WO2022222871A1

WO2022222871A1 - Heterocyclic compounds as kras g12c inhibitors

Info

Publication number: WO2022222871A1
Application number: PCT/CN2022/087197
Authority: WO
Inventors: Xiangyang Chen; Yucheng PANG
Original assignee: Beijing Innocare Pharma Tech Co., Ltd.
Priority date: 2021-04-21
Filing date: 2022-04-15
Publication date: 2022-10-27

Abstract

Provided are compounds of general Formula (I) and their pharmaceutically acceptable salts suitable for inhibiting or regulating the activity of KRAS G12C. Said compounds are useful for preventing and /or treating relevant KRAS G12C mediated cancers. Also provided are preparation methods thereof.

Description

HETEROCYCLIC COMPOUNDS AS KRAS G12C INHIBITORS

TECHNICAL FIELD

The present invention relates to heterocyclic compounds or their pharmaceutically acceptable salts thereof, suitable for regulating or inhibiting the activity of KRAS G12C. The present invention also relates to methods for preparing the heterocyclic compounds or their pharmaceutically acceptable salts thereof. The present invention further relates to methods for treating and/or preventing cancers mediated by KRAS G12C by the heterocyclic compounds or their pharmaceutically acceptable salts thereof.

BACKGROUND

Kirsten rat sarcoma 2 viral oncogene homolog (KRAS) is a small GTPase protein with a single globular structure (21 KD molecular weight) . It belongs to the RAS family of oncogenes, along with HRAS and NRAS. As a molecular switch cycling between binding to GDP and GTP, it regulates signal pathways that control cell survival, proliferation, and division. When KRAS binds GDP, it is inactive or in the off state. After receiving cellular signals from upstream tyrosine kinases, KRAS exchanges the bound GDP to GTP induced by guanine exchange factor (GEF) and then undergoes structural changes to become active or be in the on state, thereby activating downstream effectors and eventually leading to cell growth and division. As KRAS has a strong binding affinity to GTP, it is necessary for GTP-bound KRAS to interact with exogenous protein GTPase activating protein (GAP) to enable catalyzing the hydrolysis of GTP to GDP, so as to be back to the off state. However, if KRAS mutates, resulting in weakening interaction between KRAS and GAP, it would become continuously activated, which in turn would lead to excessively cell proliferating, preventing apoptosis and inducing tumor growth.

KRAS gene mutation is very common in cancers, with an incidence of about 20%, especially in tubular adenocarcinoma (98%) , rectal cancer (45%) , lung cancer (31%) and multiple myeloma (23%) . G12C mutation itself accounts for 14%in lung cancer (Cox et al., Nat. Rev. Drug Discov. 2014, 13, 828-51) . Therefore, KRAS has become an attractive anticancer drug target. However, it remains untargetable for a long period of time despite huge R&D investments. When activated, KRAS and GTP have strong binding interaction to each other, the protein is almost spherical structurally, and the GTP binding pocket is small, making it difficult to develop small molecule inhibitors. For the KRAS G12C mutant in the inactive state, its GDP binding pocket expands to a certain extent, forming a hidden "groove" adjacent to the cysteine residue from Gly-12 mutation, making it possible to target KRAS G12C mutant with irreversible inhibitors. The present invention discloses heterocyclic compounds with the structure as shown in general formula (I) which are useful in inhibiting KRAS G12C activity.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows that

Compounds

12a and 15a reduced tumor size in the Mia PaCa-2 model. The treatment with 12a, 15a and AMG-510 (positive control) demonstrated significant anti-tumor activities with tumor growth inhibition (TGI) of 76.38%, 93.29%and 56.20%, respectively.

FIG. 2 shows that

Compounds

12a and 15a at 1 mg/kg QD were well-tolerated with no significant body weight loss during the study.

DETAILED DESCRIPTION OF THE INVENTION

Definitions

Unless otherwise stated, the following terms used in this application have the following meanings.

“C _x-y” refers to a range of the number of carbon atoms, where x and y are both integers, for example, C _3-8 cycloalkyl stands for cycloalkyl having 3 to 8 carbon atoms.

“Alkyl” refers to a saturated straight-chain or branched-chain hydrocarbyl substituent containing 1 to 20 carbon atoms, for example, 1 to 8 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms. Unrestricted examples of alkyl include but are not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1, 1-dimethylpropyl, 1, 2-dimethylpropyl, 2, 2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1, 1, 2-trimethylpropyl, 1, 1-dimethylbutyl, 1, 2-dimethylbutyl, 2, 2-dimethylbutyl, 1, 3-dimethylbutyl, 2-ethylbutyl.

“Alkylene” refers to a saturated straight-chain or branched-chain hydrocarbyl divalent substituent containing 1 to 20 carbon atoms, for example, 1 to 6 carbon atoms or 1 to 4 carbon atoms. Unrestricted examples of alkylene include but are not limited to -CH ₂-, -CH (CH ₃) -, -CH ₂CH ₂-, -CH ₂CH ₂CH ₂-, -C (CH ₃) ₂-, -CH ₂CH ₂CH ₂CH ₂-and -CH ₂CH (CH ₃) CH ₂-.

“Cycloalkyl” refers to a saturated cyclic hydrocarbyl substituent containing 3 to 14 annular carbon atoms. Cycloalkyl can be a mono carbon ring substituent, typically containing 3 to 8, 3 to 7, or 3 to 6 carbon atoms. Unrestricted examples of monocyclic cycloalkyl include but are not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl. Cycloalkyl can also be a substituent with two or three mono carbon rings that are fused together, such as decahydronaphthyl.

“Heterocyclyl or heterocycle” refers to a saturated or partially unsaturated monocyclic or polycyclic group containing 3 to 20 annular atoms, for example, 3 to 14, 3 to 12, 3 to 10, 3 to 8, 3 to 6, or 5 to 6 annular atoms in which one or more of the annular atoms are selected from N, O and S (O) _m (where m is an integer from 0 to 2) . Preferably, it can have 3 to 12 annular atoms, 3 to 10 annular atoms, 4 to 7 annular atoms, and 4 to 6 annular atoms, wherein 1 to 4 are heteroatoms, 1 to 3 are heteroatoms, or 1 to 2 are heteroatoms. Heterocycle suitable for the present invention includes monocyclic and bicyclic, where one or more hydrogens of the ring is optionally substituted. For example, heterocycle includes azetidinyl, methylazetidinyl, difluoroazetidinyl, tetrahydropyranyl, tetrahydrofuranyl, oxetanyl, pyrrolidinyl, methylpyrrolidinyl, dimethylpyrrolidinyl, isopropylpyrrolidinyl, cycloalkylalkylpyrrolidinyl, hydroxypyrrolidinyl, fluoropyrrolidinyl, difluoropyrrolidinyl, methoxyethylpyrrolidinyl, dimethylaminopyrrolidinyl, piperazinyl, morpholinyl, methylmorpholinyl, 1, 4-oxazepanyl, piperidinyl, methylpiperidinyl acylpiperidinyl, cyanopiperidinyl, cycloalkylpiperidinyl, fluoropiperidinyl, difluoropiperidinyl, alkoxypiperidinyl, pyrrolidonyl, piperidinonyl, thiomorpholinyl-1, 1-dioxide, hexahydro-1H-pyrrolizinyl, 2-flurorohexahydro-1H-pyrrolizinyl, 3-azabicyclo [3.1.0] hexanyl, oxa-5-azabicyclo [2.2.1] heptan-5-yl, or azabicyclo [2.2.1] heptan-2-yl.

“Aryl or aryl ring” refers to an aromatic monocyclic or fused polycyclic group containing 6 to 14 carbon atoms, preferably 6-to 10-membered, such as phenyl and naphthyl, most preferably phenyl. The aryl ring can be fused with a heteroaryl, heterocyclyl or cycloalkyl ring, and unrestricted examples include but are not limited to:

“Heteroaryl or heteroaryl ring” refers to a heteroaromatic system containing 5 to 14 annular atoms, of which 1 to 4 annular atoms are selected from heteroatoms including O, S and N. Heteroaryl preferably is 5-to 10-membered, and more preferably 5-or 6-membered, such as furyl, thienyl, pyridyl, pyrrolyl, pyrimidyl, pyrazinyl, pyrazolyl, imidazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, quinolinyl, isoquinolinyl, indolyl and isoindolyl. The heteroaryl ring can be fused with an aryl, heterocyclyl or cycloalkyl ring, and unrestricted examples include but are not limited to:

“Halogen” refers to F, Cl, Br, or I.

“Cyano” refers to -CN.

“Oxo” refers to =O.

“Carbonyl” refers to -C (=O) -group.

“Sulfonyl” refers to a -S (O) ₂-group.

“Sulfinyl” refers to a -S (O) -group.

“Optional substitution or optionally substituted” refers to that one or more hydrogen atoms in a group, preferably 1-5, for example, 1 to 3 hydrogen atoms, are independently substituted by a corresponding number of substituents. The substituents are located only in the possible chemical positions understood by those skilled in the art. For example, amino or hydroxyl groups with free hydrogen may be unstable when bound with carbon atoms with unsaturated bonds (such as olefinic) . The substituents include but are not limited to halogen, -OH, amino, cyano, nitro, oxo, -SF ₅, C _1-4 alkyl, C _3-7 cycloalkyl, etc.

“Isomers” refer to compounds that have the same molecular formula but their atomic binding position or spatial arrangement is different. Isomers with different arrangement of their atoms in space are called “stereoisomers” . Stereoisomers include optical isomers, geometric isomers, and conformational isomers.

The compounds of the present invention can exist as optical isomers. Optical isomers include enantiomers and diastereomers. An enantiomer is one of two stereoisomers that are mirror images of each other that are non-superposable. A racemic mixture, or racemate is one that has equal amounts of left-and right-handed enantiomers of a chiral molecule. Diastereomers are stereoisomers that are not mirror images of one another and are non-superimposable on one another. Methods for preparing and separating optical isomers are known in the art. When a compound is a single isomer and its absolute configuration is determined, it is referred as a “R” or “S” isomer according to the configuration of the substituents around the chiral carbon atom; when its absolute configuration is not determined, it is referred as a (+) or (-) isomer according to its measured optical rotation value.

The compounds of the present invention may also have geometric isomers resulting from the distribution of substituents around carbon-carbon double bonds, carbon-nitrogen double bonds, cycloalkyl or heterocyclyl groups. The substituents around the carbon-carbon double bond or carbon-nitrogen bond are designated to be in a Z or E configuration, and the substituents around the cycloalkyl or heterocycle are designated to be in a cis or trans configuration.

The compounds of the present invention may also show tautomerism, such as keto-enol tautomerism.

The present invention includes any tautomeric or stereoisomeric forms and mixtures thereof and is not limited to any tautomeric or stereoisomeric forms used in the compound nomenclature or chemical structural formulae.

“Isotopes” include all isotopes of the atoms appearing in the compounds of the present invention. Isotopes include those atoms with the same atomic number but in different masses. Examples of isotopes suitable for incorporation into the compounds of the present invention are isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine, for example but not limited to ²H (D) , ³H, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O, ³¹P, ³²P, ³⁵S, ¹⁸F and ³⁶Cl. The isotopically labeled compounds of the present invention can generally be prepared by conventional techniques known to those skilled in the art or by methods similar to those described in the embodiments using appropriate isotopically labeled reagents instead of non-isotopically labeled reagents. Such compounds have various potential uses, for example, as standards and reagents in the determination of biological activities. In the case of stable isotopes such as deuterium ²H (D) , ¹³C and ¹⁵N, such compounds have the potential to beneficially alter biological, pharmacological, or pharmacokinetic properties. Deuterium ²H (D) is a preferable isotope of the present invention. For example, one or more hydrogens of -CH ₃ can be substituted by D.

The compounds of the present invention can be administered in form of prodrugs. “Prodrugs” refer to derivatives that are converted into biologically active compounds under the physiological condition in vivo, for example, by oxidation, reduction, and hydrolysis (each of which occurs with or without the participation of enzymes) . Examples of a prodrug are a compound of the present invention in which an amino is acylated, alkylated or phoshorylated, for example eicosanoyl amino, alanyl amino and pivaloyloxymethyl amino; a hydroxyl is acylated, alkylated or phoshorylated or converted into borate, for example acetoxy, palmitoyloxy, pivaloyloxy, succinyloxy, fumaroyloxy and alanyloxy; a carbonyl is esterified or amidated; and a thiol forms a disulfide bridge with a carrier molecule that selectively delivers the drug to the target and/or to the cytosol of cells, such as peptide. Prodrugs can be prepared from the compounds of the present invention according to well-known methods.

“Pharmaceutically acceptable salts” refer to the salts made from the compounds of the present invention with pharmaceutically acceptable bases or acids, including inorganic alkalis or acids and organic bases or acids, under the condition that the compounds contain one or more acidic or basic groups. Therefore, the compounds of the present invention that contain acidic groups can exist in form of salts, for example, as alkali metal salts, alkaline earth metal salts, or ammonium salts. For example, such salts include sodium salts, potassium salts, calcium salts, magnesium salts or ammonia or organic amine salts such as salts of ethylamine, ethanolamine, triethanolamine or amino acids. The compounds of the present invention that contain basic groups can exist in form of salts as inorganic or organic acid salts. Examples of suitable acids include hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, nitric acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalene disulfonic acid, oxalic acid, acetic acid, tartaric acid, lactic acid, salicylic acid, benzoic acid, formic acid, propanoic acid, pivalic acid, malonic acid, succinic acid, pimelic acid, fumaric acid, maleic acid, malic acid, sulfamic acid, phenylpropionic acid, gluconic acid, ascorbic acid, isonicotinic acid, citric acid, adipic acid and other acids known to those skilled in the art. If the compounds of the present invention contain both acidic and basic groups in the molecule, the present invention further includes internal salts in addition to the mentioned salt forms. Each salt can be obtained by conventional methods known to those skilled in the art, for example by mixing a compound of the present invention with an organic or inorganic acid or base in a solvent or dispersant, or by anion exchange or cation exchange with another salt.

“Pharmaceutical composition” refers to a composition containing one or more of the compounds described herein or pharmaceutically acceptable salts, prodrugs, stable isotope derivatives and isomers thereof, and other components such as pharmaceutically acceptable carriers and excipients.

“Cancers/tumors” include but are not limited to digestive/gastrointestinal cancer, colon cancer, liver cancer, pancreatic cancer, skin cancer (including mast cell tumor and squamous cell carcinoma) , breast cancer, ovarian cancer, prostate cancer, lymphoma, leukemia (including acute myeloid leukemia and chronic myelogenous leukemia) , kidney cancer, lung cancer, muscle cancer, bone cancer, bladder cancer, brain cancer, melanoma (including oral and metastatic melanoma) , Kaposi’s sarcoma (myeloma including multiple myeloma) , myeloproliferative diseases, proliferative diabetic retinopathy, and vascular hyperplasia-related disorders/tumors.

“Therapeutically effective amount” refers to the amount of the compound of the present invention that can effectively inhibit the functions of KRAS G12C, and/or treat or prevent the diseases mediated by KRAS G12C.

“Patients” refer to mammals, preferably humans.

The present invention provides compounds useful as inhibitors of KRAS G12C. The compounds are shown in Formula (I) , or prodrugs, stable isotope derivatives, pharmaceutically acceptable salts, and isomers thereof,

where:

W is

-C (O) CH=C=CH ₂ or -CN;

X is 4-to 12-membered heterocycle, where X is optionally substituted by 1-4 R ⁵;

R ¹ is H or halogen;

R ² is H, halogen, cyano, CF ₃, C _1-6 alkyl, C _3-6 cycloalkyl or -OR ^c;

R ³ is C _6-12 aryl or 5-to 12-membered heteroaryl, where R ³ is optionally substituted by 1-4 R ⁶;

R ⁴ is H, C _1-6 alkyl, C _3-6 cycloalkyl, 4-to 12-membered heterocyclyl or 5-to 12-membered heteroaryl, where one or more hydrogens of the alkyl, cycloalkyl, heterocyclyl and heteroaryl are optionally substituted by R ⁷;

R ⁵ is D (deuterium) , halogen, cyano, oxo, C _1-6 alkyl, C _3-6 cycloalkyl, -OC _1-6 alkyl or -OC _3-6 cycloalkyl, where one or more hydrogens of the alkyl and cycloalkyl are optionally substituted by D, halogen, cyano or -OR ^c;

R ⁶ is halogen, cyano, C _1-6 alkyl, C _3-6 cycloalkyl, 4-6 membered heterocyclyl, -OR ^c or -NR ^cR ^d, where one or more hydrogens of the alkyl, cycloalkyl and heterocyclyl are optionally substituted by D, halogen or -OR ^c;

R ⁷ is D, halogen, cyano, C _1-6 alkyl, C _3-6 cycloalkyl, -NR ^cR ^d, -C (O) NR ^cR ^d, -OR ^c, 4-to 12-membered heterocyclyl or 5-to 12-membered heteroaryl, where one or more hydrogens of the alkyl, cycloalkyl, heterocyclyl and heteroaryl are optionally substituted by halogen or C _1-6 alkyl;

L is a bond, O, S, NR ^c, alkylenyl or -C (O) NR ^c-;

R ^a is absent, H, D, cyano, halogen, C _1-6 alkyl or C _1-6 haloalkyl;

R ^b is H, D, C _1-6 alkyl, C _1-6 haloalkyl or C _3-6 cycloalkyl, where one hydrogen of the alkyl is optionally substituted by -OR ^c, -NR ^cR ^d or 4-to 6-membered heterocyclyl; and

R ^c and R ^d are independently selected from H, C _1-6 alkyl, C _1-6 haloalkyl, C _3-6 cycloalkyl or 4-6 membered heterocyclyl.

In some embodiments, the compounds shown in Formula (I) have the following Formula (II) :

where:

W is

-C (O) CH=C=CH ₂ or -CN;

R ¹ is H or halogen;

R ² is H, halogen, cyano, CF ₃, -OCF ₃, C _1-3 alkyl or -OC _1-3 alkyl;

R ³ is C _6-12 aryl or 5-to 12-membered heteroaryl, where one or more hydrogens of R ³ is optionally substituted by 1-4 R ⁶;

R ⁴ is H, C _1-6 alkyl, C _3-6 cycloalkyl or 4-to 12-membered heterocyclyl, where one or more hydrogens of the alkyl, cycloalkyl and heterocyclyl are optionally substituted by R ⁷;

R ⁵ is selected from cyano, oxo or C _1-6 alkyl, where one or more hydrogens of the alkyl is optionally substituted by D, halogen, cyano or -OR ^c;

R ⁶ is halogen, cyano, amino, hydroxy or C _1-6 alkyl, where one or more hydrogens of the alkyl is optionally substituted by D, halogen or -OR ^c;

R ⁷ is D, halogen, cyano, C _1-6 alkyl, C _3-6 cycloalkyl, -NR ^cR ^d, -OR ^c or 4-to 12-membered heterocyclyl, where one or more hydrogens of the alkyl, cycloalkyl and heterocyclyl are optionally substituted by halogen or C _1-6 alkyl;

L is absent (a bond) or O;

R ^a is absent, H, cyano, halogen or C _1-3 alkyl;

R ^b is H, D or C _1-3 alkyl, where one hydrogen of the alkyl is optionally substituted by -OR ^c, -NR ^cR ^d or 4-to 6-membered heterocyclyl; and

In one embodiment, W is -C (O) C (R ^a) =C (R ^b) ₂; R ^a is H, cyano, halogen or C _1-3 alkyl, preferably H, cyano or halogen; and R ^b is H or C _1-3 alkyl, where one hydrogen of the alkyl is optionally substituted by -NR ^cR ^d or 4-to 6-membered heterocyclyl, preferably R ^b is H.

In one embodiment, W is -C (O) C≡C (R ^b) , where R ^b is H or CH ₃.

In one embodiment, W is -S (O) ₂CH=CH ₂, -C (O) CH=C=CH ₂ or -CN.

In one preferred embodiment, W is -C (O) CH=CH ₂ or -C (O) CH=C=CH ₂.

In one preferred embodiment, R ¹ is H.

In one embodiment, R ² is H, halogen, CF ₃, C _1-3 alkyl or -OC _1-3 alkyl.

In one preferred embodiment, R ² is halogen or CF ₃.

In one embodiment, R ³ is C _6-12 aryl or 5-to 12-membered heteroaryl, where one or more hydrogens of the aryl and heteroaryl are optionally substituted by halogen, amino, hydroxy, CF ₃ or C _1-6 alkyl.

In one embodiment, R ³ is phenyl, where one hydrogen of the phenyl is substituted by -NH ₂ and the other hydrogens are optionally substituted by halogen.

In one preferred embodiment, R ³ is

In one embodiment, R ³ is 6-to 10-membered heteroaryl, where one or more hydrogens of the heteroaryl is optionally substituted by amino, halogen, CF ₃ or C _1-6 alkyl.

In one embodiment, L is a bond and R ⁴ is H.

In one embodiment, L is a bond and R ⁴ is C _1-6 alkyl, where one or more hydrogens of the alkyl is optionally substituted by halogen.

In one embodiment, L is -O-and R ⁴ is C _1-6 alkylene- (4-to 12-membered) heterocyclyl, where one or more hydrogens of the heterocyclyl is optionally substituted by halogen or C _1-6 alkyl.

In one embodiment, R ⁵ is C _1-6 alkyl, where one or more hydrogens of the alkyl is optionally substituted by cyano.

In some embodiments, the compounds shown in Formula (II) have the following Formula (III) :

where:

W is -C (O) CH=CH ₂, -S (O) ₂CH=CH ₂, or -C (O) CH=C=CH ₂;

A is N or C (R ⁶) ;

R ² is halogen, CH ₃ or CF ₃;

R ⁴ is H, C _1-6 alkyl or C _1-6 alkylene- (4-to 12-membered) heterocyclyl, where one or more hydrogens of the heterocyclyl is optionally substituted by halogen or C _1-6 alkyl;

R ⁵ is C _1-2 alkyl or -CH ₂CN;

R ⁶ is halogen, amino, hydroxy, CF ₃ or C _1-6 alkyl; and

L is a bond or O.

In some embodiments, the compounds shown in Formula (III) have the following Formula (IV) :

where:

R ² is halogen or CF ₃;

R ⁴ is H, C _1-6 alky or -OC _1-6 alkylene- (4-to 12-membered) heterocyclyl, where one or more hydrogens of the heterocyclyl is optionally substituted by halogen or C _1-6 alkyl;

R ^5a is H, CH ₃ or -CH ₂CN; and

R ^5b is H or CH ₃.

Examples of heterocyclyl or C _1-6 alkylene- (4-to 12-membered) heterocyclyl in Formula I to IV independently are azetidinyl, methylazetidinyl, difluoroazetidinyl, tetrahydropyranyl, tetrahydrofuranyl, oxetanyl, pyrrolidinyl, methylpyrrolidinyl, dimethylpyrrolidinyl, isopropylpyrrolidinyl, cycloalkylalkylpyrrolidinyl, hydroxypyrrolidinyl, fluoropyrrolidinyl, difluoropyrrolidinyl, methoxyethylpyrrolidinyl, dimethylaminopyrrolidinyl, piperazinyl, morpholinyl, methylmorpholinyl, 1, 4-oxazepanyl, piperidinyl, methylpiperidinyl, acylpiperidinyl, cyanopiperidinyl, cycloalkylpiperidinyl, fluoropiperidinyl, difluoropiperidinyl, alkoxypiperidinyl, pyrrolidonyl, piperidinonyl, thiomorpholinyl-1, 1-dioxide, hexahydro-1H-pyrrolizinyl, 2-flurorohexahydro-1H-pyrrolizinyl, 3-azabicyclo [3.1.0] hexanyl, oxa-5-azabicyclo [2.2.1] heptan-5-yl, or azabicyclo [2.2.1] heptan-2-yl. Preferred examples are N-methylpyrrolidinyl and hexahydro-1H-pyrrolizinyl. Also preferred are piperidinyl, piperazinyl, and morpholinyl.

The present invention further relates to the following Compounds 1-29, or their pharmaceutically acceptable salts, prodrugs, stable isotope derivatives, isomers, and mixtures thereof.

The compounds of the present invention effectively inhibit KRAS G12C mediated guanine nucleotide exchange, having an IC ₅₀ less than 500 nM, preferably having an IC ₅₀ less than 100 nM. The compounds of the present invention have a significant inhibitory effect on NCI-H358 cell proliferation, having an IC ₅₀ less than 500 nM, preferably having an IC ₅₀ less than 100 nM.

The present invention further relates to pharmaceutical compositions comprising compounds of Formula (I) or pharmaceutically acceptable salts, prodrugs, stable isotope derivatives, or isomers thereof, and pharmaceutically acceptable carriers or excipients. The pharmaceutical compositions are useful for the treatment or prevention of KRAS G12C mediated cancers, in particular lung cancer, colon cancer, pancreatic cancer, and ovarian cancer.

The present invention further provides a method for treating or preventing cancers mediated by KRAS G12C. The method comprises administering to a patient in need thereof a therapeutically effective amount of the compounds shown in Formula (I) or pharmaceutically acceptable salts, prodrugs, stable isotope derivatives, or isomers thereof.

The present invention provides a compound of Formula (I) , or a pharmaceutically acceptable salt, prodrug, stable isotope derivative, stereoisomer thereof, for use in a method of treating or preventing cancers mediated by KRAS G12C. According to the present invention, the pharmaceuticals can be in any dosage form, including but not limited to tablets, capsules, a solution, a freeze-drying preparation and injectable.

The pharmaceutical formulation of the present invention can be administered in form of a dosage unit containing a predetermined amount of active ingredient. Such a unit may contain 1 mg to 1 g, preferably 100 mg to 1 g of a compound of the present invention, depending on the disease being treatment, the method of administration, as well as age, weight, and condition of the patients. Furthermore, the pharmaceutical formulation can be prepared using methods well known in the pharmaceutical field, for example, by formulating the active ingredient with one or more excipients or one or more adjuvants.

The pharmaceutical formulation of the present invention is suitable for administration by any appropriate method, for example by oral (including buccal or sublingual) or parenteral (including subcutaneous, intramuscular, intravenous, or intradermal) .

The present invention further provides methods for preparing the compounds. Alternatively, the compounds can be synthesized by synthetic techniques known to those skilled in the art. The products obtained at each step of reaction are isolated by separation techniques known in the art, including but not limited to extraction, filtration, distillation, crystallization, and chromatographic separation. The starting materials and chemical reagents used for the synthesis can be conventionally made based on literature (can be searched from SciFinder) or purchased.

The heterocyclic compounds shown in Formula (I) of the present invention can be synthesized according to the route shown below: 1) activation of A1 by chlorination or sulfonation, followed by substitution by A2 to afford A3; 2) Suzuki coupling between A3 and boronic acid or borate to give A4; 3) deprotection of A4 in acid to give A5; 4) amidation of A5 to give A6 which can be further derivatized, such as by chlorination.

A1 can be synthesized according to the route shown below: 1) esterification of B1 to give B2; 2) oxidation of B2 to N-oxide B3 which is rearranged to B4 mediated by an anhydride; 3) amination of B4 to give B5; 4) cyclization reaction between B5 and formamidine to give B6; 5) bromination of B6 to give intermediate A1.

A1 can also be synthesized according to the route shown below: 1) Sandmeyer reaction of C1 to give C2 which is chemoselectively methylated to generate C3; 2) reduction of C3 by iron powder to give C4; 3) carbonylation of C4 catalyzed by palladium to give C5 which is further halogenated to give C6; 4) Sandmeyer reaction of C6 to give C7 which is further demethylated to give C8; 5) amination of C8 to give C9; 6) cyclization between C9 and formamidine to give intermediate A1.

A1 can also be synthesized according to the route shown below: 1) conversion of C9 to thiourea D1 which undergoes cyclization to give D2 under a basic condition; 2) methylation of D2 to give D3 which is further oxidized to obtain D4; 3) reaction of D4 with different nucleophiles, such as alcohols, to give intermediate A1 under a basic condition.

A1 can also be synthesized according to the route shown below: 1) conversion of B5 to thiourea E1 which is then undergoes cyclization to give E2 under a basic condition; 2) methylation of E2 to give E3; 3) reaction of E3 with different nucleophiles, such as alcohols, to give E4 under a basic condition; 4) E4 is finally brominated to give intermediate A1.

The following examples further illustrate the present invention. These examples are intended merely to be illustrative of the present invention and are not to be construed as being limiting.

EXAMPLES

The structure of a compound was determined by nuclear magnetic resonance (NMR) or mass spectrometry (MS) . NMR determination used a Bruker ASCEND-400 NMR spectrometer. The solvent for the determination was deuterated dimethyl sulfoxide (DMSO-d ₆) , deuterated chloroform (CDC1 ₃) , or deuterated methanol (CD ₃OD) . The internal standard was tetramethylsilane (TMS) , and the chemical shift was given in a unit of 10 ^-6 (ppm) . MS determination used an Agilent SQD (ESI) mass spectrometer (Agilent 6120) .

HPLC determination used Agilent 1260 DAD high pressure liquid chromatograph (column: Poroshell120 EC-C18, 50×3.0 mm, 2.7 μm) or Waters Arc high pressure liquid chromatograph (column: Sunfire C18, 150×4.6 mm, 5 μm) .

Thin layer chromatography (TLC) used GF254 silica gel plates from Qingdao Haiyang Chemical Co., Ltd. with a thickness of 0.15 to 0.2 mm, and the separation/purification of products by thin layer chromatography used silica plates with a thickness 0.4 to 0.5 mm.

Column chromatography generally used 200 to 300 mesh silica gel from Qingdao Haiyang Chemical Co., Ltd.

Known starting materials in the present invention were synthesized according to the methods known in the art, or purchased from ABCR GmbH&Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc., Beijing Ouhe Technology Co., Ltd., etc.

Unless otherwise stated in the embodiments, the reactions were carried out under an atmosphere of argon or nitrogen using a balloon with a volume of about 1 L.

Hydrogenation was carried out under an atmosphere of hydrogen using a balloon with a volume of about 1 L that was attached to the reaction vessel after being vacuumed and filled with hydrogen repeatedly for 3 times.

The microwave reaction used a CEM Discover-SP microwave reactor.

Unless otherwise stated in the embodiments, the reaction was run at room temperature.

The reaction was monitored using Agilent LCMS (1260/6120) or thin layer chromatography. The solvent eluting systems for column chromatography and TLC included a) dichloromethane/methanol, b) petroleum ether/ethyl acetate, or other systems as indicated. The ratio of the solvents was adjusted according to the polarity of the compound, and further adjusted by addition of a small amount of TEA, or an acidic or alkaline reagent as needed. The compound purification was alternatively done using Waters’ MS-guided automated preparation system (abbreviated as prep-HPLC) with a MS detector (SQD2) , eluting at a flow rate of 20 mL/min in an appropriate acetonitrile/water (containing 0.1%TFA or formic acid) or acetonitrile/water (containing 0.05%of 25-28%ammonium hydroxide) gradient (XBridge-C18, 19×150 mm, 5 μm) . Some compounds were prepared as HCl salts after prep-HPLC purification by addition of 1 N HCl to the collected fractions, followed by drying under reduced pressure.

The abbreviation TEA refers to triethylamine.

The abbreviation TFA refers to trifluoroacidic acid.

The abbreviation DMF refers to N, N-dimethylformamide.

The abbreviation THF refers to tetrahydrofuran.

The abbreviation PE refers to petroleum ether.

The abbreviation UHP refers to urea hydrogen peroxide complex.

The abbreviation TFAA refers to trifluoroacetic anhydride.

The abbreviation DIPEA refers to N, N-diisopropylethylamine.

The abbreviation DMAP refers to 4-dimethylaminopyridine.

The abbreviation NBS refers to N-bromosuccinimide.

The abbreviation NIS refers to N-iodosuccinimide.

The abbreviation NCS refers to N-chlorosuccinimide.

Pd (dppf) Cl ₂ refers to [1, 1'-bis (diphenylphosphino) ferrocene] dichloropalladium.

Example A1.7-Bromo-6- (trifluoromethyl) -3H-pyrido [2, 1-f] [1, 2, 4] triazine-4, 8-dione (Intermediate 1g)

Step 1. Methyl 4- (trifluoromethyl) picolinate (1b)

To a mixture of 4- (trifluoromethyl) picolinic acid 1a (25 g, 131 mmol) , dichloromethane (500 mL) and DMF (1 mL) at 0℃ was added a solution of oxalyl chloride (17.45 g, 137 mmol) in dichloromethane (20 mL) . After stirring at room temperature for 18 hours, to the mixture was added methanol (10 mL) . The reaction mixture was then concentrated to dryness under reduced pressure to give the title compound 1b (26.47 g, 98%) .

MS m/z (ESI) : 206 [M+1]

Step 2.2- (Methoxycarbonyl) -4- (trifluoromethyl) pyridine 1-oxide (1c)

To a mixture of 1b (26.47 g, 128.5 mmol) , UHP (24.1 g, 257 mmol) and dichloromethane (300 mL) at 0℃ was added TFAA (36 mL, 257 mmol) in portions. After stirring at room temperature for 12 hours, the reaction mixture was extracted with dichloromethane (3×200 mL) , and the combined organic phase was washed with saturated brine (200 mL) , dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure to give the title compound 1c (24.4 g, 86%) .

MS m/z (ESI) : 222 [M+1]

Step 3. Methyl 6-oxo-4- (trifluoromethyl) -1, 6-dihydropyridine-2-carboxylate (1d)

To a solution of 1c (24.4 g, 110 mmol) in DMF (100 mL) at 0℃ was added TFAA (61.6 mL, 440 mmol) dropwise. The resulting mixture was heated to 60℃ and stirred for 12 hours. After cooling to room temperature, the reaction mixture was quenched with water (100 mL) and filtered. The solid was collected to give the title compound 1d (17.4 g, 71%) .

MS m/z (ESI) : 222 [M+1]

Step 4. Methyl 1-amino-6-oxo-4- (trifluoromethyl) -1, 6-dihydropyridine-2-carboxylate (1e)

To a mixture of 1d (17.4 g, 78.4 mmol) , cesium carbonate (20.4 g, 62.7 mmol) and DMF (60 mL) was added O-diphenylphosphinylhydroxylamine (27.4 g, 117.6 mmol) . The resulting mixture was heated to 50℃, stirred for 1 hour, and then quenched with water (100 mL) and filtered. The solid was collected to give the title compound 1e (25.81 g, 69%) .

MS m/z (ESI) : 237 [M+1]

¹H NMR (400 MHz, DMSO-d ₆) δ 7.14 –7.06 (m, 1H) , 6.88 (d, J = 2.2 Hz, 1H) , 6.38 (s, 2H) , 3.90 (s, 3H) .

Step 5.6- (Trifluoromethyl) -3H-pyrido [2, 1-f] [1, 2, 4] triazine-4, 8-dione (1f)

To a solution of 1e (2.28 g, 9.65 mmol) in isopropanol (20 ml) was added formamidine acetate (4.02 g, 38.6 mmol) . The mixture was stirred at 90℃ for 12 h. After cooling to room temperature, the mixture was concentrated to dryness and the residue was purified by silica gel chromatography (dichloromethane/methanol = 10/1) to give the title compound 1f (2.0 g, 90%) .

MS m/z (ESI) : 232 [M+1]

Step 6.7-Bromo-6- (trifluoromethyl) -3H-pyrido [2, 1-f] [1, 2, 4] triazine-4, 8-dione (1g)

To a solution of 1f (2.00 g, 8.65 mmol) in DMF (20 mL) was added NBS (3.08 g, 17.3 mmol) and the resulting mixture was stirred at 50℃ for 12 h. After cooling to room temperature, the mixture was added with water (100 mL) and filtered. The filtered cake was collected to give the title compound 1g (2.00 g, solid, 75%) .

MS m/z (ESI) : 310, 312 [M+1]

Example A2.7-Bromo-6-chloro-3H-pyrido [2, 1-f] [1, 2, 4] triazine-4, 8-dione (Intermediate 6j)

Step 1.6-Bromo-3-nitropyridin-2-ol (6b)

To a solution of 6-bromo-3-nitropyridin-2-amine 6a (50 g, 229 mmol) in H ₂SO ₄ (75 mL) at 0℃ was added NaNO ₂ (32.8 g, 475 mmol) in H ₂O (400 mL) . The resulting mixture was stirred for at 0℃ for 30 minutes, and then added with water (1 L) . The mixture was stirred for another 1 hour and filtered to give the title compound 6b (47 g, 94%) .

MS m/z (ESI) : 219, 221 [M+1]

Step 2.6-Bromo-2-methoxy-3-nitropyridine (6c)

To a mixture of 6b (47 g, 215 mmol) , Ag ₂CO ₃ (71.1 g, 258 mmol) and heptane was added MeI (36.5 g, 257 mmol) . The resulting mixture was heated to 100℃ and stirred for 16 hours. The mixture was concentrated under reduced pressure, and then diluted with dichloromethane (500 mL) and H ₂O (500 mL) . The separated organic layer was dried over anhydrous Na ₂SO ₄ and filtered. The filtrate was concentrated to dryness under reduced pressure and the residue was purified by silica gel column chromatography (PE/EtOAc =17/3) to give the title compound 6c (25 g, 50%) .

MS m/z (ESI) : 233, 235 [M+1]

Step 3.6-Bromo-2-methoxypyridin-3-amine (6d)

A mixture of 6c (25 g, 107 mmol) , NH ₄Cl (58 g, 1084 mmol) , iron powder (60 g, 1080 mmol) , ethanol (250 mL) and H ₂O (250 mL) was heated to 60℃ and stirred for 1 hour. After cooling to room temperature, the mixture was filtered, and the filtrate was concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (PE/EtOAc = 17/3) to give the title compound 6d (16.3 g, 75%) .

MS m/z (ESI) : 203, 205 [M+1]

Step 4. Methyl 5-amino-6-methoxypicolinate (6e)

A mixture of 6d (16.3 g, 80.3 mmol) , Pd (dppf) Cl ₂ (1.76 g, 2.41 mmol) , triethylamine (40.4 g, 399 mmol) and methanol (900 mL) was purged with carbon monoxide for 1 hour at -40℃. The reaction mixture was then heated to 100℃ and stirred for 16 hours in a sealed tube. After cooling to room temperature, the mixture was filtered, and the filtrate was concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (PE/EtOAc = 4/1) to give the title compound 6e (13.5 g, 92%) .

MS m/z (ESI) : 183 [M+1]

Step 5. Methyl 5-amino-4-chloro-6-methoxypicolinate (6f)

A solution of 6e (11.5 g, 63.1 mmol) and NCS (9.27 g, 69.4 mmol) in DMF (150 mL) was stirred at 50℃ for 1 hour. After cooling to room temperature, to the mixture was added saturated sodium bicarbonate aqueous solution (200mL) , and the mixture was extracted with EtOAc (200 mL) . The organic phase was separated and concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (PE/EtOAc = 7/3) to give the title compound 6f (8.5 g, 62%) .

MS m/z (ESI) : 217, 219 [M+1]

Step 6. Methyl 5-bromo-4-chloro-6-methoxypicolinate (6g)

To a mixture of tert-butyl nitrite (6.07 g, 58.9 mmol) , CuBr ₂ (13.15 g, 58.9 mmol) and acetonitrile (100 mL) was added 6f (8.5 g, 39.2 mmol) in portions. The mixture was stirred at 25℃ for 30 minutes and to which was added saturated sodium bicarbonate aqueous solution (200 mL) . The mixture was extracted with EtOAc (200 mL) , and the organic phase was separated and concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (PE/EtOAc = 7/3) to give the title compound 6g (9.5 g, 86%) .

MS m/z (ESI) : 280, 282 [M+1]

Step 7. Methyl 5-bromo-4-chloro-6-oxo-1, 6-dihydropyridine-2-carboxylate (6h)

A mixture of 6g (9.5 g, 33.9 mmol) and a solution of HCl in dioxane (4 M, 100 mL) was stirred at 100℃ for 16 hours. The mixture was concentrated to dryness under reduced pressure to give the title compound 6h (8.5 g, 97%) .

MS m/z (ESI) : 266, 268 [M+1]

Step 8. Methyl 1-amino-5-bromo-4-chloro-6-oxo-1, 6-dihydropyridine-2-carboxylate (6i)

A mixture of 6h (8.5 g, 31.9 mmol) , Cs ₂CO ₃ (5.2 g, 16.0 mmol) and O-diphenylphosphinylhydroxylamine (8.93 g, 38.3 mmol) in DMF (100 mL) was stirred at 25℃ for 30 minutes. The reaction mixture was poured into HCl (1 N, 200 mL) at 0℃ and extracted with EtOAc (200 mL) . The organic phase was washed with saturated sodium bicarbonate aqueous solution (100 mL) and concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (PE/EtOAc = 3/2) to give the title compound 6i (6.5 g, 73%) .

MS m/z (ESI) : 281, 283 [M+1]

Step 9.7-Bromo-6-chloro-3H-pyrido [2, 1-f] [1, 2, 4] triazine-4, 8-dione (6j)

A solution of 6i (2.0 g, 7.11 mmol) and formamidine acetate (2.96 g, 28.4 mmol) in isopropanol (20 mL) was stirred at 90℃ for 16 hours. After cooling to room temperature, the reaction mixture was concentrated to dryness under reduced pressure and the residue was purified by silica gel column chromatography (dichloromethane/methanol = 9/1) to give the title compound 6j (1.1 g, solid, 56%) .

MS m/z (ESI) : 276, 278 [M+1]

Intermediate 8c was prepared following the procedures of step 8 and step 9 for the synthesis of intermediate 6j, except that methyl 5-bromo-6-oxo-1, 6-dihydropyridine-2-carboxylate 8a was used instead of 6h.

Example A3. Tert-butyl (S) -4- (7-bromo-6-methyl-8-oxo-8H-pyrido [2, 1-f] [1, 2, 4] triazin-4-yl) -3-methylpiperazine-1-carboxylate (Intermediate 23g)

Step 1. Methyl 5-amino-4-bromo-6-methoxypicolinate (23a)

To a solution of 6e (2 g, 11 mmol) in DMF (8 mL) at 0℃ was added NBS (1.95 g, 10.98 mmol) . The resulting mixture was stirred at 0 ℃ for 5 min and then concentrated to dryness. The residue was purified by prep-HPLC to give the title compound 23a. The above procedure was repeated and two batches of 23a were combined with a total quantity of 4.63 g (81%) .

MS m/z (ESI) : 261, 263 [M+1]

¹H NMR (400 MHz, CDCl ₃) δ 7.87 (s, 1H) , 4.53 (s, 2H) , 4.08 (s, 3H) , 3.91 (s, 3H) .

Step 2. Methyl 5-amino-6-methoxy-4-methylpicolinate (23b)

To a solution of 23a (4.63 g, 17.7 mmol) in 1, 4-dioxane (150 mL) were added water (15 mL) , K ₂CO ₃ (9.10 g, 70.9 mmol) , trimethylboroxine (4.45 g, 35.5 mmol) and Pd (dppf) Cl ₂ (649 mg, 0.886 mmol) . The resulting mixture was heated to 100℃, stirred for 18 h and cooled to room temperature. The mixture was filtered through a pad of celite, and the filtrate was concentrated to dryness. The residue was purified by silica gel column chromatography (PE/EtOAc = 100/0 to 1/1) to give the title compound 23b (3.35 g, 96%) .

MS m/z (ESI) : 197 [M+1]

¹H NMR (400 MHz, CDCl ₃) δ 7.58 (s, 1H) , 4.07 (s, 3H) , 3.91 (s, 3H) , 2.18 (s, 3H) .

Step 3. Methyl 5-bromo-6-methoxy-4-methylpicolinate (23c)

To a solution of tert-butyl nitrite (2.64 g, 25.6 mmol) in acetonitrile (66 mL) at 0℃ was added CuBr ₂ (5.72 g, 25.6 mmol) , followed by a solution of 23b (3.35 g, 17.1 mmol) in acetonitrile (33 mL) . The mixture was stirred at room temperature for 1 h and then added with a saturated sodium bicarbonate aqueous solution (200 mL) . The mixture was extracted with EtOAc (2×200 mL) . The combined organic phase was washed with saturated sodium bicarbonate aqueous solution (200 mL) and saturated brine (100 mL) sequentially. The solution was dried over anhydrous sodium sulphate and filtered. The organic phase was concentrated to dryness and the residue was purified by silica gel column chromatography (PE/EtOAc = 100/0 to 1/1) to give the title compound 23c (3.42 g, 77%) .

MS m/z (ESI) : 260, 262 [M+1]

¹H NMR (400 MHz, CDCl ₃) δ 7.60 (s, 1H) , 4.09 (s, 3H) , 3.95 (s, 3H) , 2.45 (s, 3H) .

Step 4. Methyl 5-bromo-4-methyl-6-oxo-1, 6-dihydropyridine-2-carboxylate (23d)

A mixture of 23c (1.39 g, 5.34 mmol) and a solution of HCl in 1, 4-dioxane (4 M, 30 mL) was heated to 90℃ in a 100-mL sealed tube and stirred for 18 h. After cooling to room temperature, the mixture was concentrated to dryness. The residue was suspended in a mixture of dichloromethane (100 mL) and water (20 mL) and the pH was adjusted to 7 with a saturated sodium bicarbonate aqueous solution. The precipitate was collected by filtration to give a part of product. The organic layer in the filtrate was separated, dried over anhydrous sodium sulphate and filtered. The filtrate was concentrated to dryness to give another part of product. The two parts of product were combined to give 23d (1.61 g) and it was used directly in the next step without further purification.

MS m/z (ESI) : 246, 248 [M+1]

Step 5. Methyl 1-amino-5-bromo-4-methyl-6-oxo-1, 6-dihydropyridine-2-carboxylate (23e)

To a solution of 23e (1.61 g, 5.34 mmol) in DMF (15 mL) were added with O-diphenylphosphinylhydroxylamine (1.87 g, 8.01 mmol) and Cs ₂CO ₃ (1.39 g, 4.27 mmol) . The mixture was heated to 50℃ and stirred for 1 h. After cooling to room temperature, the mixture was diluted with water (60 mL) and extracted with EtOAc (3×60 mL) . The combined organic phase was washed with water (60 mL) and saturated brine (60 mL) , dried over anhydrous sodium sulphate and filtered. The filtrate was concentrated to dryness and the residue was purified by silica gel column chromatography (dichloromethane/EtOAc = 100/0 to 1/1) to give the title compound 23e (0.86 g, 66%) .

MS m/z (ESI) : 261, 263 [M+1]

¹H NMR (400 MHz, DMSO-d ₆) δ 6.66 (s, 1H) , 6.19 (s, 2H) , 3.86 (s, 3H) , 2.30 (s, 3H) .

Step 6.7-Bromo-4-hydroxy-6-methyl-8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one (23f)

To a solution of 23e (1.86 g, 7.12 mmol) in isopropanol (30 mL) was added formimidamide acetate (1.48 g, 14.25 mmol) . The resulting mixture was heated to 90℃ and stirred for 64 h. After cooling to room temperature, the mixture was diluted with water (60 mL) and stirred for 30 min. The precipitate was collected by filtration to give the title compound 23f (1.43 g, 79%) .

MS m/z (ESI) : 256, 258 [M+1]

¹H NMR (400 MHz, DMSO-d ₆) δ 12.33 (s, 1H) , 8.01 (s, 1H) , 7.11 (s, 1H) , 2.41 (s, 3H) .

Step 7. tert-Butyl (S) -4- (7-bromo-6-methyl-8-oxo-8H-pyrido [2, 1-f] [1, 2, 4] triazin-4-yl) -3-methylpiperazine-1-carboxylate (23g)

A mixture of 23g (1.43 g, 5.58 mmol) , POCl ₃ (90 mL) and DIPEA (2.88 g, 22.32 mmol) was heated to 60℃ and stirred for 1 h. After cooling to room temperature, the mixture was concentrated to dryness. The residue was dissolved in dichloromethane (80 mL) and then added with DIPEA (3.61 g, 27.9 mmol) and tert-butyl (S) -3-methylpiperazine-1-carboxylate (1.68 g, 8.38 mmol) . The mixture was stirred for 30 min, diluted with water (80 mL) and extracted with dichloromethane (2×80 mL) . The combined organic phase was dried over anhydrous sodium sulphate and filtered. The filtrate was concentrated to dryness and the residue was purified by silica gel column chromatography (dichloromethane/EtOAc = 100/0 to 3/2) to give the title compound 23g (2.15 g, 88%) .

MS m/z (ESI) : 438, 440 [M+1]

¹H NMR (400 MHz, CDCl ₃) δ 8.24 (s, 1H) , 6.55 (s, 1H) , 4.65 (s, 1H) , 4.15 (dd, J = 24.0, 16.7 Hz, 2H) , 3.96 (s, 1H) , 3.61 -3.51 (m, 1H) , 3.20 (s, 2H) , 2.50 (s, 3H) , 1.49 (s, 9H) , 1.43 (d, J = 6.7 Hz, 3H) .

Example 1.2- (1-Acryloyl-4- (7- (2-amino-3, 5-dichloro-6-fluorophenyl) -8-oxo-6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-4-yl) piperazin-2-yl) acetonitrile

Step 1.2- (4- (7-Bromo-8-oxo-6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-4-yl) piperazin-2-yl) acetonitrile (4a)

To a solution of 1g (300 mg, 0.97 mmol) in dichloromethane (5 mL) were added DIPEA (627 mg, 4.85 mmol) , DMAP (95 mg, 0.78 mmol) , 2, 4, 6-triisopropylbenzenesulfonyl chloride (588 mg, 1.94 mmol) . The mixture was stirred at 30℃ for 5 h and then added with 2- (piperazin-2-yl) acetonitrile (243 mg, 1.94 mmol) . The mixture was stirred at 30℃ for another 12 h, added with water (10 mL) and extracted with dichloromethane (20 mL) . The organic phase was concentrated to dryness under reduced pressure and the residue was purified by silica gel column chromatography (dichloromethane/methanol = 20/1) to give the title compound 4a (240 mg, 59%) .

MS m/z (ESI) : 418 [M+1]

Step 2.2- (4- (7- (2-Amino-6-fluorophenyl) -8-oxo-6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-4-yl) piperazin-2-yl) acetonitrile (4b)

To a solution of 4a (180 mg, 0.43 mmol) in 1, 4-dioxane/H ₂O (4 mL, 20/1, v/v) were added (2-amino-6-fluorophenyl) boronic acid (133 mg, 0.86 mmol) , K ₂CO ₃ (178 mg, 1.29 mmol) and Pd (dppf) Cl ₂ (33 mg, 0.04 mmol) . The mixture was stirred at 80℃ under nitrogen atmosphere for 1.5 h. After cooling to room temperature, the mixture was filtered through a pad of anhydrous Na ₂SO _4, and the filtrate was concentrated. The residue was purified by silica gel column chromatography (dichloromethane/methanol = 50/1) to give the title compound 4b (60 mg, 31%) .

MS m/z (ESI) : 448 [M+1]

Step 3.2- (1-Acryloyl-4- (7- (2-amino-6-fluorophenyl) -8-oxo-6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-4-yl) piperazin-2-yl) acetonitrile (4c)

To a solution of 4b (60 mg, 0.134 mmol) in THF/H ₂O (5 ml) at 0℃ were added sodium bicarbonate (56 mg, 0.67 mmol) and acryloyl chloride (12 mg, 0.13 mmol) sequentially. The mixture was stirred at 0℃ for 20 min, added with water (5 mL) and extracted with dichloromethane (10 mL) . The separated organic phase was concentrated to dryness under reduced pressure and the residue was purified by silica gel column chromatography (dichloromethane/methanol = 10/1) to give the title compound 4c (26 mg, 39%) .

MS m/z (ESI) : 502 [M+1]

Step 4.2- (1-Acryloyl-4- (7- (2-amino-3, 5-dichloro-6-fluorophenyl) -8-oxo-6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-4-yl) piperazin-2-yl) acetonitrile (4)

To a solution of 4c (26 mg, 0.052 mmol) in DMF (2.5 mL) were added NCS (14 mg, 0.104 mmol) . The mixture was heat to 50℃ and stirred for 1 h. After cooling to room temperature, the mixture was added with water (2.5 mL) and the resulting mixture was purified by prep-HPLC to give the title compound 4 (2.3 mg, solid, 8%) .

MS m/z (ESI) : 570 [M+1]

¹H NMR (400 MHz, CD ₃OD) δ 8.43 (d, J = 4.2 Hz, 1H) , 7.43 (dd, J = 7.5, 1.0 Hz, 1H) , 7.29 (s, 1H) , 6.79 (s, 1H) , 6.33 (s, 1H) , 5.84 (d, J = 9.8 Hz, 1H) , 4.49 (dt, J = 64.2, 33.0 Hz, 3H) , 4.19 –3.62 (m, 4H) , 2.99 (s, 2H) .

Compound 7 was prepared according to the methods of Compound 4, except that 6j was used instead of 1g in step 1.

Example 2.4- ( (S) -4-acryloyl-2-methylpiperazin-1-yl) -7- (2-amino-3, 5-dichloro-6-fluorophenyl) -2- ( ( (S) -1-methylpyrrolidin-2-yl) methoxy) -6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one

Step 1. Methyl 1- (3-benzoylthioureido) -6-oxo-4- (trifluoromethyl) -1, 6-dihydropyridine-2-carboxylate (9a)

To a solution of 1e (10.00 g, 42.35 mmol) in THF (200 mL) was added benzoyl isothiocyanate (8.29 g, 50.8 mmol) . The mixture was stirred at room temperature for 12 h and then concentrated to dryness. The residue was added with ethyl acetate (100 mL) and stirred for 0.5 h. The mixture was filtered, and the filtered cake was collected to give the title compound 9a (14.0 g, 83%) .

MS m/z (ESI) : 400 [M+1]

Step 2.2-Mercapto-6- (trifluoromethyl) -3H-pyrido [2, 1-f] [1, 2, 4] triazine-4, 8-dione (9b)

To a solution of 9a (14.00 g, 35.1 mmol) in methanol (200 mL) at 0℃ was added sodium methoxide (30%in methanol, 50.5 g, 280 mmol) . The resulting mixture was stirred at room temperature for 12 h and then concentrated to dryness. The resulting mixture was added with water (100 mL) and adjusted to pH = 7 with 4 N HCl. The mixture was filtered, and the filter cake was collected to give the title compound 9b (7.98 g, 86%) .

MS m/z (ESI) : 264 [M+1]

Step 3.2- (Methylthio) -6- (trifluoromethyl) -3H-pyrido [2, 1-f] [1, 2, 4] triazine-4, 8-dione (9c)

To a solution of 9b (7.98 g, 30.3 mmol) in THF (100 mL) was added methyl iodide (8.61 g, 60.6 mmol) . The resulting mixture was heated to 50℃ and stirred for 12 h. After cooling to room temperature, the mixture was concentrated to dryness and added with ethyl acetate (100 mL) . The resulting mixture was stirred for 0.5 h and filtered. The filtered cake was collected to give the title compound 9c (7.90 g, 94%) .

MS m/z (ESI) : 278 [M+1]

Step 4. (S) -2- ( (1-Methylpyrrolidin-2-yl) methoxy) -6- (trifluoromethyl) -3H-pyrido [2, 1-f] [1, 2, 4] triazine-4, 8-dione (9d)

To a solution of (S) - (1-methylpyrrolidin-2-yl) methanol (4.92 g, 42.8 mmol ) in DMF (40 mL) at 0℃ was added NaH (60%) (2.28 g, 57.0 mmol) . After stirring at 0℃ for 1 h, the mixture was added with 9c (7.90 g, 28.5 mmol) and heated to 80℃ for 8 h. After cooling to room temperature, the reaction mixture was added with saturated ammonium chloride aqueous solution (40 mL) , filtered, and concentrated to dryness. The residue was purified by silica gel column chromatography (CH ₂Cl ₂/MeOH = 5/1) to give the title compound 9d (6.80 g, 69%) .

MS m/z (ESI) : 345 [M+1]

Step 5. (S) -7-Bromo-2- ( (1-methylpyrrolidin-2-yl) methoxy) -6- (trifluoromethyl) -3H-pyrido [2, 1-f] [1, 2, 4] triazine-4, 8-dione (9e)

To a solution of 9d (6.80 g, 19.8 mmol) in acetonitrile (100 mL) was added NBS (7.04 g, 39.5 mmol) . The resulting mixture was stirred at room temperature for 4 h and then concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (CH ₂Cl ₂/MeOH = 20/1) to give the title compound 9e (7.25 g, 87%) .

MS m/z (ESI) : 423, 425 [M+1]

Step 6. Tert-butyl (S) -4- (7-bromo-2- ( ( (S) -1-methylpyrrolidin-2-yl) methoxy) -8-oxo-6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-4-yl) -3-methylpiperazine-1-carboxylate (9f)

A mixture of 9e (1.00 g, 2.36 mmol) and POCl ₃ (10 mL) was stirred at 95℃ for 6 h. After cooling to room temperature, the mixture was concentrated to dryness and the residue was added with dichloromethane (10 mL) , DIPEA (1.53 g, 11.8 mmol) and tert-butyl (S) -3-methylpiperazine-1-carboxylate (0.52 g, 2.6 mmol) . The mixture was stirred for 2 h and added with water (20 mL) . The resulting mixture was extracted with DCM (3×20 mL) and washed with brine (3×60 mL) . The organic phase was dried over anhydrous Na ₂SO ₄, filtered and concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (CH ₂Cl ₂/MeOH = 25/1) to give the title compound 9e (600 mg, 41%) .

MS m/z (ESI) : 605, 607 [M+1]

Step 7. Tert-butyl (S) -4- (7- (2-amino-6-fluorophenyl) -2- ( ( (S) -1-methylpyrrolidin-2-yl) methoxy) -8-oxo-6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-4-yl) -3-methylpiperazine-1-carboxylate (9g)

To a solution of 9f (520 mg, 0.86 mmol) in a mixture of toluene and H ₂O (8.4 mL, 20/1, v/v) was added (2-amino-6-fluorophenyl) boronic acid (400 mg, 2.58 mmol) , K ₂CO ₃ (238 mg, 1.72 mmol) , sodium bicarbonate (72 mg, 0.86 mmol) and Pd (PPh ₃) ₄ (199 mg, 0.17 mmol) . The mixture was stirred at 80℃ for 16 h. After cooling to room temperature, the mixture was filtered through a pad of anhydrous Na ₂SO ₄ and concentrated to dryness under reduced pressure. The residue was purified by prep-HPLC to give the title compound 9g (60 mg, 6%) .

MS m/z (ESI) : 636 [M+1]

Step 8.7- (2-Amino-6-fluorophenyl) -4- ( (S) -2-methylpiperazin-1-yl) -2- ( ( (S) -1-methylpyrrolidin-2-yl) methoxy) -6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one hydrochloride (9h)

To a solution of 9g (30 mg, 0.05 mmol) in ethanol (2.5 mL) was added a solution of HCl in ethanol (10 N, 2.5 mL) . The mixture was stirred for 1 h and then concentrated to dryness under reduced pressure to give the title compound 9h (28.7 mg, 100%) .

MS m/z (ESI) : 536 [M+1]

Step 9.4- ( (S) -4-acryloyl-2-methylpiperazin-1-yl) -7- (2-amino-6-fluorophenyl) -2- ( ( (S) -1-methylpyrrolidin-2-yl) methoxy) -6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one (9i)

To a solution of 9h (28.7 mg, 0.05 mmol) in a mixed solvent of THF and water (5 ml, 4/1, v/v) at 0℃ were added sodium bicarbonate (20 mg, 0.25 mmol) and acryloyl chloride (4 mg, 0.05 mmol) . The resulting mixture was stirred at 0℃ for 10 min, filtered through a pad of anhydrous Na ₂SO ₄ and concentrated to dryness under reduced pressure. The residue was purified by prep-HPLC to give the title compound 9i (5 mg, 18%) .

MS m/z (ESI) : 590 [M+1]

Step 10.4- ( (S) -4-acryloyl-2-methylpiperazin-1-yl) -7- (2-amino-3, 5-dichloro-6-fluorophenyl) -2- ( ( (S) -1-methylpyrrolidin-2-yl) methoxy) -6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one monoformate (9)

To a solution of 9i (5 mg, 0.0085 mmol) in DMF (3.0 mL) was added NCS (2.3 mg, 0.017 mmol) . The mixture was then stirred at 80℃ for 8 h, added with water (2.5 mL) and purified by prep-HPLC to give the title compound 9 (3.5 mg, solid, 63%) .

MS m/z (ESI) : 658 [M+1]

¹H NMR (400 MHz, CD ₃OD) δ 8.53 (s, 1H) , 7.45 (d, J = 7.5 Hz, 1H) , 7.30 (s, 1H) , 6.82 (d, J = 13.5 Hz, 1H) , 6.31 (d, J = 16.8 Hz, 1H) , 5.83 (d, J = 12.0 Hz, 1H) , 4.77 (d, J = 13.3 Hz, 1H) , 4.67 –4.36 (m, 3H) , 4.14 (d, J = 64.9 Hz, 2H) , 3.75 (dd, J = 59.6, 12.6 Hz, 2H) , 3.50 (s, 3H) , 2.90 (dd, J = 29.5, 6.4 Hz, 4H) , 2.27 (d, J = 6.3 Hz, 1H) , 2.11 –1.89 (m, 3H) , 1.50 (d, J = 6.5 Hz, 3H) .

Example 3. (S) -4- (4-acryloyl-2-methylpiperazin-1-yl) -7- (2-amino-3, 5-dichloro-6-fluorophenyl) -6-chloro-8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one

Step 1. Tert-butyl (S) -4- (7-bromo-6-chloro-8-oxo-8H-pyrido [2, 1-f] [1, 2, 4] triazin-4-yl) -3-methylpiperazine-1-carboxylate (10a)

A solution of 6j (650 mg, 2.35 mmol) , DIPEA (1.21 g, 9.40 mmol) , 2, 4, 6- triisopropylbenzenesulfonyl chloride (1.44 g, 4.75 mmol) and DMAP (258 mg, 2.12 mmol) in dichloromethane (10 mL) at 30℃ was stirred for 5 hours. Tert-butyl (S) -3-methylpiperazine-1-carboxylate (951.33 mg, 4.75 mmol) was then added, and the reaction mixture was stirred at 30℃ for another 3 hours. The mixture was concentrated to dryness under reduced pressure and the residue was purified by silica gel column chromatography (dichloromethane/methanol = 9/1) to give the title compound 10a (867 mg, 81%) .

MS m/z (ESI) : 458, 460 [M+1]

Step 2. Tert-butyl (S) -4- (7- (2-amino-6-fluorophenyl) -6-chloro-8-oxo-8H-pyrido [2, 1-f] [1, 2, 4] triazin-4-yl) -3-methylpiperazine-1-carboxylate (10b)

A mixture of 10a (867 mg, 1.89 mmol) , K ₂CO ₃ (783.6 mg, 5.67 mmol) , Pd (dppf) Cl ₂ (138.2 mg, 0.189 mmol) , (2-amino-6-fluorophenyl) boronic acid (878.5 mg, 5.67 mmol) , dioxane (10 mL) and H ₂O (0.5 mL) at 80℃ was stirred for 1.5 hours. After cooling to room temperature, the mixture was concentrated to dryness under reduced pressure and the residue was purified by silica gel column chromatography (dichloromethane/methanol = 9/1) to give the title compound 10b (230 mg, 25%) .

MS m/z (ESI) : 489 [M+1]

Step 3. (S) -7- (2-Amino-6-fluorophenyl) -6-chloro-4- (2-methylpiperazin-1-yl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one (10c)

A mixture of 10b (230 mg, 0.471 mmol) and a solution of HCl in ethyl acetate (4 M, 2 mL) was stirred for 1.5 hours. The mixture was concentrated to dryness under reduced pressure to give the title compound 10c (183 mg, 100%) .

MS m/z (ESI) : 389 [M+1]

Step 4. (S) -4- (4-Acryloyl-2-methylpiperazin-1-yl) -7- (2-amino-6-fluorophenyl) -6-chloro-8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one (10d)

To a mixture of 10c (183 mg, 0.471 mmol) , sodium bicarbonate (198 mg, 2.35 mmol) , THF (3 mL) and H ₂O (1 mL) at 0℃ was added a solution of acryloyl chloride (42.6 mg, 0.47 mmol) in THF (1 mL) dropwise. The mixture was stirred at 0℃ for 30 minutes and concentrated to dryness under reduced pressure. The residue was purified by prep-TLC (dichloromethane/methanol = 24/1) to give the title compound 10d (120 mg, 58%) .

MS m/z (ESI) : 443 [M+1]

Step 5. (S) -4- (4-Acryloyl-2-methylpiperazin-1-yl) -7- (2-amino-3, 5-dichloro-6-fluorophenyl) -6-chloro-8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one (10)

A solution of 10d (120 mg, 0.27 mmol) and NCS (72.4 mg, 0.54 mmol) in DMF (2 mL) at 80℃ was stirred for 3 hours. After cooling to room temperature, the mixture was purified by prep-HPLC to give the title compound 10 (73 mg, solid, 53%) .

MS m/z (ESI) : 511 [M+1]

¹H NMR (400 MHz, DMSO-d ₆) δ 8.36 (d, J = 4.0 Hz, 1H) , 7.57 (d, J = 7.6 Hz, 1H) , 6.93 (d, J = 9.9 Hz, 1H) , 6.83 (s, 1H) , 6.20 (d, J = 16.5 Hz, 1H) , 5.78 –5.73 (m, 1H) , 5.60 (d, J =7.2 Hz, 2H) , 4.84 –4.67 (m, 1H) , 4.46 –4.33 (m, 1H) , 4.20 –4.10 (m, 1H) , 4.06 –4.00 (m, 1H) , 3.80 –3.57 (m, 1H) , 3.36 –3.30 (m, 1H) , 3.26 –3.16 (m, 1H) , 1.34 –1.26 (m, 3H) .

Compounds

2, 3, 5, 6, 8, 11, 14, 15 and 16 were prepared according to the methods of Compound 10, except that different compounds were used to replace 6j and tert-butyl (S) -3-methylpiperazine-1-carboxylate in step 1, respectively. Compound 3 is a byproduct when synthesizing Compound 2. For the synthesis of Compound 8, DIPEA was used to replace K ₂CO ₃ in step 2.

Compounds 1 and 13 were prepared following the procedures of step 1 through step 4 for Compound 10, except that different compounds were used to replace 6j in step 1 and (2-amino-6-fluorophenyl) boronic acid in step 2.

Compound 23 was prepared following the procedures of steps 2-5 for Compound 10, except that 23g was used instead 10a.

¹H NMR data of Compounds 1, 2, 3, 5, 6, 8, 11, 13, 14, 15, 16 and 23 are shown below:

Example 4. Chiral separation of

Compounds

5 and 15

Compound 5 (820 mg) was separated by supercritical fluid chromatography (Column: CHIRALCEL OD-H, 20 mm × 250 mm, 10 μm; fluent: methanol/carbon dioxide) to give two stereoisomers of 12a (375 mg, t _R = 9.32 min) and 12b (276 mg, t _R = 11.68 min) .

12a

MS m/z (ESI) : 545 [M+1]

¹H NMR (400 MHz, DMSO-d ₆) δ 8.47 (s, 1H) , 7.57 (d, J = 7.6 Hz, 1H) , 6.93 (s, 1H) , 6.91 –6.74 (m, 1H) , 6.20 (d, J = 16.0 Hz, 1H) , 5.75 (dd, J = 10.4, 2.1 Hz, 1H) , 5.61 (s, 2H) , 4.75 (d, J = 28.1 Hz, 1H) , 4.50 –3.95 (m, 3H) , 3.77 –3.34 (m, 2H) , 3.25 –2.96 (m, 1H) , 1.36 (s, 3H) .

12b

MS m/z (ESI) : 545 [M+1]

¹H NMR (400 MHz, DMSO-d ₆) δ 8.46 (s, 1H) , 7.57 (d, J = 7.6 Hz, 1H) , 6.96 (s, 1H) , 6.84 (t, J = 10.8 Hz, 1H) , 6.20 (dd, J = 16.8, 6.1 Hz, 1H) , 5.75 (dd, J = 10.4, 2.2 Hz, 1H) , 5.58 (s, 2H) , 4.81 (d, J = 24.4 Hz, 1H) , 4.44 –3.97 (m, 3H) , 3.90 –3.36 (m, 2H) , 3.25 –2.99 (m, 1H) , 1.34 (s, 3H) .

Compound 15 (1.77 g) was separated by supercritical fluid chromatography (Column: CHIRALCEL OD-H, 20 mm × 250 mm, 10 μm; fluent: methanol/carbon dioxide) to give two stereoisomers of 15a (1.053 g, t _R = 11.41 min) and 15b (693 mg, t _R = 12.31 min) .

15a

MS m/z (ESI) : 559 [M+1]

¹H NMR (400 MHz, CD ₃OD) δ 8.43 (s, 1H) , 7.42 (d, J = 7.5 Hz, 1H) , 7.19 (s, 1H) , 6.77 (dd, J = 16.8, 10.8 Hz, 1H) , 6.29 (ddd, J = 16.7, 6.4, 1.8 Hz, 1H) , 5.88 –5.76 (m, 1H) , 4.90 (s, 2H) , 4.55 (s, 1H) , 4.43 –4.26 (m, 2H) , 3.95 –3.77 (m, 2H) , 3.46 (dd, J = 14.0, 3.8 Hz, 1H) , 1.48 (t, J = 6.7 Hz, 3H) , 1.33 (dd, J = 37.5, 6.8 Hz, 3H) .

15b

MS m/z (ESI) : 559 [M+1]

¹H NMR (400 MHz, CD ₃OD) δ 8.44 (s, 1H) , 7.43 (d, J = 7.5 Hz, 1H) , 7.20 (d, J = 4.2 Hz, 1H) , 6.81 (d, J = 1.5 Hz, 1H) , 6.35 –6.24 (m, 1H) , 5.88 –5.78 (m, 1H) , 4.89 (d, J = 27.7 Hz, 2H) , 4.55 (s, 1H) , 4.31 (d, J = 13.9 Hz, 2H) , 3.91 (dd, J = 9.2, 3.6 Hz, 2H) , 3.45 (dd, J =13.9, 3.8 Hz, 1H) , 1.47 (d, J = 6.5 Hz, 3H) , 1.35 (dd, J = 21.4, 6.8 Hz, 3H) .

Example 5. (S) -7- (2-amino-3, 5-dichloro-6-fluorophenyl) -4- (4- (2-fluoroacryloyl) -2-methylpiperazin-1-yl) -6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one (Compound 17)

Step 1. Tert-butyl (S) -4- (7- (2-amino-3, 5-dichloro-6-fluorophenyl) -8-oxo-6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-4-yl) -3-methylpiperazine-1-carboxylate (17b)

To a solution of 17a (1.33 g, 2.55 mmol) in DMF (10 mL) was added NCS (680 mg, 5.10 mmol) . The resulting mixture was stirred at 80℃ for 2 h and then cooled to room temperature. The mixture was added with water (1 mL) and concentrated to dryness. The residue was purified by silica gel column chromatography (dichloromethane/methanol = 50/1) to give the title compound 17b (1.43 g, 95%) .

MS m/z (ESI) : 591, 593 [M+1]

Step 2. (S) -7- (2-amino-3, 5-dichloro-6-fluorophenyl) -4- (2-methylpiperazin-1-yl) -6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one hydrochloride (17c)

To a solution of 17b (150 mg, 0.254 mmol) in 1, 4-dioxane (2 ml) was added a solution of HCl in dioxane (4 M, 10 mL) . The resulting mixture was stirred for 2 h and concentrated to dryness to give the title compound 17c (143 mg) . The crude product was used directly in the next step without further purification.

MS m/z (ESI) : 491, 493 [M+1]

Step 3. (S) -7- (2-amino-3, 5-dichloro-6-fluorophenyl) -4- (4- (2-fluoroacryloyl) -2-methylpiperazin-1-yl) -6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one (17)

To a solution of 17c (191 mg, 0.338 mmol) in DMF (5 mL) were added 2-fluoroacrylic acid (30 mg, 0.338 mmol) and DIPEA (218 mg, 1.69 mmol) . The resulting mixture was stirred for 10 min and added with HATU (154 mg, 0.406 mmol) . The mixture was stirred for 1 h, added with water (10 ml) and filtered. The filter cake was collected and purified by prep-HPLC to give the title compound 17 (56 mg, 29%over two steps) .

MS m/z (ESI) : 563, 565 [M+1]

¹H NMR (400 MHz, CD ₃OD) δ 8.44 (s, 1H) , 7.42 (d, J = 7.4 Hz, 1H) , 7.16 (s, 1H) , 5.35 (dd, J = 22.4, 3.9 Hz, 1H) , 5.26 (dd, J = 9.6, 3.8 Hz, 1H) , 4.93 (s, 2H) , 4.28 (dt, J = 121.4, 41.2 Hz, 4H) , 3.88 –3.50 (m, 2H) , 3.18 (d, J = 40.7 Hz, 1H) , 1.50 (t, J = 6.5 Hz, 3H) .

Compounds 18, 19 and 20 were prepared according to the procedures for Compound 17, except different acids were used instead of 2-fluoroacrylic acid.

¹H NMR data of Compounds 18, 19 and 20 are shown below:

Example 6. (S) -7- (2-amino-3, 5-dichloro-6-fluorophenyl) -4- (2-methyl-4- (vinylsulfonyl) piperazin-1-yl) -6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one (Compound 21)

To a solution of 17c (153 mg, 0.271 mmol) in DMF (5 ml) at 0℃ were added 2-chloroethane-1-sulfonyl chloride (44 mg, 0.27 mmol) and TEA (165 mg, 1.63 mmol) . The resulting mixture was stirred at 0℃ for 2 h and concentrated to dryness. The residue was purified by prep-HPLC to give the title compound 21 (24 mg, 17%) .

MS m/z (ESI) : 581, 583 [M+1]

¹H NMR (400 MHz, CD ₃OD) δ 8.42 (s, 1H) , 7.42 (d, J = 7.5 Hz, 1H) , 7.13 (d, J = 2.1 Hz, 1H) , 6.65 (ddd, J = 16.5, 10.0, 3.6 Hz, 1H) , 6.18 (ddd, J = 13.3, 12.1, 1.8 Hz, 2H) , 4.94 (s, 1H) , 4.44 (dd, J = 21.9, 14.0 Hz, 1H) , 3.87 –3.68 (m, 2H) , 3.61 –3.45 (m, 1H) , 2.99 (ddd, J = 22.3, 12.9, 9.9 Hz, 2H) , 1.62 (t, J = 7.0 Hz, 3H) .

Example 7. (S) -4- (7- (2-amino-3, 5-dichloro-6-fluorophenyl) -8-oxo-6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-4-yl) -3-methylpiperazine-1-carbonitrile (Compound 22)

To a solution of 17c (153 mg, 0.27 mmol) in DMF (5 ml) at 0℃ were added cyanic bromide (29 mg, 0.27 mmol) and TEA (84 mg, 0.81 mmol) . The resulting mixture was stirred at 0℃ for 2 h and concentrated to dryness. The residue was purified by prep-HPLC to give the title compound 22 (24 mg, 17%) .

MS m/z (ESI) : 516, 518 [M+1]

¹H NMR (400 MHz, CD ₃OD) δ 8.45 (s, 1H) , 7.42 (d, J = 7.4 Hz, 1H) , 7.10 (t, J = 6.6 Hz, 1H) , 4.33 (t, J = 13.6 Hz, 1H) , 3.98 –3.80 (m, 1H) , 3.58 –3.33 (m, 5H) , 1.72 –1.64 (m, 3H) .

Example 8. (S) -4- (4-acryloyl-2-methylpiperazin-1-yl) -7- (2-amino-3, 5-dichloro-6-fluorophenyl) -6-fluoro-8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one (Compound 25)

Step 1.7-Bromo-4-chloro-6-fluoro-8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one (25g)

To a mixture of 25f (1.56 g, 6 mmol) and POCl ₃ (120 mL) was added DIPEA (2.33 g, 18 mmol) dropwise until 25f was completely dissolved. The resulting mixture was heated to 60℃ and stirred for 30 min. After cooling to room temperature, the mixture was concentrated to dryness under reduced pressure and the residue (crude 25g) was used directly in the next step without further purification.

Step 2. tert-Butyl (S) -4- (7-bromo-6-fluoro-8-oxo-8H-pyrido [2, 1-f] [1, 2, 4] triazin-4-yl) -3-methylpiperazine-1-carboxylate (25h)

To a mixture of 25g (crude) and dichloromethane (30 mL) were added DIPEA (3.88 g, 30 mmol) and tert-butyl (S) -3-methylpiperazine-1-carboxylate (1.80 g, 9 mmol) . The resulting mixture was stirred for 30 min, added with water (100 mL) and extracted with dichloromethane (3×100 mL) . The combined organic phase was washed with saturated brine (100 mL) , dried over anhydrous sodium sulphate and filtered. The filtrate was concentrated to dryness and the residue was purified by silica gel column chromatography (dichloromethane/EtOAc = 100/0 to 3/2) to give the title compound 25h (840 mg, 32%over two steps) .

MS m/z (ESI) : 442 [M+1]

¹H NMR (400 MHz, CDCl ₃) δ 8.28 (s, 1H) , 6.52 (d, J = 7.2 Hz, 1H) , 4.64 (s, 1H) , 4.28 -3.85 (m, 3H) , 3.57 (td, J = 13.4, 3.3 Hz, 1H) , 3.30 -3.02 (m, 2H) , 1.50 (s, 9H) , 1.45 (d, J =6.7 Hz, 3H) .

Step 3. tert-Butyl (S) -4- (7- (2-amino-6-fluorophenyl) -6-fluoro-8-oxo-8H-pyrido [2, 1-f] [1, 2, 4] triazin-4-yl) -3-methylpiperazine-1-carboxylate (25i)

To a solution of 25h (420 mg, 0.95 mmol) in 1, 4-dioxane (20 mL) were added water (1 mL) , K ₂CO ₃ (525 mg, 2.85 mmol) and Pd (dppf) Cl ₂ (70 mg, 0.095 mmol) . The resulting mixture was stirred at 100℃ under nitrogen atmosphere for 30 min. After cooling to room temperature, the mixture was filtered, and the filtrate was concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane/methanol = 100/0 to 97/3) to give the title compound 25i (412 mg, 92%) .

MS m/z (ESI) : 473 [M+1]

¹H NMR (400 MHz, CDCl ₃) δ 8.31 (d, J = 1.7 Hz, 1H) , 7.10 (dd, J = 15.0, 8.1 Hz, 1H) , 6.80 (t, J = 8.1 Hz, 1H) , 6.53 (d, J = 8.2 Hz, 1H) , 6.38 (t, J = 8.8 Hz, 1H) , 5.17 (d, J = 5.3 Hz, 2H) , 4.69 (s, 1H) , 4.08 (d, J = 13.1 Hz, 1H) , 3.97 (s, 1H) , 3.81 (d, J = 13.2 Hz, 1H) , 3.56 (dd, J = 21.4, 10.5 Hz, 1H) , 3.15 (d, J = 75.5 Hz, 2H) , 1.43 (s, 9H) , 1.30 (d, J = 6.6 Hz, 3H) .

Step 4. (S) -4- (4-acryloyl-2-methylpiperazin-1-yl) -7- (2-amino-6-fluorophenyl) -6-fluoro-8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one (25j)

To a solution of 25i (412 mg, 0.87 mmol) in dichloromethane (8 mL) was added TFA (2 mL) . The resulting mixture was stirred for 1 h and concentrated to dryness under reduced pressure. The residue was dissolved in a mixture of THF (8 mL) and water (2 mL) , and then added with sodium bicarbonate (292 mg, 3.48 mmol) followed by a solution of acryloyl chloride (90 mg, 1 mmol) in THF (1 mL) . The mixture was stirred for 15 min, diluted with water (50 mL) and extracted with EtOAc (3×50 mL) . The combined organic phase was dried over anhydrous sodium sulphate and filtered. The filtrate was concentrated to dryness under reduced pressure and the residue was purified by silica gel column chromatography (dichloromethane/methanol = 100/0 to 19/1) to give the title compound 25j (171 mg, 46%) .

MS m/z (ESI) : 427 [M+1]

Step 5. (S) -4- (4-acryloyl-2-methylpiperazin-1-yl) -7- (2-amino-3, 5-dichloro-6-fluorophenyl) -6-fluoro-8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one (25)

To a solution of 25j (171 mg, 0.40 mmol) in DMF (4 mL) was added NCS (54 mg, 0.40 mmol) . The resulting mixture was heated to 80℃, stirred for 30 min and added with NCS (27 mg, 0.20 mmol) . The mixture was stirred for another 10 min and added with NCS (14 mg, 0.10 mmol) . The resulting mixture was stirred for additional 10 min and then cooled to room temperature. The mixture was purified by prep-HPLC to give the title compound 25 (88.8 mg, 45%) .

MS m/z (ESI) : 495 [M+1]

¹H NMR (400 MHz, DMSO-d ₆) δ 8.36 (d, J = 3.4 Hz, 1H) , 7.59 (d, J = 7.6 Hz, 1H) , 6.92 -6.77 (m, 2H) , 6.19 (d, J = 17.1 Hz, 1H) , 5.75 (dd, J = 10.4, 2.3 Hz, 1H) , 5.63 (d, J = 7.2 Hz, 2H) , 4.75 (s, 1H) , 4.48 –3.94 (m, 3H) , 3.64 –3.39 (m, 2H) , 3.21 –3.01 (m, 1H) , 1.28 (d, J =5.6 Hz, 3H) .

Example 9. (S) -4- (4-acryloyl-2-methylpiperazin-1-yl) -7- (6-amino-4-methyl-3- (trifluoromethyl) pyridin-2-yl) -6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one (Compound 26)

Step 1.6-Bromo-N, N-bis (4-methoxybenzyl) -4-methylpyridin-2-amine (26b)

To a solution of 6-bromo-4-methylpyridin-2-amine 26a (5.58 g, 30 mmol) in DMF (60 mL) at 0℃ was added NaH (60%in mineral oil, 3.6 g, 90 mmol) . The resulting mixture was stirred at room temperature for 30 min and added with 1- (chloromethyl) -4-methoxybenzene. The resulting mixture was extracted with EtOAc (2×100 mL) . The combined organic phase was washed with saturated brine and dried over anhydrous sodium sulphate. The solution was filtered and the filtrated was concentrated to dryness to give the title compound 26b (11.6 g, 91%) .

MS m/z (ESI) : 427 [M+1]

Step 2. N, N-bis (4-methoxybenzyl) -4-methyl-6- (tributylstannyl) pyridin-2-amine (26b)

To a mixture of 26b (4.26 g, 10 mmol) , 1, 1, 1, 2, 2, 2-hexabutyldistannane (11.6 g, 20 mmol) , LiCl (848 mg, 20 mmol) and 1, 4-dioxane (25 mL) were added Pd ₂ (dba) ₃ (915 mg, 1 mmol) and tricyclohexylphosphane (448 mg, 2mmol) under nitrogen atmosphere. The resulting mixture was heated to 110℃ and stirred for 6h. After cooling to room temperature, the mixture was concentrated to dryness, and the residue was suspended in water (200 mL) and extracted with EtOAc (2×150 mL) . The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulphate and filtered. The filtrate was concentrated to dryness and the residue was purified by silica gel column chromatography (PE/EtOAc = 100/0 to 9/1) to give the title compound 26c (3.86 g, 61%) .

¹H NMR (400 MHz, DMSO-d ₆) δ 7.12 (t, J = 5.9 Hz, 4H) , 6.83 –6.75 (m, 4H) , 6.51 (s, 1H) , 6.22 (s, 1H) , 4.64 (s, 4H) , 3.70 (d, J = 3.5 Hz, 6H) , 2.05 (s, 3H) , 1.62 –1.45 (m, 12H) , 1.17 –1.10 (m, 6H) , 0.87 (t, J = 7.3 Hz, 9H) .

Step 3. tert-Butyl (S) -4- (7- (6- (bis (4-methoxybenzyl) amino) -4-methylpyridin-2-yl) -8-oxo-6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-4-yl) -3-methylpiperazine-1-carboxylate (26d)

A mixture of 17a (1.0 m, 2.03 mmol) , 26c (2.6 g, 4.07 mmol) , LiCl (214 mg, 5.1 mmol) , 1, 4-dioxane (25 mL) , Pd (PPh ₃) ₄ (470 mg, 0.41 mmol) and CuI (116 mg, 0.61mmol) was heated to 115℃ and stirred for 1 h under nitrogen atmosphere. After cooling to room temperature, the mixture was concentrated to dryness and the residue was purified by silica gel column chromatography (PE/EtOAc = 100/0 to 9/1) to give the title compound 26d (400 mg, 27%) .

MS m/z (ESI) : 760 [M+1]

Step 4. tert-Butyl (S) -4- (7- (6- (bis (4-methoxybenzyl) amino) -3-iodo-4-methylpyridin-2-yl) -8-oxo-6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-4-yl) -3-methylpiperazine-1-carboxylate (26e)

To a mixture of 26d (440 mg, 0.58mmol) , NIS (0.65 g, 2.89 mmol) and DMF (8 mL) was added p-toluenesulfonic acid (4 mg, 0.02 mmol) . The resulting mixture was stirred for 6 h and concentrated to dryness. The residue was suspended in water (200 mL) and extracted with EtOAc (2×100 mL) . The combined organic phase was dried over anhydrous sodium sulphate and filtered. The filtrate was concentrated to dryness and the residue was purified by silica gel column chromatography (PE/EtOAc = 100/0 to 9/1) to give the title compound 26e (100 mg, 19%) .

MS m/z (ESI) : 886 [M+1]

Step 5. tert-Butyl (S) -4- (7- (6- (bis (4-methoxybenzyl) amino) -4-methyl-3- (trifluoromethyl) pyridin-2-yl) -8-oxo-6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-4-yl) -3-methylpiperazine-1-carboxylate (26f)

A mixture of 26e (100 mg, 0.11mmol) , methyl 2, 2-difluoro-2- (fluorosulfonyl) acetate (542 mg, 2.82 mmol) , CuI (258 mg, 1.35 mmol) and DMF (2.5 mL) was heated to 90℃ and stirred for 2 h. After cooling to room temperature, the mixture was concentrated to dryness , and the residue was suspended in water (20 mL) and extracted with EtOAc (2×50 mL) . The combined organic phase was dried over anhydrous sodium sulphate and filtered. The filtrate was concentrated to dryness and the residue was purified by silica gel column chromatography (PE/EtOAc = 100/0 to 1/1) to give the title compound 26f (78.8 mg, 86%) .

MS m/z (ESI) : 828 [M+1]

Step 6. (S) -7- (6-amino-4-methyl-3- (trifluoromethyl) pyridin-2-yl) -4- (2-methylpiperazin-1-yl) -6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one (26g)

A solution of 26f (100 mg, 0.13 mmol) in TFA (2 mL) was heated to 60℃ and stirred for 1.5 h. After cooling to room temperature, the mixture was concentrated to dryness to give the title compound 26g (36 mg, 62%) .

MS m/z (ESI) : 488 [M+1]

Step 7. (S) -4- (4-acryloyl-2-methylpiperazin-1-yl) -7- (6-amino-4-methyl-3- (trifluoromethyl) pyridin-2-yl) -6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one (26)

To a mixture of 26g (25 mg, 0.05 mmol) , sodium bicarbonate (13 mg, 0.15 mmol) , THF (5 mL) and water (1 mL) at 0℃ was added acryloyl chloride (4.6 mg, 0.05 mmol) . The mixture was stirred for 15 min and concentrated to dryness. The residue was purified by prep-HPLC to give the title compound 26 (9.5 mg, 35%) .

MS m/z (ESI) : 542 [M+1]

¹H NMR (400 MHz, CD ₃OD) δ 8.38 (s, 1H) , 7.10 (s, 1H) , 6.86 –6.79 (m, 1H) , 6.57 (s, 1H) , 6.30 (d, J = 16.8 Hz, 1H) , 5.82 (dd, J = 10.6, 1.9 Hz, 1H) , 4.51 –4.39 (m, 2H) , 4.22 –4.02 (m, 1H) , 3.88 –3.33 (m, 4H) , 2.42 (d, J = 1.0 Hz, 3H) , 1.47 (d, J = 6.7 Hz, 3H) .

Example 10. (S) -4- (4-acryloyl-2-methylpiperazin-1-yl) -7- (5-chloro-6-fluoro-1H-indazol-7-yl) -6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one (Compound 27)

Step 1.4-Chloro-5-fluoro-2-methylaniline (27b)

A mixture of 5-fluoro-2-methylaniline 27a (3 g, 24 mmol) , NCS (3.2 g, 24 mmol) and acetonitrile (30 mL) was heated to 80℃ and stirred for 1 h. After cooling to room temperature, the mixture was added with water (50 mL) and extracted with EtOAc (50 mL) . The organic phase was concentrated to dryness and the residue was purified by silica gel column chromatography (PE/EtOAc = 4/1) to give the title compound 27b (2.0 g, 52%) .

MS m/z (ESI) : 160 [M+1]

Step 2.2-Bromo-4-chloro-3-fluoro-6-methylaniline (27c)

A solution of 27b (2 g, 16 mmol) and NBS (2.84 g, 16 mmol) in acetonitrile (20 mL) at 0℃ was stirred for 2 hours and then added with water (20 mL) . The mixture was extracted with EtOAc (20 mL) and the organic phase was concentrated to dryness. The residue was purified by silica gel column chromatography (PE/EtOAc = 4/1) to give the title compound 27c (2.5 g, 84%) .

MS m/z (ESI) : 238, 240 [M+1]

Step 3. (2-Amino-5-chloro-6-fluoro-3-methylphenyl) boronic acid (27d)

A mixture of 27c (2.5 g, 10.5 mmol) , 4, 4, 4', 4', 5, 5, 5', 5'-octamethyl-2, 2'-bi (1, 3, 2-dioxaborolane) (5.3 g, 21 mmol) , Pd (dppf) Cl ₂ (384 mg, 0.525 mmol) , potassium acetate (3.6 g, 37 mmol) and 1, 4-dioxane (40 mL) was heated to 110℃ and stirred for 3 h. After cooling to room temperature, the mixture was concentrated to dryness and the residue was purified by silica gel column chromatography (PE/EtOAc = 9/1) to give the title compound 27d (2.2 g, 73%) .

MS m/z (ESI) : 204 [M+1]

Step 4. tert-Butyl (S) -4- (7- (2-amino-5-chloro-6-fluoro-3-methylphenyl) -8-oxo-6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-4-yl) -3-methylpiperazine-1-carboxylate (27e)

A mixture of 27d (49.6 mg, 0.244 mmol) , 17a (120 mg, 0.244 mmol) , Pd (dppf) Cl ₂ (35.7 mg, 0.049 mmol) , potassium carbonate (101 mg, 0.732 mmol) , 1, 4-dioxane (2 mL) and water (0.02 mL) was heated to 100℃ and stirred for 1 h. After cooling to room temperature, the mixture was concentrated to dryness and the residue was purified by silica gel column chromatography (dichloromethane/methanol = 97/3) to give the title compound 27e (50 mg, 36%) .

MS m/z (ESI) : 571 [M+1]

Step 5. tert-Butyl (S) -4- (7- (5-chloro-6-fluoro-1H-indazol-7-yl) -8-oxo-6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-4-yl) -3-methylpiperazine-1-carboxylate (27f)

To a solution of 27e (50 mg, 0.088 mmol) in AcOH (1 mL) at 0℃ was added a solution of NaNO ₂ (12 mg, 0.18 mmol) in water (0.3 mL) . The mixture was warmed to 25℃ and stirred for 16 h. The mixture was added with water (5 mL) and extracted with EtOAc (5 mL) . The organic phase was concentrated to dryness and the residue was purified by prep-HPLC to give the title compound 27f (25 mg, 49%) .

MS m/z (ESI) : 582 [M+1]

Step 6. (S) -7- (5-chloro-6-fluoro-1H-indazol-7-yl) -4- (2-methylpiperazin-1-yl) -6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one (27g)

A mixture of 27f (25 mg, 0.043 mmol) and HCl solution (4 M in dioxane, 2 mL) was stirred at 25℃ for 1 hour. The mixture was concentrated to dryness to give the title compound 27g (18 mg, 87%) .

MS m/z (ESI) : 482 [M+1]

Step 7. (S) -4- (4-acryloyl-2-methylpiperazin-1-yl) -7- (5-chloro-6-fluoro-1H-indazol-7-yl) -6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one (27)

To a mixture of 27g (18 mg, 0.037 mmol) and NaHCO ₃ (15.5 mg, 0.185 mmol) , water (0.5 mL) and THF (2.5 mL) was added acryloyl chloride (3.3 mg, 0.037 mmol) . The mixture was stirred at 25℃ for 10 min, then added with water (5 mL) and extracted with EtOAc (5 mL) . The organic phase was concentrated to dryness and the residue was purified by prep- HPLC to give the title compound 27 (2.7 mg, 14%) .

MS m/z (ESI) : 536 [M+1]

¹H NMR (400 MHz, CD ₃OD) δ 8.42 (s, 1H) , 8.18 (s, 1H) , 7.68 (d, J = 5.8 Hz, 1H) , 7.20 (s, 1H) , 6.82 (dd, J = 28.3, 17.4 Hz, 1H) , 6.30 (d, J = 16.9 Hz, 1H) , 5.82 (dd, J = 10.6, 1.9 Hz, 1H) , 4.56 (d, J = 14.7 Hz, 1H) , 4.43 (d, J = 13.6 Hz, 1H) , 4.20 (d, J = 12.9 Hz, 1H) , 4.06 (d, J = 12.3 Hz, 1H) , 3.83 –3.72 (m, 1H) , 3.67 (s, 1H) , 3.51 (d, J = 23.0 Hz, 1H) , 1.48 (d, J = 6.7 Hz, 3H) .

Example 11. (S) -4- (4-acryloyl-2-methylpiperazin-1-yl) -7- (2-amino-3, 5-dichloro-6-fluorophenyl) -2- ( (tetrahydro-1H-pyrrolizin-7a (5H) -yl) methoxy) -6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one (Compound 29)

Step 1.2- (Methylsulfinyl) -6- (trifluoromethyl) -3H-pyrido [2, 1-f] [1, 2, 4] triazine-4, 8-dione (29a)

To a mixture of 9c (6.13 g, 22 mmol) , methanol (150 mL) and water (20 mL) was added oxone (37 g, 220 mmol) and the resulting mixture was stirred for 16 h. The solid was then collected by filtration to give the title compound 29a (5.4 g, 83%) .

MS m/z (ESI) : 294 [M+1]

Step 2.4-Hydroxy-2- ( (tetrahydro-1H-pyrrolizin-7a (5H) -yl) methoxy) -6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one (29b)

To a mixture of (tetrahydro-1H-pyrrolizin-7a (5H) -yl) methanol (3.1 g, 22 mmol) and DMF (60 mL) was added NaH (60%in mineral oil, 2.4 g, 60 mmol) . The resulting mixture was stirred for 1 h and then added with 29b (5.88 g, 20 mmol) . The mixture was stirred for another 1 h and concentrated to dryness. The residue was purified by silica gel column chromatography (dichloromethane /methanol = 100/0 to 9/1) to give the title compound 29b (5.4 g, 74%) .

MS m/z (ESI) : 371 [M+1]

Step 3.7-Bromo-4-hydroxy-2- ( (tetrahydro-1H-pyrrolizin-7a (5H) -yl) methoxy) -6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one (29c)

To a mixture of 29b (5.4 g, 14.7 mmol) and DMF (50 mL) was added NBS (2.6 g, 14.7 mmol) . The resulting mixture was heated to 50℃ for 1 h. After cooling to room temperature, the reaction mixture was quenched by addition of a aqueous sodium sulfite solution (0.1 mL) . The mixture was concentrated to dryness under reduced pressure and the residue was purified by silica gel column chromatography (dichloromethane/methanol = 100/0 to 9/1) to give the title compound 29c (5.4 g, 74%) .

MS m/z (ESI) : 449, 451 [M+1]

Step 4. tert-Butyl (S) -4- (7-bromo-8-oxo-2- ( (tetrahydro-1H-pyrrolizin-7a (5H) -yl )methoxy) -6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-4-yl) -3-methylpiperazine-1-carboxylate (29d)

A mixture of 29c (1.35 g, 3 mmol) , POCl ₃ (1.4 g , 9 mmol) , acetonitril (20 mL) and TEA (0.3 g, 3 mmol) was heated to 50℃ and stirred for 30 min. The mixture was added with tert-butyl (S) -3-methylpiperazine-1-carboxylate (1.2 g, 6 mmol) and stirred for 0.5 h. The mixture was then concentrated to dryness under vacuum and the residue was purified by silica gel column chromatography (dichloromethane/methanol = 100/0 to 9/1) to give the title compound 29d (0.67 g, 36%) .

MS m/z (ESI) : 631, 633 [M+1]

Step 5. tert-Butyl (S) -4- (7- (2-amino-6-fluorophenyl) -8-oxo-2- ( (tetrahydro-1H-pyrrolizin-7a (5H) -yl) methoxy) -6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-4-yl) -3-methylpiperazine-1-carboxylate (29e)

A mixture of 29d (0.3 g, 0.48 mmol) , 3-fluoro-2- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) aniline (450 mg, 1.9 mmol) , CsF (217 mg, 1.43 mmol) , Pd (dppf) Cl ₂ (39 mg, 0.05 mmol) , acetonitrile (3 mL) and water (0.3 mL) was heated to 70℃ and stirred for 1 h. After cooling to room temperature, the mixture was concentrated to dryness under vacuum and the residue was purified by silica gel column chromatography (dichloromethane/methanol = 100/0 to 9/1) to give the title compound 29e (40 mg, 13%) .

MS m/z (ESI) : 662 [M+1]

Step 6. tert-Butyl (S) -4- (7- (2-amino-3, 5-dichloro-6-fluorophenyl) -8-oxo-2- ( (tetrahydro-1H-pyrrolizin-7a (5H) -yl) methoxy) -6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-4-yl) -3-methylpiperazine-1-carboxylate (29f)

To a solution of 29e (40 mg, 0.06 mmol) in DMF (2 mL) was added NCS (16 mg, 0.12 mmol) . The resulting mixture was heated to 80℃ and stirred for 0.5 h. After cooling to room temperature, the reaction mixture was quenched with an aqueous sodium sulfite solution (0.1 mL) . The mixture was concentrated to dryness under vacuum and the residue was purified by silica gel column chromatography (dichloromethane/methanol = 100/0 to9/1) to give the title compound 29f (33.5 mg, 77%) .

MS m/z (ESI) : 730 [M+1]

Step 7. (S) -7- (2-amino-3, 5-dichloro-6-fluorophenyl) -4- (2-methylpiperazin-1-yl) -2- ( (tetrahydro-1H-pyrrolizin-7a (5H) -yl) methoxy) -6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one (29g)

To a solution of 29f (33.5 mg, 0.13 mmol) in dichloromethane (2 mL) was added TFA (26 mg, 0.23 mmol) . After stirring for 1.5 h, the reaction was concentrated to dry to give the title compound 29g (18 mg, 62%) .

MS m/z (ESI) : 630 [M+1]

Step 8. (S) -4- (4-acryloyl-2-methylpiperazin-1-yl) -7- (2-amino-3, 5-dichloro-6-fluorophenyl) -2- ( (tetrahydro-1H-pyrrolizin-7a (5H) -yl) methoxy) -6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one (29)

To a mixture of 29g (18 mg, 0.028 mmol) , sodium bicarbonate (4.7 mg, 0.056 mmol) , THF (2 mL) and water (1 mL) at 0℃ was added acryloyl chloride (2.6 mg, 0.028 mmol) . The resulting mixture was stirred for 15 min and concentrated to dryness. The residue was purified by prep-HPLC to give the title compound 29 (2.5 mg, 13%) .

MS m/z (ESI) : 684 [M+1]

¹H NMR (400 MHz, CD ₃OD) δ 8.53 (s, 1H) , 7.46 (t, J = 6.0 Hz, 1H) , 7.32 (s, 1H) , 6.82 (s, 1H) , 6.32 (d, J = 16.5 Hz, 1H) , 5.85 (dd, J = 10.6, 1.8 Hz, 1H) , 4.59 –4.58 (m, 2H) , 4.45 (s, 1H) , 3.49 (s, 2H) , 3.20 –3.07 (m, 4H) , 2.20 –2.04 (m, 12H) , 1.50 –1.44 (m, 3H)

Example 12. (S) -4- (4-acryloyl-2-methylpiperazin-1-yl) -7- (2-amino-3, 5-dichloro-6-fluorophenyl) -2-methyl-6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one (Compound 28)

Step 1.1-Amino-6-oxo-4- (trifluoromethyl) -1, 6-dihydropyridine-2-carboxamide (28a)

To a solution of 1e (3.2 g, 13.5 mmol) in THF (20 mL) was added ammonium hydroxide (5 mL) . The resulting mixture was stirred for 12 h and concentrated to dryness under reduced pressure to give the title compound 28a (crude) , which was used directly in the next step without further purification.

MS m/z (ESI) : 223 [M+1]

Step 2.2-Methyl-6- (trifluoromethyl) -3H-pyrido [2, 1-f] [1, 2, 4] triazine-4, 8-dione (28b)

To a solution of 28a (crude, about 13.5 mmol) and TEA (15 g, 145 mmol) in THF (50 mL) was added acetyl chloride (8 g, 101 mmol) . The resulting solution was stirred for 2 h, added with a sodium hydroxide aqueous solution (1N, 100 mL) at 90℃ and stirred at 100℃ for another 1h. After cooling to room temperature, the mixture was concentrated to dryness under reduced pressure and the residue was purified by prep-HPLC to give the title compound 28b (750 mg, 22%over two steps) .

MS m/z (ESI) : 246 [M+1]

Step 3.7-Bromo-2-methyl-6- (trifluoromethyl) -3H-pyrido [2, 1-f] [1, 2, 4] triazine-4, 8-dione (28c)

To a solution of 28b (780 mg, 3.2 mmol) in DMF (10 mL) was added NBS (600 mg, 3.2 mmol) . The resulting mixture was heated to 50℃ and stirred for 4 h. After cooling to room temperature, the mixture was diluted with saturated brine (50 mL) and extracted with EtOAc (2×50 mL) . The combined organic phase was concentrated to dryness and the residue was purified by silica gel column chromatography (dichloromethane/methanol = 10/0 to 4/1) to give the title compound 28c (380 mg, 37%) .

MS m/z (ESI) : 324, 326 [M+1]

Step 4. Tert-butyl (S) -4- (7-bromo-2-methyl-8-oxo-6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-4-yl) -3-methylpiperazine-1-carboxylate (28d)

To a solution of 28c (450 mg, 0.98 mmol) in POCl ₃ (5 mL) was added DIPEA (1 mL) . The resulting mixture was heated to 80℃ and stirred for 30 min. After cooling to room temperature, the mixture was concentrated to dryness under reduced pressure. The residue was diluted with dichloromethane (3 mL) , and the resulting solution was added with DIPEA (500 mg, 3.9 mmol) and a solution of tert-butyl (S) -3-methylpiperazine-1-carboxylate (450 mg, 2.2 mmol) in dichloromethane (5 mL) . The solution was stirred for 1h and concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane/methanol = 10/0 to 4/1) to give the title compound 28d (220 mg, 56%) .

MS m/z (ESI) : 506, 508 [M+1]

Step 5. Tert-butyl (S) -4- (7- (2-amino-6-fluorophenyl) -2-methyl-8-oxo-6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-4-yl) -3-methylpiperazine-1-carboxylate (28e)

A mixture of 28d (200 mg, 0.4 mmol) , (2-amino-6-fluorophenyl) boronic acid (150 mg, 0.8 mmol) , Pd (dppf) Cl ₂ (60 mg, 0.08 mmol) , K ₂CO ₃ (200 mg, 1.2 mmol) , 1, 4-dioxane (20 mL) and water (1 mL) heated to 90℃ and stirred for 1.5 h. After cooling to room temperature, the mixture was concentrated to dryness under reduced pressure and the residue was purified by silica gel column chromatography (PE/EtOAc = 100/0 to 0/100) to give the title compound 28e (200 mg, 93%) .

MS m/z (ESI) : 537 [M+1]

Step 6. Tert-butyl (S) -4- (7- (2-amino-3, 5-dichloro-6-fluorophenyl) -2-methyl-8-oxo-6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-4-yl) -3-methylpiperazine-1-carboxylate (28f)

To a solution of 28e (180 mg, 0.335 mmol) in DMF (3 mL) was added NCS (90 mg, 0.67 mmol) , and the resulting mixture was stirred at 80℃ for 3 h. After cooling to room temperature, the mixture was diluted with water (30 mL) and extracted with EtOAc (50 mL) . The combined organic phase was washed with saturated brine (20 mL) , dried over anhydrous sodium sulphate and filtered. The filtrate was concentrated to dryness under reduced pressure and the residue was purified by silica gel column chromatogrpahy (PE/EtOAc = 100/0 to 1/4) to give the title compound 28f (90 mg, 44%) .

MS m/z (ESI) : 605 [M+1]

Step 7. (S) -7- (2-amino-3, 5-dichloro-6-fluorophenyl) -2-methyl-4- (2-methylpiperazin-1-yl) -6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one (28g)

To a solution of 28f (90 mg, 0.15 mmol) in EtOAc (1 mL) was added a solution of HCl in EtOAc (4 M, 4 mL) . The resulting solution was stirred for 3 h and concentrated to dryness under reduced pressure to give the title compound 28g (crude) , which was used directly in the next step without further purification.

MS m/z (ESI) : 505 [M+1]

Step 8. (S) -4- (4-acryloyl-2-methylpiperazin-1-yl) -7- (2-amino-3, 5-dichloro-6-fluorophenyl) -2-methyl-6- (trifluoromethyl) -8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one monoformate (28)

To a mixture of 28g (crude, about 0.15 mmol) , NaHCO ₃ (40 mg, 0.6 mmol) , THF (2 mL) and water (0.5 mL) was added acryloyl chloride (8 mg, 0.09 mmol) . The resulting mixture was stirred for 20 min and concentrated to dryness under reduced pressure. The residue was purified by prep-HPLC to give the title compound 28 (20 mg, 24%) .

MS m/z (ESI) : 559 [M+1]

¹H NMR (400 MHz, DMSO-d ₆) δ 7.56 (s, J = 7.6 Hz, 1H) , 6.93 (d, J = 8.5 Hz, 1H) , 6.90 –6.75 (m, 1H) , 6.29 –6.13 (m, 2H) , 5.75 (dd, J = 10.4, 2.3 Hz, 1H) , 5.57 (d, J = 7.9 Hz, 2H) , 4.89 –4.66 (m, 1H) , 4.48 –3.95 (m, 3H) , 3.90 –3.52 (m, 1H) , 3.46 –2.92 (m, 1H) , 2.45 (s, 3H) , 1.34 (s, 3H) .

Example 13. (S) -4- (4-acryloyl-2-methylpiperazin-1-yl) -7- (2-amino-3, 5-dichloro-6-fluorophenyl) -6-methoxy-8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one (Compound 24)

Step 1.4, 6-Dimethoxypicolinic acid (24b)

To a solution of 4, 6-dichloropicolinic acid 24a (5 g, 26 mmol) in DMF (50 mmol) was added sodium methoxide (30%wt in methanol, 15 mL) . The resulting solution was heated to 120℃ for 48 h. After cooling to room temperature, the reaction solution was diluted with water (200 mL) and adjusted to pH = 7 with diluted hydrochloric acid (1 N) . The resulting mixture was extracted with EtOAc (2×150 mL) and washed with saturated brine (2×150 mL) . The combined organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to dryness under vacuum and the residue was purified by silica gel column chromatography (PE/EtOAc = 100/0 to 3/2) to give the title compound 24b (3 g, 62%) .

MS m/z (ESI) : 184 [M+1]

Step 2. Ethyl 6-hydroxy-4-methoxypicolinate (24c)

To a solution of 24b (3 g, 16.4 mmol) in ethanol (5 mL) was added a solution of HCl in EtOH (4 M, 20 mL) . The resulting solution was heated to 90℃ and stirred for 48 h. After cooling to room temperature, the solution was concentrated to dryness under reduced pressure and the residue was purified with silica gel column chromatography (PE/EtOAc =100/0 to 3/2) to give the title compound 24c (1 g, 30%) .

MS m/z (ESI) : 198 [M+1]

Step 3 to Step 5.7-bromo-4-hydroxy-6-methoxy-8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one (24f)

Intermediate 24f was prepared according to Steps 4-6 for the preparation of intermediate 1g, but replacing 1d with 24c.

MS m/z (ESI) : 272, 274 [M+1]

Step 6 to step 10. (S) -4- (4-acryloyl-2-methylpiperazin-1-yl) -7- (2-amino-3, 5-dichloro-6-fluorophenyl) -6-methoxy-8H-pyrido [2, 1-f] [1, 2, 4] triazin-8-one (24)

31 was prepared by according to Steps 4-8 for the preparation of 28, but replacing 28c with 24f.

MS m/z (ESI) : 507 [M+1]

¹H NMR (400 MHz, DMSO-d ₆) δ 8.29 (d, J = 1.1 Hz, 1H) , 7.49 (d, J = 7.5 Hz, 1H) , 6.92 –6.76 (m, 1H) , 6.59 (d, J = 7.0 Hz, 1H) , 6.27 –6.11 (m, 1H) , 5.75 (dd, J = 10.4, 2.3 Hz, 1H) , 5.29 (d, J = 6.2 Hz, 2H) , 4.83 –4.67 (m, 1H) , 4.50 –3.94 (m, 3H) , 3.91 (s, 3H) , 3.72 –2.95 (m, 3H) , 1.35 (d, J = 6.6 Hz, 3H) .

Example 14. Biological experiments

Nucleotide exchange assay

The inhibition of the compound of the present invention on the activity of recombinant KRAS G12C was assessed via detecting the nucleotide (GDP and GTP) exchange level mediated by SOS1 and RAF using a biochemical method in the HTRF assay format (Table 1) .

The experimental method is generally described below:

An assay buffer contained the following components: 25 mM HEPES, 10 mM MgCl ₂, 5mM EDTA, 0.01%Triton X-100, and 0.04%Brij35. A KRAS G12C protein solution contained 125 nM human recombinant GDP-bound KRAS G12C protein with His-tag (produced by Qinghua protein purification lab) in the assay buffer. A substrate solution contained 12.5 μM GTP and 25 nM recombinant SOS1 protein (produced by Qinghua protein purification lab) in the assay buffer. A GST-tagged RAF solution contained 62.5nM GST-tagged RAF protein (produced by Qinghua protein purification lab) in the assay buffer. A detection solution contained 90 ng/mL Eu ³⁺-labeled anti-His antibody (Cisbio, Cat. No. 61HI2KLA) and 1.2 μg/mL d2-labeled anti-GST antibody (Cisbio, Cat. No. 61GSTDLF) in the assay buffer.

The test compound was dissolved to 500 μM in DMSO, followed by a serial 4-fold dilution with DMSO to a minimum concentration of 31 nM. Each concentration is further diluted 20-fold with the reaction buffer.

To a 384-well assay plate (Corning, Cat. No. 4512) were added 2 μL of compound solution and 2 μL of KRAS G12C protein solution, and the mixture was incubated at room temperature for 30 minutes. To the mixture was added 4 μL of the substrate solution and incubation was continued for another 30 minutes, followed by addition of 2 μL of RAF protein solution. After further incubation for 30 minutes, the mixture was added with an equal volume of 10 μL detection solution and allowed to stand at room temperature for 4 hours. An Envision plate reader (Perkin Elmer) was then used to measure the absorbances of the mixture at 620 nm and 665 nm. The ratio of absorbances at 665 nm and 620 nm was positively correlated with the degree of substrate exchange, therefore the activity of KRAS G12C was detected. In the experiment, the group with 10 μM reference compound (AMG510) was treated as the 100%inhibition group, and the group with KRAS G12C protein but not the test compound was treated as the 0%inhibition group. The inhibition curve was plotted and the corresponding IC ₅₀ value of the test compound was calculated using XLfit software (ID Business Solutions Ltd., UK) .

Antiproliferative cell assay

The inhibition of the compound of the present invention on NCI-H358 cell proliferation was assessed using a Cell Titer-Glo kit (Table 1) .

The experimental method is generally described below:

The test compound was dissolved to 5 mM in DMSO, followed by a serial 4-fold dilution with DMSO to a minimum concentration of 310 nM. Each concentration was further diluted 50-fold with RPMI 1640 medium (Gibco, Cat. No. 72400-120) .

NCI-H358 (ATCC, Cat. No. CRL-5807) , a human non-small cell lung cancer (NSCLC) cell line with a KRAS G12C mutation, were cultured in RPMI 1640 medium (Gibco, Cat. No. 72400-120) containing 10%FBS and 1%penicillin-streptomycin (Gibco, Cat. No. 15070063) . The cells were seeded in a 96-well cell culture plate at a density of 8,000 cells/well and incubated overnight at 37℃/5%CO ₂ in a humidity-controlled incubator. 10 μL of the test compound solution was added to each well and mixed gently, and the plate was continuously incubated at 37℃/5%CO ₂ for 5 days. Cell proliferation was then assessed using a Cell Titer-Glo kit (Promega, Cat. No G7572) by adding 30 μL CTG reagent into each well, shaking for 10 minutes and then settling for 10 minutes at room temperature. An Envision plate reader (Perkin Elmer) was used to measure the luminescence signals. In this experiment, the group with 10 μM reference compound (MRTX849) was treated as the 100%inhibition group, and the group with 0.02%DMSO but not the test compound was treated as the 0%inhibition group. The inhibition curve was plotted and the corresponding IC ₅₀ value of the test compound was calculated using XLfit software (ID Business Solutions Ltd., UK) .

Table 1

Example 15. Efficacy study in the xenograft model

The study was conducted to evaluate in-vivo anti-tumor activity of

Compounds

12a and 15a in female BALB/c nude mice bearing Mia PaCa-2 human pancreatic ductal adenocarcinoma cell line derived xenograft tumors.

Female BALB/c nude mice of 6-8 weeks old were ordered from Beijing HFK BIOSCIENCE Co., Ltd. Mia PaCa-2 tumor cells (purchased from Cobioer) were cultured in DMEM medium (purchased from Gibco) supplemented with 10%FBS (purchased from ExCell Bio) at 37℃ under 5%CO ₂. The cells were harvested in 90%confluence and no less than 90%viability, and then counted.

Compounds

12a and 15a, and AMG 510 (a KRAS G12C inhibitor which has been approved by FDA for the treatment of KRAS G12C mutated NSCLC) were suspended in a vehicle comprised of 2%HPMC and 1%Tween-80, respectively. Mice were inoculated subcutaneously into the right flank with 150 μL of 5x10 ⁶ Mia PaCa-2 cells resuspended in 50%Matrigel (purchased from Corning) . After the tumor sizes reached 100～200 mm ³, the animals in good health condition were randomly placed into 4 groups (10 animals per group) and each group was orally dosed with vehicle, 12a, 15a, or AMG-510 at 1 mg/kg, once a day (QD) . Dosing was initiated on day 0 and effects on tumor growth were evaluated by the percentage of tumor growth inhibition (TGI) at the end of the study. Tolerability was assessed by body weight loss, lethality, and clinical signs of adverse treatment-related side effects. Tumor volume and body weight were measured three times per week during the administration period. The percentage of TGI was determined on day 44 after treatment. The difference between the mean value of tumor volume in the treatment group and that in the vehicle group was analyzed for significance using one-way ANOVA (followed by Dunnett’s test) at each time point and a P < 0.05 was considered to be statistically significant.

The efficacy results demonstrated that both

Compounds

12a and 15a dose-dependently reduced tumor size in the Mia PaCa-2 model. The treatment with 12a, 15a and AMG-510 demonstrated significant anti-tumor activities with TGI%of 76.38%, 93.29%and 56.20%, respectively (P < 0.0001 for

Compounds

12a and 15a treated groups vs. vehicle) (FIG. 1) .

Both

Compounds

12a and 15a at 1 mg/kg QD were well-tolerated with no significant body weight loss during the study (FIG. 2) .

In summary, the results of this study demonstrate that

Compounds

12a and 15a had significant anti-tumor activities against Mia PaCa-2 human pancreatic ductal adenocarcinoma xenograft model and were well-tolerated by the tumor-bearing animals. The results indicate that both

Compounds

12a and 15a are safe and efficacious anti-cancer agents.

Claims

A compound of Formula (II) , or a pharmaceutically acceptable salt thereof, or a stable isotope thereof, or a stereoisomer thereof,

wherein:

W is
-C (O) CH=C=CH ₂ or -CN;

R ¹ is H or halogen;

R ² is CF ₃, H, halogen, cyano, -OCF ₃, C _1-3 alkyl or -OC _1-3 alkyl;

R ³ is C _6-12 aryl or 5-to 12-membered heteroaryl, where 1-4 hydrogens of R ³ is optionally substituted by R ⁶;

R ⁴ is H, C _1-6 alkyl, C _3-6 cycloalkyl or 4-to 12-membered heterocyclyl, where one or more hydrogens of the alkyl, cycloalkyl and heterocyclyl are optionally substituted by R ⁷;

R ⁵ is selected from C _1-6 alkyl, cyano, or oxo , where one or more hydrogens of the alkyl is optionally substituted by D, halogen, cyano or -OR ^c;

R ⁶ is halogen, cyano, amino, hydroxy or C _1-6 alkyl, where one or more hydrogens of the alkyl is optionally substituted by D, halogen or -OR ^c;

R ⁷ is D, halogen, cyano, C _1-6 alkyl, C _3-6 cycloalkyl, -NR ^cR ^d, -OR ^c or 4-to 12-membered heterocyclyl, where one or more hydrogens of the alkyl, cycloalkyl and heterocyclyl are optionally substituted by halogen or C _1-6 alkyl;

L is a bond or O;

R ^a is H, absent, cyano, halogen or C _1-3 alkyl;

R ^b is H, D or C _1-3 alkyl, where one hydrogen of the alkyl is optionally substituted by -OR ^c, -NR ^cR ^d or 4-to 6-membered heterocyclyl; and

R ^c and R ^d are independently selected from H, C _1-6 alkyl, C _1-6 haloalkyl, C _3-6 cycloalkyl or 4-6 membered heterocyclyl.
A compound of Formula (III) , or a pharmaceutically acceptable salt thereof, or a stable isotope thereof, or a stereoisomer thereof,

wherein:

W is -C (O) CH=CH ₂, -S (O) ₂CH=CH ₂, or -C (O) CH=C=CH ₂;

A is C (R ⁶) or N;

R ² is CF ₃, halogen, or CH ₃;

R ⁴ is H, C _1-6 alkyl or C _1-6 alkylene- (4-to 12-membered) heterocyclyl, where one or more hydrogens of the heterocyclyl is optionally substituted by halogen or C _1-6 alkyl;

R ⁵ is C _1-2 alkyl or -CH ₂CN;

R ⁶ is amino, halogen, hydroxy, CF ₃ or C _1-6 alkyl; and

L is a bond or O.
A compound of Formula (IV) , or a pharmaceutically acceptable salt thereof, or a stable isotope thereof, or a stereoisomer thereof,

wherein:

R ² is CF ₃ or halogen;

R ⁴ is H, C _1-6 alky or -OC _1-6 alkylene- (4-to 12-membered) heterocyclyl, where one or more hydrogens of the heterocyclyl is optionally substituted by halogen or C _1-6 alkyl;

R ^5a is H, CH ₃ or -CH ₂CN; and

R ^5b is CH ₃ or H.
The compound of any one of Claims 1-4, wherein the heterocyclyl of R ⁴ is pyrrolidinyl and pyrrolizinyl, piperidinyl, piperazinyl, or morpholinyl.
The compound according to Claim 1, or a pharmaceutical acceptable salt, stable isotope, stereoisomer thereof, wherein the compound is:
The compound according to Claim 5, which is
A pharmaceutical composition comprising the compound of any one of Claims 1-6 and a pharmaceutically acceptable carrier or excipient thereof.
A method for preventing or treating cancers mediated by KRAS G12C, comprising administering to a patient in need thereof a therapeutically effective amount of the pharmaceutical composition of Claim 7.