; R 4 is hydrogen; X is O, CH2 or NH; Z is CR 6 or N; R 5 is hydrogen or C 1-3 alkyl; R 6 is a halogen; n1, n2, n3 and m are integers from 1 to 3, respectively.

5. The compound according to claim 1, wherein When Z is N, R 1 is independently selected from hydroxy, halogen, C 1-3 Haloalkyl, C 1-3 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, C 1-3 Phenyl or naphthyl which may be substituted or unsubstituted by one or more substituents selected from alkoxy, amino and cyano; In Z is CR 6 hour, R 1 is independently selected from hydroxy, halogen, C 1-3 Haloalkyl, C 1-3 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, C 1-3 One or more substituents selected from the group consisting of alkoxy, amino and cyano may be substituted or unsubstituted benzothiazolyl or benzothienyl.

6. The compound according to claim 1, wherein R 3 O-(L) m -A2, where A2 is They are each independently selected from halogen, C 1-3 Alkyl, C 1-3 Alkoxy and =CH2 are substituted or unsubstituted with one or more substituents; -(L) m - is methylene or 7. The compound according to claim 1, wherein R 1 is independently selected from hydroxy, halogen, C 1-3 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, amino and cyano groups may be substituted or unsubstituted with one or more substituents R 2 is a halogen; R 3 O-(L) m -A1, wherein A1 is independently selected from halogen, C 1-3 Alkyl, C 1-3 Alkoxy and =CH2 substituted or unsubstituted with one or more substituents Among them, -(L) m - is methylene or R 4 is hydrogen; X is O, CH2 or NH; Y is O; Z is CH, C(C 1-3 Alkyl) or N; R 5 is hydrogen or C 1-3 alkyl.

8. The compound according to claim 1, wherein The compound of Chemical Formula 1 is any one compound selected from the group consisting of the following compounds, or a stereoisomer, a diastereomer, an enantiomer, an atropisomer, a solvate, an isotopic variant, a tautomer, or a pharmaceutically acceptable salt thereof:

9. A pharmaceutical composition for treating cancer, wherein: The pharmaceutical composition comprises as an active ingredient the compound according to any one of claims 1 to 8, or its stereoisomer, diastereomer, enantiomer, atropisomer, solvate, isotopic variant, tautomer, or pharmaceutically acceptable salt.

10. The pharmaceutical composition according to claim 9, wherein The pharmaceutical composition exhibits KRAS protein inhibitory activity.

Images