TW202313633A - Sulfur-containing heteroaromatic tricyclic kras inhibitors - Google Patents

Abstract

The present embodiments provide compounds of Formula I, pharmaceutical compositions of the compounds, and methods for treating diseases such as cancer.

Description

Sulfur-containing heteroaromatic tricyclic KRAS inhibitors

Embodiments of the present invention provide compounds of formula I, pharmaceutical compositions of these compounds, and methods for treating diseases such as cancer.

The embodiments herein relate to compounds, compositions and methods for the treatment of RAS-mediated diseases. In particular, the embodiments herein relate to compounds and methods for treating diseases such as cancer by targeting oncogenic mutants of K-RAS isoforms.

Ras proteins are small guanine nucleotide binding proteins that act as molecular switches by cycling between active GTP-bound and inactive GDP-bound conformations. Ras signaling via activation by the guanine nucleotide exchange factor (GEF), most commonly son of sevenless (SOS), and inactivation by GTPase activating proteins (GAPs) such as neurofibromin or p120GAP to control the balance between them. Ras proteins play an important role in the regulation of cell proliferation, differentiation, and survival. Dysregulation of the Ras signaling pathway is almost certainly associated with disease. Hyperactivating somatic mutations in Ras are among the most common lesions found in human cancers. Most of these mutations have been shown to reduce the sensitivity of Ras to GAP stimulation and reduce its intrinsic GTPase activity, resulting in an increase in the active GTP-bound population. Although mutations in any of the three Ras isoforms (K-Ras, N-Ras, or H-Ras) have been shown to result in oncogenic transformation, K-Ras mutations are by far the most common in human cancers. For example, K-Ras mutations are known to be commonly associated with pancreatic, colorectal, and non-small cell lung cancers. Similarly, H-Ras mutations are common in cancers such as papillary thyroid, lung and skin cancers. Finally, N-Ras mutations frequently occur in HCC.

K-Ras is the most frequently mutated oncoprotein in human cancers. Therefore, there is a need to develop selective inhibitors of KRAS mutants. Embodiments of the present invention satisfy this and other needs.

(Ia) 其中 Z為O或S； m為1或2； p為1或2； L ¹為

其中k為0至4之整數；並且各R ¹獨立地選自甲基、及氰甲基、C ₂-C ₄烷基、氰基、環烷基、鹵基、鹵烷基、三氟甲基、及烷氧基；或任何兩個R ¹組合以形成稠環、橋、或螺環結構，視情況在橋或螺環中包含選自S、SO ₂、O或N之雜原子，並且其中橋或螺環結構視情況經側氧基取代； 各R ²獨立地選自由以下組成之群：烷基，N-烷基胺基，N,N-二烷基胺基，烷基醯胺烷基，芳基醯胺烷基，-OCH ₂CONRR’，其中R及R’獨立地選自氫、烷基、及環烷基，烷基磺醯胺基烷基，芳基磺醯胺基烷基，N-烷基胺基烷基，N,N-二烷基胺基烷基，烷氧基，烷氧基烷基，環烷基，烷基環烷基，羥基烷基，鹵素，鹵烷基，芳基，芳氧基，芳烷基，雜芳基，雜芳基烷基，雜環基，雜環基烷基，及雜芳基氧基，其中之任一者視情況經取代；或當m為2時，兩個R ²組合以形成視情況含有1至3個選自N、O、或S之雜原子的螺環3至6員環； R ³、R ⁴、R ⁵、及R ⁶獨立地選自鹵素、氫、羥基、烷氧基、烷基、環烷基、胺基、N-烷基胺基、C-醯胺(-CONRR’)、N-醯胺(-NHCOR)、脲(-NHCONHR)、醚(-OR)、磺醯胺(-NHSO ₂R或-SO ₂NHR)、及CF ₃；其中各R及R’獨立地為氫、烷基、或環烷基；或 任何兩個相鄰R ³、R ⁴、R ⁵、或R ⁶形成包含0至3個選自N、O或S之雜原子的視情況經取代之稠合5員或6員環； 限制條件為R ³、R ⁴、R ⁵、或R ⁶中之一者為連接至2-喹唑啉酮之鍵；且 R ⁷為烷基、氰基、環烷基、鹵素、鹵烷基、三氟甲基、及烷氧基。(I) In one aspect, the embodiments of the present invention provide a compound of formula (Ia) or a pharmaceutically acceptable salt thereof:

(Ia) wherein Z is O or S; m is 1 or 2; p is 1 or 2; L ¹ is

wherein k is an integer from 0 to 4; and ^each R is independently selected from methyl, and cyanomethyl, C ₂ -C ₄ alkyl, cyano, cycloalkyl, halo, haloalkyl, trifluoromethane group, and alkoxy group; or any two R ¹ combined to form a fused ring, bridge, or spiro ring structure, optionally including a heteroatom selected from S, SO ₂ , O, or N in the bridge or spiro ring, and Wherein the bridge or spiro ring structure is optionally substituted by a side oxygen group; ^each R is independently selected from the group consisting of: alkyl, N-alkylamino, N,N-dialkylamino, alkylamide Alkyl, arylamide alkyl, -OCH ₂ CONRR', wherein R and R' are independently selected from hydrogen, alkyl, and cycloalkyl, alkylsulfonamidoalkyl, arylsulfonamido Alkyl, N-Alkylaminoalkyl, N,N-Dialkylaminoalkyl, Alkoxy, Alkoxyalkyl, Cycloalkyl, Alkylcycloalkyl, Hydroxyalkyl, Halogen, Haloalkyl, aryl, aryloxy, aralkyl, heteroaryl, heteroarylalkyl, heterocyclyl, heterocyclylalkyl, and heteroaryloxy, any of which is optionally modified Substitution; or when m is 2, two R ² combine to form a spiro 3 to 6 membered ring optionally containing 1 to 3 heteroatoms selected from N, O, or S; R ³ , R ⁴ , R ⁵ , and R ⁶ are independently selected from halogen, hydrogen, hydroxyl, alkoxy, alkyl, cycloalkyl, amino, N-alkylamino, C-amide (-CONRR'), N-amide (-NHCOR), urea (-NHCONHR), ether (-OR), sulfonamide (-NHSO ₂ R or -SO ₂ NHR), and CF ₃ ; wherein each R and R' are independently hydrogen, alkyl, or cycloalkyl; or any two adjacent R ³ , R ⁴ , R ⁵ , or R ⁶ form an optionally substituted fused 5 member comprising 0 to 3 heteroatoms selected from N, O, or S, or 6-membered ring; the restriction is that one of R ³ , R ⁴ , R ⁵ , or R ⁶ is a bond connected to 2-quinazolone; and R ⁷ is alkyl, cyano, cycloalkyl, halogen , haloalkyl, trifluoromethyl, and alkoxy. (I)

In another aspect, the embodiments of the present invention provide a pharmaceutical composition comprising a pharmaceutically effective amount of a compound disclosed herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

In another embodiment, the present embodiments provide a method of treating an individual with cancer characterized by the presence of a KRAS G12C mutation, the method comprising administering to the individual a therapeutically effective amount of a compound disclosed herein, or a medicament thereof A pharmaceutically acceptable salt, or a pharmaceutical composition as disclosed herein.

In another embodiment, embodiments of the present invention provide methods for the manufacture of a medicament for treating an individual with cancer characterized by the presence of a KRAS G12C mutation using a compound comprising a compound disclosed herein, or a pharmaceutically acceptable form thereof. salt, or the drug of the pharmaceutical composition as disclosed herein.

In another embodiment, the embodiments of the present invention provide the use of the compound disclosed herein, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition disclosed herein for the manufacture of a medicament for treating cancer in an individual, the cancer It is characterized by the presence of the KRAS G12C mutation.

In another embodiment, embodiments of the invention provide a compound disclosed herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as disclosed herein, for use in treating a cancer in a subject characterized by KRAS G12C mutation.

I. General

The present embodiments provide inhibitors of KRAS G12C that exhibit good selectivity over wild-type KRAS and are useful in the treatment of cancers characterized by KRAS G12C mutations. II. Definition

Unless specifically indicated otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the embodiments belong. In addition, any methods or materials similar or equivalent to those described herein can be used in the practice of embodiments of the invention. For the purposes of the embodiments of the present invention, the following terms are defined.

As used herein, "a (a/an)" or "the" includes not only aspects having one member, but also aspects having more than one member. For example, the singular forms "a (a/an)" and "the" include plural referents unless the context clearly requires otherwise. Thus, for example, reference to "a cell" includes a plurality of such cells and reference to "an agent" includes reference to one or more agents known to those skilled in the art, and so forth.

"Alkyl" means a straight or branched chain, saturated, aliphatic group having the indicated number of carbon atoms. The alkyl group may contain any number of carbons, such as C _1-2 , C _1-3 , C _1-4 , C _1-5 , C _1-6 , C _1-7 , C _1-8 , C _1-9 , C _1-10 , C _2-3 , C _2-4 , C _2-5 , C _2-6 , C _3-4 , C _3-5 , C _3-6 , C _4-5 , C _4-6 and C _5-6 . For example, C _1-6 alkyl includes, but is not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, second butyl, third butyl, pentyl, isopentyl, hexyl wait. Alkyl may also refer to alkyl groups having up to 20 carbon atoms, such as, but not limited to, heptyl, octyl, nonyl, decyl, and the like. Alkyl groups can be substituted or unsubstituted.

"Alkylene" refers to a straight or branched, saturated, aliphatic group having the indicated number of carbon atoms and linked to at least two other groups, ie , a divalent hydrocarbon group. The two moieties attached to the alkylene group may be attached to the same atom or different atoms of the alkylene group. For example, the linear alkylene group can be a divalent group of -(CH ₂ ) _n -, wherein n is 1, 2, 3, 4, 5 or 6. Representative alkylene groups include, but are not limited to, methylene, ethylidene, propylidene, isopropylidene, butylene, isobutylene, sec-butylene, pentylene, and hexylene. Alkylene groups can be substituted or unsubstituted.

"Alkenyl" means a straight or branched chain hydrocarbon having at least 2 carbon atoms and at least one double bond. Alkenyl can contain any number of carbons, such as C ₂ , C _2-3 , C _2-4 , C _2-5 , C _2-6 , C _2-7 , C _2-8 , C _2-9 , C _{2 -10} , C ₃ , C _3-4 , C _3-5 , C _3-6 , C ₄ , C _4-5 , C _4-6 , C ₅ , C _5-6 , and C ₆ . An alkenyl group can have any suitable number of double bonds, including but not limited to 1, 2, 3, 4, 5 or more. Examples of alkenyl include, but are not limited to, vinyl/ethenyl, propenyl, isopropenyl, 1-butenyl, 2-butenyl, isobutenyl, butadienyl, 1-pentenyl, 2-butenyl, -pentenyl, isopentenyl, 1,3-pentadienyl, 1,4-pentadienyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 1,3- Hexadienyl, 1,4-hexadienyl, 1,5-hexadienyl, 2,4-hexadienyl, or 1,3,5-hexatrienyl. Alkenyl groups can be substituted or unsubstituted.

"Alkenylene" means an alkenyl group as defined above linked to at least two other groups, ie , a divalent hydrocarbon group. The two moieties attached to the alkenylene group can be attached to the same atom or different atoms of the alkenylene group. Alkenylene groups include, but are not limited to, ethenyl, propenyl, isopropenyl, butenyl, isobutenyl, sec-butenyl, pentenyl, and hexenyl. Alkenylene groups can be substituted or unsubstituted.

"Alkynyl" means a straight or branched chain hydrocarbon having at least 2 carbon atoms and at least one triple bond. Alkynyl groups may contain any number of carbons, such as C ₂ , C _2-3 , C _2-4 , C _2-5 , C _2-6 , C _2-7 , C _2-8 , C _2-9 , C _{2 -10} , C ₃ , C _3-4 , C _3-5 , C _3-6 , C ₄ , C _4-5 , C _4-6 , C ₅ , C _5-6 , and C ₆ . Examples of alkynyl include, but are not limited to, ethynyl, propynyl, 1-butynyl, 2-butynyl, butadiynyl, 1-pentynyl, 2-pentynyl, isopentynyl, 1 ,3-pentadiynyl, 1,4-pentadiynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 1,3-hexadiynyl, 1,4-hexanedi Alkynyl, 1,5-hexadiynyl, 2,4-hexadiynyl, or 1,3,5-hexatriynyl. Alkynyl groups can be substituted or unsubstituted.

"Alkynylene" means an alkynyl group as defined above linked to at least two other groups, ie , a divalent hydrocarbon group. The two moieties attached to the alkynylene group can be attached to the same atom or different atoms of the alkynylene group. Alkynyl groups include, but are not limited to, ethynyl, propynyl, isopropynyl, butynyl, second butynyl, pentynyl, and hexynyl. Alkynylene groups can be substituted or unsubstituted.

"Alkoxy" refers to an alkyl group having an oxygen atom connecting the alkyl group to the point of attachment: alkyl-O-. Like alkyl, alkoxy may have any suitable number of carbon atoms, such as C _1-6 . Alkoxy includes, for example, methoxy, ethoxy, propoxy, isopropoxy, butoxy, 2-butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentyloxy, Oxygen, Hexyloxy, etc. The alkoxy group may be further substituted with various substituents described therein. Alkoxy groups can be substituted or unsubstituted.

"Alkoxyalkyl" means a group having an alkyl component and an alkoxy component, wherein the alkyl component links the alkoxy component to the point of attachment. The alkyl component is as defined above except that the alkyl component is at least divalent, is an alkylene, is attached to the alkoxy component and is attached to the point of attachment. The alkyl component may contain any number of carbons, such as C _0-6 , C _1-2 , C _1-3 , C _1-4 , C _1-5 , C _1-6 , C _2-3 , C _{2- 4.} C _2-5 , C _2-6 , C _3-4 , C _3-5 , C _3-6 , C _4-5 , C _4-6 and C _5-6 . The alkoxy component is as defined above. Examples of alkoxyalkyl include, but are not limited to, 2-ethoxy-ethyl and methoxymethyl.

"Alkylhydroxy" or "hydroxyalkyl" refers to an alkyl group as defined above wherein at least one of the hydrogen atoms is replaced by a hydroxy group. Like an alkyl group, an alkylhydroxy group can have any suitable number of carbon atoms, such as C _1-6 . Exemplary alkyl hydroxy groups include, but are not limited to, hydroxy-methyl, hydroxyethyl (wherein the hydroxy is in the 1- or 2-position), hydroxypropyl (wherein the hydroxy is in the 1-, 2-, or 3-position), hydroxybutyl (wherein the hydroxyl is at the 1-, 2-, 3- or 4-position), hydroxypentyl (wherein the hydroxyl is at the 1-, 2-, 3-, 4- or 5-position), hydroxyhexyl (wherein the hydroxyl is at the 1- , 2-, 3-, 4-, 5- or 6-position), 1,2-dihydroxyethyl, and similar groups.

"Halogen" or "halo" refers to fluorine, chlorine, bromine and iodine.

"Haloalkyl" means an alkyl group as defined above in which some or all of the hydrogen atoms are replaced by halogen atoms. Like alkyl groups, haloalkyl groups can have any suitable number of carbon atoms, such as C _1-6 . For example, haloalkyl includes trifluoromethyl, fluoromethyl, and the like. In some instances, the term "perfluoro" may be used to define a compound or group in which all hydrogens are replaced with fluorine. For example, perfluoromethyl refers to 1,1,1-trifluoromethyl.

"Haloalkoxy" refers to an alkoxy group in which some or all of the hydrogen atoms are replaced by halogen atoms. Like alkyl, haloalkoxy may have any suitable number of carbon atoms, such as C _1-6 . Alkoxy can be substituted with 1, 2, 3, or more halogens. A compound is persubstituted, eg, perfluorinated, when all hydrogens are replaced by halogen, eg, fluorine. Haloalkoxy includes, but is not limited to, trifluoromethoxy, 2,2,2,-trifluoroethoxy, perfluoroethoxy, and the like.

"Cycloalkyl" means a saturated or partially unsaturated, monocyclic, fused bicyclic or bridged polycyclic combination containing from 3 to 12 ring atoms, or the number of atoms indicated. Cycloalkyl can include any number of carbons, such as C _3-6 , C _4-6 , C _5-6 , C _3-8 , C _4-8 , C _5-8 , C _6-8 , C _3-9 , C _3-10 , C _3-11 , and C _3-12 . Saturated monocyclic cycloalkyl rings include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cyclooctyl. Saturated bicyclic and polycyclic cycloalkyl rings include, for example, norbornane, [2.2.2]bicyclooctane, decahydronaphthalene and adamantane. Cycloalkyl groups can also be partially unsaturated, having one or more double or triple bonds in the ring. Representative cycloalkyl groups that are partially unsaturated include, but are not limited to, cyclobutene, cyclopentene, cyclohexene, cyclohexadiene (1,3- and 1,4-isomers), cycloheptene, cycloheptene Diene, cyclooctene, cyclooctadiene (1,3-, 1,4-, and 1,5-isomers), norcamphene, and norcamphene. When the cycloalkyl is a saturated monocyclic C _3-8 cycloalkyl, exemplary groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. When the cycloalkyl is a saturated monocyclic C _3-6 cycloalkyl, exemplary groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. Cycloalkyl groups can be substituted or unsubstituted.

"Cycloalkylene" refers to a cycloalkyl group having the indicated number of carbon atoms and linked to at least two other groups, ie , a divalent group. The two moieties attached to the cycloalkylene can be attached to the same atom or different atoms of the cycloalkylene. Examples of cycloalkylene rings include, inter alia, cyclopropyl, cyclobutylene, cyclopentylene and cyclohexylene. The cycloalkylene group can be attached 1,1, 1,2, 1,3, or 1,4. The cyclohexylene ring, for example, can adopt a number of configurations, including boat and chair configurations. The chair configuration of the cyclohexylene group may have substituents in the axial or equatorial orientation. The divalent nature of the cycloalkylene leads to cis and trans structures, where cis means that the two substituents are on the same side (top or bottom) of the cycloalkylene ring, and where trans means the substituents are on the Opposite side of the cycloalkyl ring. For example, cis -1,2- and cis -1,4-cyclohexylene can have one substituent in the axial orientation and the other in the equatorial orientation, while trans -1,2- and The trans -1,4-cyclohexylene has two substituents in the axial or equatorial orientation. cis -1,3-cyclohexylene has two substituents in axial or equatorial orientation, and trans -1,3-cyclohexylene can have one substituent in axial orientation and equatorial orientation Another substituent. Cycloalkylene groups can be substituted or unsubstituted.

"Alkyl-cycloalkyl" means a group having an alkyl component and a cycloalkyl component, wherein the alkyl component links the cycloalkyl component to a point of attachment. The alkyl component is as defined above except that the alkyl component is at least divalent, is an alkylene, is attached to the cycloalkyl component and is attached to the point of attachment. In some cases, the alkyl component may be absent. The alkyl component can include any number of carbons, _such as C _1-6 , C _1-2 , C 1-3 , C _1-4 , C _1-5 , C _2-3 , C _2-4 , C _{2- 5.} C _2-6 , C _3-4 , C _3-5 , C _3-6 , C _4-5 , C _4-6 and C _5-6 . The cycloalkyl component is as defined therein. Exemplary alkyl-cycloalkyl groups include, but are not limited to, methyl-cyclopropyl, methyl-cyclobutyl, methyl-cyclopentyl, and methyl-cyclohexyl.

"Heterocycloalkyl" or "heterocyclyl" refers to a saturated ring system having 3 to 12 ring members and 1 to 4 N, O and S heteroatoms. Additional heteroatoms may also be useful, including, but not limited to, B, Al, Si, and P. Heteroatoms can also be oxidized, such as but not limited to -S(O)- and -S(O) _2- . Heterocycloalkyl groups can include any number of ring atoms, such as 3 to 6, 4 to 6, 5 to 6, 3 to 8, 4 to 8, 5 to 8, 6 to 8, 3 to 9, 3 to 10, 3 to 11, or 3 to 12 ring members. Any suitable number of heteroatoms may be included in the heterocycloalkyl, such as 1, 2, 3, or 4, or 1 to 2, 1 to 3, 1 to 4, 2 to 3, 2 to 4, or 3 to 4 . Heterocycloalkyl groups may include groups such as aziridine, azetidine, pyrrolidine, piperidine, nitrogen, azacyclooctane, quinuclidine, pyrazolidine, imidazolidine, piperazine (1 , 2-, 1,3- and 1,4-isomers), ethylene oxide, oxetane, tetrahydrofuran, oxane (tetrahydropyran), hexane oxide, sulfide, Thietane, thiolane (tetrahydrothiophene), thiane (tetrahydrothiopyran), oxazolidine, isoxazolidine, thiazolidine, isothiazolidine, dioxolane, dithia Cyclopentane, morpholine, thiomorpholine, dioxane, dithiane, and hexahydro-1H-pyrrolazine. Heterocycloalkyl groups can also be fused to aromatic or non-aromatic ring systems to form members including, but not limited to, indolines. A heterocycloalkyl group can be unsubstituted or substituted. For example, heterocycloalkyl groups can be substituted with C _1-6 alkyl or pendant oxy (=O), among many others.

A heterocycloalkyl group can be attached via any position on the ring. For example, aziridine can be 1- or 2-aziridine, azetidine can be 1- or 2-azetidine, and pyrrolidine can be 1-, 2-, or 3-pyrrolidine , piperidine can be 1-, 2-, 3- or 4-piperidine, pyrazolidine can be 1-, 2-, 3-, or 4-pyrazolidine, and imidazolidine can be 1-, 2-, 3- or 4-imidazolidine, piperazine can be 1-, 2-, 3- or 4-piperazine, tetrahydrofuran can be 1- or 2-tetrahydrofuran, oxazolidine can be 2-, 3-, 4- or 5-oxazolidine, isoxazolidine can be 2-, 3-, 4- or 5-isoxazolidine, thiazolidine can be 2-, 3-, 4- or 5-thiazolidine, isothiazolidine can be is 2-, 3-, 4-, or 5-isothiazolidine, and the morpholine can be 2-, 3-, or 4-morpholine.

When the heterocycloalkyl group includes 3 to 8 ring members and 1 to 3 heteroatoms, representative members include, but are not limited to, pyrrolidine, piperidine, tetrahydrofuran, oxane, tetrahydrothiophene, thiane, pyrazolidine, Imidazolidine, piperazine, oxazolidine, isoxazolidine, thiazolidine, isothiazolidine, morpholine, thiomorpholine, dioxane and dithiane. Heterocycloalkyl groups can also form rings having 5 to 6 ring members and 1 to 2 heteroatoms, representative members include, but are not limited to, pyrrolidine, piperidine, tetrahydrofuran, tetrahydrothiophene, pyrazolidine, imidazolidine, Piperazine, oxazolidine, isoxazolidine, thiazolidine, isothiazolidine, morpholine, and hexahydro-1H-pyrrolizine.

"Heterocycloalkylene" refers to a heterocycloalkyl group as defined above linked to at least two other groups. The two moieties attached to the heterocycloalkylene can be attached to the same atom or different atoms of the heterocycloalkylene. A heterocycloalkylene group can be substituted or unsubstituted.

"Alkyl-heterocycloalkyl" means a group having an alkyl component and a heterocycloalkyl component, wherein the alkyl component links the heterocycloalkyl component to the point of attachment. The alkyl component is as defined above except that the alkyl component is at least divalent, is an alkylene, is attached to the heterocycloalkyl component and is attached to the point of attachment. The alkyl component can include any number of carbons, such as C _0-6 , C _1-2 , C _1-3 , C _1-4 , C _1-5 , C _1-6 , C _2-3 , C _{2- 4.} C _2-5 , C _2-6 , C _3-4 , C _3-5 , C _3-6 , C _4-5 , C _4-6 and C _5-6 . In some cases, the alkyl component may be absent. The heterocycloalkyl component is as defined above. Alkyl-heterocycloalkyl groups can be substituted or unsubstituted.

"Aryl" means an aromatic ring system having any suitable number of ring atoms and any suitable number of rings. The aryl group may comprise any suitable number of ring atoms, such as 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16 ring atoms, and 6 to 10, 6 to 12, or 6 to 14 ring members. Aryl groups can be monocyclic, fused to form bicyclic or tricyclic groups, or linked by bonds to form biaryl groups. Representative aryl groups include phenyl, naphthyl and biphenyl. Other aryl groups include benzyl, which has a methylene linking group. Some aryl groups have 6 to 12 ring members, such as phenyl, naphthyl or biphenyl. Other aryl groups have 6 to 10 ring members, such as phenyl or naphthyl. Some other aryl groups have 6 ring members, such as phenyl. Aryl groups can be substituted or unsubstituted.

"Alkyl-aryl" means a group having an alkyl component and an aryl component, wherein the alkyl component links the aryl component to a point of attachment. The alkyl component is as defined above except that the alkyl component is at least divalent, is an alkylene, is attached to the aryl component and is attached to the point of attachment. The alkyl component may include any number of carbons, such as C _0-6 , C _1-2 , C _1-3 , C _1-4 , C _1-5 , C _1-6 , C _2-3 , C _{2- 4.} C _2-5 , C _2-6 , C _3-4 , C _3-5 , C _3-6 , C _4-5 , C _4-6 and C _5-6 . In some cases, the alkyl component may be absent. The aryl component is as defined above. Examples of alkyl-aryl groups include, but are not limited to, benzyl and ethyl-benzene. Alkyl-aryl groups can be substituted or unsubstituted.

"Arylenyl" means an aryl group as defined above linked to at least two other groups. The two moieties attached to the aryl group may be attached to the same atom or different atoms of the aryl group. Arylene groups can be substituted or unsubstituted.

"Heteroaryl" means a combination of monocyclic or fused bicyclic or tricyclic aromatic rings containing 5 to 16 ring atoms, wherein 1 to 5 of the ring atoms are heteroatoms such as N, O or S. Additional heteroatoms may also be useful, including but not limited to B, Al, Si, and P. Heteroatoms can also be oxidized, such as but not limited to -S(O)- and -S(O) _2- . A heteroaryl group can include any number of ring atoms, such as 5 to 6, 5 to 8, 6 to 8, 5 to 9, 5 to 10, 5 to 11, or 5 to 12 ring members. Any suitable number of heteroatoms may be included in the heteroaryl, such as 1, 2, 3, 4, or 5, or 1 to 2, 1 to 3, 1 to 4, 1 to 5, 2 to 3, 2 to 4 , 2 to 5, 3 to 4, or 3 to 5. The heteroaryl group can have 5 to 8 ring members and 1 to 4 heteroatoms, or 5 to 8 ring members and 1 to 3 heteroatoms, or 5 to 6 ring members and 1 to 4 heteroatoms, or 5 to 6 ring members and 1 to 3 heteroatoms. Heteroaryl groups may include groups such as pyrrole, pyridine, imidazole, pyrazole, triazole, tetrazole, pyrazine, pyrimidine, pyrazine, triazine (1,2,3-, 1,2,4- and 1, 3,5-isomer), thiophene, furan, thiazole, isothiazole, oxazole, and isoxazole. Heteroaryl groups may also be fused to aromatic ring systems such as phenyl rings to form members including, but not limited to, benzopyrroles such as indole and isoindole, benzopyridines such as quinoline and isoquinoline, benzopyrazine (quinoxaline), benzopyrimidine (quinazoline), benzopyridazines such as phthalazine and phenoline, benzothiophene, and benzofuran. Other heteroaryl groups include heteroaryl rings joined by bonds, such as bipyridyl. Heteroaryl groups can be substituted or unsubstituted.

A heteroaryl group can be attached via any position on the ring. For example, pyrrole includes 1-, 2- and 3-pyrrole, pyridine includes 2-, 3- and 4-pyridine, imidazole includes 1-, 2-, 4- and 5-imidazole, and pyrazole includes 1-, 3-, 4- and 5-pyrazoles, triazoles include 1-, 4- and 5-triazoles, tetrazoles include 1- and 5-tetrazoles, pyrimidines include 2-, 4-, 5- and 6-pyrimidines, pyrazines Including 3- and 4-pyridazine, 1,2,3-triazine including 4- and 5-triazine, 1,2,4-triazine including 3-, 5- and 6-triazine, 1,3, 5-triazine includes 2-triazine, thiophene includes 2- and 3-thiophene, furan includes 2- and 3-furan, thiazole includes 2-, 4-, and 5-thiazole, and isothiazole includes 3-, 4-, and 5- - Isothiazole, oxazole includes 2-, 4- and 5-oxazole, isoxazole includes 3-, 4- and 5-isoxazole, indole includes 1-, 2- and 3-indole, isoindole Indole includes 1- and 2-isoindole, quinoline includes 2-, 3-, and 4-quinoline, isoquinoline includes 1-, 3-, and 4-isoquinoline, and quinazoline includes 2- and 4- Quinazoline, phenoline include 3- and 4-phenoline, benzothiophene includes 2- and 3-benzothiophene, and benzofuran includes 2- and 3-benzofuran.

Some heteroaryl groups include heteroaryl groups having 5 to 10 ring members and 1 to 3 ring atoms including N, O or S, such as pyrrole, pyridine, imidazole, pyrazole, triazole, pyrazine, pyrimidine, pyridyl oxazine, triazine (1,2,3-, 1,2,4- and 1,3,5-isomers), thiophene, furan, thiazole, isothiazole, oxazole, isoxazole, indole, iso Indole, quinoline, isoquinoline, quinoxaline, quinazoline, phthalazine, phenoline, benzothiophene, and benzofuran. Other heteroaryl groups include heteroaryl groups having 5 to 8 ring members and 1 to 3 heteroatoms such as pyrrole, pyridine, imidazole, pyrazole, triazole, pyrazine, pyrimidine, pyrazine, triazine (1, 2,3-, 1,2,4- and 1,3,5-isomers), thiophene, furan, thiazole, isothiazole, oxazole, and isoxazole. Some other heteroaryl groups include heteroaryl groups having 9 to 12 ring members and 1 to 3 heteroatoms such as indole, isoindole, quinoline, isoquinoline, quinoxaline, quinazoline, phthalazine , phenoline, benzothiophene, benzofuran and bipyridine. Other heteroaryl groups include those having 5 to 6 ring members and 1 to 2 ring atoms including N, O or S, such as pyrrole, pyridine, imidazole, pyrazole, pyrazine, pyrimidine, pyrazine, thiophene , furan, thiazole, isothiazole, oxazole, and isoxazole.

Some heteroaryl groups include 5 to 10 ring members and only nitrogen heteroatoms, such as pyrrole, pyridine, imidazole, pyrazole, triazole, pyrazine, pyrimidine, pyrazine, triazine (1,2,3-, 1, 2,4- and 1,3,5-isomers), indole, isoindole, quinoline, isoquinoline, quinoxaline, quinazoline, phthalazine, and phenoline. Other heteroaryl groups include 5 to 10 ring members and only oxygen heteroatoms, such as furan and benzofuran. Some other heteroaryl groups include 5 to 10 ring members and only sulfur heteroatoms, such as thiophene and benzothiophene. Other heteroaryl groups include 5 to 10 ring members and at least two heteroatoms, such as imidazole, pyrazole, triazole, pyrazine, pyrimidine, pyrazine, triazine (1,2,3-, 1,2,4 - and 1,3,5-isomers), thiazole, isothiazole, oxazole, isoxazole, quinoxaline, quinazoline, phthalazine, and phenoline.

"Heteroarylylene" refers to a heteroaryl group as defined above linked to at least two other groups. The two moieties attached to the heteroaryl are attached to different atoms of the heteroaryl. A heteroarylylene group can be substituted or unsubstituted.

"Alkyl-heteroaryl" refers to a group having an alkyl component and a heteroaryl component, wherein the alkyl component links the heteroaryl component to a point of attachment. The alkyl component is as defined above except that the alkyl component is at least divalent, is an alkylene group, is attached to the heteroaryl component and is attached to the point of attachment. The alkyl component may include any number of carbons, such as C _0-6 , C _1-2 , C _1-3 , C _1-4 , C _1-5 , C _1-6 , C _2-3 , C _{2- 4.} C _2-5 , C _2-6 , C _3-4 , C _3-5 , C _3-6 , C _4-5 , C _4-6 and C _5-6 . In some cases, the alkyl component may be absent. The heteroaryl component is as defined therein. Alkyl-heteroaryl groups can be substituted or unsubstituted.

The groups defined above may optionally be substituted with any suitable number and type of substituents. Representative substituents include, but are not limited to, halo, haloalkyl, haloalkoxy, -OR', =O, -OC(O)R', -(O)R', _-O2R ', -ONR'R",-OC(O)NR'R",=NR',=N-OR',-NR'R",-NR"C(O)R',-NR'-(O)NR"R"',-NR”C(O)OR', -NH-(NH ₂ )=NH, -NR'C(NH ₂ )=NH, -NH-(NH ₂ )=NR', -SR', -S (O)R', -S(O) ₂ R', -S(O) ₂ NR'R", -NR'S(O) ₂ R", -N ₃ and -NO ₂ . R', R" and R "' each independently refers to hydrogen, unsubstituted alkyl, such as unsubstituted C _1-6 alkyl. Alternatively, when attached to the same nitrogen, R' and R", or R" and R"' are attached to it The nitrogens combine to form a heterocycloalkyl or heteroaryl ring as defined above.

"Salt" refers to acid or base salts of compounds useful in the methods disclosed herein. Illustrative examples of pharmaceutically acceptable salts are salts of inorganic acids (hydrochloric, hydrobromic, phosphoric, and the like), organic acids (acetic, propionic, glutamic, citric, and the like) , Quaternary ammonium (methyl iodide, ethane iodide, and similar ammonium) salts. It is understood that pharmaceutically acceptable salts are non-toxic. Additional information on suitable pharmaceutically acceptable salts can be found in Remington's Pharmaceutical Sciences, 17th Edition, Mack Publishing Company, Easton, Pa., 1985, which is incorporated herein by reference.

The pharmaceutically acceptable salts of the acidic compounds disclosed herein are salts with bases, i.e., cationic salts such as alkali metal and alkaline earth metal salts, such as sodium, lithium, potassium, calcium, magnesium, and ammonium salts, such as ammonium, tris Methylammonium, diethylammonium, and ginseng-(hydroxymethyl)-methyl-ammonium salts.

Similarly, acid addition salts such as mineral acids, organic carboxylic acids and organic sulfonic acids, e.g. hydrochloric acid, methanesulfonic acid, maleic acid are also possible as long as a basic group such as pyridyl constitutes the structure. part.

The neutral form of the compound can be regenerated by contacting the salt with a base or acid and isolating the parent compound in conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents, but otherwise the salts are equivalent to the parent form for purposes of the present embodiments.

Certain compounds disclosed herein possess asymmetric carbon atoms (optical centers) or double bonds; racemates, diastereomers, geometric isomers, and individual isomers are all intended to be encompassed within the scope of the present embodiments .

"Hydrate" means a compound complexed with at least one water molecule. The compounds disclosed herein can complex with 1 to 10 water molecules.

A "composition" as used herein is intended to encompass a product comprising the specified ingredients in the specified amounts, as well as any product resulting, directly or indirectly, from the combination of the specified ingredients in the specified amounts. "Pharmaceutically acceptable" means that the carrier, diluent or excipient must be compatible with the other ingredients of the formulation and not harmful to the recipient thereof.

A "pharmaceutically acceptable excipient" refers to a substance that facilitates administration of an active agent to and absorption by a subject. Pharmaceutical excipients that can be used in the embodiments of the present invention include but are not limited to binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavoring agents and pigments. Those skilled in the art will recognize that other pharmaceutical excipients may be used in embodiments of the invention.

"Treat/treating/treatment" means the successful treatment or amelioration of any sign of an injury, pathology, condition or symptom (e.g. pain), including any objective or subjective parameter such as alleviation; relief; To make the injury, pathology or condition more tolerable to the patient; to reduce the frequency or duration of the symptoms or condition; or, in some cases, to prevent the onset of symptoms. Treatment or amelioration of symptoms can be based on any objective or subjective parameter; including, for example, the results of a physical examination.

"Administration" means oral administration, administration by suppository, topical contact, parenteral, intravenous, intraperitoneal, intramuscular, intralesional, intranasal or subcutaneous administration, intrathecal administration, or A slow release device such as a miniature osmotic pump is implanted in the individual.

A "therapeutically effective amount or dose" or "therapeutically sufficient amount or dose" or "effective or sufficient amount or dose" refers to a dose that produces the therapeutic effect for which its administration is intended. The precise dosage depends on the purpose of the treatment and can be determined by one skilled in the art using known techniques ( see , e.g. , Lieberman, Pharmaceutical Dosage Forms (Volumes 1-3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999); and Remington: The Science and Practice of Pharmacy , 20th ed., 2003, Gennaro, Ed., Lippincott, Williams & Wilkins). In sensitized cells, the therapeutically effective dose may generally be lower than the conventional therapeutically effective dose for non-sensitized cells.

"Individual" refers to animals such as mammals, including but not limited to primates ( eg, humans), cows, sheep, goats, horses, dogs, cats, rabbits, rats, mice, and the like. In certain embodiments, the individual is human. III. Compounds

(I) 其中 m為1或2； p為1或2； L ¹為

其中k為0至4之整數；並且各R ¹獨立地選自甲基、及氰甲基、C ₂-C ₄烷基、氰基、環烷基、鹵基、鹵烷基、三氟甲基、及烷氧基；或任何兩個R ¹組合以形成稠環、橋或螺環結構，視情況在橋或螺環中包含選自S、SO ₂、O或N之雜原子，並且其中橋或螺環結構視情況經側氧基取代； R ²選自由以下組成之群：烷基、N-烷基胺基、N,N-二烷基胺基、烷基醯胺烷基、芳基醯胺烷基、烷基磺醯胺基烷基、芳基磺醯胺基烷基、N-烷基胺基烷基、N,N-二烷基胺基烷基、烷氧基、烷氧基烷基、環烷基、烷基環烷基、羥基烷基、鹵素、鹵烷基、芳基、芳烷基、雜芳基、雜芳基烷基、雜環基、及雜環基烷基，其中之任一者視情況經取代；或當m為2時，兩個R ²組合以形成視情況含有1至3個選自N、O、或S之雜原子的螺環3至6員環； R ³、R ⁴、R ⁵、及R ⁶獨立地選自鹵素、氫、羥基、烷氧基、烷基、環烷基、胺基、N-烷基胺基、C-醯胺(-CONRR’)、N-醯胺(-NHCOR)、脲(-NHCONHR)、醚(-OR)、磺醯胺(-NHSO ₂R或-SO ₂NHR)、及CF ₃；其中各R及R’獨立地為氫、烷基、或環烷基；或 任何兩個相鄰R ³、R ⁴、R ⁵、或R ⁶形成包含0至3個選自N、O或S之雜原子的視情況經取代之稠合5員或6員環； 限制條件為R ³、R ⁴、R ⁵、或R ⁶中之一者為連接至2-喹唑啉酮之鍵；且 R ⁷為烷基、氰基、環烷基、鹵素、鹵烷基、三氟甲基、及烷氧基。 Embodiments of the present invention provide compounds of formula (I) and pharmaceutically acceptable salts thereof:

(I) where m is 1 or 2; p is 1 or 2; L ¹ is

wherein k is an integer from 0 to 4; and ^each R is independently selected from methyl, and cyanomethyl, C ₂ -C ₄ alkyl, cyano, cycloalkyl, halo, haloalkyl, trifluoromethane group, and alkoxy group; or any two R ¹ combined to form a fused ring, bridge or spiro ring structure, optionally including a heteroatom selected from S, SO ₂ , O or N in the bridge or spiro ring, and wherein The bridge or spiro ring structure is optionally substituted by a side oxygen group; R ^is selected from the group consisting of: alkyl, N-alkylamino, N,N-dialkylamino, alkylamidoalkyl, aromatic Aminoalkyl, Alkylsulfonamidoalkyl, Arylsulfonamidoalkyl, N-Alkylaminoalkyl, N,N-Dialkylaminoalkyl, Alkoxy, Alkyl Oxyalkyl, cycloalkyl, alkylcycloalkyl, hydroxyalkyl, halogen, haloalkyl, aryl, aralkyl, heteroaryl, heteroarylalkyl, heterocyclyl, and heterocyclyl Alkyl, any of which is optionally substituted; or when m is 2, two R combined to form a spiro ring optionally containing 1 to 3 heteroatoms selected from ^N , O, or S 3 to 6-membered ring; R ³ , R ⁴ , R ⁵ , and R ⁶ are independently selected from halogen, hydrogen, hydroxyl, alkoxy, alkyl, cycloalkyl, amino, N-alkylamino, C-acyl Amine (-CONRR'), N-amide (-NHCOR), urea (-NHCONHR), ether (-OR), sulfonamide (-NHSO ₂ R or -SO ₂ NHR), and CF ₃ ; wherein each R And R' is independently hydrogen, alkyl, or cycloalkyl; or any two adjacent R ³ , R ⁴ , R ⁵ , or R ⁶ form 0 to 3 heteroatoms selected from N, O, or S An optionally substituted fused 5- or 6-membered ring; provided that one of R ³ , R ⁴ , R ⁵ , or R ⁶ is a bond to a 2-quinazolinone; and R ⁷ is Alkyl, cyano, cycloalkyl, halogen, haloalkyl, trifluoromethyl, and alkoxy.

(IIa) 其中Ar ¹為C連接芳基、雜芳基、雜環、或碳環； n為0至3之整數；且 各R ¹⁰獨立地選自烷基、胺基、氰基鹵素、三氟甲基、雜環基，或兩個R ¹⁰組合以形成雙環稠合雜環。 In an embodiment, provided herein is a compound having formula (IIa): a pharmaceutically acceptable salt thereof:

(IIa) wherein Ar is C ^- linked aryl, heteroaryl, heterocycle, or carbocycle; n is an integer from 0 to 3; and ^each R is independently selected from alkyl, amine, cyanohalogen, tris Fluoromethyl, heterocyclyl, or two R ¹⁰ combined to form a bicyclic fused heterocycle.

(IIb) 其中Ar ²為N連接雜芳基或雜環； n為0至3之整數；且 各R ¹⁰獨立地選自烷基、胺基、氰基鹵素、三氟甲基、雜環基，或兩個R ¹⁰組合以形成雙環稠合雜環。 In an embodiment, provided herein is a compound having Formula (IIb), or a pharmaceutically acceptable salt thereof:

(IIb) wherein Ar is N ^- linked heteroaryl or heterocycle; n is an integer from 0 to 3; and ^each R is independently selected from alkyl, amino, cyanohalogen, trifluoromethyl, heterocyclyl , or two R ¹⁰ combine to form a bicyclic fused heterocycle.

(IIc) 其中Ar ³為芳基或雜芳基； n為0至3之整數；且 各R ¹⁰獨立地選自烷基、胺基、氰基鹵素、三氟甲基、雜環基，或兩個R ¹⁰組合以形成雙環稠合雜環。 In an embodiment, provided herein is a compound having Formula (IIc), or a pharmaceutically acceptable salt thereof:

(IIc) wherein Ar is aryl or heteroaryl; n is an integer from 0 to 3; and ^{each R} is independently selected from alkyl, amine, cyanohalogen, trifluoromethyl, heterocyclyl, or Two R ¹⁰ combine to form a bicyclic fused heterocycle.

In an embodiment, n is 1. In an embodiment, n is 2. In an embodiment, n is 3. In an embodiment, n is 4.

In an embodiment, p is 1. In an embodiment, p is 2.

、

、

、

、

、

、

、

、

，其中

經由L ¹之兩個氮原子中之任一者來連接。 In an embodiment, ^L is selected from:

,

,

,

,

,

,

,

,

,in

Linkage is via either of the two nitrogen atoms of L ¹ .

、

、

、

、

、

、

、

、

、

、及

、 C-連接芳基或雜芳基、 N-連接雜芳基或雜環基，其中R及R’獨立地選自氫、烷基、及環烷基。 In an embodiment, R ² is selected from methoxy, amine, MeOCH ₂ -, EtOCH ₂ -, MeO(CH ₂ ) ₂ NH-,

,

,

,

,

,

,

,

,

,

,and

, C -linked aryl or heteroaryl, N -linked heteroaryl or heterocyclyl, wherein R and R' are independently selected from hydrogen, alkyl, and cycloalkyl.

、

、

、及

； 其中各W、X、Y、及Z獨立地選自C=O、NH、O、S、CH、C-Q，其中Q為胺基、鹵素、甲基、-O-烷基、-O-環烷基、或三氟甲基。 In an embodiment, R ³ , R ⁴ , R ⁵ , and R ⁶ are defined as fused thiophenes selected from the following:

,

,

,and

; Wherein each W, X, Y, and Z are independently selected from C=O, NH, O, S, CH, CQ, wherein Q is amino, halogen, methyl, -O-alkyl, -O-ring Alkyl, or trifluoromethyl.

、

、

、

，其中X為氫、氯、甲基、或CF ₃，

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、及

。 In an embodiment, R ³ , R ⁴ , R ⁵ , and R ⁶ are defined as thiophenes selected from the following:

,

,

,

, wherein X is hydrogen, chlorine, methyl, or CF ₃ ,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,and

.

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、

、或

。 In embodiments, the compounds disclosed herein include the following compounds or pharmaceutically acceptable salts thereof:

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,or

.

In embodiments, the compounds disclosed herein include the following further compounds, or pharmaceutically acceptable salts thereof:

The compounds disclosed herein may exist in salt form. The embodiments of the present invention include such salts, which may be pharmaceutically acceptable salts. Examples of suitable salt forms include hydrochloride, hydrobromide, sulfate, methanesulfonate, nitrate, maleate, acetate, citrate, fumarate, tartrate (e.g. (+) -tartrates, (-)-tartrates or mixtures thereof including racemic mixtures, succinates, benzoates and salts with amino acids such as glutamic acid. These salts can be obtained by those skilled in the art Prepared by known methods. Also include base addition salts such as sodium, potassium, calcium, ammonium, organic amino groups, or magnesium salts, or similar salts. When the compounds disclosed herein contain relatively basic functional groups, acid addition salts Can be obtained by contacting the neutral form of these compounds with a sufficient amount of the desired acid, neat or in a suitable inert solvent. Examples of acceptable acid addition salts include those obtained from inorganic acids such as hydrochloric acid, hydrobromic acid , nitric acid, carbonic acid, monohydrogencarbonic acid, phosphoric acid, monohydrogenphosphoric acid, dihydrogenphosphoric acid, sulfuric acid, monohydrogensulfuric acid, hydroiodic acid or phosphoric acid, etc., and those derived from organic acids such as acetic acid, propionic acid, isobutyric acid, Maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid Salts derived from acids, etc. Also include salts of amino acids such as arginine and similar acids, and organic acids such as glucuronic acid or galacturonic acid and salts of similar acids. Certain specific compounds disclosed herein contain allowable Basic and acidic functional groups that convert compounds into base or acid addition salts.

Other salts include acid or base salts of compounds used in the methods of the embodiments of the invention. Illustrative examples of pharmaceutically acceptable salts are salts of inorganic acids (hydrochloric, hydrobromic, phosphoric, and the like), organic acids (acetic, propionic, glutamic, citric, and the like) , and quaternary ammonium (methyl iodide, ethane iodide, and similar ammonium) salts. It is understood that pharmaceutically acceptable salts are non-toxic. Additional information on suitable pharmaceutically acceptable salts can be found in Remington's Pharmaceutical Sciences, 17th Edition, Mack Publishing Company, Easton, Pa., 1985, which is incorporated herein by reference.

Pharmaceutically acceptable salts include salts of the active compounds which are prepared with relatively non-toxic acids or bases, depending on the particular substituents present on the compounds described herein. When compounds disclosed herein contain relatively acidic functionalities, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, neat or in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, or magnesium salts, or similar salts. When compounds disclosed herein contain relatively basic functionalities, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, neat or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include those obtained from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrogencarbonic acid, phosphoric acid, monohydrogenphosphoric acid, dihydrogenphosphoric acid, sulfuric acid, monohydrogensulfuric acid, hydroiodic acid or Those derived from phosphoric acid, etc., as well as from relatively non-toxic organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, almond Salts derived from acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid, etc. Also included are salts of amino acids such as arginine and similar acids, and salts of organic acids such as glucuronic acid or galacturonic acid and similar acids ( see , e.g., Berge et al ., "Pharmaceutical Salts", Journal of Pharmaceutical Science , 1977 , 66 , 1-19). Certain specific compounds disclosed herein contain basic and acidic functional groups that allow conversion of the compounds into base or acid addition salts.

The neutral form of the compound is preferably regenerated by contacting the salt with a base or acid and isolating the parent compound in conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents.

Certain compounds disclosed herein can exist in unsolvated forms as well as solvated forms, including hydrated forms. In general, the solvated forms are equivalent to unsolvated forms and are within the scope of the embodiments of this invention. Certain compounds disclosed herein can exist in multiple crystalline or amorphous forms. In general, all physical forms are equivalent for the uses contemplated by the embodiments of the invention and are intended to be within the scope of the embodiments of the invention.

Certain compounds disclosed herein have asymmetric carbon atoms (optical centers) or double bonds; enantiomers, racemates, diastereomers, tautomers, geometric isomers, amino acid Stereoisomeric forms defined as (R)- or (S)- or (D)- or (L)- in terms of absolute stereochemistry, and individual isomers are encompassed within the scope of the present embodiments. Compounds disclosed herein do not include compounds known in the art to be too unstable to be synthesized and/or isolated. The present embodiments are intended to include the compounds in racemic and optically pure forms. Optically active (R)- and (S)-, or (D)- and (L)-isomers can be prepared using chiral synthons or chiral reagents, or resolved using known techniques. Compounds disclosed herein may be provided as a mixture of atropisomers or may be pure atropisomers.

Isomers include compounds that have the same number and kind of atoms, and thus the same molecular weight, but differ in the structural arrangement or configuration of the atoms.

Unless otherwise indicated, structures depicted herein are also intended to include all stereochemical forms of the structure; that is, the R and S configurations of each asymmetric center. Thus, single stereochemical isomers as well as enantiomerically and diastereomeric mixtures of the compounds of the present invention are within the scope of the embodiments.

Unless otherwise indicated, the compounds disclosed herein may also contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the compounds disclosed herein can be used with radioactive or stable isotopes such as, for example, deuterium ( ² H), deuterium ( ³ H), iodine-125 ( ¹²⁵ I), fluorine-18 ( ¹⁸ F), nitrogen-15 ( ¹⁵ N), Oxygen-17 ( ¹⁷ O), Oxygen-18 ( ¹⁸ O), Carbon-13 ( ¹³ C), or Carbon-14 ( ¹⁴ C). All isotopic variations of the compounds disclosed herein, whether radioactive or not, are encompassed within the scope of the present embodiments.

In addition to salt forms, embodiments of the present invention provide compounds that are in prodrug form. Prodrugs of the compounds described herein are those compounds that readily undergo chemical changes under physiological conditions to provide the compounds disclosed herein. In addition, prodrugs can be converted to the compounds disclosed herein by chemical or biochemical methods in an ex vivo environment. For example, prodrugs can be slowly converted to compounds disclosed herein when placed in a transdermal patch reservoir with a suitable enzyme or chemical reagent.

The compounds disclosed herein can be prepared by various methods described in the illustrative synthetic reaction schemes shown and described below. Starting materials and reagents for the preparation of these compounds are generally available from commercial suppliers, such as Aldrich Chemical Co., or can be prepared by methods known to those skilled in the art, following the procedures set forth in references such as the following : Fieser and Fieser's Reagents for Organic Synthesis ; Wiley & Sons: New York, vol. 1-21; RC LaRock, Comprehensive Organic Transformations , 2nd ed. Wiley-VCH, New York 1999 ; Comprehensive Organic Synthesis , B. Trost and I. Fleming (ed.) Volumes 1-9 Pergamon, Oxford, 1991 ; Comprehensive Heterocyclic Chemistry , AR Katritzky and CW Rees (eds.) Pergamon, Oxford 1984 , volumes 1-9; Comprehensive Heterocyclic Chemistry II , AR Katritzky and CW Rees (eds. ) Pergamon, Oxford 1996 , vol. 1-11; and Organic Reactions , Wiley & Sons: New York, 1991 , vol. 1-40. The following synthetic reaction schemes illustrate only some of the methods by which the compounds disclosed herein may be synthesized, and various modifications of these synthetic reaction schemes can be made and will occur to those skilled in the art having reference to the disclosure contained herein.

For exemplary purposes, the following reaction schemes provide routes to the synthesis of compounds disclosed herein as well as key intermediates. For a more detailed description of individual reaction steps, see the Examples section below. Those skilled in the art recognize that other synthetic routes can be used. Although some specific starting materials and reagents are described in the schemes and discussed below, other starting materials and reagents can be substituted in order to provide various derivatives or reaction conditions. In addition, many of the compounds prepared by the methods described below can be further modified in light of the present disclosure using conventional chemistry well known to those skilled in the art.

Starting materials and intermediates of the synthetic reaction schemes can be isolated and purified if necessary using conventional techniques including, but not limited to, filtration, distillation, crystallization, chromatography, and the like. Such materials can be characterized using conventional means including physical constants and spectral data.

Unless specified to the contrary, the reactions described herein are preferably under an inert atmosphere, at atmospheric pressure, at a reaction temperature range of from about −78°C to about 150°C, more preferably from about 0°C to about 125°C, and most preferably And conveniently at about room (or ambient) temperature or about 20°C.

Some of the compounds in the following schemes are described as having common substituents; however, one skilled in the art immediately recognizes that the nature of the substituents can be varied in order to provide a variety of compounds encompassed in the examples of the invention. In addition, reaction conditions are exemplary and alternative conditions are well known. The reaction series in the following examples are not intended to limit the scope of the examples as set forth in the claims. IV. Pharmaceutical formulations

In some embodiments, a pharmaceutical composition comprises a compound of any of the compounds disclosed herein and a pharmaceutically acceptable excipient.

In some embodiments, a pharmaceutical composition is provided, which comprises a pharmaceutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

In some embodiments, the pharmaceutical compositions further comprise additional therapeutic agents.

In some embodiments, the additional therapeutic agent is a chemotherapeutic agent. In some embodiments, the chemotherapeutic agent is an antimicrotubule agent, a platinum coordination complex, an alkylating agent, an antibiotic agent, a topoisomerase II inhibitor, an antimetabolite, a topoisomerase I inhibitor, a hormone, or Hormone analogs, signal transduction pathway inhibitors, non-receptor tyrosine kinase angiogenesis inhibitors, immunotherapeutics, pro-apoptotic agents, LDH-A inhibitors, fatty acid biosynthesis inhibitors, cell cycle signal transduction Inhibitors, HDAC inhibitors, proteasome inhibitors, or inhibitors of cancer metabolism. In some embodiments, the chemotherapeutic agent is cisplatin, carboplatin, doxorubicin, ionizing radiation, docetaxel, or paclitaxel.

The compounds disclosed herein can be prepared and administered in a variety of oral, parenteral, and topical dosage forms. Oral formulations include tablets, pills, powders, dragees, capsules, liquids, lozenges, gels, syrups, slurries, suspensions and the like suitable for ingestion by the patient. The compounds disclosed herein can also be administered by injection, ie, intravenous, intramuscular, intradermal, subcutaneous, intraduodenal, or intraperitoneal. Also, the compounds described herein can be administered by inhalation, eg, intranasally. Additionally, the compounds disclosed herein can be administered transdermally. The compounds disclosed herein can also be administered by intraocular, intravaginal, and intrarectal routes, including suppository, insufflation, powder, and aerosol formulations (eg, steroid inhalation, see Rohatagi, J. Clin. Pharmacol. 35:1187-1193, 1995; Tjwa, Ann. Allergy Asthma Immunol. 75:107-111, 1995). Therefore, the embodiment of the present invention also provides a pharmaceutical composition, which comprises one or more pharmaceutically acceptable carriers and/or excipients and the compound of formula I, or a pharmaceutically acceptable salt of the compound of formula I.

For preparing pharmaceutical compositions from the compounds disclosed herein, pharmaceutically acceptable carriers can be solid or liquid. Solid form preparations include powders, lozenges, pills, capsules, cachets, suppositories, and dispersible granules. A solid carrier can be one or more substances which may also act as diluents, flavoring agents, surface active agents, binders, preservatives, tablet disintegrating agents, or an encapsulating material. Details of formulation and administration techniques are well described in the scientific and patent literature, see, eg, the latest edition of Remington's Pharmaceutical Sciences, Maack Publishing Co, Easton PA ("Remington's").

In powders, the carrier is a finely divided solid, which is in admixture with the finely divided active component. In tablets, the active component is admixed with the carrier having the necessary binding properties and, if necessary, additional excipients in suitable proportions and compacted in the shape and size desired.

Powders, capsules and lozenges preferably contain from 5% or 10% to 70% of the active compound. Suitable carriers are magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, a low melting wax, cocoa butter , and similar substances. The term "preparation" is intended to include the formulation of the active compound with encapsulating material as carrier, providing a capsule in which the active component is surrounded by carrier, with or without other excipients, thereby being in association therewith. Similarly, cachets and lozenges are included. Tablets, powders, capsules, pills, cachets, and lozenges can be used as solid dosage forms suitable for oral administration.

Suitable solid excipients are carbohydrate or protein fillers, including but not limited to sugars, including lactose, sucrose, mannitol, or sorbitol; starches from corn, wheat, rice, potatoes, or other plants; cellulose such as methylcellulose, hydroxypropylmethylcellulose, or sodium carboxymethylcellulose; and gums including acacia and tragacanth; and proteins such as gelatin and collagen. If necessary, a disintegrating or dissolving agent, such as cross-linked polyvinylpyrrolidone, agar, alginic acid or a salt thereof, such as sodium alginate, may be added.

Dragee cores are provided with suitable coatings such as concentrated sugar solutions, which may also contain gum arabic, talc, polyvinylpyrrolidone, carbomer gel, polyethylene glycol, and/or titanium dioxide, varnish solutions, and suitable organic solvents or solvent mixture. Dyestuffs or pigments can be added to tablets or dragee coatings for product identification or to characterize the amount (ie, dose) of active compound. Pharmaceutical preparations which can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a coating, such as glycerol or sorbitol. Push-fit capsules can contain a compound disclosed herein in admixture with filler or binders, such as lactose or starches, lubricants, such as talc or magnesium stearate, and, optionally, stabilizers. In soft capsules, the compounds disclosed herein may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols, with or without stabilizers.

For preparing suppositories, a low-melting wax, such as a mixture of fatty acid glycerides or cocoa butter, is first melted and the active ingredient is dispersed uniformly therein, such as by stirring. The molten homogeneous mixture is then poured into conveniently sized molds, allowed to cool, and thereby solidify.

Liquid form preparations include solutions, suspensions, and emulsions, for example, water or water/propylene glycol solutions. For parenteral injection, liquid preparations can be formulated in solution in aqueous polyethylene glycol solution.

Aqueous solutions suitable for oral use can be prepared by dissolving the active component in water and adding suitable colorants, flavours, stabilizing, and thickening agents, as desired. Aqueous suspensions suitable for oral use can be prepared by dispersing the finely divided active component in water with viscous material, such as natural or synthetic gums, resins, methylcellulose, and dispersing or wetting agents. sodium carboxymethylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth, and gum arabic, and such dispersing or wetting agents as naturally occurring phospholipids (e.g., lecithin), condensation products of alkylene oxides and fatty acids (for example, polyoxyethylene stearate), condensation products of ethylene oxide and long-chain fatty alcohols (for example, heptadecanyloxycetyl alcohol), epoxy Condensation products of ethane with partial esters derived from fatty acids and hexitols (for example, polyoxyethylene sorbitan monooleate), or ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides products (eg, polyoxyethylene sorbitan monooleate). Aqueous suspensions may also contain one or more preservatives such as ethyl or n-propyl paraben, one or more coloring agents, one or more flavoring agents and one or more sweetening agents such as sucrose, aspartame Sweet or saccharin. Formulations can be adjusted for osmolarity.

Also included are solid form preparations which are intended to be converted, shortly before use, to liquid form preparations for oral administration. Such liquid forms include solutions, suspensions, and emulsions. Such preparations may contain, in addition to the active ingredient, coloring agents, flavoring agents, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizers, and the like.

Oily suspensions can be formulated by suspending a compound disclosed herein in a vegetable oil, such as arachis oil, olive oil, sesame oil, or coconut oil, or in a mineral oil such as liquid paraffin; or mixtures of these substances. The oily suspensions may contain a thickening agent, such as beeswax, hard paraffin or cetyl alcohol. Sweetening agents can be added to provide a palatable oral preparation, such as glycerol, sorbitol or sucrose. These formulations can be preserved by the addition of antioxidants such as ascorbic acid. For examples of injectable oil vehicles, see Minto, J. Pharmacol. Exp. Ther. 281:93-102, 1997. Pharmaceutical formulations may also be in the form of oil-in-water emulsions. The oily phase may be a vegetable or mineral oil as described above, or a mixture of these. Suitable emulsifiers include naturally occurring gums such as acacia and tragacanth, naturally occurring phospholipids such as soybean lecithin, esters or partial esters derived from fatty acids and hexitol anhydrides such as sorbitan monooleate, and condensation products of such partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. The emulsions may also contain sweetening and flavoring agents, as in the preparation of syrups and elixirs. These formulations may also contain a demulcent, a preservative, or a coloring agent.

The compounds disclosed herein can be delivered transdermally by topical routes, formulated as applicators, solutions, suspensions, emulsions, gels, creams, ointments, pastes, colloids, paints, powders, and aerosols.

The compounds disclosed herein can also be delivered as microspheres for slow release in vivo. For example, microspheres can be administered via intradermal injection of drug-containing microspheres that are slowly released subcutaneously (see Rao, J. Biomater Sci. Polym. Ed . 7:623-645, 1995; as biodegradable and injectable gel formulations (see, for example, Gao Pharm. Res . 12:857-863, 1995); or as microspheres for oral administration (see, for example, Eyles, J. Pharm. Pharmacol. 49:669-674, 1997). Transdermal and intradermal routes provide constant delivery for weeks or months.

Pharmaceutical formulations of the compounds disclosed herein can be provided as salts and can be formed with many acids including, but not limited to, hydrochloric, sulfuric, acetic, lactic, tartaric, malic, succinic, and the like. Salts tend to be more soluble in aqueous or other protic solvents as the corresponding free base form. In other cases, the formulation may be a lyophilized powder in 1 mM-50 mM histidine, 0.1%-2% sucrose, 2%-7% mannitol at a pH range of 4.5 to 5.5, which is given prior to use. Combine with buffer.

Pharmaceutical formulations of the compounds disclosed herein can be provided as salts and can be formed with bases, i.e., cationic salts such as alkali metal and alkaline earth metal salts, such as sodium, lithium, potassium, calcium, magnesium, and ammonium salts, such as ammonium, trimethyl ammonium, diethylammonium, and ginseng-(hydroxymethyl)-methyl-ammonium salts.

In some embodiments, formulations of compounds disclosed herein can be delivered using liposomes that fuse with cell membranes or are endocytosed, that is, by using attachment to liposomes, or directly to oligonucleotides ligands that bind to cell surface membrane protein receptors, leading to endocytosis. By using liposomes, one can aim to deliver GR modulators to target cells in vivo, especially when the liposome surface bears a ligand specific for the target cell, or is otherwise preferentially directed to a specific organ. (See e.g. Al-Muhammed, J. Microencapsul. 13:293-306, 1996; Chonn, Curr. Opin. Biotechnol. 6:698-708, 1995; Ostro, Am. J. Hosp. Pharm. 46:1576-1587 , 1989).

The pharmaceutical formulations are preferably in unit dosage form. In such form, the preparation is subdivided into unit doses containing appropriate quantities of the active component. The unit dosage form can be a packaged preparation, the package containing discrete quantities of preparation, such as packeted tablets, capsules, and powders in vials or ampoules. Also, the unit dosage form can be a capsule, tablet, cachet, or lozenge itself, or it can be the appropriate number of any of these in packaged form.

Depending on the particular application and potency of the active ingredient, the amount of active ingredient in a unit dosage formulation may be varied or adjusted from 0.1 mg to 10000 mg, more usually 1.0 mg to 1000 mg, most usually 10 mg to 500 mg. The compositions may also contain other compatible therapeutic agents, if desired.

Dosage regimens also take into account pharmacokinetic parameters well known in the art, that is, absorption rate, bioavailability, metabolism, clearance, and the like (see, e.g., Hidalgo-Aragones (1996) J. Steroid Biochem. Mol. Biol. 58:611-617; Groning (1996) Pharmazie 51:337-341; Fotherby (1996) Contraception 54:59-69; Johnson (1995) J. Pharm. Sci . 84:1144-1146; Rohatagi (1995) Pharmazie 50:610-613; Brophy (1983) Eur. J. Clin. Pharmacol . 24:103-108; latest Remington's, supra ). The state of the art allows the clinician to determine the dosage regimen for each individual patient, GR and/or MR modulator, and disease or condition being treated.

Single or multiple administrations of the compound formulations disclosed herein can be administered depending on the dosage and frequency as needed and tolerated by the patient. The formulation should provide a sufficient amount of active agent to effectively treat the disease state. Thus, in one embodiment, pharmaceutical formulations for oral administration of a compound disclosed herein are a daily total amount of between about 0.5 to about 30 mg per kilogram of body weight per day. In an alternative embodiment, a dose of about 1 mg to about 20 mg/kg body weight per patient per day is used. Lower doses may be used compared to oral administration into the bloodstream, into a body cavity, or into the lumen of an organ, especially when the drug is administered to an anatomically hidden site, such as the cerebrospinal fluid (CSF) space. Substantially higher doses may be used in topical administration. Actual methods of preparing formulations comprising the compounds disclosed herein for parenteral administration are known, or will be apparent, to those skilled in the art and are described in more detail in publications such as Remington's supra. See also Nieman, "Receptor Mediated Antisteroid Action", eds. Agarwal et al., De Gruyter, New York (1987).

The compounds described herein may be used in combination with each other, with other active agents known to be useful in modulating glucocorticoid receptors, or with adjuvants that may not be effective alone, but may contribute to the efficacy of the active agents.

In some embodiments, co-administration comprises administering one active agent within 0.5, 1, 2, 4, 6, 8, 10, 12, 16, 20, or 24 hours of a second active agent. Co-administration includes simultaneous, approximately simultaneous (eg, within about 1, 5, 10, 15, 20, or 30 minutes of each other), or sequential administration of the two active agents in any order. In some embodiments, co-administration can be accomplished by co-formulation, ie, the preparation of a single pharmaceutical composition comprising both active agents. In some embodiments, the active agents may be formulated separately. In some embodiments, active and/or adjuvants may be linked or coupled to each other.

After a pharmaceutical composition comprising a compound disclosed herein has been formulated in one or more acceptable carriers, it can be placed in a suitable container and labeled for treatment of an indicated condition. For administration of a compound of formula I, such labeling includes, for example, instructions as to the amount, frequency and method of administration.

In some embodiments, the compositions disclosed herein are useful for parenteral administration, such as intravenous (IV) administration or administration into a body cavity or lumen of an organ. Formulations for administration generally comprise solutions of the compositions disclosed herein dissolved in one or more pharmaceutically acceptable carriers. Acceptable vehicles and solvents that may be employed include water and Ringer's solutions, isotonic sodium chloride. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid may likewise be used in the preparation of injectables. Such solutions are sterile and generally free of inappropriate substances. These formulations can be sterilized by conventional, well-known sterilization techniques. The formulations may contain pharmaceutically acceptable auxiliary substances such as pH adjusting and buffering agents, tonicity adjusting agents such as sodium acetate, sodium chloride, potassium chloride, calcium chloride, sodium lactate and the like as required to approximate physiological conditions. The concentrations of the components in such formulations can vary widely and are selected primarily based on fluid volumes, viscosities, body weight, and the like, depending on the particular mode of administration chosen and the needs of the patient. For IV administration, the formulation may be a sterile injectable preparation, such as a sterile injectable aqueous or oleaginous suspension. This suspension may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be in the form of a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example a solution in 1,3-butanediol.

In some embodiments, formulations of the compositions disclosed herein can be delivered using liposomes that fuse with cell membranes or are endocytosed, i.e., by using attachment to liposomes, or directly to oligonucleotides Ligands for acids that bind to cell surface membrane protein receptors, leading to endocytosis. By using liposomes, especially when the liposome surface bears ligands specific to the target cell, or is otherwise preferentially directed to a particular organ, one can aim to deliver the compositions disclosed herein to the target in vivo. cell. (See e.g. Al-Muhammed, J. Microencapsul. 13:293-306, 1996; Chonn, Curr. Opin. Biotechnol. 6:698-708, 1995; Ostro, Am. J. Hosp. Pharm. 46:1576-1587 , 1989). V. method

In some embodiments, there is provided a method of treating a disorder or disorder in a subject comprising administering to a human a therapeutically effective amount of a compound disclosed herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as disclosed herein.

In some embodiments, there is provided a method of inhibiting KRAS G12C activity in a cell comprising contacting a cell in need of inhibition of KRAS G12C activity with an effective amount of a compound disclosed herein or a pharmaceutically acceptable salt thereof.

In some embodiments, there is provided a method of inhibiting KRAS G12C activity in a cell comprising contacting a cell in need of inhibition of KRAS G12C activity with a pharmaceutical composition disclosed herein.

In some embodiments, a method of treating KRAS G12C-related cancer is provided, comprising administering a therapeutically effective amount of a compound disclosed herein, or a pharmaceutically acceptable salt thereof, to a patient in need thereof.

In some embodiments, methods of treating KRAS G12C-related cancers are provided, comprising administering a pharmaceutical composition disclosed herein to a patient in need thereof.

In some embodiments, there is provided a method of treating an individual with a cancer characterized by the presence of a KRAS G12C mutation, the method comprising administering to the human a therapeutically effective amount of any of Formula (I) or Formula (II) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as disclosed herein.

In some embodiments, methods are provided for the manufacture of a medicament comprising a compound of formula (I) or formula (II), or a pharmaceutical medicament thereof, for treating an individual with cancer characterized by the presence of a KRAS G12C mutation. acceptable salts, or pharmaceutical compositions.

In some embodiments, use of a compound of formula (I) or formula (II), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as disclosed herein for the manufacture of a medicament for treating a human suffering from cancer is provided , the cancer is characterized by the presence of a KRAS G12C mutation.

In some embodiments, there is provided a compound of Formula (I) or Formula (II), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as disclosed herein, for use in the treatment of an individual having a cancer characterized by in the presence of the KRAS G12C mutation.

In some embodiments, there is provided a method of treating cancer in a patient in need thereof, the method comprising (a) determining that the cancer is associated with a KRAS G12C mutation ( e.g. , a KRAS G12C-associated cancer); and (b) administering to the patient a therapeutically effective Quantities of compounds disclosed herein.

In some embodiments, there are provided methods of treating cancer in a patient in need thereof comprising (a) determining that the cancer is associated with a KRas G12C mutation ( e.g. , a KRAS G12C-associated cancer); and (b) administering to the patient a Pharmaceutical composition.

In some embodiments, the cancer is heart: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma, and teratoma; lung: bronchial carcinoma (squamous cell, Undifferentiated small cell, undifferentiated large cell, adenocarcinoma), alveolar (bronchiole) carcinoma, bronchial adenoma, sarcoma, lymphoma, enchondroma hamartoma, mesothelioma; gastrointestinal tract: esophagus (squamous cell carcinoma, Adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumor, VIP tumor), Small bowel (adenocarcinoma, lymphoma, carcinoid, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large bowel (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma); urogenital tract: kidney (adenocarcinoma, Wilm's tumor (Wilms tumor), lymphoma, leukemia), bladder and urethra (squamous cell Carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testis (seminoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, stromal cell carcinoma, fibroma, fibroadenoma, adenomatous tumor, lipoma); liver: hepatocellular carcinoma (liver cell carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma; biliary tract: gallbladder carcinoma, amputation Abdominal cancer, cholangiocarcinoma; bone: osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulocyte sarcoma), multiple myeloma malignant giant cell tumor, chordoma, osteochondroma (exostoses of osteochondral), benign chondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma, and giant cell tumor; nervous system: skull (bone tumor, hemangioma, granuloma, xanthoma, osteitis deformans), meninges (meningioma, meningiosarcoma, gliomatosis), brain (astrocytoma, medulloblastoma, glioma, ependyma multiforme glioblastoma, oligodendroglioma, schwannoma, retinoblastoma, congenital tumor), neurofibroma of the spinal cord, meningioma, glial tumor, sarcoma); gynecology: uterine (endometrial carcinoma (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma), granulosa cell-ootheca cell tumor, Sertoli-stromal cell tumor (Sertoli- Leydig cell tumor), dysgerminoma, malignant teratoma), vulva (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, botryoid Sarcoma (embryonal rhabdomyosarcoma), fallopian tube (carcinoma); hematology: blood (myeloid leukemia (acute and chronic), acute lymphoblastic leukemia, chronic lymphocytic leukemia, myeloproliferative disorders, multiple myeloma, bone marrow Dysplastic syndrome), Hodgkin's disease, non-Hodgkin's lymphoma (malignant lymphoma); skin: malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, nevus dysplastic nevus , lipoma, hemangioma, dermatofibroma, keloid, psoriasis; or adrenal gland: neuroblastoma.

In some embodiments, the cancer is non-small cell lung cancer, small cell lung cancer, colorectal cancer, rectal cancer, or pancreatic cancer.

In certain embodiments, treatment may be administered after one or more symptoms have occurred. In other embodiments, treatment may be administered in the absence of symptoms. For example, treatment can be administered to a predisposed individual prior to the onset of symptoms (eg, in view of a history of symptoms and/or in view of genetic or other predisposition factors). Treatment can also be continued after symptoms have subsided, for example, to prevent or delay their recurrence.

The compound of formula (I) or formula (II), or a pharmaceutically acceptable salt thereof, can be an inhibitor of KRAS G12C. For example, compounds disclosed herein may have an inhibition constant (Ki) of less than about 50 µM, or less than about 40, 30, 20, 10, 9, 8, 7, 6, 5, 4, 3, 2, or less than About 1 µM. Compounds disclosed herein may have inhibition constants (Ki) of less than about 1,000 nM, or less than about 900, 800, 700, 600, 500, 400, 300, 200, 100, 90, 80, 70, 60, 50, 40 , 30, 20, 10, 9, 8, 7, 6, 5, 4, 3, 2, or less than about 1 nM. Compounds disclosed herein may have an inhibition constant (Ki) of less than about 1 nM, or less than about 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, or less than about 0.1 nM.

Compounds of formula (I) or formula (II), or pharmaceutically acceptable salts thereof, may be selective inhibitors of KRAS G12C. For example, the KRAS G12C inhibition constant (IC50) of the compounds disclosed herein may be at least 2-fold lower, or at least 3, 4, 5, 6, 7 times lower than the inhibition constant of one or more of KRAS wild-type, or NRAS, or HRAS , 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90 or 100 times. The KRAS G12C inhibition constant (Ki) of the compounds disclosed herein may also be at least 100-fold lower, or at least 200, 300, 400, 500, 600, 700, 800, 900, 1000, or 10,000 times. A. Combination Therapies for Cancer

The compounds disclosed herein, or salts thereof, can be used in therapy alone or in combination with other agents. For example, the second agent of the pharmaceutical combination formulation or dosing regimen may have complementary activities to the compounds disclosed herein such that they do not deleteriously affect each other. The compounds can be administered together in a single pharmaceutical composition or separately. In one embodiment, the compounds or pharmaceutically acceptable salts may be co-administered with cytotoxic agents for the treatment of proliferative diseases and cancers.

The term "co-administration" refers to the simultaneous administration, or any manner of separate sequential administration, of a compound disclosed herein, or a salt thereof, and one or more further active pharmaceutical ingredients, including cytotoxic agents and radiation therapy. If not administered simultaneously, the compounds are administered in close proximity to each other. Furthermore, it does not matter whether the compounds are administered in the same dosage form, eg, one compound may be administered topically and the other may be administered orally.

Such additional agents may be administered separately from the compositions containing the compounds of this invention as part of a multiple dosage regimen. Alternatively, these agents may be part of a single dosage form, mixed together with the compounds disclosed herein in a single composition. If administered as part of a multiple dose regimen, the two active agents may be provided simultaneously, sequentially, or within a period of each other, typically within five hours of each other.

As used herein, the term "combination/combined" and related terms refer to simultaneous or sequential administration of the therapeutic agents according to the embodiments herein. For example, a compound disclosed herein can be administered with another therapeutic agent simultaneously or sequentially in separate unit dosage forms or co-administered in a single unit dosage form. Accordingly, embodiments of the invention provide single unit dosage forms comprising a compound of formula I , an additional therapeutic agent, and a pharmaceutically acceptable carrier, adjuvant, or vehicle.

The amount of a compound of the invention and additional therapeutic agent (in a composition comprising an additional therapeutic agent as described above) that can be combined with a carrier material to produce a single dosage form will vary depending upon the host treated and the particular mode of administration. In certain embodiments, compositions disclosed herein are formulated such that doses of the invention of between 0.01-100 mg/kg body weight/day can be administered.

Generally, any agent that is active against the disease or condition being treated can be co-administered. Examples of such agents can be found in Cancer Principles and Practice of Oncology, V.T. Devita and S. Hellman (eds.), 6th edition (February 15, 2001), Lippincott Williams & Wilkins Publishers. Those skilled in the art are able to discern which combinations of agents will be useful based on the particular properties of the drugs and the disease involved.

In one embodiment, the method of treatment comprises co-administering a compound disclosed herein, or a pharmaceutically acceptable salt thereof, and at least one cytotoxic agent. The term "cytotoxic agent" as used herein refers to a substance that inhibits or prevents cellular function and/or causes cell death or destruction. Cytotoxic agents include, but are not limited to, radioisotopes (e.g., radioisotopes of At ²¹¹ , I ¹³¹ , I ¹²⁵ , Y ⁹⁰ , Re ¹⁸⁶ , Re ¹⁸⁸ , Sm ¹⁵³ , Bi ²¹² , P ³² , Pb ²¹² , and Lu); chemotherapy growth inhibitors; enzymes and fragments thereof such as nuclease; and toxins such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including fragments and/or variants thereof.

Exemplary cytotoxic agents may be selected from antimicrotubule agents, platinum coordination complexes, alkylating agents, antibiotic agents, topoisomerase II inhibitors, antimetabolites, topoisomerase I inhibitors, hormones, and hormone-like agents Drugs, signal transduction pathway inhibitors, non-receptor tyrosine kinase angiogenesis inhibitors, immunotherapeutic agents, pro-apoptotic agents, inhibitors of LDH-A; inhibitors of fatty acid biosynthesis; cell cycle signal transduction inhibition agents; HDAC inhibitors, proteasome inhibitors; and inhibitors of cancer metabolism.

"Chemotherapeutic agents" include compounds useful in the treatment of cancer. Examples of chemotherapeutic agents include erlotinib ( ^TARCEVA® , Genentech/OSI Pharm.), bortezomib ( ^VELCADE® , Millennium Pharm.), disulfiram, epigalloc Catechin gallate, salinosporamide A, carfilzomib, 17-AAG (geldanamycin), radicicol, Lactate dehydrogenase A (LDH-A), fulvestrant (FASLODEX ^® , AstraZeneca), sunitinib (SUTENT ^® , Pfizer/Sugen), letrozole (FEMARA ^® , Novartis), imatinib mesylate (GLEEVEC ^® , Novartis), finasunate (VATALANIB ^® , Novartis), oxaliplatin (ELOXATIN ^® , Sanofi), 5 -FU (5-fluorouracil), leucovorin, rapamycin (Sirolimus, RAPAMUNE ^® , Wyeth), lapatinib (TYKERB ^® , GSK572016, Glaxo Smith Kline), lonafamib (SCH 66336), sorafenib (NEXAVAR ^® , Bayer Labs), gefitinib (IRESSA ^® , AstraZeneca), AG1478, alkylating agent , such as thiotepa and ^CYTOXAN® cyclophosphamide; alkyl sulfonates, such as busulfan, improsulfan and piposulfan; aziridine, Such as benzodopa (benzodopa), carboquone (carboquone), meturedopa (meturedopa) and uretepa (uredopa); ethyleneimine and methylmelamine, including hexamethylmelamine (altretamine), Triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide and trimethylolmelamine; acetogenin (especially bullatacin and bullatacin) bullatacinone); camptothecin (including topotecan and irinotecan); bryostatin; callystatin; CC-1065 (including its adozelesin, carzelesin, and bizelesin synthetic analogues); cryptophycin (especially nodocin 1 and nodocin 8); adrenal cortex Steroids (including prednisone and prednisolone); cyproterone acetate; 5α-reductase, which includes finasteride and dutasteride); vorino vorinostat; romidepsin; panobinostat; valproic acid; mocetinostat; dolastatin; aldesleukin; Talc; duocarmycin (including synthetic analogues KW-2189 and CB1-TM1); eleutherobin; pancratistatin; sarcodictyn; spongostatin (spongistatin); nitrogen mustards such as chlorambucil, chlormaphazine, chlorophosphamide, estramustine, ifosfamide, Mechlorethamine, Dichloromethyldiethylamine Oxide Hydrochloride, Melphalan, Novembichin, Cholesterol Phenesterine, Turpentine Mustard (prednimustine), trofosfamide, uracil mustard; nitrosoureas such as carmustine, chlorozotocin, fotemustine , lomustine (lomustine), nimustine (nimustine) and ramustine (ranimnustine); antibiotics, such as enediyne antibiotics (such as calicheamicin (calicheamicin), especially calicheamicin γ1I and calicheamicin ω1I ( Angew Chem. Intl. Ed. Engl. 1994 33:183-186); dynemicins, including dynemicin A; bisphosphonates, such as clodronate (clodronate; esperamicin; and neocarzinostatin (neocarzinostatin) chromophore and related chromoproteins enediyne antibiotic chromophore), aclacinomysin (aclacinomysin), actinomycin ( actinomycin), antramycin (authramycin), azaserine (azaserine), bleomycin (bleomycin), actinomycin C (cactinomycin), carabicin (carabicin), carmine Carzinophilin, chromomycinis, dactinomycin, daunomycin, detorubicin, 6-diazo-5-oxo-L-n Leucine, ADRIAMYCIN ^® (doxorubicin), N-morpholino doxorubicin, cyano(N-morpholino)-doxorubicin, 2-(N-pyrrolyl)-doxorubicin and deoxy Adriamycin, epirubicin, esorubicin, idarubicin, marcellomycin, mitomycin (such as mitomycin C), mycophenolic acid, nogalamycin, olivomycin, peplomycin, porfiromycin, puromycin, triiron doxorubicin (quelamycin), rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin , zorubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs such as denopterin, methotrexate, terra Pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs, Such as ancitabine, azacitidine, 6-azuridine, carmofur, cytarabine, dideoxyuridine, doxifluridine ), enocitabine, floxuridine; mepitiostane, testolactone; anti-adrenergics such as aminoglutethimide, mitotane, trilostane; folic acid supplements such as frolinic acid ); aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; betrabucil ; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elfomithine; elliptinium acetate; epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids such as maytansinoids maytansine and ansamitocin; mitoguazone; mitoxantrone; mopidamnol; nitraerine; pentostatin; Phenomet; pirarubicin; losoxantrone; podophyllinic acid; 2-ethylhydrazine; procarbazine; PSK ^® polysaccharide complex (JHS Natural Products, Eugene, Oreg.); Razoxane; Rhizoxin; Sizofuran; Spirogermanium; Tenuazonic acid ; triaziquone; 2,2',2"-trichlorotriethylamine; trichothecene (especially T-2 toxin, verracurin A, bacitracin A (roridin A and anguidine); urethan; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C");cyclophosphamide;thiotepa; taxoid, For example, TAXOL (paclitaxel; Bristol-Myers Squibb Oncology, Princeton, NJ), ABRAXANE ^® (Cremophor-free), albumin-engineered nanoparticle formulations of paclitaxel ( American Pharmaceutical Partners, Schaumberg, Ill.) and TAXOTERE ^® (docetaxel, doxetaxel; Sanofi-Aventis); chlorambucil (chloranmbucil); GEMZAR ^® (gemcitabine); 6- Thioguanine; Mercaptopurine; Methotrexate; Platinum analogues, such as cisplatin and carboplatin; Vinblastine; Etoposide (VP-16); Ifosfamide; Mitoxantrone; vincristine; NAVELBINE ^® (vinorelbine); novantrone; teniposide; edatrexate; daunomycin; Capecitabine (XELODA ^® ); ibandronate; CPT-11; topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); Retinoids, such as retinoic acid; and pharmaceutically acceptable salts, acids and derivatives of any of the above.

Chemotherapeutic agents also include: (i) Antihormonal agents used to modulate or inhibit the effects of hormones on tumors, such as antiestrogens and selective estrogen receptor modulators (SERMs), which include, for example, tamoxifen ) (including NOLVADEX ^® ; tamoxifen citrate), raloxifene, droloxifene, iodoxyfene, 4-hydroxytamoxifen, travoxifen ( trioxifene), keoxifene, LY117018, onapristone, and FARESTON ^® (toremifine citrate); (ii) aromatase inhibitors that inhibit aromatase, which regulate Estrogen production in the adrenal glands such as 4(5)-imidazole, amine glutethimide, MEGASE ^® (megestrol acetate), AROMASIN ^® (exemestane; Pfizer), formestane (formestanie), fadrozole, RIVISOR ^® (vorozole), FEMARA ^® (letrozole; Novartis), and ARIMIDEX ^® (anastrozole; AstraZeneca); (iii) antiandrogenic Hormones such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; buserelin, Tripterelin, medroxyprogesterone acetate, diethylstilbestrol, premarin, fluoxymesterone, all-trans retinoic acid, fenretinide, and troxastat (troxacitabine) (1,3-dioxolane nucleoside cytosine analog); (iv) protein kinase inhibitors; (v) lipid kinase inhibitors; (vi) antisense oligonucleotides, especially Inhibition of expression of genes involved in signaling pathways of abnormal cell proliferation, such as PKC-α, Raf and H-Ras; (vii) ribonucleases, such as VEGF expression inhibitors (eg ANGIOZYME ^® ) and HER2 expression inhibitors; ( viii) vaccines, such as gene therapy vaccines, such as ^ALLOVECTIN® , ^LEUVECTIN® , and ^VAXID® ; ^PROLEUKIN® ; rIL-2; topoisomerase 1 inhibitors, such as ^LURTOTECAN® ; ^ABARELIX® rmRH; and (ix) any of the above The pharmaceutically acceptable salts, acids and derivatives thereof.

Chemotherapeutic agents also include antibodies such as alemtuzumab (Campath), bevacizumab (AVASTIN®, Genentech); cetuximab (ERBITUX®, Imclone); panitumumab (VECTIBIX®, Amgen), rituximab (RITUXAN®, Genentech/Biogen Idec), pertuzumab (OMNITARG®, 2C4, Genentech), trastuzumab (HERCEPTIN®, Genentech), tositumomab ( Bexxar, Corixia) and an antibody-drug conjugate, gemtuzumab ozogamicin (MYLOTARG®, Wyeth). Other humanized monoclonal antibodies that have therapeutic potential as agents in combination with the compounds disclosed herein include: apolizumab, aselizumab, atlizumab, Pilizumab, bivatuzumab-maytansine, cantuzumab-maytansine, cedelizumab, certolizumab- Polyethylene glycol (certolizumab pegol), cidfusituzumab, cidtuzumab, daclizumab, eculizumab, efalizumab ), epratuzumab, erlizumab, felvizumab, fontolizumab, gemtuzumab-calicheamicin, Inotuzumab-inotuzumab ozogamicin, ipilimumab, labetuzumab, lintuzumab, matuzumab, Mepolizumab, motavizumab, motovizumab, natalizumab, nimotuzumab, navelizumab (nolovizumab), numavizumab, ocrelizumab, omalizumab, palivizumab, pascolizumab, Pafo Pecfusituzumab, pectuzumab, pexelizumab, ralivizumab, ranibizumab, reslivizumab , Reslizumab, Resyvizumab, Rovelizumab, Ruplizumab, Sibrotuzumab, Siplizumab, Sontuzumab, tacatuzumab tetraxetan, tadocizumab, talizumab, tefibazumab, Tocilizumab, toralizumab, tucotuzumab celmoleukin, tucusituzumab, umavizumab, umavizumab Monoclonal antibody (urtoxazumab), ustekinumab (ustekinumab), visilizumab (visilizumab) and anti-interleukin-12 (ABT-874/J695, Wyeth Research and Abbott Laboratories), which is a genetically modified Recombinant obligate human sequence full-length IgG ₁ λ antibody that recognizes interleukin-12 p40 protein.

Chemotherapeutic agents also include "EGFR inhibitors," which refer to compounds that bind to or otherwise directly interact with EGFR and prevent or reduce its signaling activity, and are alternatively referred to as "EGFR antagonists." Examples of such agents include antibodies and small molecules that bind to EGFR. Examples of antibodies that bind to EGFR include MAb 579 (ATCC CRL HB 8506), MAb 455 (ATCC CRL HB8507), MAb 225 (ATCC CRL 8508), MAb 528 (ATCC CRL 8509) (see U.S. Patent No. 4,943,533, Mendelsohn et al ) and variants thereof such as chimeric 225 (C225 or cetuximab; ^ERBUTIX® ) and reshaped human 225 (H225) (see WO 96/40210, Imclone Systems Inc.); IMC-11F8, A fully human EGFR targeting antibody (Imclone); an antibody that binds type II mutant EGFR (US Patent No. 5,212,290); a humanized and chimeric antibody that binds EGFR as described in US Patent No. 5,891,996; and a human that binds EGFR Antibodies, such as ABX-EGF or panitumumab (see WO98/50433, Abgenix/Amgen); EMD 55900 (Stragliotto et al. , Eur. J. Cancer 32A:636-640 (1996)); EMD7200 (matuzumab anti), a humanized EGFR antibody against EGFR that competes with both EGF and TGF-α for EGFR binding (EMD/Merck); human EGFR antibody, HuMax-EGFR (GenMab); designated E1.1, E2.4 , E2.5, E6.2, E6.4, E2.11, E6.3, and E7.6.3 and the fully human antibodies described in US 6,235,883; MDX-447 (Medarex Inc); and mAb 806 or humanized mAb 806 (Johns et al. , J. Biol. Chem . 279(29):30375-30384 (2004)). Anti-EGFR antibodies can be conjugated with cytotoxic agents to generate immunoconjugates (see eg EP659,439A2, Merck Patent GmbH). EGFR antagonists include small molecules such as those described in U.S. Pat. 84, 5,770,599, 6,140,332, 5,866,572, 6,399,602, 6,344,459, 6,602,863, 6,391,874, 6,344,455, 5,760,041, 6,002,008, and 5,747,498, and compounds described in the following PCT publications: WO98/14451, WO98/50038, WO99/09016, and WO99/24037. Specific small molecule EGFR antagonists include OSI-774 (CP-358774, erlotinib, TARCEVA ^® Genentech/OSI Pharmaceuticals); PD 183805 (CI 1033, N-[4-[(3-chloro-4-fluorophenyl )amino]-7-[3-(4-morpholinyl)propoxy]-6-quinazolinyl]-2-acrylamide dihydrochloride, Pfizer Inc.); ZD1839, gefitinib IRESSA® (4-(3'-chloro-4'-fluoroaniline)-7-methoxy-6-(3-N-morpholinopropoxy)quinazoline, AstraZeneca); ZM 105180 ((6-amino-4-(3-methylphenyl-amino)-quinazoline, Zeneca); BIBX-1382 (N8-(3-chloro-4-fluoro-phenyl)-N2-( 1-methyl-piperidin-4-yl)-pyrimido[5,4-d]pyrimidine-2,8-diamine, Boehringer Ingelheim); PKI-166 ((R)-4-[4-[( 1-phenylethyl)amino]-1H-pyrrolo[2,3-d]pyrimidin-6-yl]-phenol); (R)-6-(4-hydroxyphenyl)-4-[( 1-phenylethyl)amino]-7H-pyrrolo[2,3-d]pyrimidine); CL-387785 (N-[4-[(3-bromophenyl)amino]-6-quinazole Linyl]-2-butynamide); EKB-569 (N-[4-[(3-chloro-4-fluorophenyl)amino]-3-cyano-7-ethoxy-6- Quinolinyl]-4-(dimethylamino)-2-butyramide) (Wyeth); AG1478 (Pfizer); AG1571 (SU 5271; Pfizer); dual EGFR/HER2 tyrosine kinase inhibitors such as Lapatinib (TYKERB®, GSK572016 or N-[3-chloro-4-[(3-fluorophenyl)methoxy]phenyl]-6[5[[[2-methylsulfonyl)ethyl base] amino] methyl] -2-furyl] -4-quinazolinamine).

Chemotherapeutic agents also include "tyrosine kinase inhibitors", which include the EGFR targeting drugs mentioned in the previous paragraph; small molecule HER2 tyrosine kinase inhibitors, such as TAK165 available from Takeda; CP-724,714, a Oral selective inhibitors of ErbB2 receptor tyrosine kinase (Pfizer and OSI); dual HER inhibitors, such as EKB-569 (commercially available from Wyeth), which bind preferentially to EGFR but inhibit HER2-overexpressing cells as well as EGFR-overexpressing cells Both; lapatinib (GSK572016; available from Glaxo-SmithKline), an oral HER2 and EGFR tyrosine kinase inhibitor; PKI-166 (available from Novartis); pan-HER inhibitors such as canertinib ( canertinib, CI-1033; Pharmacia); Raf-1 inhibitors, such as the antisense reagent ISIS-5132 available from ISIS Pharmaceuticals, which inhibits Raf-1 signaling; non-HER-targeting TK inhibitors, such as ISIS mesylate Matinib (GLEEVEC®, available from Glaxo SmithKline); multi-targeted tyrosine kinase inhibitors, such as sunitinib (SUTENT®, available from Pfizer); VEGF receptor tyrosine kinase inhibitors, Such as vatalanib (PTK787/ZK222584, available from Novartis/Schering AG); MAPK extracellular regulated kinase I inhibitor CI-1040 (available from Pharmacia); quinazolines, such as PD 153035, 4-( 3-chloroanilino)quinazoline; pyridopyrimidine; pyrimidopyrimidine; pyrrolopyrimidine, such as CGP 59326, CGP 60261 and CGP 62706; pyrazolopyrimidine, 4-(phenylamino)-7H-pyrrolo [2,3-d]pyrimidine; curcumin (diaferuloylmethane, 4,5-bis(4-fluoroanilino)phthalimide); tyrosine phosphorylation inhibitor containing nitrothiophene moiety PD-0183805 (Warner-Lamber); antisense molecules ( such as molecules that bind to HER-encoding nucleic acids); quinoxalines (US Patent No. 5,804,396); tyrosine phosphorylation inhibitors (US Patent No. 5,804,396) ; ZD6474 (Astra Zeneca); PTK-787 (Novartis/Schering AG); pan-HER inhibitors such as CI-1033 (Pfizer); Affinitac (ISIS 3521; Isis/Lilly); Matinib (GLEEVEC®); PKI 166 (Novartis); GW2016 (Glaxo SmithKline); CI-1033 (Pfizer); EKB-569 (Wyeth); Sematinib (Pfizer); ZD6474 (AstraZeneca); 787 (Novartis/ Schering AG); INC-1C11 (Imclone), rapamycin (sirolimus, RAPAMUNE®); or a substance as described in any of the following patent publications: U.S. Patent No. 5,804,396 No., WO 1999/09016 (American Cyanamid), WO 1998/43960 (American Cyanamid), WO 1997/38983 (Warner Lambert), WO 1999/06378 (Warner Lambert), WO 1999/06396 (Warner Lambert), WO 1996/ 30347 (Pfizer, Inc), WO 1996/33978 (Zeneca), WO 1996/3397 (Zeneca), and WO 1996/33980 (Zeneca).

Chemotherapeutic agents also include dexamethasone, interferon, colchicine, metoprine, cyclosporine, amphotericin, metronidazole ), alemtuzumab, alitretinoin, allopurinol, amifostine, arsenic trioxide, asparaginase, BCG live, bevacizimab, bevacizimab Bexarotene, cladribine, clofarabine, darbepoetin alfa, denileukin, dexrazoxane, alfa Epoetin alfa, elotinib, filgrastim, histrelin acetate, ibritumomab, interferon alpha-2a, interferon Alpha-2b, lenalidomide, levamisole, mesna, methoxsalen, nandrolone, nelarabine, normomab (nofetumomab), oprelvekin, palifermin, pamidronate, pegademase, pegaspargase, pegfilgrastim (pegfilgrastim), pemetrexed disodium, plicamycin, porfimer sodium, quinacrine, rasburicase, sargrax Sargramostim, temozolomide, VM-26, 6-TG, toremifene, tretinoin, all-trans retinoic acid (ATRA), valrubicin , zoledronate, zoledronic acid and pharmaceutically acceptable salts thereof.

Chemotherapeutic agents also include hydrocortisone, hydrocortisone acetate, cortisone acetate, tixocortol pivalate, triamcinolone acetonide ), triamcinolone alcohol, mometasone, amcinonide, budesonide, desonide, fluocinonide, fluocinolone acetonide, betamethasone, betamethasone sodium phosphate, dexamethasone, dexamethasone sodium phosphate, fluocortolone, hydrocortisone -17-butyrate, hydrocortisone-17-valerate, alclometasone dipropionate, betamethasone valerate, betamethasone dipropionate, Prednicarbate, clobetasone-17-butyrate, clobetasol-17-propionate, fluocorolone caproate ( fluorocortolone caproate), fluorocortolone pivalate, and fluprednidene acetate; immunoselective anti-inflammatory peptides (ImSAID) such as phenylalanine-glutamine-glycine (FEG) and Its D-isomeric form (feG) (IMULAN Bio Therapeutics, LLC); antirheumatic drugs such as azathioprine, ciclosporin (cyclosporin A), D-penicillamine , gold salt, hydroxychloroquine, leflunomideminocycline, sulfasalazine, tumor necrosis factor alpha (TNFα) blockers such as etanercept (Enbrel) , infliximab (infliximab; Remicade), adalimumab (adalimumab; Humira), certolizumab-pegol (certolizumab pegol; Cimzia), golimumab (golimumab; Simponi), mediated Interleukin 1 (IL-1) blockers such as anakinra (Kineret), T cell co-stimulation blockers such as abatacept (Orencia), interleukin 6 (IL-6) blockers Blockers such as tocilizumab (ACTEMERA®); Interleukin 13 (IL-13) blockers such as lebrikizumab; Interferon alpha (IFN) blockers such as Rontalizumab ); β7 integrin blockers such as rhuMAb β7; IgE pathway blockers such as anti-M1 prime; secretion homotrimeric LTa3 and membrane bound heterotrimeric LTa1/β2 blockers such as anti-lymphotoxin alpha (LTa ); radioisotopes (for example, At ²¹¹ , I ¹³¹ , I ¹²⁵ , Y ⁹⁰ , Re ¹⁸⁶ , Re ¹⁸⁸ , Sm ¹⁵³ , Bi ²¹² , P ³² , Pb ²¹² and radioisotopes of Lu); various research agents such as thioplatinum, PS-341, phenylbutyrate, ET-18-OCH ₃ , or farnesyltransferase inhibitors (L-739749, L-744832); polyphenols such as quercetin, resveratrol ), piceatannol, epigallocatechin gallate, theaflavin, flavanol, procyanidin, betulinic acid and its derivatives autophagy inhibitors such as chloroquine; delta-9-tetrahydrocannabinol (dronabinol, MARINOL®); beta-lapachone; lapachol; colchicine; betulinic acid; acetylcamptothecin, scopolectin, and 9-aminocamptothecin; podophyllotoxin; fluridine (tegafur; UFTORAL®); bexarotene (TARGRETIN®); bisphosphonates such as clodronate (eg, BONEFOS® or OSTAC®), etidronate (etidronate; DIDROCAL®), NE -58095, ZOLEDRONATE/ZOLEDRONATE (ZOMETA®), ALENDRONATE (ALENDRONATE; FOSAMAX®), PAMIDRONATE (AREDIA®), TILUDRONATE (TILUDRONATE; SKELID®) , or risedronate (risedronate; ACTONEL®); and epidermal growth factor receptor (EGF-R); vaccines such as THERATOPE® vaccine; perifosine, a COX-2 inhibitor ( eg , Celex celecoxib or etoricoxib), proteosome inhibitors ( eg PS341); CCI-779; tipifarnib (R11577); sorafenib, ABT510; Bcl-2 inhibitors such as Oblimersen sodium (GENASENSE®); pixantrone; farnesyltransferase inhibitors such as lonafarnib (SCH 6636, SARASAR ^™ ); and any of the above above acceptable salts, acids or derivatives; and a combination of two or more of the above such as CHOP, which is an abbreviation for combination therapy of cyclophosphamide, doxorubicin, vincristine, and presulcin and FOLFOX, which is an abbreviation for the treatment regimen with oxaliplatin (ELOXATIN ^™ ) in combination with 5-FU and folinic acid.

Chemotherapeutic agents also include non-steroidal anti-inflammatory drugs that have analgesic, antipyretic and anti-inflammatory effects. NSAIDs include non-selective inhibitors of cyclooxygenases. Specific examples of NSAIDs include aspirin, propionic acid derivatives such as ibuprofen, fenoprofen, ketoprofen, flurbiprofen, oxaprozin ( oxaprozin and naproxen, acetic acid derivatives such as indomethacin, sulindac, etodolac, diclofenac, enolic acid derivatives such as piroxicam Piroxicam, meloxicam, tenoxicam, droxicam, lornoxicam and isoxicam, fenamic acid derivatives Such as mefenamic acid (mefenamic acid), meclofenamic acid (meclofenamic acid), flufenamic acid (flufenamic acid), tolfenamic acid (tolfenamic acid), and COX-2 inhibitors such as celecoxib (celecoxib) etoricoxib, lumiracoxib, parecoxib, rofecoxib, rofecoxib, and valdecoxib. NSAIDs may be needed for symptom relief in conditions such as: rheumatoid arthritis, osteoarthritis, inflammatory joint disease, ankylosing spondylitis, psoriatic arthritis, Reiter's syndrome, acute gout, dysmenorrhea, metastatic Bone pain, headache and migraine, postoperative pain, mild to moderate pain due to inflammation and tissue damage, fever, intestinal obstruction, and renal colic.

In certain embodiments, chemotherapeutic agents include, but are not limited to, doxorubicin, dexamethasone, vincristine, cyclophosphamide, fluorouracil, topotecan, interferon, platinum derivatives, taxanes ( eg, paclitaxel, docetaxel), vinca alkaloids (eg, vinblastine), anthracyclines (eg, doxorubicin), epipodophyllotoxins (eg, etoposide), cisplatin, mTOR inhibition (eg, rapamycin), methotrexate, actinomycin D, dolastatin 10, colchicine, trimetrexate, chloraniline, cyclosporine, daunorubicin, tenipol Glycosides, amphotericins, alkylating agents (eg, chlorambucil), 5-fluorouracil, camptothecin, cisplatin, metronidazole, and imatinib mesylate. In other embodiments, compounds disclosed herein are administered in combination with biologics such as bevacizumab or panitumumab.

In certain embodiments, a compound disclosed herein, or a pharmaceutically acceptable composition thereof, is administered in combination with an antiproliferative or chemotherapeutic agent selected from any one or more of the following: Interleukin, alemtuzumab, alitretinoin, allopurinol, hexamethylmelamine, amifostine, anastrozole, arsenic trioxide, asparaginase, azacitidine, BCG live, bevacizumab, Fluorouracil, bexarotene, bleomycin, bortezomib, busulfan, dimethyltestosterone, capecitabine, camptothecin, carboplatin, carmustine, cetuximab, phenylbutyrate mustard, cladribine, clofarabine, cyclophosphamide, cytarabine, actinomycin D, darbepoetin alfa, daunorubicin, denileukin, dexrazoxane, docetaxel, multi Ruubicin (neutral), doxorubicin hydrochloride, drotandrosterone propionate, epirubicin, epoetin alfa, erlotinib, estramustine, etoposide phosphate, etoposide Poside, exemestane, filgrastim, floxuridine, fludarabine, fulvestrant, gefitinib, gemcitabine, gemtuzumab, ralin acetate, acetate group Amurelin, hydroxyurea, icomomab, idarubicin, ifosfamide, imatinib mesylate, interferon α-2a, interferon α-2b, irinotecan, lenalidomide , letrozole, leucovorin, leuprolide acetate, levamisole, lomustine, megestrol, melphalan, mercaptopurine, 6-MP, mesna, methotrexate, methoxyxa Lin, Mitomycin C, Mitotane, Mitoxantrone, Nandrolone, Nelarabine, Normomab, Opreinterleukin, Oxaliplatin, Paclitaxel, Palifermin, Pamidron salt, pegasin, pegaspargase, pefilgrastim, pemetrexed disodium, pentostatin, pipepobromide, plicamycin, porfimer sodium, procarbazine, albino Deping, rasburicase, rituximab, sargragrastim, sorafenib, streptozocin, sunitinib maleate, talc, tamoxifen, temozolomide, teniposide, VM-26, testrolactone, thioguanine, 6-TG, thiotepa, topotecan, toremifene, tositumomab, trastuzumab, tretinoin, ATRA, uracil nitrogen mustard, valrubicin, vinblastine, vincristine, vinorelbine, zoledronate, or zoledronic acid.

Chemotherapeutic agents also include those for Alzheimer's disease such as donepezil hydrochloride and rivastigmine; those for Parkinson's disease such as levodopa (L-DOPA)/Carbon Carbidopa, entacapone, ropinrole, pramipexole, bromocriptine, pergolide, trihexafendi, and Vajra Amines (amantadine); agents for the treatment of multiple sclerosis (MS) such as beta interferon (for example, Avonex ^® and Rebif ^® ), glatiramer acetate, and mitoxantrone; agents for the treatment of asthma such as albuterol ( albuterol) and montelukast sodium; drugs for the treatment of schizophrenia such as zyprexa, risperdal, seroquel, and haloperidol; Anti-inflammatory agents such as corticosteroids, TNF blockers, IL-1RA, azathioprine, cyclophosphamide, and sulfasalazine; immunomodulators and immunosuppressants such as cyclosporine, tacrolimus, rapamycin mycophenolate mofetil, interferon, corticosteroids, cyclophosphamide, azathioprine, and sulfasalazine; neurotrophic factors such as acetylcholinesterase inhibitors, MAO inhibitors, interferons, anticonvulsants agents, ion channel blockers, riluzole, and anti-Parkinson's disease agents; agents for the treatment of cardiovascular diseases such as beta-blockers, ACE inhibitors, diuretics, nitrates, calcium channel blockers, and Statins; agents for the treatment of liver diseases such as corticosteroids, cholestyramine, interferon, and antiviral agents; agents for the treatment of blood disorders such as corticosteroids, anti-leukemic agents, and growth factors; and agents for the treatment of immunodeficiency disorders such as gamma globulin.

In addition, chemotherapeutic agents include pharmaceutically acceptable salts, acids or derivatives of any of the chemotherapeutic agents described herein, as well as combinations of two or more thereof. VI. Example synthesis program General program

流程II

Compounds of formula I can be prepared from commercially available reagents using the synthetic methods and reaction schemes herein, or using other reagents and conventional methods well known to those skilled in the art. For example, compounds of the invention can be prepared according to the general reactions in Schemes I and II, using conventional crosslinking chemistry: Scheme I Compounds of Formula (I)

Process II

(E1) In Scheme II, X is a halogen or pseudohalogen such as mesylate, triflate or the like. Example 1 : (S)-8-((3S,5R)-4- acryloyl -3,5- dimethylpiperazin -1- yl )-11-(4- chlorothien- 2- yl )- 3- Methoxy -10-( trifluoromethyl )-3,4- dihydro -2H,6H-[1,4] thiazeza [2,3,4-ij] quinazoline -6- Ketones (E1) .

(E1)

At 0°C, to (S)-11-(4-chlorothiophen-2-yl)-8-((3S,5R)-3,5-dimethylpiperazin-1-yl)-3-methanol Oxygen-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-6-one (0.095 mmol) in dichloromethane (1 mL), triethylamine (0.57 mmol) and acryloyl chloride (0.19 mmol) were added. The reaction mixture was stirred at room temperature for 2 hours. The solvent was removed in vacuo to afford a residue which was purified by preparative HPLC to afford the title compound as a yellow solid in 39% yield

m/z (ESI, +ve)= 599.0 [M+H]+; 1H NMR (400 MHz, DMSO-d6) δ 8.00 (s, 1H), 7.88 (s, 1H), 7.15 (s, 1H), 6.81 (dd, J = 16.0, 8.0 Hz, 1H), 6.19 (dd, J = 16.0, 4.0 Hz, 1H), 5.74 (dd, J = 8.0, 4.0 Hz, 1H), 4.66 - 4.45 (m, 3H) , 4.05 - 3.94 (m, 2H), 3.89 - 3.81 (m, 1H), 3.32 (s, 5H), 3.28 - 3.24 (m, 2H), 3.17 (s, 1H), 1.39 (s, 6H). Step 1: (3-Chlorothiophen-2-yl)triisopropylsilane

To a solution of 3-chlorothiophene (73.4 mmol) in THF (50 mL) was added LDA (73.4 mmol) at -78°C and after one hour at -78°C triisopropylchlorosilane was added. The mixture was stirred at room temperature for 16 hours, quenched with saturated aqueous ammonium chloride and extracted with ethyl acetate. The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography to afford the title compound in 90% yield. Step 2: (3-Chloro-5-iodothiophen-2-yl)triisopropylsilane

To a solution of (3-chlorothien-2-yl)triisopropylsilane (8.4 mmol) in THF (20 mL) was added LDA (10.08 mol) at -78°C. The reaction mixture was stirred at -78°C for 2 hours. A solution of iodine (10.92 mmol) in THF (5 mL) was added and the reaction mixture was stirred at room temperature for 16 hours. The reaction was diluted with water and extracted three times with ethyl acetate. The combined organic layers were washed three _times with saturated aqueous _Na2S2O3 , dried over sodium sulfate and _concentrated to provide a residue which was purified by silica gel chromatography to provide the title compound as a colorless oil in 91% yield. Step 3: Methyl 2-amino-4-(4-chloro-5-(triisopropylsilyl)thiophen-2-yl)-5-(trifluoromethyl)benzoate

2-Amino-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-(trifluoromethyl)benzoic acid Methyl ester (0.14 mol), (3-chloro-5-iodothiophen-2-yl)triisopropylsilane (0.11 mol), Pd(dppf)Cl ₂ (24.5 mmol) and K ₃ PO ₄ (0.37 mol) The mixture in 1,4-dioxane: _H2O (300 mL:30 mL) was stirred at 100 °C for 16 h. The mixture was cooled to room temperature and the solid was filtered. The filtrate was concentrated to provide a residue which was purified by silica gel chromatography to provide the title compound in 63% yield as a yellow solid; m/z (ESI, +ve) = 492.1 [M+H]+. Step 4: Methyl 2-amino-4-(4-chlorothien-2-yl)-5-(trifluoromethyl)benzoate

TBAF (14.7 mmol) was added to methyl 2-amino-4-(4-chloro-5-(triisopropylsilyl)thiophen-2-yl)-5-(trifluoromethyl)benzoate ( 4.9 mmol) in THF (30 mL). The mixture was stirred at room temperature for three hours, diluted with water and extracted three times with ethyl acetate. The combined organic layers were dried over sodium sulfate and the volatiles were removed under reduced pressure to provide a residue which was purified by silica gel chromatography to provide the title compound in 97% yield as a white solid; m/z (ESI, +ve)= 336.0 [M+H]+. Step 5: Methyl 2-amino-4-(4-chlorothien-2-yl)-3-iodo-5-(trifluoromethyl)benzoate

To a mixture of methyl 2-amino-4-(4-chlorothien-2-yl)-5-(trifluoromethyl)benzoate (27.1 mmol) in AcOH (100 mL) was added N-iodosuccinyl Amine (44.4 mol). The reaction was stirred at room temperature for 36 hours and volatiles were removed under reduced pressure. The resulting residue was redissolved in ethyl acetate and _{washed sequentially} with saturated aqueous _Na2S2O3 , water and brine. The organic layer was dried over sodium sulfate and concentrated to provide a residue which was purified by silica gel chromatography (0-15% ethyl acetate in hexanes) to provide the title compound in 35% yield as a white solid; m /z (ESI, +ve)= 461.9 [M+H]+. Step 6: 2-Amino-4-(4-chlorothien-2-yl)-3-iodo-5-(trifluoromethyl)benzoic acid

To 2-amino-4-(4-chlorothiophen-2-yl)-3-iodo-5-(trifluoromethyl)benzoic acid methyl ester (23.8 mmol) in MeOH:THF (30 mL:30 mL) To the mixture was added 0.01M aqueous NaOH (0.238 mol) and the reaction was stirred at room temperature for 30 minutes. The pH of the mixture was adjusted to 5 by adding 6M HCl and the solution was extracted three times with ethyl acetate. The combined organic layers were washed with water, brine, dried over sodium sulfate and concentrated to afford the title compound as a yellow solid in 96% yield; m/z (ESI, +ve)=447.9 [M+H]+. Step 7: 7-(4-Chlorothien-2-yl)-8-iodo-6-(trifluoromethyl)quinazoline-2,4(1H,3H)-dione

A mixture of 2-amino-4-(4-chlorothiophen-2-yl)-3-iodo-5-(trifluoromethyl)benzoic acid (20.4 mmol) and urea (1.67 mol) was heated at 200°C 2 hours. The reaction was cooled to 90 °C and diluted with 600 mL of methanol:ethyl acetate (1:1). The mixture was allowed to cool slowly to room temperature and stirred for an additional 3 hours. After filtration, volatiles were removed under reduced pressure and the crude material was purified by reverse phase silica gel chromatography to afford the title compound as a white solid in 50% yield; m/z (ESI, +ve)=472.8 [M+H ]+. Step 8: (S)-7-(4-Chlorothiophen-2-yl)-8-((3-hydroxy-2-methoxypropyl)thio)-6-(trifluoromethyl)quinazole Phenyl-2,4(1H,3H)-dione

To 7-(4-chlorothiophen-2-yl)-8-iodo-6-(trifluoromethyl)quinazoline-2,4(1H,3H)-dione (0.0124 mol), cuprous iodide (470 mg, 0.0024 mol) and potassium carbonate (5.14 g, 00.0372 mol) in isopropanol (30 ml) and

To a solution in ethylene glycol (60 ml) was added (S)-3-mercapto-2-methoxypropan-1-ol (0.0372 mol). The reaction mixture was stirred at 85°C for 36 hours. The mixture was concentrated under reduced pressure and the crude material was purified by reverse phase column to afford the title compound in 39% yield as a white solid; m/z (ESI, +ve) = 467.0 [M+H]+. Step 9: (S)-11-(4-Chlorothiophen-2-yl)-3-methoxy-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4 ]Thiazaza[2,3,4-ij]quinazoline-6,8(7H)-dione

To a solution of triphenylphosphine (6.4 mmol) in THF (10 ml) cooled to 0° C. was added N,N -diisopropylethylamine (6.4 mmol) and the mixture was stirred for 30 minutes. Add (S)-7-(4-chlorothiophen-2-yl)-8-((3-hydroxy-2-methoxypropyl)thio)-6-(trifluoromethyl)quinazoline- 2,4(1H,3H)-dione (6.4 mmol) and stirred at 0 °C for 1 hour. The mixture was concentrated under reduced pressure and the residue was purified by reverse phase column chromatography to afford the title compound as a white solid in 77% yield; m/z (ESI, +ve) = 449.0 [M+H]+. Step 10: (S)-11-(4-Chlorothiophen-2-yl)-8-((3S,5R)-3,5-dimethylpiperazin-1-yl)-3-methoxy- 10-(Trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-6-one

(E2) To (S)-11-(4-chlorothiophen-2-yl)-3-methoxy-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]sulfur To a solution of azaza[2,3,4-ij]quinazoline-6,8(7H)-dione (0.05 mmol) in toluene (1 mL) was added N,N -diisopropylethylamine ( 1.0 mmol) and POCl3 (1 mL). The reaction mixture was stirred at 120 °C for 1.5 hours, cooled to room temperature and concentrated to afford a residue which was dissolved in dichloroethane (1 mL) and added to (2R,6S)-2,6-dimethyl A solution of piperazine (0.35 mmol) and N,N diisopropylethylamine (1.0 mmol) in dichloroethane (1 mL). The reaction mixture was stirred at room temperature for 1 hour, concentrated and the resulting solid was purified by silica gel chromatography to provide the title compound as a yellow solid in 51% yield; m/z (ESI, +ve) = 545.1 [M+H ]+. Example 2 : ( S )-7-(( 3S , 5R )-4- acryloyl -3,5- dimethylpiperazin -1- yl )-10-(4- chlorothiophen -2- yl )-3-( methoxymethyl )-9-( trifluoromethyl )-2,3- dihydro -5H-[1,4] thiazino [2,3,4-ij] quinazoline -5- keto (E2)

(E2)

To (12 S )-8-(4-chloro-2-thienyl)-4-[(3 S ,5 R )-3,5-dimethylpiperazin-1-yl]-12-(methoxy Methyl)-7-(trifluoromethyl)-10-aza-1,3-diazatricyclo[7.3.1.05,13]thirteen-3,5(13),6,8-tetra En-2-one; 2,2,2-Trifluoroacetic acid (0.05 mmol) in acetonitrile (2 mL) at 0 °C Add diisopropylethylamine (0.46 mmol) followed by acryloyl chloride (0.14 mmol) . The reaction was stirred at 0 °C for 20 minutes, diluted with ethyl acetate and washed with water. The organic layer was dried over sodium sulfate and concentrated under reduced pressure to provide a residue which was purified by HPLC. The title compound was isolated as a white solid in 36% yield. MS (ESI) m/z : 599.1 [M+H] ⁺ . 1H NMR (400 MHz, MeOD) δ 8.02 (s, 1H), 7.50 (s, 1H), 6.90 (broad singlet, 1H), 6.74 ( dd, J = 16.7, 10.6 Hz, 1H), 6.19 (d, J = 16.7, 2.0 Hz, 1H), 5.71 (d, J = 10.6, 2.0 Hz, 1H), 5.25 (s, 1H), 4.69–4.47 (m, 2H), 4.24 (d, 16 Hz, 1H), 4.15 (d, 16 Hz, 1H), 3.67-3.62 (m, 1H), 3.54-3.50 (m, 1H), 3.40 (m, 1H) , 3.36–3.23 (m, 5H), 3.02 (dd, J= 14, 3.0 Hz, 1H), 1.47 (d, J= 6.9 Hz, 6H), 1.31 (d, J= 6.9 Hz, 3H). Step 1: Methyl 2-Acetamido-4-chloro-5-iodobenzoate

To a mixture of methyl 2-amino-4-chloro-5-iodobenzoate (161 mmol) in AcOH (500 mL) was added _Ac2O (193 mmol). The mixture was stirred at 100 °C for 16 hours, cooled to room temperature, filtered and washed with hexanes to afford the title compound as a white solid in 62% yield. MS (ESI) m/z : 353.9 [M+H] ⁺ . Step 2: Methyl 2-Acetamido-4-chloro-5-(trifluoromethyl)benzoate

To a mixture of methyl 2-acetamido-4-chloro-5-iodobenzoate (99 mmol) in DMF (350 mL) was added methyl 2,2-difluoro-2-(fluorosulfonyl)acetate Ester (396 mmol), HMPA (396 mmol) and CuI (79 mmol). The mixture was stirred at 90°C for 16 hours, poured into water and extracted three times with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by silica gel chromatography (1-5% ethyl acetate in hexanes) to afford the title compound as a white solid in 84% yield. MS (ESI) m/z : 296.0 [M+H] ⁺ . Step 3: Methyl 2-amino-4-chloro-5-(trifluoromethyl)benzoate

A mixture of methyl 2-acetamido-4-chloro-5-(trifluoromethyl)benzoate (68 mmol) in HCl/MeOH (200 mL) was stirred at 70 °C for 2 h. The mixture was concentrated and the residue was treated with saturated aqueous NaHCO ₃ (100 mL). The resulting mixture was extracted three times with ethyl acetate and the combined organic layers were washed with brine and dried over sodium sulfate. The volatiles were evaporated under reduced pressure to provide the title compound as a yellow solid in quantitative yield. MS (ESI) m/z : 254.0 [M+H] ⁺ Step 4: Methyl 2-amino-4-chloro-3-iodo-5-(trifluoromethyl)benzoate

To a solution of methyl 2-amino-4-chloro-5-(trifluoromethyl)benzoate (110 mmol) in acetic acid (280 mL) was added N -iodosuccinimide (143 mmol). The mixture was stirred at 50°C for 16 hours. The reaction mixture was poured into water, extracted three times with ethyl acetate and the combined organic layers were washed with brine, dried over sodium sulfate, filtered and concentrated. The resulting residue was washed with hexanes to provide the title compound as a white solid in quantitative yield. MS (ESI) m/z : 379.9 [M+H] ⁺ Step 5: 2-Amino-4-chloro-3-iodo-5-(trifluoromethyl)benzoic acid

To a solution of methyl 2-amino-4-chloro-3-iodo-5-(trifluoromethyl)benzoate (66 mmol) in dioxane (200 mL) and water (200 mL) was added solid hydrogen Sodium oxide (132 mmol). The mixture was stirred at 90°C for 3 hours. After completion, the solution was poured into water and the pH was adjusted to 4-5. The mixture was extracted three times with ethyl acetate and the combined organic layers were washed with brine, dried over sodium sulfate, filtered and concentrated to provide the title compound as a yellow solid in 95% yield. MS (ESI) m/z : 365.9 [M+H] ⁺ Step 6: 7-Chloro-8-iodo-6-(trifluoromethyl)quinazoline-2,4(1H,3H)-dione

A mixture of 2-amino-4-chloro-3-iodo-5-(trifluoromethyl)benzoic acid (14 mmol) and urea (274 mmol) was stirred at 200 °C for 5 hours, cooled to 80 °C and water (100 mL) for treatment. The mixture was stirred for an additional hour, cooled to room temperature and extracted three times with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, filtered and concentrated. The resulting residue was purified by silica gel chromatography (20% ethyl acetate in hexanes) to afford the title compound as a white solid in 33% yield. MS (ESI) m/z : 388.8 [MH]- Step 7: ( R )-7-Chloro-8-((2-hydroxy-3-methoxypropyl)thio)-6-(trifluoromethyl base) quinazoline-2,4(1H,3H)-dione

To a solution of 7-chloro-8-iodo-6-(trifluoromethyl)quinazoline-2,4(1H,3H)-dione (51.2 mmol) in dioxane (800 mL) was added carbonic acid Potassium (153.6 mmol), ( R )-1-mercapto-3-methoxypropan-2-ol (92.1 mmol), 4,5-bis(diphenyl-phosphino)-9,9-dimethyl Dibenzopyran (10.24 mmol) and ginseng(dibenzylideneacetone)dipalladium (5.1 mmol). The mixture was stirred at 55°C for 18 hours. After completion, insoluble material was filtered off, the filtrate was concentrated and the pH was adjusted to 4 with acetic acid, extracted with ethyl acetate and washed with brine. Evaporation of volatiles gave a residue which was crystallized (dichloromethane/methanol = 1/10) to afford the title compound as a white solid in 73% yield. MS (ESI) m/z: 385.0 [M+H]+. Step 8: ( S )-10-Chloro-3-(methoxymethyl)-9-(trifluoromethyl)-2H-[1,4]thiazino[2,3,4-ij]quinone Azoline-5,7(3H,6H)-dione

At 0°C, to ( R )-7-chloro-8-((2-hydroxy-3-methoxypropyl)thio)-6-(trifluoromethyl)quinazoline-2,4( A mixture of 1H,3H)-diketone (89.5 mmol) and triphenylphosphoranylidene (134.3 mmol) in tetrahydrofuran (500 mL) was added diethyl azodicarboxylate (134.3 mmol). The mixture was stirred at 0 °C for 45 min. After completion, the mixture was poured into ice water (300 mL) and extracted three times with ethyl acetate. After concentration, the residue was recrystallized (dichloromethane/methanol=1/10) to provide the title compound as a white solid in 73% yield. MS (ESI): m/z 367.0 [M+H] ⁺ . Step 9: ( 2S , 6R )-4-((S)-10-Chloro-3-(methoxymethyl)-5-oxo-9-(trifluoromethyl)-3,5 -Dihydro-2H-[1,4]thiazino[2,3,4-ij]quinazolin-7-yl)-2,6-dimethylpiperazine-1-carboxylic acid tertiary butyl ester

At 0°C, to ( S )-10-chloro-3-(methoxymethyl)-9-(trifluoromethyl)-2H-[1,4]thiazino[2,3,4- ij] A mixture of quinazoline-5,7(3H,6H)-dione (27.32 mmol) and potassium carbonate (273.2 mmol) in acetonitrile (500 mL) was added 4-methylbenzenesulfonic anhydride (40.98 mmol). The mixture was stirred at 0 °C for 30 min and at 30 °C for 4 h, after which time tert-butyl ( 2S , 6R )-2,6-dimethylpiperazine- 1 -carboxylate ( 54.64 mmol) and the resulting mixture was stirred at 0 °C for a further 2 hours. The mixture was filtered through a pad of celite, and the filtrate was concentrated. The residue was purified by silica gel chromatography (1-3% methanol in dichloromethane) to provide the title compound as a white solid in quantitative yield. This material was used in the next step without further purification. MS (ESI) m/z 563.5 [M+H] ⁺ . Step 10: ( 2S , 6R )-4-(( S )-10-(4-chlorothiophen-2-yl)-3-(methoxymethyl)-5-oxo-9-( Trifluoromethyl)-2,3-dihydro-5H-[1,4]thiazino[2,3,4-ij]quinazolin-7-yl)-2,6-dimethylpiperazine -1-Tertiary butyl carboxylate

(2 S ,6 R )-4-[(12 S )-8-chloro-12-(methoxymethyl)-2-oxo-7-(trifluoromethyl)-10-aza -1,3-diazatricyclo[7.3.1.05,13]trideca-3,5(13),6,8-tetraen-4-yl]-2,6-dimethyl-piperazine- Tert-butyl 1-carboxylate (0.07 mmol), RuPhos G4 (0.01 mmol), potassium phosphate (0.22 mmol) and (4-chloro-2-thienyl)boronic acid (0.22 mmol) were dissolved in dioxane (2 mL) and water (0.5 mL) and the mixture was degassed with nitrogen for 2 minutes. The reaction was stirred at 80 °C for 1 hour, cooled to room temperature, diluted with ethyl acetate and washed with water and brine. The organic layer was dried over sodium sulfate and concentrated to provide a residue which was purified by HPLC to provide the title compound as a tan solid in 62% yield. Step 11: ( S )-10-(4-chlorothien-2-yl)-7-(( 3S , 5R )-3,5-dimethylpiperazin-1-yl)-3-(methyl Oxymethyl)-9-(trifluoromethyl)-2,3-dihydro-5H-[1,4]thiazino[2,3,4-ij]quinazolin-5-one

(E3) 反應流程 1

(R)-1- 巰基 -3- 甲氧基丙 -2- 醇 (2) To (2 S ,6 R )-4-[(12 S )-8-(4-chloro-2-thienyl)-12-(methoxymethyl)-2-methylene-7-(tri Fluoromethyl)-10-aza-1,3-diazatricyclo[7.3.1.05,13]trideca-3,5(13),6,8-tetraen-4-yl]-2, A stirred solution of tert-butyl 6-dimethyl-piperazine-1-carboxylate (0.05 mmol) in dichloromethane (3 mL) was added TFA (0.3 mL) and the reaction was stirred at room temperature for 45 minutes. Volatiles were removed under reduced pressure to afford an orange residue which was used in the next step without further purification. Example 3 : (S)-7-((3S,5R)-4- acryloyl -3,5- dimethylpiperazin -1- yl )-3-( methoxymethyl )-10-( 5- methylthiophen- 2- yl )-9-( trifluoromethyl )-2H-[1,4] thiazino [2,3,4-ij] quinazolin -5(3H) -one ( E3)

(E3) Reaction scheme 1

(R)-1- Mercapto -3- methoxypropan -2- ol (2)

To a mixture of (S)-2-(methoxymethyl)oxirane (50 g, 568.2 mmol) in tetrahydrofuran (800 mL) was added 1,1,1,3,3, 3-Hexamethyldisilathane (119.4 g, 681.8 mmol) and tetrabutylammonium fluoride (1.0 M in tetrahydrofuran, 170.6 mL, 170.5 mmol). The mixture was stirred at room temperature for 2 hours. After completion, the mixture was poured into water (3000 mL), extracted with ethyl acetate (3 x 800 mL). The combined organic phases were washed with brine (800 mL) and dried over anhydrous sodium sulfate. After filtration and concentration, the residue was purified by silica gel column with petroleum ether/ethyl acetate=3/1 to provide (R)-1-mercapto-3-methoxypropan-2-ol as colorless oil (70.0 g, crude product). ¹ H NMR (300 MHz, CDCl ₃ ) δ 3.86-3.82 (m, 1H), 3.49-3.46 (m, 1H), 3.41-3.37 (m, 4H), 2.71-2.64 (m, 1H), 1.55-1.50 (m, 1H). (R)-7- chloro -8-((2- hydroxy -3- methoxypropyl ) thio )-6-( trifluoromethyl ) quinazoline -2,4(1H,3H) -di Ketones (4)

To 7-chloro-8-iodo-6-(trifluoromethyl)quinazoline-2,4(1H,3H)-dione ( 3 ) (20 g, 51.2 mmol) in dioxane (800 mL) In the solution, add potassium carbonate (21.2 g, 153.6 mmol), (R)-1-mercapto-3-methoxypropan-2-ol (11.26 g, 92.1 mmol), 4,5-bis(diphenyl -phosphino)-9,9-dimethyldibenzopyran (5.93 g, 10.24 mmol) and para(dibenzylideneacetone)dipalladium (4.7 g, 5.1 mmol). The mixture was stirred at 55°C under nitrogen atmosphere for 18 hours. After completion, the insolubles were filtered off. The filtrate was then concentrated, the residue was adjusted to pH=4 with acetic acid and extracted with ethyl acetate (1000 mL), washed with brine (500 mL). The organic phase was concentrated and recrystallized (dichloromethane/methanol=1/10). The mixture was filtered and the filter cake collected to afford (R)-7-chloro-8-((2-hydroxy-3-methoxypropyl)thio)-6-(trifluoromethyl)quinoline as a white solid Azoline-2,4(1H,3H)-dione (4) (14 g, yield: 71%). MS (ESI) m/z 385.0 [M+H] ⁺ . (S)-10- Chloro -3-( methoxymethyl )-9-( trifluoromethyl )-2H-[1,4] thiazino [ 2,3,4-ij] quinazoline- 5,7(3H,6H) -diketone (5)

At 0°C, to (R)-7-chloro-8-((2-hydroxy-3-methoxypropyl)thio)-6-(trifluoromethyl)quinazoline-2,4( A mixture of 1H,3H)-diketone ( 4 ) (34.4 g, 89.5 mmol) and triphenylphosphoranylidene (35.2 g, 134.3 mmol) in THF (500 mL) was added with diethyl azodicarboxylate (23.3 g, 134.3 mmol). The mixture was stirred at 0 °C for 45 min. After completion, the mixture was poured into ice water (300 mL) and extracted with ethyl acetate (3 x 500 mL). After concentration, the residue was recrystallized (dichloromethane/methanol=1/10), the mixture was filtered and the filter cake was collected to afford (S)-10-chloro-3-(methoxymethyl) as a white solid -9-(Trifluoromethyl)-2H-[1,4]thiazino[2,3,4-ij]quinazoline-5,7(3H,6H)-dione (24.0 g, 73% Yield). MS (ESI): m/z 367.0 [M+H] ⁺ . (2S,6R)-4-((S)-10- chloro -3-( methoxymethyl )-5- oxo -9-( trifluoromethyl )-3,5- dihydro -2H -[1,4] thiazino [2,3,4-ij] quinazolin -7- yl )-2,6 -dimethylpiperazine -1- carboxylic acid tert-butyl ester (6)

At 0°C, to (S)-10-chloro-3-(methoxymethyl)-9-(trifluoromethyl)-2H-[1,4]thiazino[2,3,4- ij] A mixture of quinazoline-5,7(3H,6H)-dione (10 g, 27.32 mmol) and potassium carbonate (37.7 g, 273.2 mmol) in acetonitrile (500 mL) was added with 4-methylbenzenesulfonate Anhydride (13.4 g, 40.98 mmol). The mixture was stirred at 0°C for 30 min and at 30°C for 4 hours. After completion, tert-butyl (2S,6R)-2,6-dimethylpiperazine-1-carboxylate (11.7 g, 54.64 mmol) was added to the reaction solution. The reaction mixture was stirred at 0 °C for 2 hours. Upon completion, the mixture was filtered through a pad of celite, and the filtrate was concentrated. The residue was purified by chromatography column (dichloromethane/methanol=100/1 to 30/1) to afford (2S,6R)-4-((S)-10-chloro-3 as a pale solid -(methoxymethyl)-5-oxo-9-(trifluoromethyl)-3,5-dihydro-2H-[1,4]thiazino[2,3,4-ij] Quinazolin-7-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester (15 g, crude product). MS (ESI) m/z 563.5 [M+H] ⁺ . (2S,6R)-4-((S)-3-( Methoxymethyl )-10-(5- methylthiophen -2- yl )-5- oxo -9-( trifluoromethyl )-3,5- dihydro -2H-[1,4] thiazino [2,3,4-ij] quinazolin -7- yl )-2,6- dimethylpiperazine -1- carboxylic acid Tertiary butyl ester (7)

To (2S,6R)-4-((S)-10-chloro-3-(methoxymethyl)-5-oxo-9-(trifluoromethyl)-3,5-dihydro- 2H-[1,4]thiazino[2,3,4-ij]quinazolin-7-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester ( 6 )(200 mg, 0.35 mmol) in 1,4-dioxane (6 mL) and water (1 mL), add tripotassium phosphate (780 mg, 3.5 mmol), (5-methylthiophene-2- base) boronic acid (497 mg, 3.5 mmol), and chloro(2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)[2-(2'-amine yl-1,1'-biphenyl)]palladium(II) (41 mg, 0.52 mmol). The mixture was stirred at 85°C for 4 hours under nitrogen atmosphere. After completion, the mixture was concentrated. The residue was purified by silica gel column chromatography (dichloromethane/methanol=100/1 to 30/1) to provide (2S,6R)-4-((S)-3-(methoxy Methyl)-10-(5-methylthiophen-2-yl)-5-oxo-9-(trifluoromethyl)-3,5-dihydro-2H-[1,4]thiazine [2,3,4-ij]quinazolin-7-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester ( 7 ) (200 mg, crude product). MS (ESI) m/z 625.5 [M+H] ⁺ . (S)-7-((3S,5R)-3,5- Dimethylpiperazin -1- yl )-3-( methoxymethyl )-10-(5- methylthiophen -2- yl )-9-( trifluoromethyl )-2H-[1,4] thiazino [2,3,4-ij] quinazolin -5(3H) -one (8)

At 0°C, to (2S,6R)-4-((S)-3-(methoxymethyl)-10-(5-methylthiophen-2-yl)-5-oxo-9 -(trifluoromethyl)-3,5-dihydro-2H-[1,4]thiazino[2,3,4-ij]quinazolin-7-yl)-2,6-dimethyl To a mixture of tert-butyl piperazine-1-carboxylate (200 mg, 0.32 mmol) in dichloromethane (3 ml) was added trifluoroacetic acid (1 mL). The reaction solution was stirred at room temperature for 30 minutes. After completion, the mixture was concentrated. The residue was redissolved in dichloromethane and _dried over _Na2SO4 . After concentration, the residue was purified by column (dichloromethane/methanol=15:1) to provide (S)-7-((3S,5R)-3,5-dimethylpiperazine as a yellow solid -1-yl)-3-(methoxymethyl)-10-(5-methylthiophen-2-yl)-9-(trifluoromethyl)-2H-[1,4]thiazino[ 2,3,4-ij]quinazolin-5(3H)-one ( 8 ) (150 mg, yield: 89%). MS (ESI) m/z 525.3 [M+H] ⁺ . (S)-7-((3S,5R)-4- acryloyl -3,5- dimethylpiperazin -1- yl )-3-( methoxymethyl )-10-(5- methyl ylthiophen -2- yl )-9-( trifluoromethyl )-2H-[1,4] thiazino [2,3,4-ij] quinazolin -5(3H) -one ( Example 3)

(E4)

(2S,6R)-4-((S)-10-(5- 氯噻吩 -2- 基 )-3-( 甲氧基甲基 )-5- 側氧基 -9-( 三氟甲基 )-3,5- 二氫 -2H-[1,4] 噻嗪并 [2,3,4-ij] 喹唑啉 -7- 基 )-2,6- 二甲基哌嗪 -1- 甲酸第三丁酯 (2) At 0°C, to (S)-7-((3S,5R)-3,5-dimethylpiperazin-1-yl)-3-(methoxymethyl)-10-(5-methyl ylthiophen-2-yl)-9-(trifluoromethyl)-2H-[1,4]thiazino[2,3,4-ij]quinazolin-5(3H)-one (100 mg, 0.19 mmol) and triethylamine (28 mg, 0.28 mmol) in dichloromethane (3 ml) was added acrylic anhydride (36 mg, 0.28 mmol). The mixture was stirred at 0°C for 30 minutes. After completion, the mixture was poured into ice water (1 mL) and extracted with ethyl acetate (3 x 5 mL). After concentration, the residue was purified by chromatography column (dichloromethane/methanol=60/1 to 30/1) to provide (S)-7-((3S,5R)-4-propene as white powder Acyl-3,5-dimethylpiperazin-1-yl)-3-(methoxymethyl)-10-(5-methylthiophen-2-yl)-9-(trifluoromethyl) -2H-[1,4]thiazino[2,3,4-ij]quinazolin-5(3H)-one ( Example 3 ) (48 mg yield: 44%). MS (ESI) m/z 579.3 [M+H]+; ¹ H NMR (400 MHz, CDCl ₃ ) δ8.03 (s, 1H), 6.82 (s, 2H), 6.62 (dd, J = 10.4 Hz, J = 16.4 Hz, 1H), 6.40 (dd, J = 1.6 Hz, J = 16.4 Hz, 1H), 5.77 (dd, J = 1.6 Hz, J = 10.4 Hz, 1H), 5.42-5.38 (m, 1H) , 4.78-4.54 (m, 2H), 4.20-4.15 (m, 2H), 3.72-3.62 (m ,2H), 3.40 (s, 3H), 3.38-3.28 (m, 3H), 2.96 (dd, J = 2.8 Hz, J = 13.6 Hz, 1H), 2.57 (s, 3H), 1.61 (d, J = 6.8 Hz, 3H), 1.46 (d, J = 6.8 Hz, 3H). Example 4 : (S)-7-((3S,5R)-4- acryloyl -3,5- dimethylpiperazin -1- yl )-10-(5- chlorothien- 2- yl )- 3-( methoxymethyl )-9-( trifluoromethyl )-2H-[1,4] thiazino [2,3,4-ij] quinazolin -5(3H) -one (E4 )

(E4)

(2S,6R)-4-((S)-10-(5- chlorothiophen -2- yl )-3-( methoxymethyl )-5- oxo -9-( trifluoromethyl ) -3,5- dihydro -2H-[1,4] thiazino [2,3,4-ij] quinazolin -7- yl )-2,6- dimethylpiperazine -1- carboxylic acid Tributyl ester (2)

To (2S,6R)-4-((S)-10-chloro-3-(methoxymethyl)-5-oxo-9-(trifluoromethyl)-3,5-dihydro- 2H-[1,4]thiazino[2,3,4-ij]quinazolin-7-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester (400 mg, 0.71 mmol) in a solution of 1,4-dioxane (15 mL) and water (2 mL), add tripotassium phosphate (1.5 g, 7.1 mmol), (5-chlorothiophen-2-yl) boronic acid ( 497 mg, 3.5 mmol), and chloro(2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)[2-(2'-amino-1, 1'-biphenyl)]palladium(II) (83 mg, 0.1 mmol). The mixture was stirred at 85°C for 4 hours under nitrogen atmosphere. After completion, the mixture was concentrated. The residue was purified by silica gel column chromatography (dichloromethane/methanol=100/1 to 30/1) to provide (2S,6R)-4-((S)-10-(5- Chlorothiophen-2-yl)-3-(methoxymethyl)-5-oxo-9-(trifluoromethyl)-3,5-dihydro-2H-[1,4]thiazino [2,3,4-ij]quinazolin-7-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester ( 2 ) (450 mg, crude product). MS (ESI) m/z 645.5 [M+H] ⁺ . (S)-10-(5- chlorothien -2- yl )-7-((3S,5R)-3,5- dimethylpiperazin -1- yl )-3-( methoxymethyl ) -9-( trifluoromethyl )-2H-[1,4] thiazino [2,3,4-ij] quinazolin -5(3H) -one (3)

At 0°C, to (2S,6R)-4-((S)-10-(5-chlorothien-2-yl)-3-(methoxymethyl)-5-oxo-9- (Trifluoromethyl)-3,5-dihydro-2H-[1,4]thiazino[2,3,4-ij]quinazolin-7-yl)-2,6-dimethylpiper To a mixture of tert-butyloxine-1-carboxylate ( 2 ) (450 mg, 0.69 mmol) in dichloromethane (3 ml) was added trifluoroacetic acid (1 mL). The reaction solution was stirred at room temperature for 30 minutes. After completion, the mixture was concentrated and the residue was purified by column (dichloromethane/methanol=15:1) to provide (S)-10-(5-chlorothiophen-2-yl)-7- as a yellow solid ((3S,5R)-3,5-Dimethylpiperazin-1-yl)-3-(methoxymethyl)-9-(trifluoromethyl)-2H-[1,4]thiazine and[2,3,4-ij]quinazolin-5(3H)-one ( 3 ) (350 mg, crude product). MS (ESI) m/z 544.9 [M+H] ⁺ . (S)-7-((3S,5R)-4- acryloyl -3,5- dimethylpiperazin -1- yl )-10-(5- chlorothiophen -2- yl )-3-( Methoxymethyl )-9-( trifluoromethyl )-2H-[1,4] thiazino [2,3,4-ij] quinazolin -5(3H) -one (4)

步驟1：(2S,6R)-4-((S)-11-(4-氯-5-(三異丙基矽基)噻吩-2-基)-6-側氧基-3-(嘧啶-2-基氧基)-10-(三氟甲基)-3,4-二氫-2H,6H-[1,4]硫氮呯并[2,3,4-ij]喹唑啉-8-基)-2,6-二甲基哌嗪-1-甲酸第三丁酯

At 0°C, to (S)-10-(5-chlorothiophen-2-yl)-7-((3S,5R)-3,5-dimethylpiperazin-1-yl)-3-( Methoxymethyl)-9-(trifluoromethyl)-2H-[1,4]thiazino[2,3,4-ij]quinazolin-5(3H)-one (350 mg, 0.64 mmol) and triethylamine (97 mg, 0.96 mmol) in dichloromethane (3 ml) was added acrylic anhydride (121 mg, 0.96 mmol). The mixture was stirred at 0°C for 30 minutes. After completion, the mixture was poured into ice water (1 mL) and extracted with ethyl acetate (3 x 5 mL). The organic layer was dried over Na2So4 and concentrated. The residue was purified by chromatography column (dichloromethane/methanol=60/1 to 30/1) to provide (S)-10-(5-chlorothien-2-yl)-7- ((3S,5R)-3,5-Dimethylpiperazin-1-yl)-3-(methoxymethyl)-9-(trifluoromethyl)-2H-[1,4]thiazine And[2,3,4-ij]quinazolin-5(3H)-one ( 4 ) (88 mg yield: 24%). MS (ESI) m/z 599.9 [M+H]+; ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.03 (s, 1H), 6.98 (d, J = 4.0 Hz, 1H), 6.81 (d, J = 3.2 Hz, 1H), 6.61 (dd, J = 10.8 Hz, J = 16.8 Hz, 1H), 6.40 (dd, J = 2.0 Hz, J = 16.8 Hz, 1H), 5.77 (dd, J = 2.0 Hz, J = 10.4 Hz, 1H), 5.46-5.41 (m, 1H), 4.82-4.48 (m, 2H), 4.17 (t, J = 13.6 Hz, 2H), 3.70-3.62 (m, 2H), 3.40-3.28 (m, 6H), 2.96 (dd, J = 2.8 Hz, J = 13.2 Hz, 1H), 1.61 (d, J = 6.8 Hz, 3H), 1.46 (d, J = 7.2 Hz, 3H). Example 23 : (S)-8-((3S,5R)-4- acryloyl -3,5- dimethylpiperazin -1- yl )-11-(4- chlorothiophen -2- yl )- 3-( pyrimidin -2- yloxy )-10-( trifluoromethyl )-3,4- dihydro -2H,6H-[1,4] thiazepine [2,3,4-ij] Quinazolin -6- one

Step 1: (2S,6R)-4-((S)-11-(4-Chloro-5-(triisopropylsilyl)thiophen-2-yl)-6-oxo-3-(pyrimidine -2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazoline- 8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester

(2S,6R)-4-((S)-11-chloro-6-oxo-3-(pyrimidin-2-yloxy)-10-(trifluoromethyl)-3,4-di Hydrogen-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester (0.32 mmol), (4-chloro-5-(triisopropylsilyl)thiophen-2-yl)boronic acid (1.28 mmol), potassium phosphate (0.96 mmol) and Ruphos Pd G4 (0.03 mmol) in dioxane (5 mL) and water (0.1 mL) was degassed and purged three times with nitrogen. The mixture was stirred at 80 °C for 30 minutes and the volatiles were removed under reduced pressure to afford a residue which was purified by preparative TLC (65% ethyl acetate in hexanes). The title compound was isolated in 63% yield as a white solid. MS (ESI) m/z: 865.3 [M+1]+. Step 2: (2S,6R)-4-((S)-11-(4-chlorothiophen-2-yl)-6-oxo-3-(pyrimidin-2-yloxy)-10-( Trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl)-2,6-dimethyl Tributyl piperazine-1-carboxylate

To (2S,6R)-4-((S)-11-(4-chloro-5-(triisopropylsilyl)thiophen-2-yl)-6-oxo-3-(pyrimidine-2 -yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazoline-8- To a solution of tert-butyl)-2,6-dimethylpiperazine-1-carboxylate (0.20 mmol) in THF (5 mL) was added a 1M solution of tetrabutylammonium fluoride in THF (0.3 mL) . The mixture was stirred at room temperature for 30 minutes, the volatiles were removed under reduced pressure and the resulting residue was purified by preparative TLC (60% ethyl acetate in hexanes) to afford the title as a colorless semi-solid in 72% yield compound. MS (ESI) m/z: 709.2 [M+1]+ Step 3: (S)-11-(4-chlorothien-2-yl)-8-((3S,5R)-3,5-dimethyl Piperazin-1-yl)-3-(pyrimidin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine [2,3,4-ij]quinazolin-6-one

Add trifluoroacetic acid (2 mL) to (2S,6R)-4-((S)-11-(4-chlorothien-2-yl)-6-oxo-3-(pyrimidin-2-yl Oxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl) - A solution of tert-butyl 2,6-dimethylpiperazine-1-carboxylate (0.14 mmol) in dichloromethane (6 mL). After 30 minutes, the volatiles were removed under reduced pressure to afford the title compound as a yellow oil in 99% yield. This material was used in the next step without further purification. MS (ESI) m/z: 609.2 [M+1]+. Step 4: (S)-8-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-11-(4-chlorothien-2-yl)- 3-(pyrimidin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3,4-ij] Quinazolin-6-one

反應流程

(S)-1-(1-( 苄氧基 )-3-( 三苯甲硫基 ) 丙 -2- 基 )-1H- 吡唑并 [3,4-b] 吡啶 (2) Triethylamine (0.43 mmol) and prop-2-enyl chloride (0.21 mmol) were added to (S)-11-(4-chlorothien-2-yl)-8-((3S,5R)-3 ,5-Dimethylpiperazin-1-yl)-3-(pyrimidin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4 A solution of ]thiazeza[2,3,4-ij]quinazolin-6-one (0.14 mmol) in dichloromethane (2 mL). After 30 minutes, the volatiles were removed under reduced pressure to provide a residue which was purified by preparative TLC (100% ethyl acetate in hexanes). The title compound was isolated in 67% yield as a white solid. MS (ESI) m/z: 663.4 [M+1]+. 1H NMR (400 MHz, CDCl3) δ 8.52 (d, J = 4.8 Hz, 2H), 8.06 (s, 1H), 7.34 (s, 1H) , 7.00 (t, J = 4.8 Hz, 1H), 6.92 (s, 1H), 6.63 (dd, J = 10.4, 16.4 Hz, 1H), 6.47 - 6.37 (m, 1H), 5.84 - 5.69 (m, 2H ), 4.95 - 4.53 (m, 4H), 4.21 (d, J = 13.2 Hz, 2H), 3.60 (dd, J = 2.8, 13.6 Hz, 1H), 3.45 - 3.32 (m, 2H), 3.19 - 3.06 ( m, 1H), 1.64 - 1.60 (m, 3H), 1.48 (d, J = 6.4 Hz, 3H) Example 24 : (S)-8-((3S,5R)-4- acryloyl -3,5 -Dimethylpiperazin -1- yl )-11-(4- chlorothien -2- yl )-3-(1H- pyrazolo [3,4-b] pyridin -1- yl )-10- ( Trifluoromethyl )-3,4- dihydro -2H,6H-[1,4] thiazeza [2,3,4-ij] quinazolin- 6- one

Reaction flow

(S)-1-(1-( Benzyloxy )-3-( tritylthio ) propan -2- yl )-1H- pyrazolo [3,4-b] pyridine (2)

At 0°C, (R)-1-(benzyloxy)-3-(tritylthio)propan-2-ol (30 g, 68.18 mmol), 1H-pyrazolo[3,4- b] A mixture of pyridine (9.74 g, 81.85 mmol) and triphenylphosphine (53.6 g, 204.6 mmol) in tetrahydrofuran (340 mL) was added with (E)-diethyldiazene-1,2-dicarboxylic acid Salt (35.6 g, 204.6 mmol). The mixture was stirred at room temperature under nitrogen atmosphere for 12 hours. After completion, the mixture was quenched with water (300 mL) and extracted with dichloromethane (100 mLx3). The organic phase was concentrated under reduced pressure. The residue was purified by silica gel column with petroleum ether/ethyl acetate=10/1 to afford (S)-1-(1-(benzyloxy)-3-(tritylthio) as colorless oil )prop-2-yl)-1H-pyrazolo[3,4-b]pyridine ( 2 ) (23.36 g, 63% yield). ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.51-8.49 (m, 1H), 8.02-8.00 (m, 2H), 7.43-7.32 (m, 6H), 7.24-6.99 (m, 15H), 4.36-4.26 (m, 3H), 3.79-3.64 (m, 2H), 3.20-3.15 (m, 1H), 2.72-2.68 (m, 1H). (S)-3-( Benzyloxy )-2-(1H- pyrazolo [3,4-b] pyridin -1- yl ) propane -1- thiol (3)

Under nitrogen atmosphere at 0°C, to (S)-1-(1-(benzyloxy)-3-(tritylthio)prop-2-yl)-1H-pyrazolo[3,4 -b] A mixture of pyridine ( 2 ) (23.36 g, 43.18 mmol) in dichloromethane (90 mL) and 2,2,2-trifluoroacetic acid (30 mL) was added with triethylsilane (15 g, 129.3 mmol ). The mixture was stirred at 0 °C for 10 min. After completion, the mixture was concentrated under reduced pressure and adjusted to pH=8 with NH ₃ ^.MeOH . The mixture was then extracted with ethyl acetate (60 mL x 3). After concentration, the residue was purified by silica gel column with petroleum ether/ethyl acetate=10/1 to afford (S)-3-(benzyloxy)-2-(1H-pyrazolo [3,4-b]pyridin-1-yl)propane-1-thiol ( 3 ) (11 g, 85% yield). ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.56-8.64 (m, 1H), 8.08-8.05 (m, 2H), 7.29-7.12 (m, 6H), 5.39-5.35 (m, 1H), 4.53-4.45 (m, 2H), 4.01-3.93 (m, 2H), 3.35-3.13 (m, 2H), 1.27-1.22 (m, 1H). (S)-8-((3-( Benzyloxy )-2-(1H- pyrazolo [3,4-b] pyridin -1- yl ) propyl ) thio )-7- chloro -6- ( Trifluoromethyl ) quinazoline -2,4- diol (4)

To 7-chloro-8-iodo-6-(trifluoromethyl)quinazoline-2,4-diol ( 3 ) (8 g, 20.51 mmol) in 1,4-dioxane (100 mL) Potassium carbonate (8.5 g, 61.59 mmol), (S)-3-(benzyloxy)-2-(1H-pyrazolo[3,4-b]pyridin-1-yl)propane-1- Thiol (11 g, 36.79 mmol), 4,5-bis(diphenyl-phosphino)-9,9-dimethyldibenzopyran (1.78 g, 3.08 mmol) and ginseng (dibenzylidene acetone) dipalladium (1.87 g, 2.04 mmol). The mixture was stirred at 60°C under nitrogen atmosphere for 18 hours. After completion, the mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane/methanol=50/1) to provide (S)-8-((3-(benzyl Oxygen)-2-(1H-pyrazolo[3,4-b]pyridin-1-yl)propyl)thio)-7-chloro-6-(trifluoromethyl)quinazoline-2, 4-diol ( 4 ) (15 g, crude product). MS (ESI): m/z 563 [MH] ^- . (S)-7- chloro -8-((3- hydroxy -2-(1H- pyrazolo [3,4-b] pyridin -1- yl ) propyl ) thio )-6-( trifluoromethyl base ) quinazoline -2,4- diol (5)

(S)-8-((3-(Benzyloxy)-2-(1H-pyrazolo[3,4-b]pyridin-1-yl)propyl)thio)-7-chloro-6- A solution of (trifluoromethyl)quinazoline-2,4-diol ( 4 ) (15 g, 26.69 mmol) in trifluoroacetic acid (120 mL). The mixture was stirred at 80°C for 18 hours. After completion, the mixture was concentrated and adjusted to PH=7~8 at 0°C. After concentration, the residue was purified by a silica gel column with dichloromethane/methanol=40/1 to provide (S)-7-chloro-8-((3-hydroxy-2-(1H- Pyrazolo[3,4-b]pyridin-1-yl)propyl)thio)-6-(trifluoromethyl)quinazoline-2,4-diol ( 5 ) (7.67 g, for two steps, 26% yield). MS (ESI): m/z 472 [MH] ^- . (S)-11- chloro -8- hydroxyl -3-(1H- pyrazolo [3,4-b] pyridin -1- yl )-10-( trifluoromethyl )-3,4 - dihydro- [1,4] Thiazaza [2,3,4-ij] quinazolin -6(2H) -one (6)

At 0°C, to (S)-7-chloro-8-((3-hydroxy-2-(1H-pyrazolo[3,4-b]pyridin-1-yl)propyl)thio)- A mixture of 6-(trifluoromethyl)quinazoline-2,4-diol ( 5 ) (7.57 g, 16.07 mmol) and triphenylphosphine (12.6 g, 48.09 mmol) in THF (300 mL) was added Diethyl azodicarboxylate (8.4 g, 48.28 mmol). The mixture was stirred at 0 °C for 45 min. After completion, the mixture was poured into ice water (100 mL) and extracted with ethyl acetate (100 mL x 3). After concentration, the residue was purified by C18 with 30-95% acetonitrile in water to afford (S)-11-chloro-8-hydroxy-3-(1H-pyrazolo[3,4-b ]pyridin-1-yl)-10-(trifluoromethyl)-3,4-dihydro-[1,4]thiazazo[2,3,4-ij]quinazoline-6(2H) - Ketone (6) (6 g, 82% yield). ¹ H NMR (400 MHz, CDCl ₃ ) δ 12.12 (s, 1H), 8.61-8.60 (m, 1H), 8.32-8.23 (m, 2H), 8.09 (s, 1H), 7.32-7.29 (m, 1H ), 5,69-5,63 (m, 1H), 5.14-4.22 (m, 3H), 3.59 (s, 1H). (2S,6R)-4-((S)-11- chloro -6- oxo -3-(1H- pyrazolo [3,4-b] pyridin -1- yl )-10-( trifluoro Methyl )-2,3,4,6 - tetrahydro- [1,4] thiazazo [2,3,4-ij] quinazolin -8- yl )-2,6- dimethylpiper tert -Butyloxine - 1- carboxylate (7)

To (S)-11-chloro-8-hydroxyl-3-(1H-pyrazolo[3,4-b]pyridin-1-yl)-10-(trifluoromethyl)-3,4-dihydro -[1,4]thiazeza[2,3,4-ij]quinazolin-6(2H)-one ( 6 ) (3 g, 6.62 mmol) and potassium carbonate (9.2 g, 66.7 mmol) in To a mixture in acetonitrile (120 mL) and dichloromethane (80 mL) was added 2,4,6-triisopropylphenyl 4-tosylate (4 g, 13.2 mmol). The mixture was stirred at 35°C for 5 hours. After completion, (2S,6R)-tert-butyl 2,6-dimethylpiperazine-1-carboxylate (2.2 g, 10.3 mmol) was added to the reaction solution. The reaction mixture was stirred at 35°C for 1 hour. After completion, the mixture was poured into ice water (200 mL) and extracted with ethyl acetate (100 mL x 3). After concentration, the residue was purified by C18 column with 20-95% acetonitrile in water to afford (2S,6R)-4-((S)-11-chloro-6-oxo-3 as a yellow solid -(1H-pyrazolo[3,4-b]pyridin-1-yl)-10-(trifluoromethyl)-2,3,4,6-tetrahydro-[1,4]thiazepine [2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester ( 7 ) (3.92 g, 91% yield). MS (ESI) m/z 650 [M+H] ⁺ . (2S,6R)-4-((S)-11-(4- Chloro -5-( triisopropylsilyl ) thiophen -2- yl )-6- oxo -3-(1H- pyrazole And [3,4-b] pyridin -1- yl )-10-( trifluoromethyl )-3,4- dihydro -2H,6H-[1,4] thiazepine [2,3,4 -ij] quinazolin -8- yl )-2,6- dimethylpiperazine -1- carboxylic acid tert-butyl ester (8)

To (2S,6R)-4-((S)-11-chloro-6-oxo-3-(1H-pyrazolo[3,4-b]pyridin-1-yl)-10-(tri Fluoromethyl)-2,3,4,6-tetrahydro-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl)-2,6-dimethyl Tripotassium phosphate (491 mg, 1.85 mmol), (4-chloro-5-(triisopropylsilyl)thiophen-2-yl)boronic acid (586 mg, 1.84 mmol), and chloro(2-dicyclohexylphosphino-2',6'- Diisopropoxy-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (49 mg, 0.062 mmol). The mixture was stirred at 80°C for 2 hours under nitrogen atmosphere. After completion, the mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane/methanol=60/1) to provide (2S,6R)-4-((S) -11-(4-Chloro-5-(triisopropylsilyl)thiophen-2-yl)-6-oxo-3-(1H-pyrazolo[3,4-b]pyridine-1- Base)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl)- tert-butyl 2,6-dimethylpiperazine-1-carboxylate ( 8 ) (200 mg, 37% yield). MS (ESI) m/z 888.2 [M+H] ⁺ . (2S,6R)-4-((S)-11-(4- chlorothien -2- yl )-6- oxo -3-(1H- pyrazolo [3,4-b] pyridine -1 -yl )-10-( trifluoromethyl )-3,4- dihydro -2H , 6H-[1,4] thiazezo [2,3,4-ij] quinazolin -8- yl ) -2,6- Dimethylpiperazine -1- carboxylic acid tert-butyl ester (9)

At 0°C, to (2S,6R)-4-((S)-11-(4-chloro-5-(triisopropylsilyl)thiophen-2-yl)-6-oxo-3 -(1H-pyrazolo[3,4-b]pyridin-1-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine [2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester (8) (200 mg, 0.23 mmol) in tetrahydrofuran (5 mL ) was added tetrabutylammonium fluoride (0.35 mL, 1.0 M solution in THF). The reaction solution was stirred at 0°C for 1 hour. After completion, the mixture was concentrated and extracted with ethyl acetate (50 mL×3). The organic phase was concentrated and the residue was purified by a C18 column with 20%-95% acetonitrile in water to provide (2S,6R)-4-((S)-11-(4-chlorothiophene as a light yellow solid -2-yl)-6-oxo-3-(1H-pyrazolo[3,4-b]pyridin-1-yl)-10-(trifluoromethyl)-3,4-dihydro- 2H, 6H-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester (9 ) (130 mg, 77% yield). MS (ESI) m/z 732.2 [M+H] ⁺ . (S)-11-(4- chlorothien -2- yl )-8-((3S,5R)-3,5- dimethylpiperazin -1- yl )-3-(1H- pyrazolo [ 3,4-b] pyridin -1- yl )-10-( trifluoromethyl )-3,4- dihydro -2H,6H-[1,4] thiazepine [2,3,4-ij ] quinazolin -6- one (10)

At 0°C, to (2S,6R)-4-((S)-11-(4-chlorothien-2-yl)-6-oxo-3-(1H-pyrazolo[3,4 -b]pyridin-1-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazole To a mixture of tertiary butyl-2,6-dimethylpiperazine-1-carboxylate ( 9 ) (130 mg, 0.178 mmol) in dichloromethane (5 ml) was added trifluoroacetic acid ( 2 mL). The mixture was stirred at room temperature for 1 hour. After completion, the mixture was concentrated and adjusted to pH=7~8 at 0°C. After concentration, the residue was purified by a silica gel column with dichloromethane/methanol=30/1 to provide (S)-11-(4-chlorothiophen-2-yl)-8-(( 3S,5R)-3,5-Dimethylpiperazin-1-yl)-3-(1H-pyrazolo[3,4-b]pyridin-1-yl)-10-(trifluoromethyl) -3,4-Dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-6-one ( 10 ) (92 mg, 82% yield). MS (ESI) m/z 632.2 [M+H] ⁺ . (S)-8-((3S,5R)-4- acryloyl -3,5- dimethylpiperazin -1- yl )-11-(4- chlorothiophen -2- yl )-3-( 1H- pyrazolo [3,4-b] pyridin -1- yl )-10-( trifluoromethyl )-3,4- dihydro -2H,6H-[1,4] thiazepine [2 ,3,4-ij] quinazolin- 6- one (11)

反應流程

(R)-1-( 苄氧基 )-3-( 三苯甲硫基 ) 丙 -2- 醇 (2) At 0°C, to (S)-11-(4-chlorothiophen-2-yl)-8-((3S,5R)-3,5-dimethylpiperazin-1-yl)-3-( 1H-pyrazolo[3,4-b]pyridin-1-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2 ,3,4-ij]quinazolin-6-one ( 10 ) (92 mg, 0.146 mmol) and triethylamine (29 mg, 0.292 mmol) in dichloromethane (2 ml) was added acrylic anhydride (27 mg, 0.219 mmol). The mixture was stirred at 0°C for 1 hour. After completion, the mixture was poured into ice water (30 mL) and extracted with ethyl acetate (10 mL x 3). Concentration and the residue was purified by preparative high performance liquid chromatography (20% to 95% acetonitrile in water) to afford (S)-8-((3S,5R)-4-acryloyl-3 as a white solid ,5-Dimethylpiperazin-1-yl)-11-(4-chlorothien-2-yl)-3-(1H-pyrazolo[3,4-b]pyridin-1-yl)-10 -(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-6-one ( 11 ) (28 mg , 28% yield). MS (ESI) m/z 686.1 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.53 (d, J = 4.0 Hz, 1H), 8.10-8.04 (m, 3H), 7.34 (s, 1H), 7.18-7.15 (dd, J = 8.0 Hz, 4.4 Hz, 1H), 7.02-6.88 (m, 1H), 6.67-6.60 (m, 1H), 6.42 (dd, J = 16.8 Hz, 2.0 Hz, 1H), 5.78 (dd, J = 10.4 Hz, 2.0 Hz , 2H), 5.54-5.28 (m, 1H), 5.10 (d, J = 13.6 Hz, 1H), 4.80-4.44 (m, 2H), 4.17 (d, J = 13.2 Hz, 2H), 3.91-3.57 ( m, 1H), 3.40-3.33 (m, 3H), 1.59-1.58 (m, 6H). Example 25 : (S)-8-((3S,5R)-4- acryloyl -3,5- dimethylpiperazin -1- yl )-11-(4- chlorothiophen -2- yl )- 3-(3 -oxo- [1,2,4] triazolo [4,3-a] pyridin -2(3H) -yl )-10-( trifluoromethyl )-3,4- di Hydrogen -2H,6H-[1,4] thiazeza [2,3,4-ij] quinazolin -6- one

Reaction flow

(R)-1-( Benzyloxy )-3-( tritylthio ) propan -2- ol (2)

To a mixture of (S)-2-((benzyloxy)methyl)oxirane (25 g, 152.4 mmol) and potassium fluoride (17.7 g, 304.8 mmol) in methanol (250 mL) was added triphenyl Methylmercaptan (42 g, 152.4 mmol). The mixture was stirred at room temperature for 18 hours. After completion, the mixture was concentrated under reduced pressure and purified by silica gel column with petroleum ether/ethyl acetate=5/1 to provide (R)-1-(benzyloxy)-3-( Tritylthio)propan-2-ol (2) (60 g, 90% yield). ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.44-7.38 (m, 6H), 7.35-7.18 (m, 14H), 4.46 (s, 2H), 3.58-3.48 (m, 1H), 3.37-3.26 (m , 2H), 2.45-2.35 (m, 2H). (S)-2-(1-( Benzyloxy )-3-( tritylthio ) propan -2- yl )-[1,2,4] triazolo [4,3-a ] pyridine- 3(2H) -Kone (3)

[1,2,4]triazolo[4,3-a]pyridin-3(2H)-one (5.5 g, 41 mmol) and triphenylphosphine (13.4 g , 51 mmol) in tetrahydrofuran (400 mL) was added diethyl azodicarboxylate (8.8 g, 51 mmol). The mixture was stirred at room temperature for 10 min, then (R)-1-(benzyloxy)-3-(tritylthio)propan-2-ol (2) (15 g, 34 mmol) was added. The mixture was stirred for 2 hours. After completion, the mixture was poured into ice water (300 mL) and extracted with ethyl acetate (150 mL x 3). After concentration, the residue was purified by silica gel column with petroleum ether/ethyl acetate=50/1 to afford crude (S)-2-(1-(benzyloxy)-3-(tris) as white solid Benzylthio)propan-2-yl)-[1,2,4]triazolo[4,3-a]pyridin-3(2H)-one (3) (16.5 g, crude product). MS (ESI): m/z 558.2 [M+H] ⁺ . (S)-2-(1-( Benzyloxy )-3- mercaptopropan- 2- yl )-[1,2,4] triazolo [4,3-a] pyridin -3(2H) -one (4)

To (S)-2-(1-(benzyloxy)-3-(tritylthio)prop-2-yl)-[1,2,4]triazolo[4,3-a]pyridine - To a mixture of 3(2H)-one (3) (165 g, 30 mmol) and triethylsilane (17.4 g, 150 mmol) in dichloromethane (300 mL) was added trifluoroacetic acid (30 mL). The mixture was stirred at room temperature for 2 hours. After completion, the mixture was concentrated at 0 °C and adjusted to pH=7~8. After concentration, the residue was purified by a silica gel column with petroleum ether/ethyl acetate=2/1 to provide (S)-2-(1-(benzyloxy)-3-mercaptopropan-2-yl) -[1,2,4]triazolo[4,3-a]pyridin-3(2H)-one (4) (7.9 g, 85% yield) MS (ESI): m/z 316.1[M+ H] ⁺ . (S)-2-(1-( benzyloxy )-3-((7- chloro -2,4- dihydroxy -6-( trifluoromethyl ) quinazolin -8- yl ) thio ) propane -2- yl )-[1,2,4] triazolo [4,3-a] pyridin -3(2H) -one (5)

To a solution of 7-chloro-8-iodo-6-(trifluoromethyl)quinazoline-2,4-diol (4.5 g, 11 mmol) in 1,4-dioxane (200 mL), Potassium carbonate (4.6 g, 33 mmol), (S)-2-(1-(benzyloxy)-3-mercaptopropan-2-yl)-[1,2,4]triazolo[4,3 -a]pyridin-3(2H)-one (4) (55 g, 17 mmol), 4,5-bis(diphenyl-phosphino)-9,9-dimethyldibenzopyran (1.2 g, 1.03 mmol) and ginseng (dibenzylideneacetone) dipalladium (0.92 g, 1.0 mmol). The mixture was stirred at 60°C under nitrogen atmosphere for 18 hours. After completion, the mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane/methanol=20/1) to provide (S)-2-(1-(benzyloxy Base)-3-((7-chloro-2,4-dihydroxy-6-(trifluoromethyl)quinazolin-8-yl)thio)propan-2-yl)-[1,2,4 ]Triazolo[4,3-a]pyridin-3(2H)-one (5) (6.2 g, 93% yield) MS (ESI): m/z 578.1 [M+H] ⁺ . (S)-2-(1-((7- chloro -2,4- dihydroxy -6-( trifluoromethyl ) quinazolin -8- yl ) thio )-3- hydroxypropan -2- yl )-[1,2,4] triazolo [4,3-a] pyridin -3(2H) -one (6)

(S)-2-(1-(benzyloxy)-3-((7-chloro-2,4-dihydroxy-6-(trifluoromethyl)quinazolin-8-yl)thio)propane -2-yl)-[1,2,4]triazolo[4,3-a]pyridin-3(2H)-one (5) (6.2 g, 10 mmol) in trifluoroacetic acid (50 mL) The solution was stirred at 80°C for 18 hours. After completion, the mixture was concentrated at 0 °C and adjusted to pH=7~8. After concentration, the residue was purified by a silica gel column with dichloromethane/methanol=10/1 to provide (S)-2-(1-((7-chloro-2,4-dihydroxy -6-(trifluoromethyl)quinazolin-8-yl)thio)-3-hydroxypropan-2-yl)-[1,2,4]triazolo[4,3-a]pyridine- 3(2H)-Kone (6) (3.6 g, 69% yield). MS (ESI): m/z 488.1 [M+H] ⁺ . (S)-11- chloro -8- hydroxy -3-(3- oxo- [1,2,4] triazolo [4,3-a] pyridin -2(3H) -yl )-10- ( Trifluoromethyl )-3,4- dihydro- [1,4] thiazazo [2,3,4-ij] quinazolin -6(2H) -one (7)

Under nitrogen atmosphere at 0°C, to (S)-2-(1-((7-chloro-2,4-dihydroxy-6-(trifluoromethyl)quinazolin-8-yl)thio) -3-Hydroxypropan-2-yl)-[1,2,4]triazolo[4,3-a]pyridin-3(2H)-one (6) (3.5 g, 7.1 mmol) and triphenyl A mixture of phosphine (7.5 g, 28.6 mmol) in tetrahydrofuran (130 mL) was added diethyl azodicarboxylate (4.9 g, 28.6 mmol). The mixture was stirred at room temperature for 3 hours. After completion, the mixture was poured into ice water (100 mL) and extracted with ethyl acetate (100 mL x 3). Concentration and the residue was purified by C18 with 30-95% acetonitrile in water to afford (S)-11-chloro-3-(4-fluorophenoxy)-8-hydroxy-10-(tris) as a yellow solid Fluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-6-one (7) (2.6 g, 78% Yield). MS (ESI): m/z 470. [M+H] ⁺ . (2S,6R)-4-((S)-11- Chloro -6- oxo -3-(3 -oxo- [1,2,4] triazolo [4,3-a] pyridine -2(3H)-yl ) -10-( trifluoromethyl )-2,3,4,6 - tetrahydro- [1,4] thiazazo [2,3,4-ij] quinazoline -8- yl )-2,6- dimethylpiperazine -1- carboxylic acid tert-butyl ester (8)

To ((S)-11-chloro-3-(4-fluorophenoxy)-8-hydroxyl-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4] A mixture of thiazeza[2,3,4-ij]quinazolin-6-one (7) (2.4 g, 5.1 mmol) and potassium carbonate (7.0 g, 51 mmol) in acetonitrile (100 mL) was added 4-Methylbenzenesulfonic anhydride (4.9 g, 15.3 mmol). At 35° C., the mixture was stirred for 8 hours. After completion, (2S,6R)-2,6-dimethylpiperazine-1-carboxylic acid was Tributyl ester (3.8 g, 17.8 mmol) was added to the reaction solution. The reaction mixture was stirred overnight at 35 °C. After completion, the mixture was poured into ice water (200 mL) and extracted with ethyl acetate (100 mL x 3 ). Concentrate and the residue is purified by a C18 column with 20-95% acetonitrile in water to afford (2S,6R)-4-((S)-11-chloro-6-oxo as a light yellow solid -3-(3-oxo-[1,2,4]triazolo[4,3-a]pyridin-2(3H)-yl)-10-(trifluoromethyl)-2,3, 4,6-tetrahydro-[1,4]thiazepine[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl Ester (8) (2.3 g, 72% yield). MS (ESI) m/z 666.2 [M+H] ⁺ . (2S,6R)-4-((S)-11-(4- chloro -5 -( Triisopropylsilyl ) thiophen -2- yl )-6- oxo -3-(3 -oxo- [1,2,4] triazolo [4,3-a ] pyridine- 2(3H) -yl )-10-( trifluoromethyl )-3,4- dihydro -2H,6H-[1,4] thiazeza [ 2,3,4-ij] quinazoline- 8- yl )-2,6- dimethylpiperazine -1- carboxylic acid tert-butyl ester (9)

To (2S,6R)-4-((S)-11-chloro-6-oxo-3-(3-oxo-[1,2,4]triazolo[4,3-a] Pyridin-2(3H)-yl)-10-(trifluoromethyl)-2,3,4,6-tetrahydro-[1,4]thiazazo[2,3,4-ij]quinazole Phyllin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester (8) (350 mg, 0.53 mmol) in 1,4-dioxane (5 mL) and water (0.5 mL) was added tripotassium phosphate (337 mg, 1.59 mmol), (4-chloro-5-(triisopropylsilyl)thiophen-2-yl)boronic acid (507 mg, 1.59 mmol), and chlorine ( 2-Dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II ) (41 mg, 0.053 mmol). The mixture was stirred at 80°C for 1 hour under nitrogen atmosphere. After completion, the mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane/methanol=60/1) to provide (2S,6R)-4-((S)- 11-(4-Chloro-5-(triisopropylsilyl)thiophen-2-yl)-6-oxo-3-(3-oxo-[1,2,4]triazolo[ 4,3-a]pyridin-2(3H)-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3, 4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester (9) (250 mg, crude product). MS (ESI) m/z 904.1 [M+H] ⁺ . (2S,6R)-4-((S)-11-(4- chlorothien- 2- yl )-6- oxo -3-(3 -oxo- [1,2,4] triazole And [4,3-a] pyridin -2(3H)-yl ) -10-( trifluoromethyl )-3,4- dihydro -2H,6H-[1,4] thiazepine [2, 3,4-ij] quinazolin -8- yl )-2,6- dimethylpiperazine -1- carboxylic acid tert-butyl ester (10)

At 0°C, to (2S,6R)-4-((S)-11-(4-chloro-5-(triisopropylsilyl)thiophen-2-yl)-6-oxo-3 -(3-oxo-[1,2,4]triazolo[4,3-a]pyridin-2(3H)-yl)-10-(trifluoromethyl)-3,4-dihydro -2H,6H-[1,4]thiazazo[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester ( 9) To a mixture of (250 mg, 0.28 mmol) in THF (5 mL) was added tetrabutylammonium fluoride (0.34 mL, 1.0 M solution in THF). The reaction solution was stirred at 0°C for 1 hour. After completion, the mixture was concentrated and extracted with ethyl acetate (50 mL×3). Concentrated and the residue was purified by C18 column with 20%-95% acetonitrile in water to provide (2S,6R)-4-((S)-11-(4-chlorothiophene-2- Base)-6-oxo-3-(3-oxo-[1,2,4]triazolo[4,3-a]pyridin-2(3H)-yl)-10-(trifluoro Methyl)-3,4-dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiper Tert-butyloxazine-1-carboxylate (10) (150 mg, 38% yield for two steps). MS (ESI) m/z 748.2 [M+H] ⁺ . (S)-11-(4- chlorothien - 2- yl )-8-((3S,5R)-3,5- dimethylpiperazin -1- yl )-3-(3- oxo- [1,2,4] triazolo [4,3-a] pyridin -2(3H)-yl ) -10-( trifluoromethyl )-3,4- dihydro -2H,6H-[1, 4] Thiaazepine [2,3,4-ij] quinazolin- 6- one (11)

At 25°C, to (2S,6R)-4-((S)-11-(4-chlorothien-2-yl)-6-oxo-3-(3-oxo-[1, 2,4]triazolo[4,3-a]pyridin-2(3H)-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]sulfur Azo[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester (10) (150 mg, 0.20 mmol) in di To the mixture in methyl chloride (6 mL) was added trifluoroacetic acid (2 mL). The mixture was stirred at 25°C for 1 hour. After completion, the mixture was concentrated at 0 °C and the pH was adjusted to 8 with ammonia in methanol. The mixture was concentrated and purified by silica gel column chromatography (dichloromethane/methanol=30/1) to provide (S)-11-(4-chlorothiophen-2-yl)-8-( (3S,5R)-3,5-Dimethylpiperazin-1-yl)-3-(3-oxo-[1,2,4]triazolo[4,3-a]pyridine-2 (3H)-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazoline-6 - Ketone (11) (80 mg, 62% yield). MS (ESI) m/z 648.2 [M+H] ⁺ . (S)-8-((3S,5R)-4- acryloyl -3,5- dimethylpiperazin -1- yl )-11-(4- chlorothiophen -2- yl )-3-( 3 -oxo- [1,2,4] triazolo [4,3-a] pyridin -2(3H) -yl )-10-( trifluoromethyl )-3,4- dihydro -2H ,6H-[1,4] thiazeza [2,3,4-ij] quinazolin -6- one (12)

反應流程

(3- 氯噻吩 -2- 基 ) 三異丙基矽烷 (2) At 0°C, to (S)-11-(4-chlorothiophen-2-yl)-8-((3S,5R)-3,5-dimethylpiperazin-1-yl)-3-( 3-oxo-[1,2,4]triazolo[4,3-a]pyridin-2(3H)-yl)-10-(trifluoromethyl)-3,4-dihydro-2H , A mixture of 6H-[1,4]thiazeza[2,3,4-ij]quinazolin-6-one (11) (80 mg, 0.12 mmol) in dichloromethane (6 mL) was added Triethylamine (24 mg, 0.24 mmol) and acrylic anhydride (23 mg, 0.18 mmol). The mixture was stirred at 25°C for 1 hour. After completion, methanol was added to the mixture at 25 °C and concentrated. The mixture was purified by preparative high performance liquid chromatography (20% to 95% acetonitrile in water) to provide (S)-8-((3S,5R)-4-acryloyl-3,5 as a light yellow powder -Dimethylpiperazin-1-yl)-11-(4-chlorothien-2-yl)-3-(3-oxo-[1,2,4]triazolo[4,3-a ]pyridin-2(3H)-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeb[2,3,4-ij]quinone Azolin-6-one (12) (24 mg, 0.034 mmol, 28% yield). MS (ESI) m/z 702.3 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.09 (s, 1H), 7.75 (d, J = 7.2 Hz, 1H), 7.34 (s, 1H), 7.10-7.00 (m, 2H), 6.96-6.95 ( m, 1H), 6.65-6.59 (m, 1H), 6.49 (t, J = 7.2 Hz, 1H), 6.41 (dd, J = 16.8 Hz, 2.0 Hz, 1H), 5.78 (dd, J = 10.4 Hz, 2.0 Hz, 1H), 5.35-5.27 (m, 1H), 5.25-5.08 (m, 1H), 5.02-4.90 (m, 1H), 4.90-4.50 (m, 2H), 4.15 (d, J = 13.2 Hz , 2H), 3.85-3.00 (m, 4H), 1.60-1.40 (m, 6H). Example 26 : (S)-8-((3S,5R)-4- acryloyl -3,5- dimethylpiperazin -1- yl )-11-(4- chlorothiophen -2- yl )- 3-(1- oxoisoindol -2 - yl )-10-( trifluoromethyl )-3,4- dihydro -2H,6H-[1,4] thiazepine [2,3 ,4-ij] quinazolin- 6- one

Reaction flow

(3- Chlorothiophen -2- yl ) triisopropylsilane (2)

Lithium diisopropylamide (221.37 mL, 442.74 mmol, 2.0 M solution in tetrahydrofuran/n-heptane) was slowly added to 3-chlorothiophene (50 g, 421.66 mmol) at -78 °C under nitrogen atmosphere. Solution in THF (1.05 L). The mixture was stirred at -78°C for 1 hour, then triisopropylsilyl chloride (85.36 g, 442.74 mmol) was added. The mixture was allowed to warm slowly to room temperature and stirred overnight. After completion, saturated aqueous ammonium chloride (800 mL) was slowly added and extracted with ethyl acetate (1 L x 3). After concentration, the residue was purified on a silica gel column (with petroleum ether/ethyl acetate=500/1) to provide (3-chlorothiophen-2-yl)triisopropylsilane (2) (57 g , 49% yield). ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.52 (d, J = 4.8 Hz, 1H), 7.04 (d, J = 4.8 Hz, 1H), 1.57-1.51 (m, 3H), 1.12 (d, J = 8.0 Hz, 18H). (4- Chloro -5-( triisopropylsilyl ) thiophen -2- yl ) boronic acid (3)

(25) Lithium diisopropylamide (10.91 mL, 21.83 mmol, 2.0 M solution in THF/n-heptane) was slowly added to (3-chlorothiophen-2-yl ) A solution of triisopropylsilane (2) (5 g, 18.19 mmol) in tetrahydrofuran (90 mL). The mixture was stirred at -78°C for 1 hour, then dimethyl borate (2.27 g, 21.83 mmol) was added. The mixture was allowed to warm slowly to room temperature and stirred for 1 hour. After completion, saturated aqueous ammonium chloride (10 mL) was slowly added. After concentration, the residue was purified by silica gel column (with dichloromethane/methanol=50/1) to provide (4-chloro-5-(triisopropylsilyl)thiophene-2- base) boronic acid (3) (3.5 g, 60% yield). ¹ H NMR (400 MHz, CD3OD) δ 7.53 (s, 1H), 1.59-1.52 (m, 3H), 1.13 (d, J = 8.0 Hz, 18H). (R)-1-( Benzyloxy )-3-( tritylthio ) propan -2- ol (5)

To a mixture of (S)-2-((benzyloxy)methyl)oxirane (25 g, 152.4 mmol) and potassium fluoride (17.7 g, 304.8 mmol) in methanol (250 mL) was added triphenyl Methylmercaptan (42 g, 152.4 mmol). The mixture was stirred at room temperature for 18 hours. After completion, the mixture was concentrated under reduced pressure and purified by silica gel column with petroleum ether/ethyl acetate=5/1 to provide (R)-1-(benzyloxy)-3-( Tritylthio)propan-2-ol (5) (60 g, 90% yield). ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.44-7.38 (m, 6H), 7.35-7.18 (m, 14H), 4.46 (s, 2H), 3.58-3.48 (m, 1H), 3.37-3.26 (m , 2H), 2.45-2.35 (m, 2H). (S)-2-(1-( Benzyloxy )-3-( tritylthio ) propan -2- yl ) isoindoline -1,3- dione (6)

To a mixture of isoindoline-1,3-dione (6.0 g, 40.9 mmol) and triphenylphosphine (13.3 g, 51.1 mmol) in tetrahydrofuran (400 mL) was added diisoindoline-1,3-dione (6.0 g, 40.9 mmol) at 0°C under nitrogen atmosphere. Diethyl nitrogen dicarboxylate (8.9 g, 51.1 mmol). The mixture was stirred at room temperature for 10 min, then (R)-1-(benzyloxy)-3-(tritylthio)propan-2-ol (5) (15 g, 34.1 mmol) was added. The mixture was stirred for 2 hours. After completion, the mixture was poured into ice water (300 mL) and extracted with ethyl acetate (150 mL x 3). After concentration, the residue was purified by silica gel column with petroleum ether/ethyl acetate=50/1 to afford crude (S)-2-(1-(benzyloxy)-3-(tris) as colorless oil Benzylthio)propan-2-yl)isoindoline-1,3-dione (6) (19.0 g, crude product). MS (ESI) m/z 570.2 [M+H] ⁺ . (S)-2-(1-( Benzyloxy )-3- mercaptopropan -2- yl ) isoindolin -1- one (7)

At 0°C, to (S)-2-(1-(benzyloxy)-3-(tritylthio)prop-2-yl)isoindoline-1,3-dione (6) (18.0 g, 31.66 mmol) in acetic acid (250 mL) was added zinc (30.8 g, 474 mmol). The mixture was stirred at 100°C for 4 hours. After completion, the mixture was concentrated at 0°C and adjusted to pH=7~8 and extracted with dichloroethane. After concentration, the residue was purified by a silica gel column with petroleum ether/ethyl acetate=3/1 to provide (S)-2-(1-(benzyloxy)-3-mercaptopropane- 2-yl)isoindolin-1-one (7) (3.7 g, 37% yield). MS (ESI) m/z 314.1 [M+H] ⁺ . (S)-2-(1-( benzyloxy )-3-((7- chloro -2,4- dihydroxy -6-( trifluoromethyl ) quinazolin -8- yl ) thio ) propane -2- yl ) isoindolin -1- one (8)

To a solution of 7-chloro-8-iodo-6-(trifluoromethyl)quinazoline-2,4-diol (4.6 g, 12 mmol) in 1,4-dioxane (200 mL) was added Potassium carbonate (11.8 g, 36 mmol), (S)-2-(1-(benzyloxy)-3-mercaptopropan-2-yl)isoindolin-1-one (7) (4.7 g, 15 mmol), 4,5-bis(diphenyl-phosphino)-9,9-dimethyldibenzopyran (1.4 g, 1.6 mmol) and ginseng(dibenzylideneacetone)dipalladium (1.1 g , 1.2 mmol). The mixture was stirred at 60°C under nitrogen atmosphere for 18 hours. After completion, the mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane/ethyl acetate=3/1) to provide (S)-2-(1- (Benzyloxy)-3-((7-chloro-2,4-dihydroxy-6-(trifluoromethyl)quinazolin-8-yl)thio)propan-2-yl)isoindoline -1-one (8) (4.6 g, 66% yield). MS (ESI) m/z 576.1 [M+H] ⁺ . (S)-2-(1-((7- chloro -2,4- dihydroxy -6-( trifluoromethyl ) quinazolin -8- yl ) thio )-3- hydroxypropan -2- yl ) isoindolin -1- one (9)

(S)-2-(1-(benzyloxy)-3-((7-chloro-2,4-dihydroxy-6-(trifluoromethyl)quinazolin-8-yl)thio)propane -2-yl)isoindolin-1-one (8) (4.5 g, 7.8 mmol) in trifluoroacetic acid (100 mL). The mixture was stirred at 80°C for 18 hours. After completion, the mixture was concentrated at 0 °C and adjusted to pH = 7~8. After concentration, the residue was purified by a silica gel column with dichloromethane/ethyl acetate=1/2 to provide (S)-2-(1-((7-chloro-2,4- Dihydroxy-6-(trifluoromethyl)quinazolin-8-yl)thio)-3-hydroxypropan-2-yl)isoindoline-1-one (9) (2.8 g, 75% yield Rate). MS (ESI): m/z 486.1 [M+H] ⁺ . (S)-11- chloro -8- hydroxy -3-(1- oxoisoindol -2- yl )-10-( trifluoromethyl )-3,4- dihydro- [1,4] Thiazeza [2,3,4-ij] quinazolin -6(2H) -one (10)

Under nitrogen atmosphere at 0°C, to (S)-2-(1-((7-chloro-2,4-dihydroxy-6-(trifluoromethyl)quinazolin-8-yl)thio) A mixture of -3-hydroxypropan-2-yl)isoindolin-1-one (9) (2.6 g, 5.3 mmol) and triphenylphosphine (5.6 g, 21.4 mmol) in tetrahydrofuran (100 mL) was added Diethyl azodicarboxylate (3.7 g, 21.4 mmol). The mixture was stirred at room temperature for 3 hours. After completion, the mixture was poured into ice water (100 mL) and extracted with ethyl acetate (100 mL x 3). Concentration and the residue was purified by C18 with 30-95% acetonitrile in water to afford (S)-11-chloro-8-hydroxy-3-(1-oxoisoindol-2-yl as a yellow solid )-10-(trifluoromethyl)-3,4-dihydro-[1,4]thiazeza[2,3,4-ij]quinazolin-6(2H)-one (10) ( 2.0 g, 83% yield). MS (ESI) m/z 468.2 [M+H] ⁺ . (2S,6R)-4-((S)-11- chloro -6- oxo -3-(1 -oxoisoindol- 2- yl )-10-( trifluoromethyl )-2 , 3,4,6 - tetrahydro- [1,4] thiazazo [2,3,4-ij] quinazolin- 8- yl )-2,6- dimethylpiperazine -1- carboxylic acid Tertiary butyl ester (11)

To (S)-11-chloro-8-hydroxyl-3-(1-oxoisoindol-2-yl)-10-(trifluoromethyl)-3,4-dihydro-[1,4 ] Thiazeza[2,3,4-ij]quinazolin-6(2H)-one (10) (500 mg, 1.06 mmol) and DIEA (690.38 mg, 5.34 mmol) in toluene (20 mL) To the mixture was added phosphonyl trichloride (1.31 g, 8.547 mmol). The mixture was stirred at 120°C for 4 hours. After completion, the mixture was concentrated and the residue was added to the solution. The reaction mixture was stirred at 35°C for 1 hour. After completion, the mixture was poured into ice water (100 mL) and extracted with ethyl acetate (50 mL x 3). Concentrate and the residue is purified by C18 column with 20-95% acetonitrile in water to afford (2S,6R)-4-((S)-11-chloro-6-oxo-3 as a light yellow solid -(1-oxoisoindol-2-yl)-10-(trifluoromethyl)-2,3,4,6-tetrahydro-[1,4]thiazepine[2,3, 4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester (11) (532 mg, 75% yield). MS (ESI) m/z 664.1 [M+H] ⁺ . (2S,6R)-4-((S)-11-(4- Chloro -5-( triisopropylsilyl ) thiophen -2- yl )-6- oxo -3-(1- oxo Isoindol -2- yl )-10-( trifluoromethyl )-3,4- dihydro -2H,6H-[1,4] thiazeza [2,3,4-ij] quinazole Lin -8- yl )-2,6- dimethylpiperazine -1- carboxylic acid tert-butyl ester (12)

To (2S,6R)-4-((S)-11-chloro-6-oxo-3-(1-oxoisoindol-2-yl)-10-(trifluoromethyl)- 2,3,4,6-tetrahydro-[1,4]thiazazo[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1- To a solution of tert-butyl formate (11) (350 mg, 0.53 mmol) in 1,4-dioxane (5 mL) and water (0.5 mL) was added tripotassium phosphate (337 mg, 1.59 mmol), (4 -Chloro-5-(triisopropylsilyl)thiophen-2-yl)boronic acid (507 mg, 1.59 mmol), and chloro(2-dicyclohexylphosphino-2',6'-diisopropoxy -1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (41 mg, 0.053 mmol). The mixture was stirred at 80°C for 1 hour under nitrogen atmosphere. After completion, the mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane/methanol=60/1) to provide (2S,6R)-4-((S) -11-(4-Chloro-5-(triisopropylsilyl)thiophen-2-yl)-6-oxo-3-(1-oxoisoindol-2-yl)-10- (Trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl)-2,6-di Methylpiperazine-1-carboxylic acid tert-butyl ester (12) (190 mg, 40% yield). MS (ESI) m/z 902.1 [M+H] ⁺ . (2S,6R)-4-((S)-11-(4- Chlorothiophen -2- yl )-6- oxo -3-(1 -oxoisoindol -2- yl )-10 -( trifluoromethyl )-3,4- dihydro -2H,6H-[1,4] thiazeza [2,3,4-ij] quinazolin- 8- yl )-2,6- tert-butyl dimethylpiperazine -1- carboxylate (13)

At 0°C, to (2S,6R)-4-((S)-11-(4-chloro-5-(triisopropylsilyl)thiophen-2-yl)-6-oxo-3 -(1-oxoisoindol-2-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3, A mixture of 4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester (12) (190 mg, 0.21 mmol) in THF (5 mL) was added Tetrabutylammonium fluoride (0.23 mL, 1.0 M solution in tetrahydrofuran). The reaction solution was stirred at 0°C for 1 hour. After completion, the mixture was concentrated and extracted with ethyl acetate (100 mL×3). Concentrated and the residue was purified by C18 column with 20%-95% acetonitrile in water to provide (2S,6R)-4-((S)-11-(4-chlorothiophene-2- Base)-6-oxo-3-(1-oxoisoindol-2-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4 ]Thiazaza[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester (13) (110 mg, 70% yield Rate). MS (ESI) m/z 746.2 [M+H] ⁺ . (S)-11-(4- chlorothiophen -2- yl )-8-((3S,5R)-3,5- dimethylpiperazin -1- yl )-3-(1- oxoiso Indol -2- yl )-10-( trifluoromethyl )-3,4- dihydro -2H , 6H-[1,4] thiazazo [2,3,4-ij] quinazoline- 6- keto (14)

At 0°C, to (2S,6R)-4-((S)-11-(4-chlorothien-2-yl)-6-oxo-3-(1-oxoisoindole- 2-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazolin-8-yl To a mixture of tert-butyl )-2,6-dimethylpiperazine-1-carboxylate (13) (110 mg, 0.15 mmol) in dichloromethane (6 mL) was added trifluoroacetic acid (2 mL). The reaction solution was stirred at room temperature for 1 hour. After completion, the mixture was concentrated and the residue was purified by silica gel column chromatography (dichloromethane/methanol=15/1) to provide (S)-11-(4-chlorothiophene-2- Base)-8-((3S,5R)-3,5-dimethylpiperazin-1-yl)-3-(1-oxoisoindol-2-yl)-10-(trifluoromethyl base)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-6-one (14) (62 mg, 64% yield ). MS (ESI) m/z 646.2 [M+H] ⁺ . (S)-8-((3S,5R)-4- acryloyl -3,5- dimethylpiperazin -1- yl )-11-(4- chlorothiophen -2- yl )-3-( 1- oxoisoindol -2- yl )-10-( trifluoromethyl )-3,4- dihydro -2H,6H-[1,4] thiazepine [2,3,4- ij] quinazolin- 6- one (15)

反應流程

(R)-1-( 苄氧基 )-3-( 三苯甲硫基 ) 丙 -2- 醇 (2) At 0°C, to (S)-11-(4-chlorothiophen-2-yl)-8-((3S,5R)-3,5-dimethylpiperazin-1-yl)-3-( 1-oxoisoindol-2-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3,4- ij] A mixture of quinazolin-6-one (14) (62 mg, 0.096 mmol) and triethylamine (19 mg, 0.192 mmol) in dichloromethane (3 ml) was added with acrylic anhydride (18 mg, 0.144 mmol). The mixture was stirred at 0°C for 1 hour. After completion, the mixture was poured into ice water (10 mL) and extracted with ethyl acetate (10 mL x 3). Concentration and the residue was purified by preparative high performance liquid chromatography (20% to 95% acetonitrile in water) to afford (S)-8-((3S,5R)-4-acryloyl-3 as a white solid ,5-Dimethylpiperazin-1-yl)-11-(4-chlorothien-2-yl)-3-(1-oxoisoindol-2-yl)-10-(trifluoroform base)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-6-one (15) (20 mg, 30% yield ). MS (ESI) m/z 700.3 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.09 (s, 1H), 7.86 (d, J = 7.6 Hz, 1H), 7.57-7.52 (m, 1H), 7.47 (t, J = 7.6 Hz, 2H) , 7.36 (s, 1H), 7.00-6.93 (m, 1H), 6.66-6.60 (m, 1H), 6.44-6.40 (m, 1H), 5.79 (dd, J = 10.4 Hz, 2.0 Hz, 1H), 5.20-4.15 (m, 10H), 3.43-3.37 (m, 3H), 1.57-1.56 (m, 6H). Example 27 : (S)-8-((3S,5R)-4- acryloyl -3 ,5- Dimethylpiperazin -1- yl )-3-(1H- benzo [d] imidazol -1- yl )-11-(4- chlorothien -2- yl )-10-( trifluoromethane base )-3,4- dihydro- [1,4] thiazepine [2,3,4-ij] quinazolin -6(2H) -one

Reaction flow

(R)-1-( Benzyloxy )-3-( tritylthio ) propan -2- ol (2)

To a mixture of (S)-2-((benzyloxy)methyl)oxirane (25 g, 152.4 mmol) and potassium fluoride (17.7 g, 304.8 mmol) in methanol (250 mL) was added triphenyl Methylmercaptan (42 g, 152.4 mmol). The mixture was stirred at room temperature for 18 hours. After completion, the mixture was concentrated under reduced pressure and purified by silica gel column with petroleum ether/ethyl acetate=5/1 to provide (R)-1-(benzyloxy)-3-( Tritylthio)propan-2-ol (2) (60 g, 90% yield). ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.44-7.38 (m, 6H), 7.35-7.18 (m, 14H), 4.46 (s, 2H), 3.58-3.48 (m, 1H), 3.37-3.26 (m , 2H), 2.45-2.35 (m, 2H). (S)-1-(1-( Benzyloxy )-3-( tritylthio ) propan -2- yl )-1H- benzo [d] imidazole (3)

To a mixture of 1H-benzo[d]imidazole (8.45 g, 71.59 mmol) and triphenylphosphine (25.0 g, 95.46 mmol) in tetrahydrofuran (470 mL) was added azodicarboxylate at 0 °C under nitrogen atmosphere diethyl ester (16.6 g, 95.46 mmol). The mixture was stirred at room temperature for 10 min, then (R)-1-(benzyloxy)-3-(tritylthio)propan-2-ol (2) (21 g, 47.73 mmol) was added. The mixture was stirred for 2 hours. After completion, the mixture was poured into ice water (300 mL) and extracted with ethyl acetate (150 mL x 3). After concentration, the residue was purified by a silica gel column with petroleum ether/ethyl acetate=30/1 to provide (S)-1-(1-(benzyloxy)-3-(triphenyl Methylthio)propan-2-yl)-1H-benzo[d]imidazole (3) (12.4 g, 48% yield). MS (ESI) m/z 541.0 [M+H] ⁺ . (S)-2-(1H- Benzo [d] imidazol -1- yl )-3-( benzyloxy ) propane -1- thiol (4)

To (S)-1-(1-(benzyloxy)-3-(tritylthio)prop-2-yl)-1H-benzo[d]imidazole (3) (12.47 g, 23.1 mmol) and a mixture of triethylsilane (6.7 g, 57.7 mmol) in dichloromethane (230 mL) was added trifluoroacetic acid (26.3 g, 231 mmol). The mixture was stirred at room temperature for 3 hours. After completion, the mixture was concentrated at 0 °C and adjusted to pH = 7~8. After concentration, the residue was purified by silica gel column with petroleum ether/ethyl acetate=10/1 to provide (S)-2-(1H-benzo[d]imidazol-1-yl) as yellow oil - 3-(Benzyloxy)propane-1-thiol (4) (3.95 g, 57% yield). MS (ESI) m/z 299.2 [M+H] ⁺ . (S)-8-((2-(1H- Benzo [d] imidazol -1 -yl )-3-( benzyloxy ) propyl ) thio )-7- chloro - 6-( trifluoromethyl ) quinazoline -2,4- diol (5)

To a solution of 7-chloro-8-iodo-6-(trifluoromethyl)quinazoline-2,4-diol (2.87 g, 7.36 mmol) in 1,4-dioxane (73 mL) was added Potassium carbonate (3.0 g, 22.08 mmol), (S)-2-(1H-benzo[d]imidazol-1-yl)-3-(benzyloxy)propane-1-thiol (4) (3.95 g , 13.26 mmol), 4,5-bis(diphenyl-phosphino)-9,9-dimethyldibenzopyran (639 mg, 1.10 mmol) and ginseng(dibenzylideneacetone)dipalladium ( 674 mg, 0.736 mmol). The mixture was stirred at 60°C under nitrogen atmosphere for 18 hours. After completion, the mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane/ethyl acetate=3/1) to provide (S)-8-((2 -(1H-Benzo[d]imidazol-1-yl)-3-(benzyloxy)propyl)thio)-7-chloro-6-(trifluoromethyl)quinazoline-2,4- Diol (5) (3.8 g, 93% yield). MS (ESI) m/z 561 [M+H] ⁺ . (S)-8-((2-(1H- Benzo [d] imidazol -1- yl )-3- hydroxypropyl ) thio )-7- chloro - 6-( trifluoromethyl ) quinazoline -2,4- diol (6)

(S)-8-((2-(1H-Benzo[d]imidazol-1-yl)-3-(benzyloxy)propyl)thio)-7-chloro-6-(trifluoromethyl ) A solution of quinazoline-2,4-diol (5) (3.8 g, 6.78 mmol) in trifluoroacetic acid (55 mL). The mixture was stirred at 80°C for 18 hours. After completion, the mixture was concentrated at 0 °C and adjusted to pH = 7~8. After concentration, the residue was purified by a silica gel column with dichloromethane/ethyl acetate=1/2 to provide (S)-8-((2-(1H-benzo[d]imidazole -1-yl)-3-hydroxypropyl)thio)-7-chloro-6-(trifluoromethyl)quinazoline-2,4-diol (6) (2.5 g, 78% yield) . MS (ESI): m/z 471.0 [M+H] ⁺ . (S)-3-(1H- Benzo [d] imidazol -1- yl )-11- chloro -8- hydroxy -10-( trifluoromethyl )-3,4- dihydro- [1,4] Thiazeza [2,3,4-ij] quinazolin -6(2H) -one (7)

At 0°C, to (S)-8-((2-(1H-benzo[d]imidazol-1-yl)-3-hydroxypropyl)thio)-7-chloro-6-(trifluoro A mixture of methyl)quinazoline-2,4-diol (6) (2.56 g, 5.45 mmol) and triphenylphosphine (5.7 g, 21.78 mmol) in THF (750 mL) was added with azodicarboxylic acid Diethyl ester (3.79 g, 21.78 mmol). The mixture was stirred at 0 °C for 45 min. After completion, the mixture was poured into ice water (100 mL) and extracted with ethyl acetate (100 mL x 3). Concentrate and the residue is purified by C18 with 30-95% acetonitrile in water to afford (S)-3-(1H-benzo[d]imidazol-1-yl)-11-chloro-8- as a yellow solid. Hydroxy-10-(trifluoromethyl)-3,4-dihydro-[1,4]thiazeza[2,3,4-ij]quinazolin-6(2H)-one (7) ( 840 mg, 34% yield). MS (ESI) m/z 451.3 [MH] ^- . (2S,6R)-4-((S)-3-(1H- Benzo [d] imidazol -1- yl )-11- chloro -6- oxo -10-( trifluoromethyl )-2 ,3,4,6 - tetrahydro- [1,4] thiazepine [2,3,4-ij] quinazolin -8- yl )-2,6- dimethylpiperazine -1- carboxylic acid Tertiary butyl ester (8)

To (S)-3-(1H-benzo[d]imidazol-1-yl)-11-chloro-8-hydroxyl-10-(trifluoromethyl)-3,4-dihydro-[1,4 ] Thiazeza[2,3,4-ij]quinazolin-6(2H)-one (7) (680 mg, 1.5 mmol) and potassium carbonate (2.07 g, 15.0 mmol) in acetonitrile (50 mL) To the mixture in was added 2,4,6-triisopropylbenzenesulfonyl chloride (908 mg, 3.0 mmol). The mixture was stirred at 35°C for 4 hours. After completion, (2S,6R)-tert-butyl 2,6-dimethylpiperazine-1-carboxylate (802 mg, 3.75 mmol) was added to the reaction solution. The reaction mixture was stirred at 35°C for 1 hour. After completion, the mixture was poured into ice water (200 mL) and extracted with ethyl acetate (100 mL x 3). Concentrated and the residue was purified by C18 column with 20-95% acetonitrile in water to provide (2S,6R)-4-((S)-3-(1H-benzo[d]imidazole as a light yellow solid -1-yl)-11-chloro-6-oxo-10-(trifluoromethyl)-2,3,4,6-tetrahydro-[1,4]thiazepine[2,3, 4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester (8) (486 mg, 50% yield). MS (ESI) m/z 649.2 [M+H] ⁺ . (2S,6R)-4-((S)-3-(1H- Benzo [d] imidazol -1- yl )-11-(4- chloro- 5-( triisopropylsilyl ) thiophene -2 -yl )-6- oxo -10-( trifluoromethyl )-2,3,4,6 - tetrahydro- [1,4] thiazeb [2,3,4-ij] quinazole Lin -8- yl )-2,6- dimethylpiperazine -1- carboxylic acid tert-butyl ester (9)

To (2S,6R)-4-((S)-3-(1H-benzo[d]imidazol-1-yl)-11-chloro-6-oxo-10-(trifluoromethyl)- 2,3,4,6-tetrahydro-[1,4]thiazazo[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1- To a solution of tert-butyl formate (8) (350 mg, 0.54 mmol) in 1,4-dioxane (5 mL) and water (0.5 mL) was added tripotassium phosphate (343 mg, 1.62 mmol), (4 -Chloro-5-(triisopropylsilyl)thiophen-2-yl)boronic acid (515 mg, 1.62 mmol), and chloro(2-dicyclohexylphosphino-2',6'-diisopropoxy -1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (42 mg, 0.054 mmol). The mixture was stirred at 80°C for 1 hour under nitrogen atmosphere. After completion, the mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane/methanol=60/1) to provide (2S,6R)-4-((S) -3-(1H-Benzo[d]imidazol-1-yl)-11-(4-chloro-5-(triisopropylsilyl)thiophen-2-yl)-6-oxo-10- (Trifluoromethyl)-2,3,4,6-tetrahydro-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl)-2,6-di Methylpiperazine-1-carboxylic acid tert-butyl ester (9) (197 mg, 41% yield). MS (ESI) m/z 888 [M+H] ⁺ . (2S,6R)-4-((S)-3-(1H- Benzo [d] imidazol -1- yl )-11-(4- chlorothien -2- yl )-6- oxo -10 -( trifluoromethyl )-2,3,4,6- tetrahydro- [1,4] thiazeza [2,3,4-ij] quinazolin -8- yl )-2,6- tert-butyl dimethylpiperazine -1- carboxylate (10)

At 0°C, to (2S,6R)-4-((S)-3-(1H-benzo[d]imidazol-1-yl)-11-(4-chloro-5-(triisopropyl Silyl)thiophen-2-yl)-6-oxo-10-(trifluoromethyl)-2,3,4,6-tetrahydro-[1,4]thiazepine[2,3, A mixture of 4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester (9) (197 mg, 0.22 mmol) in THF (5 mL) was added Tetrabutylammonium fluoride (0.24 mL). The reaction solution was stirred at 0°C for 1 hour. After completion, the mixture was concentrated and extracted with ethyl acetate (100 mL×3). Concentrated and the residue was purified by C18 column with 20%-95% acetonitrile in water to provide (2S,6R)-4-((S)-3-(1H-benzo[d] as a pale yellow solid Imidazol-1-yl)-11-(4-chlorothien-2-yl)-6-oxo-10-(trifluoromethyl)-2,3,4,6-tetrahydro-[1,4 ]Thiazaza[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester (10) (81 mg, 50% yield Rate). MS (ESI) m/z 731.2 [M+H] ⁺ . (S)-3-(1H- Benzo [d] imidazol -1- yl )-11-(4- chlorothien -2- yl )-8-((3S,5R)-3,5- dimethyl Piperazin -1- yl )-10-( trifluoromethyl )-3,4- dihydro- [1,4] thiazeza [2,3,4-ij] quinazoline -6(2H) -ketone (11 )

At 0°C, to (2S,6R)-4-((S)-3-(1H-benzo[d]imidazol-1-yl)-11-(4-chlorothien-2-yl)-6 -Oxy-side-10-(trifluoromethyl)-2,3,4,6-tetrahydro-[1,4]thiazezo[2,3,4-ij]quinazolin-8-yl To a mixture of tert-butyl )-2,6-dimethylpiperazine-1-carboxylate (10) (81 mg, 0.11 mmol) in dichloromethane (3 mL) was added trifluoroacetic acid (1 mL). The reaction solution was stirred at room temperature for 1 hour. After completion, the mixture was concentrated and the residue was purified by silica gel column chromatography (dichloromethane/methanol=20/1) to provide (S)-3-(1H-benzo[d]imidazole as light yellow solid -1-yl)-11-(4-chlorothiophen-2-yl)-8-((3S,5R)-3,5-dimethylpiperazin-1-yl)-10-(trifluoromethyl )-3,4-dihydro-[1,4]thiazeza[2,3,4-ij]quinazolin-6(2H)-one (11) (60 mg, crude product). MS (ESI) m/z 631.2 [M+H] ⁺ . (S)-8-((3S,5R)-4- acryloyl - 3,5- dimethylpiperazin -1- yl )-3-(1H- benzo [d] imidazol -1- yl ) -11-(4- Chlorothiophen -2- yl )-10-( trifluoromethyl )-3,4- dihydro- [1,4] thiazazo [2,3,4-ij] quinazole Lin -6(2H) -one (12)

反應流程

(S)-(2- 疊氮基 -3-( 苄氧基 ) 丙基 )( 三苯甲基 ) 硫烷 (2) At 0°C, to (S)-3-(1H-benzo[d]imidazol-1-yl)-11-(4-chlorothien-2-yl)-8-((3S,5R)-3 ,5-Dimethylpiperazin-1-yl)-10-(trifluoromethyl)-3,4-dihydro-[1,4]thiazazo[2,3,4-ij]quinazole To a mixture of lin-6(2H)-one (11) (60 mg, crude product) and triethylamine (56 mg, 0.55 mmol) in dichloromethane (5 ml) was added acrylic anhydride (42 mg, 0.33 mmol ). The mixture was stirred at 0°C for 1 hour. After completion, the mixture was poured into ice water (30 mL) and extracted with ethyl acetate (30 mL x 3). Concentration and the residue was purified by preparative high performance liquid chromatography (20% to 95% acetonitrile in water) to provide (S)-8-((3S,5R)-4-acryloyl-3 as a white solid ,5-Dimethylpiperazin-1-yl)-3-(1H-benzo[d]imidazol-1-yl)-11-(4-chlorothien-2-yl)-10-(trifluoromethane yl)-3,4-dihydro-[1,4]thiazeza[2,3,4-ij]quinazolin-6(2H)-one (12) (2.5 mg, 3% yield) . MS (ESI) m/z 685.8 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.21 (s, 1H), 8.13 (s, 1H), 7.83-7.79 (m, 1H), 7.60-7.54 (m, 1H), 7.40-7.32 (m, 4H ), 6.66-6.59 (m, 1H), 6.43 (dd, J = 16.4 Hz, 1.6 Hz, 1H), 5.81 (dd, J = 10.8 Hz, 1.6 Hz, 1H), 5.39-5.32 (m, 4H), 5.12-4.95 (m, 3H), 4.76-4.61 (m, 2H), 4.24-4.17 (m, 2H), 2.01-2.00 (m, 6H). Example 28 : (S)-8-((3S,5R )-4- acryloyl -3,5- dimethylpiperazin-1-yl ) -11- (4- chlorothien -2- yl )-3-(1H-1,2,3 - triazole- 1- yl )-10-( trifluoromethyl )-3,4- dihydro- [1,4] thiazeza [2,3,4-ij] quinazolin -6(2H) -one

Reaction flow

(S)-(2- Azido -3-( benzyloxy ) propyl )( trityl ) sulfane (2)

Under a nitrogen atmosphere at 0°C, (R)-1-(benzyloxy)-3-(tritylthio)propan-2-ol ( 1 ) (20 g, 45.45 mmol) and triphenylphosphine (30 g, 113.6 mmol) in tetrahydrofuran (400 mL) was added diethyl azodicarboxylate (30 mL, 113.6 mmol). The mixture was stirred at room temperature for 10 min, then DPPA (11 mL, 47.73 mmol) was added. The mixture was stirred for 2 hours. After completion, the mixture was poured into ice water (300 mL) and extracted with ethyl acetate (150 mL x 3). After concentration, the residue was purified by silica gel column with petroleum ether/ethyl acetate=20/1 to afford crude (S)-(3-(benzyloxy)-2-(4-fluoro) as yellow oil Phenoxy)propyl)(trityl)sulfane (2) (16.5 g, crude product). ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.43-7.19 (m, 20H), 4.52-4.49 (m, 1H), 4.45 (s, 2H), 3.39-3.35 (m, 2H), 2.41-2.36 (m , 2H). (S)-1-(1-( Benzyloxy )-3-( tritylthio ) propan -2- yl )-5-( trimethylsilyl )-1H-1,2,3- tri Azole (3)

To a solution of (S)-(2-azido-3-(benzyloxy)propyl)(trityl)sulfane (2) (16.5 g, 35.48 mmol) in toluene (70 mL) was added Ethynyltrimethylsilane (13 mL, 88.71 mmol). The mixture was stirred at 100°C for 16 hours. After completion, the mixture was concentrated and purified by silica gel column with petroleum ether/ethyl acetate=15/1 to afford the product (S)-1-(1-(benzyloxy)-3-(tri Benzylthio)propan-2-yl)-5-(trimethylsilyl)-1H-1,2,3-triazole (3) (14.1 g, 55% yield for two steps ). ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.40-7.19 (m, 20H), 7.12-7.09 (m, 1H), 4.43 (s, 2H), 4.13-4.09 (m, 1H), 3.70-3.59 (m , 2H), 2.87-2.83 (m, 2H), 1.26 (s, 9H). (S)-1-(1-( Benzyloxy )-3-( tritylthio ) propan -2- yl )-1H-1,2,3- triazole (4)

To (S)-1-(1-(benzyloxy)-3-(tritylthio)prop-2-yl)-5-(trimethylsilyl)-1H-1,2,3- To a solution of triazole (3) (14.1 g 24.87 mmol) in tetrahydrofuran (70 mL) was added tetrabutylammonium fluoride (9.2 g, 35 mmol). The mixture was stirred at 40°C for 2 hours. After completion, the mixture was poured into ice water (100 mL) and extracted with ethyl acetate (100 mL x 3). After concentration, the residue was purified by a silica gel column with petroleum ether/ethyl acetate=10/1 to provide (S)-1-(1-(benzyloxy)-3-(tritylthio) Propan-2-yl)-1H-1,2,3-triazole (4) (12 g, 98% yield). ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.61 (s, 1H), 7.40-7.19 (m, 20H), 7.13-7.11 (m, 1H), 4.35 (s, 2H), 4.15-4.01 (m, 1H ), 3.75-3.57 (m, 2H), 2.87-2.84 (m, 2H). (S)-3-( Benzyloxy )-2-(1H-1,2,3- triazol -1- yl ) propane -1- thiol (5)

To (S)-1-(1-(benzyloxy)-3-(tritylthio)propan-2-yl)-1H-1,2,3-triazole (4) (12 g, 24.87 mmol) and triethylsilane (2.9 g, 24.87 mmol) in dichloromethane (40 mL) was added trifluoroacetic acid (40 mL). The mixture was stirred at room temperature for 2 hours. After completion, the mixture was concentrated at 0 °C and adjusted to pH=7~8. After concentration, the residue was purified by silica gel column with petroleum ether/ethyl acetate=5/1 to afford (S)-3-(benzyloxy)-2-(1H-1,2 ,3-triazol-1-yl)propane-1-thiol (5) (5 g, 83% yield). ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.73-7.71 (m, 2H), 7.38-7.28 (m, 4H), 7.25-7.23 (m, 1H), 4.82-4.80 (m, 1H), 4.55-4.47 (m, 2H), 3.99-3.86 (m, 2H), 3.17-3.09 (m, 2H), 2.05 (s, 1H). (S)-8-((3-( Benzyloxy )-2-(1H-1,2,3- triazol -1- yl ) propyl ) thio )-7- chloro - 6-( trifluoro Methyl ) quinazoline -2,4- diol (6)

To a solution of 7-chloro-8-iodo-6-(trifluoromethyl)quinazoline-2,4-diol (5.3 g, 13.39 mmol) in 1,4-dioxane (100 mL) was added Potassium carbonate (5.5 g, 40.17 mmol), (S)-3-(benzyloxy)-2-(1H-1,2,3-triazol-1-yl)propane-1-thiol (5) ( 5 g, 20.08 mmol), 4,5-bis(diphenyl-phosphino)-9,9-dimethyldibenzopyran (1.17 g, 2.01 mmol) and ginseng (dibenzylideneacetone) di Palladium (1.23 g, 1.34 mmol). The mixture was stirred at 60°C under nitrogen atmosphere for 18 hours. After completion, the mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane/ethyl acetate=4/1) to provide (S)-8-((3 -(Benzyloxy)-2-(1H-1,2,3-triazol-1-yl)propyl)thio)-7-chloro-6-(trifluoromethyl)quinazoline-2, 4-diol (6) (6.8 g, 97% yield). MS (ESI) m/z 512.0 [M+H] ⁺ . (S)-7- chloro -8-((3- hydroxy -2-(1H-1,2,3- triazol -1- yl ) propyl ) thio )-6-( trifluoromethyl ) quinone Oxazoline -2,4- diol (7)

(S)-8-((3-(Benzyloxy)-2-(1H-1,2,3-triazol-1-yl)propyl)thio)-7-chloro-6-(trifluoro A solution of methyl)quinazoline-2,4-diol (6) (6.8 g, 13.70 mmol) in trifluoroacetic acid (12 mL). The mixture was stirred at 80°C for 18 hours. After completion, the mixture was concentrated at 0 °C and adjusted to pH=7~8. After concentration, the residue was purified by silica gel column with dichloromethane/ethyl acetate=1/2 to provide (S)-7-chloro-8-((3-hydroxy-2-( 1H-1,2,3-triazol-1-yl)propyl)thio)-6-(trifluoromethyl)quinazoline-2,4-diol (7) (6.0 g, crude product) . MS (ESI): m/z 422.1 [M+H] ⁺ . (S)-11- chloro -8- hydroxyl -3-(1H-1,2,3 - triazol -1- yl )-10-( trifluoromethyl )-3,4- dihydro- [1, 4] Thiazaza [2,3,4-ij] quinazolin -6(2H) -one (8)

At 0°C, to (S)-7-chloro-8-((3-hydroxyl-2-(1H-1,2,3-triazol-1-yl)propyl)thio)-6-( A mixture of trifluoromethyl)quinazoline-2,4-diol (7) (6.0 g, 13.54 mmol) and triphenylphosphine (11.0 g, 40.62 mmol) in tetrahydrofuran (400 mL) was added with azobis Diethyl carboxylate (11 mL, 40.62 mmol). The mixture was stirred at 0 °C for 45 min. After completion, the mixture was poured into ice water (100 mL) and extracted with ethyl acetate (100 mL x 3). The organic phase was concentrated and the residue was purified by C18 with 30-95% acetonitrile in water to afford (S)-11-chloro-8-hydroxy-3-(1H-1,2,3-triazole as a yellow solid -1-yl)-10-(trifluoromethyl)-3,4-dihydro-[1,4]thiazeza[2,3,4-ij]quinazolin-6(2H)-one (8) (4.2 g, 76% yield for two steps). MS (ESI) m/z 402.0 [MH] ⁺ . (2S,6R)-4-((S)-11- chloro -6- oxo -3-(1H-1,2,3- triazol -1- yl )-10-( trifluoromethyl ) -2,3,4,6 - tetrahydro- [1,4] thiazazo [2,3,4-ij] quinazolin- 8- yl )-2,6- dimethylpiperazine -1 - Tertiary butyl formate (9)

To (S)-11-chloro-8-hydroxyl-3-(1H-1,2,3-triazol-1-yl)-10-(trifluoromethyl)-3,4-dihydro-[1 ,4]Thiazeza[2,3,4-ij]quinazolin-6(2H)-one (8) (2.4 g, 5.96 mmol) and potassium carbonate (8.23 g, 59.6 mmol) in acetonitrile (50 mL) was added 2,4,6-paras(prop-2-yl)benzenesulfonyl chloride (7.22 g, 23.82 mmol). The mixture was stirred at 35°C for 4 hours. After completion, (2S,6R)-tert-butyl 2,6-dimethylpiperazine-1-carboxylate (5.1 g, 23.82 mmol) was added to the reaction solution. The reaction mixture was stirred at 35°C for 1 hour. After completion, the mixture was poured into ice water (200 mL) and extracted with ethyl acetate (100 mL x 3). Concentrate and the residue is purified by C18 column with 20-95% acetonitrile in water to afford (2S,6R)-4-((S)-11-chloro-6-oxo-3 as a light yellow solid -(1H-1,2,3-triazol-1-yl)-10-(trifluoromethyl)-2,3,4,6-tetrahydro-[1,4]thiazepine[2, tert-butyl 3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylate (9) (1.5 g, 43% yield). MS (ESI) m/z 600.3 [M+H] ⁺ . (2S,6R)-4-((S)-11-(4- Chloro -5-( triisopropylsilyl ) thiophen -2- yl )-6- oxo -3-(1H-1, 2,3- triazol -1- yl )-10-( trifluoromethyl )-2,3,4,6- tetrahydro- [1,4] thiazepine [2,3,4-ij] Quinazolin -8- yl )-2,6- dimethylpiperazine -1- carboxylic acid tert-butyl ester (10)

To (2S,6R)-4-((S)-11-chloro-6-oxo-3-(1H-1,2,3-triazol-1-yl)-10-(trifluoromethyl )-2,3,4,6-tetrahydro-[1,4]thiazazo[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine- To a solution of tert-butyl 1-carboxylate (9) (200 mg, 0.33 mmol) in 1,4-dioxane (3 mL) and water (0.5 mL) was added tripotassium phosphate (270 mg, 1.0 mmol), (4-Chloro-5-(triisopropylsilyl)thiophen-2-yl)boronic acid (430 mg, 1.34 mmol), and chloro(2-dicyclohexylphosphino-2',6'-diisopropyl Oxy-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (40 mg, 0.05 mmol). The mixture was stirred at 80°C for 1 hour under nitrogen atmosphere. After completion, the mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane/methanol=20/1) to provide (2S,6R)-4-((S)- 11-(4-Chloro-5-(triisopropylsilyl)thiophen-2-yl)-6-oxo-3-(1H-1,2,3-triazol-1-yl)-10 -(trifluoromethyl)-2,3,4,6-tetrahydro-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl)-2,6- Dimethylpiperazine-1-carboxylic acid tert-butyl ester (10) (175 mg, 63% yield). MS (ESI) m/z 838.0 [M+H] ⁺ . (2S,6R)-4-((S)-11-(4- chlorothien- 2- yl )-6- oxo -3-(1H-1,2,3- triazol -1- yl ) -10-( trifluoromethyl )-2,3,4,6- tetrahydro- [1,4] thiazeza [2,3,4-ij] quinazolin- 8- yl )-2, 6- Dimethylpiperazine -1- carboxylic acid tert-butyl ester (11)

To (2S,6R)-4-((S)-11-(4-chloro-5-(triisopropylsilyl)thiophen-2-yl)-6-oxo-3-(1H-1 ,2,3-triazol-1-yl)-10-(trifluoromethyl)-2,3,4,6-tetrahydro-[1,4]thiazepine[2,3,4-ij ]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester (10) (150 mg, 0.2 mmol) in tetrahydrofuran (1 mL) was added tetrabutyl Ammonium fluoride (130 mg, 0.5 mmol). The mixture was stirred at 40°C for 2 hours. After completion, the mixture was poured into ice water (5 mL) and extracted with ethyl acetate (10 mL x 3). After concentration, the residue was purified by a silica gel column with dichloromethane/methanol=80/1 to provide (2S,6R)-4-((S)-11-(4-chlorothiophene- 2-yl)-6-oxo-3-(1H-1,2,3-triazol-1-yl)-10-(trifluoromethyl)-2,3,4,6-tetrahydro- [1,4]Thiazaza[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester (11) (150 mg , crude product). MS (ESI) m/z 682.2 [M+H] ⁺ . (S)-11-(4- chlorothien -2- yl )-8-((3S,5R)-3,5- dimethylpiperazin -1- yl )-3-(1H-1,2, 3- triazol -1- yl )-10-( trifluoromethyl )-3,4- dihydro- [1,4] thiazezo [2,3,4-ij] quinazoline -6( 2H) -ketone (12)

At 0°C, to (2S,6R)-4-((S)-11-(4-chlorothiophen-2-yl)-6-oxo-3-(1H-1,2,3-tri Azol-1-yl)-10-(trifluoromethyl)-2,3,4,6-tetrahydro-[1,4]thiazazo[2,3,4-ij]quinazoline-8 To a mixture of tert-butyl)-2,6-dimethylpiperazine-1-carboxylate (11) (150 mg, crude product) in dichloromethane (1 mL) was added trifluoroacetic acid (1 mL) . The reaction solution was stirred at room temperature for 1 hour. After completion, the mixture was concentrated and the residue was purified by silica gel column chromatography (dichloromethane/methanol=15/1) to provide (S)-11-(4-chlorothiophen-2-yl as light yellow solid )-8-((3S,5R)-3,5-Dimethylpiperazin-1-yl)-3-(1H-1,2,3-triazol-1-yl)-10-(trifluoro Methyl)-3,4-dihydro-[1,4]thiazazo[2,3,4-ij]quinazolin-6(2H)-one (12) (80 mg, for two steps In terms of 69% yield). MS (ESI) m/z 582.3 [M+H] ⁺ . (S)-8-((3S,5R)-4- acryloyl -3,5- dimethylpiperazin -1- yl )-11-(4- chlorothiophen -2- yl )-3-( 1H-1,2,3- triazol -1- yl )-10-( trifluoromethyl )-3,4- dihydro- [1,4] thiazepine [2,3,4-ij] Quinazolin -6(2H) -one (13)

步驟1：(S)-2-((1-((第三丁基二苯基矽烷基)氧基)-3-(三苯甲硫基)丙-2-基)氧基)吡嗪

At 0°C, to (S)-11-(4-chlorothiophen-2-yl)-8-((3S,5R)-3,5-dimethylpiperazin-1-yl)-3-( 1H-1,2,3-triazol-1-yl)-10-(trifluoromethyl)-3,4-dihydro-[1,4]thiazepine[2,3,4-ij] A mixture of quinazolin-6(2H)-one (12) (80 mg, 0.138 mmol) and triethylamine (28 mg, 0.275 mmol) in dichloromethane (2 ml) was added with acrylic anhydride (26 mg, 0.207 mmol). The mixture was stirred at 0°C for 1 hour. After completion, the mixture was poured into ice water (10 mL) and extracted with ethyl acetate (10 mL x 3). Concentration and the residue was purified by preparative high performance liquid chromatography (20% to 95% acetonitrile in water) to provide (S)-8-((3S,5R)-4-acryloyl-3 as a white solid ,5-Dimethylpiperazin-1-yl)-11-(4-chlorothien-2-yl)-3-(1H-1,2,3-triazol-1-yl)-10-(tri Fluoromethyl)-3,4-dihydro-[1,4]thiazazo[2,3,4-ij]quinazolin-6(2H)-one (13) (12 mg, 14% yield Rate). MS (ESI) m/z 636.2 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.11 (s, 1H), 7.87-7.77 (m, 2H), 7.36 (s, 1H), 6.98 (s, 1H), 6.67-6.59 (m, 1H), 6.42 (dd, J = 16.4 Hz, 2.0 Hz, 1H), 5.79 (dd, J = 10.4 Hz, 1.6 Hz 1H), 5.64-5.42 (m, 1H), 5.29-5.17 (m, 1H), 5.11-4.95 (m, 1H), 4.79-4.64 (m, 2H), 4.21-4.18 (m, 2H), 3.80-3.67 (m, 1H), 3.41-3.38 (m, 3H), 1.55-1.54 (m, 6H) . Example 29 : (S)-8-((3S,5R)-4- acryloyl -3,5- dimethylpiperazin -1- yl )-11-(4- chlorothiophen -2- yl )- 3-( pyrazin -2- yloxy )-10-( trifluoromethyl )-3,4- dihydro -2H,6H-[1,4] thiazeza [2,3,4-ij ] quinazolin -6- one

Step 1: (S)-2-((1-((tertyldiphenylsilyl)oxy)-3-(tritylthio)propan-2-yl)oxy)pyrazine

Sodium hydride (42 mmol) was added to (S)-1-((tert-butyldiphenylsilyl)oxy)-3-(tritylthio)propan-2-ol (17 mmol) in 0°C solution in THF (100 mL). After 30 minutes, 2-fluoropyrazine (25.5 mmol) was added, and the mixture was stirred at room temperature for 16 hours. The reaction was diluted with water, extracted with ethyl acetate and the combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to provide the title compound as a white solid in 93% yield. MS (ESI) m/z: 667.0 [M+1]+. Step 2: (S)-3-((tert-butyldiphenylsilyl)oxy)-2-(pyrazin-2-yloxy)propane-1-thiol

Add triethylsilane (37 mmol) to (S)-2-((1-((tert-butyldiphenylsilyl)oxy)-3-(tritylthio)propane-2- (1)oxy)pyrazine (19 mmol) in TFA (30 mL) and dichloromethane (90 mL) at 0 °C. The reaction was allowed to reach room temperature over 60 minutes and water was added at this point. The mixture was extracted three times with dichloromethane and the combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to provide a residue which was analyzed by silica gel chromatography (0-10 in dichloromethane % methanol) to provide the title compound as a yellow oil in 90% yield. MS (ESI) m/z: 425.0 [M+1]+. Step 3: (S)-8-((3-((tert-butyldiphenylsilyl)oxy)-2-(pyrazin-2-yloxy)propyl)thio)-7- Chloro-6-(trifluoromethyl)quinazoline-2,4(1H,3H)-dione

7-chloro-8-iodo-6-(trifluoromethyl)quinazoline-2,4(1H,3H)-dione (6.10 mmol), (S)-3-((tertiary butyldi Phenylsilyl)oxy)-2-(pyrazin-2-yloxy)propane-1-thiol (9.1 mmol), Xantphos (1.2 mmol), diisopropylethylamine (24.4 mmol) and Pd2 A mixture of (dba)3 (0.06 mmol) in dioxane (20 mL) was stirred at 100 °C for two hours. The reaction was allowed to cool to room temperature and the volatiles were removed under reduced pressure to provide a residue which was purified by reverse phase chromatography (0-100% acetonitrile in water). The title compound was isolated in 93% yield as a yellow solid. MS (ESI) m/z: 681.0 [M+1]+. Step 4: (S)-7-Chloro-8-((3-hydroxy-2-(pyrazin-2-yloxy)propyl)thio)-6-(trifluoromethyl)quinazoline- 2,4(1H,3H)-Diketone

A 1M solution of TBAF in THF (15 mL) was added to (S)-8-((3-((tert-butyldiphenylsilyl)oxy)-2-(pyrazin-2-yloxy A solution of (5.0 mmol) in THF (1 mL). After one hour, the reaction mixture was diluted with water, extracted with ethyl acetate and the combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to provide the title compound as a brown solid in 44% yield. MS (ESI) m/z: 449.0 [M+1]+. Step 5: (S)-11-Chloro-3-(pyrazin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]sulfur Aza[2,3,4-ij]quinazoline-6,8(7H)-dione

A 0 °C solution of DIAD (3.81 mmol) in THF (5 mL) was added to a 0 °C solution of triphenylphosphine (3.61 mmol) in THF (5 mL). After 10 minutes, (S)-7-chloro-8-((3-hydroxy-2-(pyrazin-2-yloxy)propyl)thio)-6-(trifluoromethyl)quinazole was added Phenyl-2,4(1H,3H)-dione (10 mL) in 0.2 M THF and the mixture was stirred at room temperature for 16 hours. Volatiles were removed under reduced pressure to provide a residue which was purified by reverse phase chromatography (0-100% acetonitrile in water). The title compound was isolated in 44% yield as a white solid. MS (ESI) m/z: 431.0 [M+1]+. Step 6: (2S,6R)-4-((S)-11-Chloro-6-oxo-3-(pyrazin-2-yloxy)-10-(trifluoromethyl)-3, 4-Dihydro-2H, 6H-[1,4]thiazazo[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tributyl ester

(S)-11-chloro-3-(pyrazin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine And[2,3,4-ij]quinazoline-6,8(7H)-dione (0.77 mmol), N,N'-diisopropylethylamine (7.66 mmol) and POCl3 (4 mL) in The solution in toluene (4 mL) was stirred at 120 °C for 90 min. The volatiles were removed under reduced pressure and the residue was redissolved in dichloroethane (5 mL). This solution was added to tert-butyl (2R,6S)-2,6-dimethylpiperazine-1-carboxylate (3.10 mmol) and N,N'-diisopropylethylamine (7.66 mmol) in 2 solution in ethyl chloride at 0°C and the mixture was stirred for an additional hour at room temperature. Volatiles were removed under reduced pressure to provide a residue which was purified by silica gel chromatography. The title compound was isolated in 51% yield as a yellow solid. MS (ESI) m/z: 627.0 [M+1]+. Step 7: (2S,6R)-4-((S)-11-(4-Chloro-5-(triisopropylsilyl)thiophen-2-yl)-6-oxo-3-(pyridine Oxyzin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazoline -8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester

The title compound was prepared analogously to Example 23, Step 1, wherein (2S,6R)-4-((S)-11-chloro-6-oxo-3-(pyrimidin-2-yloxy)-10- (Trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl)-2,6-di Methylpiperazine-1-butyl carboxylate was replaced by (2S,6R)-4-((S)-11-chloro-6-oxo-3-(pyrazin-2-yloxy)- 10-(Trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl)-2,6 - tert-butyl dimethylpiperazine-1-carboxylate. The title compound was isolated in 88% yield as a brown solid. MS (ESI) m/z = 865.0 [M+H]+. Step 8: (S)-11-(4-chloro-5-(triisopropylsilyl)thiophen-2-yl)-8-((3S,5R)-3,5-dimethylpiperazine- 1-yl)-3-(pyrazin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2, 3,4-ij]quinazolin-6-one

The title compound was prepared analogously to Example 23, Step 3, wherein (2S,6R)-4-((S)-11-(4-chlorothien-2-yl)-6-oxo-3-(pyrimidine- 2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazoline-8 -yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester was replaced by (2S,6R)-4-((S)-11-(4-chloro-5-(triisopropyl Silyl)thiophen-2-yl)-6-oxo-3-(pyrazin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[ 1,4]Thiazaza[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester. The title compound was isolated in 80% yield as a yellow oil. MS (ESI) m/z = 765.0 [M+H]+. Step 9: (S)-11-(4-Chlorothiophen-2-yl)-8-((3S,5R)-3,5-dimethylpiperazin-1-yl)-3-(pyrazine- 2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazoline-6 -ketone

The title compound was prepared analogously to Example 23, step 2, wherein (2S,6R)-4-((S)-11-(4-chloro-5-(triisopropylsilyl)thiophen-2-yl)- 6-oxo-3-(pyrimidin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2, 3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester is replaced by (S)-11-(4-chloro-5-(triiso Propylsilyl)thiophen-2-yl)-8-((3S,5R)-3,5-dimethylpiperazin-1-yl)-3-(pyrazin-2-yloxy)-10 -(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-6-one. The title compound was isolated in 83% yield as a yellow oil. MS (ESI) m/z = 609.0 [M+H]+. Step 10: (S)-8-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-11-(4-chlorothien-2-yl)- 3-(pyrazin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij ]quinazolin-6-one

步驟1：(S)-3-(三苯甲硫基)丙烷-1,2-二醇

The title compound was prepared analogously to Example 23, step 4, wherein (S)-11-(4-chlorothien-2-yl)-8-((3S,5R)-3,5-dimethylpiperazine-1 -yl)-3-(pyrimidin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3, 4-ij] quinazolin-6-one was replaced by (S)-11-(4-chlorothiophen-2-yl)-8-((3S,5R)-3,5-dimethylpiperazine-1 -yl)-3-(pyrazin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3 ,4-ij]quinazolin-6-one. The title compound was isolated in 10% yield as a white solid. MS (ESI) m/z = 663.0 [M+H]+. 1H NMR (400 MHz, CDCl3) δ 8.27 - 8.22 (m, 1H), 8.20 - 8.15 (m, 1H), 8.09 - 8.06 (m, 2H), 7.36 - 7.35 (m, 1H), 7.00 - 6.87 (m , 1H), 6.67 - 6.57 (m, 1H), 6.45 - 6.38 (m, 1H), 5.82 - 5.58 (m, 2H), 5.01 - 4.78 (m, 1H), 4.71 - 4.58 (m, 2H), 4.27 - 4.09 (m, 2H), 3.70 - 3.26 (m, 4H), 3.19 - 3.02 (m, 1H), 1.64 - 1.50 (m, 6H). Example 31 : (S)-8-((3S,5R)-4- acryloyl -3,5- dimethylpiperazin -1- yl )-11-(4- chlorothiophen -2- yl )- 3-( pyridin -2- yloxy )-10-( trifluoromethyl )-3,4- dihydro -2H,6H-[1,4] thiazepine [2,3,4-ij] Quinazolin -6- one

Step 1: (S)-3-(Tritylthio)propane-1,2-diol

Potassium tert-butoxide (498 mmol) was added to a 0 °C solution of trityl mercaptan (452 mmol) in DMF (800 mL). After 30 minutes, a solution of (2S)-3-chloropropane-1,2-diol (452 mmol) in DMF (200 mL) was added and the mixture was allowed to slowly reach room temperature and stirred overnight. The reaction mixture was diluted with water, extracted with ethyl acetate and the combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to provide a residue which was analyzed by silica gel chromatography (10% ethyl acetate in hexane ester) to afford the title compound as a yellow oil in 97% yield. MS (ESI) m/z = 351.1 [M+H]+. Step 2: (S)-1-((tert-butyldiphenylsilyl)oxy)-3-(tritylthio)propan-2-ol

Imidazole (228 mmol) was added to a 0° C. solution of (S)-3-(tritylthio)propane-1,2-diol (143 mmol) in dichloroethane (500 mL). After 30 minutes, tert-butylbiphenylchlorosilane (171 mmol) was added and the reaction was stirred at room temperature overnight. The reaction was filtered and the volatiles were removed under reduced pressure to provide a residue which was purified by silica gel chromatography (0-10% ethyl acetate in hexanes) to provide β-R as a colorless oil in 58% yield. title compound. MS (ESI) m/z = 589.3 [M+H]+. Step 3: (S)-2-((1-((tertyldiphenylsilyl)oxy)-3-(tritylthio)propan-2-yl)oxy)pyridine

The title compound was prepared analogously to Example 29, Step 1, wherein 2-fluoropyrazine was replaced by 2-fluoropyridine. The title compound was isolated in 75% yield as a colorless oil. MS (ESI) m/z = 666.3 [M+H]+. Step 4: (S)-2-(pyridin-2-yloxy)-3-(tritylthio)propan-1-ol

The title compound was prepared analogously to Example 23, step 2, wherein (2S,6R)-4-((S)-11-(4-chloro-5-(triisopropylsilyl)thiophen-2-yl)- 6-oxo-3-(pyrimidin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2, 3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tertiary butyl ester is replaced by (S)-2-((1-((tertiary butyl Diphenylsilyl)oxy)-3-(tritylthio)propan-2-yl)oxy)pyridine. The title compound was isolated in 82% yield as a colorless oil. MS (ESI) m/z = 428.2 [M+H]+. Step 5: (S)-3-Mercapto-2-(pyridin-2-yloxy)propan-1-ol

The title compound was prepared analogously to Example 29, step 2, wherein (S)-2-((1-((tert-butyldiphenylsilyl)oxy)-3-(tritylthio)propane- 2-yl)oxy)pyrazine was replaced by (S)-2-(pyridin-2-yloxy)-3-(tritylthio)propan-1-ol. The title compound was isolated in quantitative yield as a colorless solid. Step 6: (S)-7-Chloro-8-((3-hydroxy-2-(pyridin-2-yloxy)propyl)thio)-6-(trifluoromethyl)quinazoline-2 ,4(1H,3H)-Diketone

The title compound was prepared analogously to Example 29, step 3, wherein (S)-3-((tert-butyldiphenylsilyl)oxy)-2-(pyrazin-2-yloxy)propane-1 Displacement of -thiol by (S)-3-mercapto-2-(pyridin-2-yloxy)propan-1-ol. The title compound was isolated in 42% yield as a pale green solid. MS (ESI) m/z = 447.9 [M+H]+. Step 7: (S)-11-Chloro-3-(pyridin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]sulfuride Qua[2,3,4-ij]quinazoline-6,8(7H)-dione

The title compound was prepared analogously to Example 29, Step 5, wherein (S)-7-chloro-8-((3-hydroxy-2-(pyrazin-2-yloxy)propyl)thio)-6- (Trifluoromethyl)quinazoline-2,4(1H,3H)-dione was replaced by (S)-7-chloro-8-((3-hydroxy-2-(pyridin-2-yloxy) Propyl)thio)-6-(trifluoromethyl)quinazoline-2,4(1H,3H)-dione. The title compound was isolated in 28% yield as a white solid. MS (ESI) m/z = 430.0 [M+H]+. Step 8: (2S,6R)-4-((S)-11-Chloro-6-oxo-3-(pyridin-2-yloxy)-10-(trifluoromethyl)-3,4 -Dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid third Butyl ester

The title compound was prepared analogously to Example 29, step 6, wherein (S)-11-chloro-3-(pyrazin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro- 2H,6H-[1,4]thiazaza[2,3,4-ij]quinazoline-6,8(7H)-dione was replaced by (S)-11-chloro-3-(pyridine- 2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazoline-6 ,8(7H)-diketone. The title compound was isolated in 78% yield as a pale yellow solid. MS (ESI) m/z = 626.2 [M+H]+. Step 9: (2S,6R)-4-((S)-11-(4-Chloro-5-(triisopropylsilyl)thiophen-2-yl)-6-oxo-3-(pyridine -2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazoline- 8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester

The title compound was prepared analogously to Example 23, Step 1, wherein (2S,6R)-4-((S)-11-chloro-6-oxo-3-(pyrimidin-2-yloxy)-10- (Trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl)-2,6-di Methylpiperazine-1-carboxylic acid tert-butyl ester was replaced by (2S,6R)-4-((S)-11-chloro-6-oxo-3-(pyridin-2-yloxy)-10 -(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl)-2,6- Dimethylpiperazine-1-carboxylic acid tert-butyl ester. The title compound was isolated in 36% yield as a yellow oil. MS (ESI) m/z = 865.6 [M+H]+. Step 10: (2S,6R)-4-((S)-11-(4-Chlorothiophen-2-yl)-6-oxo-3-(pyridin-2-yloxy)-10-( Trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl)-2,6-dimethyl Tributyl piperazine-1-carboxylate

The title compound was prepared analogously to Example 23, step 2, wherein (2S,6R)-4-((S)-11-(4-chloro-5-(triisopropylsilyl)thiophen-2-yl)- 6-oxo-3-(pyrimidin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2, 3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester is replaced by (2S,6R)-4-((S)-11-( 4-Chloro-5-(triisopropylsilyl)thiophen-2-yl)-6-oxo-3-(pyridin-2-yloxy)-10-(trifluoromethyl)-3, 4-Dihydro-2H, 6H-[1,4]thiazazo[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tributyl ester. The title compound was isolated in 87% yield as a white solid. MS (ESI) m/z = 708.2 [M+H]+. Step 11: (S)-11-(4-Chlorothiophen-2-yl)-8-((3S,5R)-3,5-dimethylpiperazin-1-yl)-3-(pyridine-2 -yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazoline-6- ketone

The title compound was prepared analogously to Example 23, Step 3, wherein (2S,6R)-4-((S)-11-(4-chlorothien-2-yl)-6-oxo-3-(pyrimidine- 2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazoline-8 -yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester was replaced by (2S,6R)-4-((S)-11-(4-chlorothien-2-yl)-6 -Oxy-3-(pyridin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3 ,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester. The title compound was isolated in 87% yield as a white solid. MS (ESI) m/z = 608.2 [M+H]+. Step 12: (S)-8-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-11-(4-chlorothien-2-yl)- 3-(pyridin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3,4-ij] Quinazolin-6-one

The title compound was prepared analogously to Example 23, step 4, wherein (S)-11-(4-chlorothien-2-yl)-8-((3S,5R)-3,5-dimethylpiperazine-1 -yl)-3-(pyrimidin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3, 4-ij] quinazolin-6-one was replaced by (S)-11-(4-chlorothiophen-2-yl)-8-((3S,5R)-3,5-dimethylpiperazine-1 -yl)-3-(pyridin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3, 4-ij] quinazolin-6-one. The title compound was isolated in 13% yield as a white solid. MS (ESI) m/z = 662.2 [M+H]+. 1H NMR (400MHz, CDCl3) δ 8.14 - 8.05 (m, 2H), 7.59 (ddd, J = 2.0, 7.2, 8.4 Hz, 1H), 7.34 (s, 1H), 7.01 - 6.86 (m, 2H), 6.77 (d, J = 8.4 Hz, 1H), 6.63 (dd, J = 10.4, 16.8 Hz, 1H), 6.42 (dd, J = 1.6, 16.8 Hz, 1H), 5.84 - 5.70 (m, 2H), 5.00 - 4.57 (m, 4H), 4.20 - 4.10 (m, 2H), 3.52 - 3.33 (m, 4H), 1.55 (br s, 6H). Example 32 : (S)-8-((3S,5R)-4- acryloyl -3,5- dimethylpiperazin -1- yl )-11-(4- chlorothiophen -2- yl )- 3-( Pyridazin -3- yloxy )-10-( trifluoromethyl )-3,4- dihydro -2H,6H-[1,4] thiazepine [2,3,4-ij ] quinazolin -6- one

The title compound was prepared analogously to Example 29, wherein in Step 1 2-fluoropyrazine was replaced by 2-chloropyrazine. The title compound was isolated as a white solid. MS (ESI) m/z: 663.0 [M+H]+. 1H NMR (400 MHz, CDCl3) δ 8.93 (d, J = 4.0 Hz, 1H), 8.09 (s, 1H), 7.66 - 7.56 (m, 1H), 7.35 (d, J = 1.2 Hz, 1H), 7.21 (d, J = 8.4 Hz, 1H), 7.03 - 6.87 (m, 1H), 6.68 - 6.58 (m, 1H), 6.42 (dd, J = 2.0, 16.6 Hz, 1H), 5.89 - 5.76 (m, 2H ), 4.97 - 4.89 (m, 1H), 4.83 (s, 2H), 4.75 - 4.70 (m, 1H), 4.27 - 4.15 (m, 2H), 3.48 - 3.39 (m, 2H), 3.38 - 3.31 (m , 1H), 3.17 (dd, J = 2.4, 13.6 Hz, 1H), 1.61 (d, J = 7.2 Hz, 3H), 1.50 (d, J = 7.2 Hz, 3H). Example 33 : (S)-8-((3S,5R)-4- acryloyl -3,5- dimethylpiperazin -1- yl )-11-(4- chlorothiophen -2- yl )- 3-( pyrimidin -4- yloxy )-10-( trifluoromethyl )-3,4- dihydro -2H,6H-[1,4] thiazepine [2,3,4-ij] Quinazolin -6- one

步驟1：2,4,7-三氯-8-碘-6-(三氟甲基)喹唑啉

The title compound was prepared analogously to Example 29, wherein in Step 1 2-fluoropyrazine was replaced by 4-chloropyrimidine. The title compound was isolated as a yellow solid. MS (ESI) m/z: 690.0 [M+Na]+. Example 35 : (S)-8-((3S,5R)-4- acryloyl -3,5- dimethylpiperazin -1- yl )-11-(4- chlorothiophen -2- yl )- 3-( Pyridin -4- yl )-10-( trifluoromethyl )-3,4- dihydro -2H,6H-[1,4] thiazazo [2,3,4-ij] quinazole Lin -6- one

Step 1: 2,4,7-Trichloro-8-iodo-6-(trifluoromethyl)quinazoline

To 7-chloro-8-iodo-6-(trifluoromethyl)quinazoline-2,4(1H,3H)-dione (5.1 mmol) in N,N-diisopropylethylamine (17.9 mmol ) was added phosphoryl trichloride (12 mL). The mixture was stirred at 110°C for 12 hours and the reaction mixture was concentrated under reduced pressure to provide a residue which was dissolved in cold water and extracted three times with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The resulting crude material was purified by silica gel chromatography (0-15% ethyl acetate in petroleum ether) to afford the title compound as a white solid in 44% yield. MS (ESI) m/z: 427.0 [M+1]+. Step 2: (2S,6R)-4-(2,7-Dichloro-8-iodo-6-(trifluoromethyl)quinazolin-4-yl)-2,6-dimethylpiperazine- 1-tert-butyl carboxylate

Add 2,4,7-trichloro-8-iodo-6-(trifluoromethyl)quinazoline (2.04 mmol) and triethylamine (6.11 mmol) in dichloromethane (10 mL) at 0°C To the solution in was added tert-butyl (2S,6R)-2,6-dimethylpiperazine-1-carboxylate (1.83 mmol). The mixture was stirred at room temperature for 1 hour and the volatiles were removed under reduced pressure. The residue was purified by silica gel column chromatography (5-15% ethyl acetate in hexanes) to afford the title compound as a white solid in 82% yield. MS (ESI) m/z: 605.2 [M+H]+. Step 3: (2S,6R)-4-(7-Chloro-8-iodo-2-oxo-6-(trifluoromethyl)-1,2-dihydroquinazolin-4-yl)- 2,6-Dimethylpiperazine-1-carboxylic acid tert-butyl ester

To (2S,6R)-4-(2,7-dichloro-8-iodo-6-(trifluoromethyl)quinazolin-4-yl)-2,6-dimethylpiperazine-1- To a solution of tert-butyl formate (1.73 mmol) in acetonitrile (50 mL) was added 2-(methylsulfonyl)ethan-1-ol (3.47 mmol), cesium carbonate (3.47 mmol) and 1,4-diazepine Heterobicyclo[2.2.2]octane (0.17 mmol). The mixture was stirred at 80 °C for 2 hours and the volatiles were removed under reduced pressure to afford a solid which was redissolved in dichloromethane, washed with water, dried over sodium sulfate, filtered and concentrated under reduced pressure. The resulting residue was triturated with 60 mL of a 10:1 mixture of tert-butylmethyl ether and ethyl acetate to afford a yellow solid, which was filtered and dried to afford the title compound in 88% yield. MS (ESI) m/z: 587.2 [M+H]+. Step 4: 2-(Pyridin-4-yl)propane-1,3-diol

A mixture of 4-picoline (430 mmol) in 37% formaldehyde (1.72 mol) was stirred at 100°C for 12 hours. The reaction mixture was concentrated under reduced pressure to provide a residue which was purified by silica gel chromatography (1-10% methanol in ethyl acetate). The title compound was isolated in 88% yield as a colorless oil. 1H NMR (400 MHz, methanol-d4) δ ppm 2.99 (q, J = 6.4 Hz, 1H) 3.77-3.91 (m, 4 H) 7.29-7.47 (m, 2 H) 8.33-8.54 (m, 2 H) . Step 5: 3-Hydroxy-2-(pyridin-4-yl)propyl 4-tosylate

To a 0 °C solution of 2-(pyridin-4-yl)propane-1,3-diol (65.3 mmol) in dichloromethane (100 mL) was added p -toluenesulfonyl chloride (65.3 mmol), N, N'-dimethylaminopyridine (6.53 mmol) and pyridine (65.3 mmol). The mixture was stirred at room temperature for 3 hours, quenched with water and extracted three times with ethyl acetate. The organic layers were combined and dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to provide a residue which was used in the next step without further purification. Step 6: S-(3-Hydroxy-2-(pyridin-4-yl)propyl)thioacetic acid

To a solution of 3-hydroxy-2-(pyridin-4-yl)propyl 4-tosylate (55.3 mmol) in dimethylformamide (150 mL) was added potassium thioacetate (81.9 mmol). The mixture was stirred at 50°C for 2 hours, quenched with water and extracted three times with ethyl acetate. The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to provide a residue which was purified by silica gel chromatography (50-100% ethyl acetate in hexanes). The title compound was isolated in 15% yield as a yellow oil. 1H NMR (400 MHz, CDCl3) δ ppm 2.35 (s, 3 H) 2.99 (quin, J= 6.4 Hz, 1 H) 3.15-3.24 (m, 1 H), 3.37 (dd, J= 14.0, 7.2 Hz, 1H) 3.79-3.90 (m, 2H) 7.14-7.24 (m, 2H), 8.48-8.59 (m, 2H). Step 7: (2S,6R)-4-(7-Chloro-8-((3-hydroxy-2-(pyridin-4-yl)propyl)thio)-2-oxo-6-(tri Fluoromethyl)-1,2-dihydroquinazolin-4-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester

To S-(3-hydroxy-2-(pyridin-4-yl)propyl)thioacetic acid (0.95 mmol) and (2S,6R)-4-(7-chloro-8-iodo-2-oxo -6-(trifluoromethyl)-1,2-dihydroquinazolin-4-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester (1.42 mmol) in 1,2 - solution in ethylene glycol (2 mL) and isopropanol (2 mL), addition of potassium carbonate (2.84 mmol) and cuprous iodide (0.47 mmol). The mixture was stirred at 85°C for 2 hours, quenched with water and extracted three times with ethyl acetate. The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to provide a residue which was purified by preparative TLC. The title compound was isolated in 65% yield as a yellow solid. MS (ESI) m/z: 627.9 [M+1]+. Step 8: (2S,6R)-4-((R)-11-Chloro-6-oxo-3-(pyridin-4-yl)-10-(trifluoromethyl)-3,4-di Hydrogen-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester (Int-1a) and (2S,6R)-4-((S)-11-chloro-6-oxo-3-(pyridin-4-yl)-10-(trifluoromethyl)-3, 4-Dihydro-2H, 6H-[1,4]thiazazo[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid Tributyl ester (Int-1b)

To (2S,6R)-4-(7-chloro-8-((3-hydroxy-2-(pyridin-4-yl)propyl)thio)-2-oxo-6-(trifluoromethane A solution of tert-butyl)-1,2-dihydroquinazolin-4-yl)-2,6-dimethylpiperazine-1-carboxylate (2.11 mmol) in tetrahydrofuran (100 mL) at 0°C Triphenylphosphine (10.6 mmol) was added. The mixture was stirred at 0°C for 30 minutes and diethyl azodicarboxylate (10.6 mmol) was added. After 12 hours, the volatiles were removed under reduced pressure to provide a residue which was purified by silica gel chromatography (0-10% methanol in ethyl acetate). Compounds Int-1a and Int-1b were isolated in 65% yield as a 1:1 mixture of diastereomers. Pure diastereomers Int-1a and Int-1b were obtained by purifying the mixture of diastereomers by semi-preparative reverse-phase HPLC (column=Daicel Chiracel OD 250mm*30mm, 10um; mobile phase B=methanol; Gradient = 50% for 6.4 min execution time) and characterized by SFC (column = Chiracel OD-3, 50x4.6 mm ID, 3 um, stabilized at 35°C with a back pressure of 100 bar E. Mobile phase A=CO2 and mobile phase B=methanol containing 0.05% diethylamine; gradient=40%; flow rate=3 mL/min. detector=photodiode array). Int-1a: SFC Rt= 1.03 min; MS (ESI) m/z: 609.9 [M+1]+. Int-1b: SFC Rt= 1.40 min; MS (ESI) m/z: 609.9 [M+1]+ Step 9: (2S,6R)-4-((S)-11-(4-chloro-5- (Triisopropylsilyl)thiophen-2-yl)-6-oxo-3-(pyridin-4-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H -[1,4]thiazazo[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester

The title compound was prepared analogously to Example 23, Step 1, wherein (2S,6R)-4-((S)-11-chloro-6-oxo-3-(pyrimidin-2-yloxy)-10- (Trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl)-2,6-di Methylpiperazine-1-carboxylic acid tert-butyl ester was replaced by (2S,6R)-4-((S)-11-chloro-6-oxo-3-(pyridin-4-yl)-10-( Trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl)-2,6-dimethyl tert-butyl piperazine-1-carboxylate. The title compound was isolated in 38% yield as a yellow solid. MS (ESI) m/z: 848.1 [M+H]+. Step 10: (2S,6R)-4-((S)-11-(4-chlorothien-2-yl)-6-oxo-3-(pyridin-4-yl)-10-(trifluoro Methyl)-3,4-dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiper tertiary butyl oxazine-1-carboxylate

The title compound was prepared analogously to Example 23, step 2, wherein (2S,6R)-4-((S)-11-(4-chloro-5-(triisopropylsilyl)thiophen-2-yl)- 6-oxo-3-(pyrimidin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2, 3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester is replaced by (2S,6R)-4-((S)-11-( 4-Chloro-5-(triisopropylsilyl)thiophen-2-yl)-6-oxo-3-(pyridin-4-yl)-10-(trifluoromethyl)-3,4- Dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tertiary butyl ester. The title compound was isolated in 98% yield as a white solid. MS (ESI) m/z: 692.2 [M+H]+. Step 11: (S)-11-(4-Chlorothien-2-yl)-8-((3S,5R)-3,5-dimethylpiperazin-1-yl)-3-(pyridine-4 -yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeb[2,3,4-ij]quinazolin-6-one

The title compound was prepared analogously to Example 23, Step 3, wherein (2S,6R)-4-((S)-11-(4-chlorothien-2-yl)-6-oxo-3-(pyrimidine- 2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazoline-8 -yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester was replaced by (2S,6R)-4-((S)-11-(4-chlorothien-2-yl)-6 -Oxy-3-(pyridin-4-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3,4 -ij] quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester. The title compound was isolated in 99% yield as a yellow solid. MS (ESI) m/z: 592.2 [M+H]+. Step 12: (S)-8-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-11-(4-chlorothien-2-yl)- 3-(pyridin-4-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazole Lin-6-one

步驟1：(2S,6R)-4-((S)-11-氯-6-側氧基-3-(吡啶-2-基)-10-(三氟甲基)-3,4-二氫-2H,6H-[1,4]硫氮呯并[2,3,4-ij]喹唑啉-8-基)-2,6-二甲基哌嗪-1-羧第三丁酸酯(Int-2a)及(2S,6R)-4-((R)-11-氯-6-側氧基-3-(吡啶-2-基)-10-(三氟甲基)-3,4-二氫-2H,6H-[1,4]硫氮呯并[2,3,4-ij]喹唑啉-8-基)-2,6-二甲基哌嗪-1-甲酸第三丁酯(Int-2b)

The title compound was prepared analogously to Example 23, step 4, wherein (S)-11-(4-chlorothien-2-yl)-8-((3S,5R)-3,5-dimethylpiperazine-1 -yl)-3-(pyrimidin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3, 4-ij] quinazolin-6-one was replaced by (S)-11-(4-chlorothiophen-2-yl)-8-((3S,5R)-3,5-dimethylpiperazine-1 -yl)-3-(pyridin-4-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3,4- ij] quinazolin-6-one. The title compound was isolated in 33% yield as a white solid. MS (ESI) m/z: 646.2 [M+H]+. ¹ H NMR (400 MHz, CDCl ₃ -d) δ = 8.62 (d, J = 5.6 Hz, 2H), 8.13 (s, 1H), 7.39 - 7.32 (m, 3H), 7.12 - 6.89 (m, 1H) , 6.70 - 6.61 (m, 1H), 6.44 (dd, J = 1.6, 16.8 Hz, 1H), 5.83 - 5.79 (m, 1H), 4.88 - 4.61 (m, 4H), 4.19 (br dd, J = 9.2 , 11.4 Hz, 2H), 3.83 - 3.66 (m, 2H), 3.46 - 3.37 (m, 2H), 3.22 - 3.09 (m, 1H), 1.61 (br d, J = 6.8 Hz, 3H), 1.54 (br d, J = 7.2 Hz, 3H). Example 37 : (R)-8-((3S,5R)-4- acryloyl -3,5- dimethylpiperazin -1- yl )-11-(4- chlorothiophen -2- yl )- 3-( pyridin -2- yl )-10-( trifluoromethyl )-3,4- dihydro -2H,6H-[1,4] thiazazo [2,3,4-ij] quinazole Lin -6- one

Step 1: (2S,6R)-4-((S)-11-Chloro-6-oxo-3-(pyridin-2-yl)-10-(trifluoromethyl)-3,4-di Hydrogen-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxytributanoic acid Ester (Int-2a) and (2S,6R)-4-((R)-11-chloro-6-oxo-3-(pyridin-2-yl)-10-(trifluoromethyl)-3 ,4-Dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid Tertiary butyl ester (Int-2b)

Analogously to Example 35, Steps 1-8, the title compound was prepared as a 1:1 mixture of diastereomers, wherein in Step 4, 2-(pyridin-4-yl)propane-1,3-diol was replaced by 2-(pyridin-2-yl)propane-1,3-diol. Int-2a and Int-2b were obtained as a single diastereomer by semi-preparative reverse phase HPLC to purify the aforementioned mixture (column: Diacel Chiracel OD 250mmx30mm, 10um; mobile phase B: methanol; for 6.4 minutes run time and , gradient = 50%) and characterized by SFC (column: Chiracel OD-3, 50x4.6mm, ID = 3 um, stabilized at 35°C with a back pressure of 100 bar. Mobile phase B = containing 0.05% diethylamine in 40% methanol; gradient=40%, flow rate=3 mL/min. detector: photodiode array) Int-1a: SFC Rt= 1.39 min; MS (ESI) m/z : 610.2 [M+H]+. Int-1b: SFC Rt = 2.25 min; MS (ESI) m/z: 610.2 [M+H]+. Step 2: (2S,6R)-4-((R)-11-(4-Chloro-5-(triisopropylsilyl)thiophen-2-yl)-6-oxo-3-(pyridine -2-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazoline-8- base)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester

The title compound was prepared analogously to Example 23, Step 1, wherein (2S,6R)-4-((S)-11-chloro-6-oxo-3-(pyrimidin-2-yloxy)-10- (Trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl)-2,6-di Methylpiperazine-1-carboxylic acid tert-butyl ester was replaced by (2S,6R)-4-((R)-11-chloro-6-oxo-3-(pyridin-2-yl)-10-( Trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl)-2,6-dimethyl tert-butyl piperazine-1-carboxylate. The title compound was isolated in 72% yield as a yellow solid. MS (ESI) m/z: 848.3 [M+H]+. Step 3: (2S,6R)-4-((R)-11-(4-chlorothiophen-2-yl)-6-oxo-3-(pyridin-2-yl)-10-(trifluoro Methyl)-3,4-dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiper tertiary butyl oxazine-1-carboxylate

The title compound was prepared analogously to Example 23, step 2, wherein (2S,6R)-4-((S)-11-(4-chloro-5-(triisopropylsilyl)thiophen-2-yl)- 6-oxo-3-(pyrimidin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2, 3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester is replaced by (2S,6R)-4-((R)-11-( 4-Chloro-5-(triisopropylsilyl)thiophen-2-yl)-6-oxo-3-(pyridin-2-yl)-10-(trifluoromethyl)-3,4- Dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tertiary butyl ester. The title compound was isolated in 95% yield as a yellow solid. MS (ESI) m/z: 692.2 [M+H]+. Step 4: (R)-11-(4-Chlorothiophen-2-yl)-8-((3S,5R)-3,5-dimethylpiperazin-1-yl)-3-(pyridine-2 -yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeb[2,3,4-ij]quinazolin-6-one

The title compound was prepared analogously to Example 23, Step 3, wherein (2S,6R)-4-((S)-11-(4-chlorothien-2-yl)-6-oxo-3-(pyrimidine- 2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazoline-8 -yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester is replaced by (2S,6R)-4-((R)-11-(4-chlorothien-2-yl)-6 -Oxy-3-(pyridin-2-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3,4 -ij] quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester. The title compound was isolated in 99% yield as a yellow solid. MS (ESI) m/z: 592.2 [M+H]+. Step 5: (R)-8-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-11-(4-chlorothien-2-yl)- 3-(pyridin-2-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazole Lin-6-one

步驟1：2-(吡啶-3-基)丙-2-烯-1-醇

The title compound was prepared analogously to Example 23, step 4, wherein (S)-11-(4-chlorothien-2-yl)-8-((3S,5R)-3,5-dimethylpiperazine-1 -yl)-3-(pyrimidin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3, 4-ij] quinazolin-6-one was replaced by (R)-11-(4-chlorothiophen-2-yl)-8-((3S,5R)-3,5-dimethylpiperazine-1 -yl)-3-(pyridin-2-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3,4- ij] quinazolin-6-one. The title compound was isolated in 33% yield as a yellow solid. MS (ESI) m/z: 616.2 [M+H]+. 1H NMR (400 MHz, CDCl3) δ, 8.54 - 8.45 (m, 1H), 8.01 (s, 1H), 7.70 - 7.56 (m, 1H) , 7.30 - 7.22 (m, 2H), 7.10 (s, 1H), 6.98 - 6.78 (m, 1H), 6.62 - 6.50 (m, 1H), 6.35 (dd, J = 2.0, 16.8 Hz, 1H), 5.71 (dd, J = 2.0, 10.4 Hz, 1H), 5.02 - 4.85 (m, 2H), 4.72 - 4.45 (m, 2H), 4.20 - 4.03 (m, 2H), 3.96 - 3.74 (m, 1H), 3.63 - 3.41 (m, 2H), 3.34 - 3.19 (m, 2H), 1.51 (br s, 3H), 1.46 - 1.38 (m, 3H). Example 43 : (S)-8-((3S,5R)-4- acryloyl -3,5- dimethylpiperazin -1- yl )-11-(4- chlorothiophen -2- yl )- 3-( pyridin -3- yl )-10-( trifluoromethyl )-3,4- dihydro -2H,6H-[1,4] thiazazo [2,3,4-ij] quinazole Lin -6- one

Step 1: 2-(Pyridin-3-yl)prop-2-en-1-ol

At 60°C, pyridin-3-ylboronic acid (41 mmol), 2-bromoprop-2-en-1-ol (49 mmol), potassium phosphate (122 mmol) and XPhosPd G2 (1.0 mmol) were dissolved in THF ( 50 mL) and water (50 mL) was stirred for 12 hours. The reaction mixture was diluted with water, extracted three times with ethyl acetate and the combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to provide a residue which was analyzed by silica gel chromatography (35-80 in hexane % ethyl acetate) for purification. The title compound was isolated in 53% yield as a colorless oil. MS (ESI) m/z: 136.1 [M+H]+. Step 2: 2-(Pyridin-3-yl)propane-1,3-diol

A 10 M solution of borane dimethyl sulfide complex in THF (8.6 mL) was added to 2-(pyridin-3-yl)prop-2-en-1-ol (22 mmol) in THF (30 mL) solution at 0°C. After 5 minutes, 1M aqueous NaOH (6.5 mL) was added dropwise, followed by 35% aqueous hydrogen peroxide (86 mmol). After 2 hours, methanol (50 mL) was added and the resulting reaction was stirred at 70 °C for 12 hours. The reaction mixture was diluted with water, extracted three times with ethyl acetate and the combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to provide a residue which was analyzed by silica gel chromatography (0- 20% methanol) purification. The title compound was isolated in 73% yield as a colorless oil. 1H NMR (400 MHz, CDCl3) δ 8.58 - 8.48 (m, 2H), 7.68 - 7.59 (m, 1H), 7.32 - 7.28 (m, 1H), 4.10 - 3.96 (m, 4H), 3.16 - 3.07 (m , 1H). Step 3: 3-((tert-butyldiphenylsilyl)oxy)-2-(pyridin-3-yl)propan-1-ol

The title compound was prepared analogously to Example 31, Step 1, wherein (S)-3-(tritylthio)propane-1,2-diol was replaced by 2-(pyridin-3-yl)propane-1,3 -diol. The title compound was isolated in 47% yield as a colorless oil. 1H NMR (400 MHz, CDCl3) δ 8.49 (d, J = 4.4 Hz, 1H), 8.45 (s, 1H), 7.61 (dd, J = 7.6, 11.2 Hz, 4H), 7.54 (d, J = 8.0 Hz , 1H), 7.49 - 7.33 (m, 6H), 7.21 (dd, J = 4.8, 7.6 Hz, 1H), 4.11 - 4.05 (m, 1H), 3.99 - 3.89 (m, 3H), 3.08 (quin, J = 6.0 Hz, 1H), 2.14 (s, 1H), 1.05 (s, 9H). Step 4: S-(3-((tert-butyldiphenylsilyl)oxy)-2-(pyridin-3-yl)propyl)thioacetic acid

A solution of triphenylphosphine (8.20 mmol) and DIAD (8.17 mmol) in THF (50 mL) was stirred at 0 °C for 30 min. Add 3-((tert-butyldiphenylsilyl)oxy)-2-(pyridin-3-yl)propan-1-ol (4.09 mmol) and ethylthio S-acid (9.81 mmol) and in The mixture was stirred at room temperature for one hour. Evaporation of the volatiles under reduced pressure provided a residue which was purified by silica gel chromatography (10-25% ethyl acetate in hexanes). The title compound was isolated in 98% yield as a colorless oil. MS (ESI) m/z: 450.1 [M+H]+. Step 5: (2S,6R)-4-(8-((3-((tert-butyldiphenylsilyl)oxy)-2-(pyridin-3-yl)propyl)thio)- 7-Chloro-2-oxo-6-(trifluoromethyl)-1,2-dihydroquinazolin-4-yl)-2,6-dimethylpiperazine-1-carboxylic acid tertiary butyl ester

The title compound was prepared analogously to Example 35, Step 7, wherein S-(3-hydroxy-2-(pyridin-4-yl)propyl)thioacetic acid was replaced by S-(3-((tert-butyldiphenyl (ylsilyl)oxy)-2-(pyridin-3-yl)propyl)thioacetic acid. The title compound was isolated in 97% yield as a brown solid. MS (ESI) m/z: 866.3 [M+H]+. Step 6: (2S,6R)-4-(7-Chloro-8-((3-hydroxy-2-(pyridin-3-yl)propyl)thio)-2-oxo-6-(tri Fluoromethyl)-1,2-dihydroquinazolin-4-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester

The title compound was prepared analogously to Example 23, step 2, wherein (2S,6R)-4-((S)-11-(4-chloro-5-(triisopropylsilyl)thiophen-2-yl)- 6-oxo-3-(pyrimidin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2, 3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester is replaced by (2S,6R)-4-(8-((3-( (Tertiary butyldiphenylsilyl)oxy)-2-(pyridin-3-yl)propyl)thio)-7-chloro-2-oxo-6-(trifluoromethyl)- 1,2-dihydroquinazolin-4-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester. The title compound was isolated in 80% yield as a brown solid. MS (ESI) m/z: 628.2 [M+H]+. Step 7: (2S,6R)-4-((S)-11-Chloro-6-oxo-3-(pyridin-3-yl)-10-(trifluoromethyl)-3,4-di Hydrogen-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester (int-3a) and (2S,6R)-4-((R)-11-chloro-6-oxo-3-(pyridin-3-yl)-10-(trifluoromethyl)-3, 4-Dihydro-2H, 6H-[1,4]thiazazo[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid Tributyl ester (int-3b)

Similar to Example 29, Step 5, the title compound was isolated as a 1:1 mixture of diastereomers, where (S)-7-chloro-8-((3-hydroxy-2-(pyrazin-2-yloxy Substitution of (2S,6R)-4-(7-chloro-8-( (3-Hydroxy-2-(pyridin-3-yl)propyl)thio)-2-oxo-6-(trifluoromethyl)-1,2-dihydroquinazolin-4-yl) -Tertiary butyl 2,6-dimethylpiperazine-1-carboxylate. Int-3a and Int-3b were purified by semi-preparative reverse-phase HPLC to obtain a single diastereomer (column: Daicel Chiralcel OD 250 mm x 30 mm, 10 um); mobile phase B = methanol; Gradient = 50% for 6.4 min execution time) and characterized by SFC (column = Chiracel OD-3, 50x4.6 mm, ID = 3 um stabilized at 35°C with 100 bar back pressure Mobile phase B = methanol containing 0.05% diethylamine; gradient = 40%, flow rate = 3 mL/min. detector: photodiode array). Int-3a: SFC Rt = 1.94 min; MS (ESI) m/z: 610.1 [M+H]+. Int-3b: SFC Rt = 2.42 min; MS (ESI) m/z: 610.1 [M+H]+. Step 8: (2S,6R)-4-((S)-11-(4-Chloro-5-(triisopropylsilyl)thiophen-2-yl)-6-oxo-3-(pyridine -3-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazoline-8- base)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester

The title compound was prepared analogously to Example 23, Step 1, wherein (2S,6R)-4-((S)-11-chloro-6-oxo-3-(pyrimidin-2-yloxy)-10- (Trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl)-2,6-di Methylpiperazine-1-carboxylic acid tert-butyl ester was replaced by (2S,6R)-4-((S)-11-chloro-6-oxo-3-(pyridin-3-yl)-10-( Trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl)-2,6-dimethyl tert-butyl piperazine-1-carboxylate. The title compound was isolated in 72% yield as a yellow oil. MS (ESI) m/z: 848.2 [M+H]+. Step 9: (2S,6R)-4-((S)-11-(4-Chlorothiophen-2-yl)-6-oxo-3-(pyridin-3-yl)-10-(trifluoro Methyl)-3,4-dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiper tertiary butyl oxazine-1-carboxylate

The title compound was prepared analogously to Example 23, step 2, wherein (2S,6R)-4-((S)-11-(4-chloro-5-(triisopropylsilyl)thiophen-2-yl)- 6-oxo-3-(pyrimidin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2, 3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester is replaced by (2S,6R)-4-((S)-11-( 4-Chloro-5-(triisopropylsilyl)thiophen-2-yl)-6-oxo-3-(pyridin-3-yl)-10-(trifluoromethyl)-3,4- Dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tertiary butyl ester. The title compound was isolated in 92% yield as a white semi-solid. MS (ESI) m/z: 692.2 [M+H]+. Step 10: (S)-11-(4-Chlorothiophen-2-yl)-8-((3S,5R)-3,5-dimethylpiperazin-1-yl)-3-(pyridine-3 -yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeb[2,3,4-ij]quinazolin-6-one

The title compound was prepared analogously to Example 23, Step 3, wherein (2S,6R)-4-((S)-11-(4-chlorothien-2-yl)-6-oxo-3-(pyrimidine- 2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazoline-8 -yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester was replaced by (2S,6R)-4-((S)-11-(4-chlorothien-2-yl)-6 -Oxy-3-(pyridin-3-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3,4 -ij] quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester. The title compound was isolated in 94% yield as a yellow solid. MS (ESI) m/z: 592.3 [M+H]+. Step 11: (S)-8-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-11-(4-chlorothien-2-yl)- 3-(pyridin-3-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazole Lin-6-one

步驟1：2-(嘧啶-2-基)丙二酸二甲酯

The title compound was prepared analogously to Example 23, step 4, wherein (S)-11-(4-chlorothien-2-yl)-8-((3S,5R)-3,5-dimethylpiperazine-1 -yl)-3-(pyrimidin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3, 4-ij] quinazolin-6-one was replaced by (S)-11-(4-chlorothiophen-2-yl)-8-((3S,5R)-3,5-dimethylpiperazine-1 -yl)-3-(pyridin-3-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3,4- ij] quinazolin-6-one. The title compound was isolated in 41% yield as a white solid. MS (ESI) m/z: 646.1 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 8.58 (s, 1H), 8.48 (d, J = 4.4 Hz, 1H), 8.06 (s, 1H), 7.88 (s, 1H), 7.84 - 7.68 (m, 1H), 7.37 (s, 1H), 7.33 - 6.98 (m, 1H), 6.82 (dd, J = 10.4, 16.4 Hz, 1H), 6.20 (dd, J = 2.4, 16.4 Hz, 1H), 5.78 - 5.71 (m, 1H), 4.86 - 4.47 (m, 4H), 4.08 (d, J = 13.2 Hz, 2H), 3.97 - 3.68 (m, 1H ), 3.67 - 3.35 (m, 2H), 3.29 - 3.22 (m, 2H), 1.40 (s, 6H). Example 45 and Example 46 : (S)-8-((3S,5R)-4- acryloyl -3,5- dimethylpiperazin -1- yl )-11-(4- chlorothiophene -2- Base )-3-( pyrimidin -2- yl )-10-( trifluoromethyl )-3,4- dihydro -2H,6H-[1,4] thiazepine [2,3,4-ij ] quinazolin- 6- one and (R)-8-((3S,5R)-4- acryloyl -3,5- dimethylpiperazin -1- yl )-11-(4- chlorothiophene -2- yl )-3-( pyrimidin -2- yl )-10-( trifluoromethyl )-3,4- dihydro -2H,6H-[1,4] thiazepine [2,3, 4-ij] quinazolin- 6- one

Step 1: Dimethyl 2-(pyrimidin-2-yl)malonate

At 100°C, 2-chloropyrimidine (87.0 mmol), 1,3-diethylmalonate (175 mmol), copper(I) iodide (17 mmol), cesium carbonate (262 mmol) and A mixture of corinic acid (35 mmol) in dioxane (300 mL) was stirred for 16 hours. The reaction mixture was diluted with water, extracted three times with ethyl acetate and the combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to provide a residue which was analyzed by silica gel chromatography (0-30 in hexane % ethyl acetate) purification. The title compound was isolated in 25% yield as a yellow solid. Mass Spectrum (ESI) m/z = 211.0 (M+H)+. Step 2: 2-(Pyrimidin-2-yl)propane-1,3-diol

A 1M solution of DIBALH in THF (380 mL) was added to a -40 °C solution of 1,3-dimethyl 2-(pyrimidin-2-yl)malonate (95.2 mmol) in THF (200 mL) . After 1 hour, the reaction was quenched with sodium sulfate decahydrate, diluted with water and extracted three times with ethyl acetate. The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford the title compound as a yellow oil. MS (ESI) m/z = 155.1 (M+H)+. Step 3: 3-((tert-butyldiphenylsilyl)oxy)-2-(pyrimidin-2-yl)propan-1-ol

The title compound was prepared analogously to Example 31, Step 2, wherein (S)-3-(tritylthio)propane-1,2-diol was replaced by 2-(pyrimidin-2-yl)propane-1,3 -diol. The title compound was isolated in 55% yield as a yellow solid. MS (ESI) m/z = 393.0 (M+H)+. Step 4: S-(3-((tert-butyldiphenylsilyl)oxy)-2-(pyrimidin-2-yl)propyl)thioacetic acid

The title compound was prepared analogously to Example 43, Step 4, wherein 3-((tert-butyldiphenylsilyl)oxy)-2-(pyridin-3-yl)propan-1-ol was replaced by 3-( (tert-butyldiphenylsilyl)oxy)-2-(pyrimidin-2-yl)propan-1-ol. The title compound was isolated in 99% yield as a yellow oil. MS (ESI) m/z = 451.2 (M+H)+. Step 5: 3-((tert-butyldiphenylsilyl)oxy)-2-(pyrimidin-2-yl)propane-1-thiol

S-(3-((tert-butyldiphenylsilyl)oxy)-2-(pyrimidin-2-yl)propyl)thioacetic acid (48.8 mmol) in hydrazine monohydrate (60 mL), A solution in THF (90 mL) and methanol (90 mL) was stirred at room temperature for 1 hour. The mixture was diluted with water and most volatiles were removed under reduced pressure. The resulting aqueous solution was extracted with ethyl acetate and concentrated to provide a residue which was purified by silica gel chromatography (0-15% ethyl acetate in hexanes). The title compound was isolated in 67% yield as a colorless oil. MS (ESI) m/z = 409.2 [M+H]+. Step 6: 8-((3-((tert-butyldiphenylsilyl)oxy)-2-(pyrimidin-2-yl)propyl)thio)-7-chloro-6-(trifluoro Methyl)quinazoline-2,4(1H,3H)-dione

The title compound was prepared analogously to Example 29, step 3, wherein (S)-3-((tert-butyldiphenylsilyl)oxy)-2-(pyrazin-2-yloxy)propane-1 Replacement of -thiol with 3-((t-butyldiphenylsilyl)oxy)-2-(pyrimidin-2-yl)propane-1-thiol. The title compound was isolated in 86% yield as a white solid. MS (ESI) m/z = 671.0 [M+H]+. Step 7: 7-Chloro-8-((3-hydroxy-2-(pyrimidin-2-yl)propyl)thio)-6-(trifluoromethyl)quinazoline-2,4(1H,3H )-diketone

The title compound was prepared analogously to Example 23, step 2, wherein (2S,6R)-4-((S)-11-(4-chloro-5-(triisopropylsilyl)thiophen-2-yl)- 6-oxo-3-(pyrimidin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2, 3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tertiary butyl ester is replaced by 8-((3-((tertiary butyldiphenylsilane yl)oxy)-2-(pyrimidin-2-yl)propyl)thio)-7-chloro-6-(trifluoromethyl)quinazoline-2,4(1H,3H)-dione. The title compound was isolated in 92% yield as a white solid. MS (ESI) m/z: 433.0 [M+H]+. Step 8: 11-Chloro-3-(pyrimidin-2-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3 ,4-ij]quinazoline-6,8(7H)-dione

The title compound was prepared analogously to Example 29, Step 5, wherein (S)-7-chloro-8-((3-hydroxy-2-(pyrazin-2-yloxy)propyl)thio)-6- (Trifluoromethyl)quinazoline-2,4(1H,3H)-dione replaced by 7-chloro-8-((3-hydroxy-2-(pyrimidin-2-yl)propyl)thio) -6-(Trifluoromethyl)quinazoline-2,4(1H,3H)-dione. The title compound was isolated in 55% yield as a yellow solid. MS (ESI) m/z: 415.0 [M+H]+. Step 9: (2S,6R)-4-(11-Chloro-6-oxo-3-(pyrimidin-2-yl)-10-(trifluoromethyl)-3,4-dihydro-2H, 6H-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester

The title compound was prepared analogously to Example 29, step 6, wherein (S)-11-chloro-3-(pyrazin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro- 2H,6H-[1,4]thiazeza[2,3,4-ij]quinazoline-6,8(7H)-dione replaced by 11-chloro-3-(pyrimidin-2-yl) -10-(Trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazoline-6,8(7H)- Diketones. The title compound was isolated in 61% yield as a yellow solid. MS (ESI) m/z: 611.2 [M+H]+. Step 10: (2S,6R)-4-(11-(4-Chloro-5-(triisopropylsilyl)thiophen-2-yl)-6-oxo-3-(pyrimidin-2-yl )-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3,4-ij]quinazolin-8-yl)-2 ,6-Dimethylpiperazine-1-carboxylate tertiary butyl ester

The title compound was prepared analogously to Example 23, Step 1, wherein (2S,6R)-4-((S)-11-chloro-6-oxo-3-(pyrimidin-2-yloxy)-10- (Trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl)-2,6-di Methylpiperazine-1-carboxylic acid tert-butyl ester was replaced by (2S,6R)-4-(11-chloro-6-oxo-3-(pyrimidin-2-yl)-10-(trifluoromethyl )-3,4-dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine- 1-tert-butyl carboxylate. The title compound was isolated in 45% yield as a brown solid. MS (ESI) m/z: 849.2 [M+H]+. Step 11: 11-(4-Chloro-5-(triisopropylsilyl)thiophen-2-yl)-8-((3S,5R)-3,5-dimethylpiperazin-1-yl) -3-(Pyrimidin-2-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazaza[2,3,4-ij]quinone Azolin-6-one

The title compound was prepared analogously to Example 23, step 3, wherein (2S,6R)-4-((S)-11-(4-chlorothien-2-yl)-6-oxo-3-(pyrimidine- 2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazoline-8 -yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester is replaced by (2S,6R)-4-(11-(4-chloro-5-(triisopropylsilyl)thiophene -2-yl)-6-oxo-3-(pyrimidin-2-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine And[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester. The title compound was isolated in 95% yield as a yellow oil. MS (ESI) m/z: 749.2 [M+H]+. Step 12: 11-(4-Chlorothiophen-2-yl)-8-((3S,5R)-3,5-dimethylpiperazin-1-yl)-3-(pyrimidin-2-yl)- 10-(Trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-6-one

The title compound was prepared analogously to Example 23, step 2, wherein (2S,6R)-4-((S)-11-(4-chloro-5-(triisopropylsilyl)thiophen-2-yl)- 6-oxo-3-(pyrimidin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2, 3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester was replaced by 11-(4-chloro-5-(triisopropylsilyl )thiophen-2-yl)-8-((3S,5R)-3,5-dimethylpiperazin-1-yl)-3-(pyrimidin-2-yl)-10-(trifluoromethyl) -3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-6-one. The title compound was isolated in 95% yield as a yellow solid. MS (ESI) m/z: 593.1 [M+H]+. Step 13: (S)-8-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-11-(4-chlorothien-2-yl)- 3-(pyrimidin-2-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazole Lin-6-one and (R)-8-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-11-(4-chlorothiophene-2- Base)-3-(pyrimidin-2-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3,4-ij ]quinazolin-6-one

The title compound was prepared analogously to Example 23, step 4, wherein (S)-11-(4-chlorothien-2-yl)-8-((3S,5R)-3,5-dimethylpiperazine-1 -yl)-3-(pyrimidin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3, 4-ij] quinazolin-6-one is replaced by 11-(4-chlorothiophen-2-yl)-8-((3S,5R)-3,5-dimethylpiperazin-1-yl)- 3-(pyrimidin-2-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazole Lin-6-one. The title compound was isolated as a 1:1 mixture of diastereomers in 34% yield. Examples 45 and 46 were obtained as single diastereomers by SFC purification of the mixture (column = Daicel Chiralpak AS 250mmx30mm, 10um; phase A: 60% ethanol in acetonitrile, phase B: CO2; performed for 3 minutes In terms of time, gradient: 60% A in B) and characterized by SFC (column: Chirappak AS-3, 50x4.6 mm, I.D.=3 um, stabilized at 35° C. with a back pressure of 100 bar .Mobile Phase B: Methanol with 0.05% Diethylamine; Gradient = 5-40% B in A, Flow Rate = 3 mL/min. Detector: Photodiode Array).

Example 45 : SFC Rt= 1.95 min; MS (ESI) m/z: 647.1 [M+H]+.1H NMR (400 MHz, CDCl3) δ 8.72 (s, 2H), 8.09 (s, 1H), 7.32 ( m, 1H), 7.20 (m, 1H), 6.93 (m, 1H), 6.65 (m, 1H), 6.43 (m, 1H), 5.81 (m, 1H), 5.32 – 4.43 (m, 4H), 4.25 – 3.65 (m, 4H), 3.50 – 3.10 (m, 3H), 1.47 – 1.45 (m, 6H).

步驟1：(2S,6R)-4-((R)-11-氯-3-(5-氟吡啶-3-基)-6-側氧基-10-(三氟甲基)-3,4-二氫-2H,6H-[1,4]硫氮呯并[2,3,4-ij]喹唑啉-8-基)-2,6-二甲基哌嗪-1-甲酸第三丁酯(int-4a)及(2S,6R)-4-((S)-11-氯-3-(5-氟吡啶-3-基)-6-側氧基-10-(三氟甲基)-3,4-二氫-2H,6H-[1,4]硫氮呯并[2,3,4-ij]喹唑啉-8-基)-2,6-二甲基哌嗪-1-甲酸第三丁酯(int-4b)

Example 46 : SFC Rt= 2.36 min; MS (ESI) m/z: 647.1 [M+H]+. 1H NMR (400 MHz, CDCl3) δ 8.73 (s, 2H), 8.10 (s, 1H), 7.32 ( m, 1H), 7.24 (m, 1H), 6.92 (m, 1H), 6.66 (m, 1H), 6.45 (m, 1H), 5.82 (m, 1H), 5.34 – 4.45 (m, 4H), 4.26 – 3.66 (m, 4H), 3.52 – 3.12 (m, 3H), 1.50 – 1.47 (m, 6H). Example 49 : (S)-8-((3S,5R)-4- acryloyl -3,5- dimethylpiperazin -1- yl )-11-(4- chlorothiophen -2- yl )- 3-(5- fluoropyridin -3- yl )-10-( trifluoromethyl )-3,4- dihydro -2H,6H-[1,4] thiazepine [2,3,4-ij ] quinazolin -6- one

Step 1: (2S,6R)-4-((R)-11-chloro-3-(5-fluoropyridin-3-yl)-6-oxo-10-(trifluoromethyl)-3, 4-Dihydro-2H, 6H-[1,4]thiazazo[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid Tributyl ester (int-4a) and (2S,6R)-4-((S)-11-chloro-3-(5-fluoropyridin-3-yl)-6-oxo-10-(trifluoro Methyl)-3,4-dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiper tertiary butyl oxazine-1-carboxylate (int-4b)

The title compound was prepared analogously to Example 43, steps 1-7, wherein in step 1 pyridine-3-boronic acid was replaced by (5-fluoropyridin-3-yl)boronic acid. The title compound was isolated as a 1:1 mixture of diastereomers. Int-4a and Int-4b were obtained as a single diastereomer by SFC to purify the mixture (column = Daicel Chiralpak IC 250mmx30mm, 10um; phase A: ethanol, phase B: CO2; for 7.45 min run time , gradient: 65% A in B) and characterized by SFC (column: Chiralpak IC-3, 50x4.6 mm, ID=3 um, stabilized at 35°C with a back pressure of 100 bar. Mobile Phase: 60% ethanol with 0.05% diethylamine in CO2, flow rate = 3 mL/min. Detector: photodiode array). Int-4a: SFC Rt = 1.58 min; MS (ESI) m/z: 628.2 [M+H]+. Int-4b: SFC Rt = 2.87 min; MS (ESI) m/z: 628.2 [M+H]+. Step 2: (2S,6R)-4-((S)-11-(4-chloro-5-(triisopropylsilyl)thiophen-2-yl)-3-(5-fluoropyridine-3- Base)-6-oxo-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazoline -8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester

The title compound was prepared analogously to Example 23, Step 1, wherein (2S,6R)-4-((S)-11-chloro-6-oxo-3-(pyrimidin-2-yloxy)-10- (Trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl)-2,6-di Methylpiperazine-1-carboxylic acid tert-butyl ester was replaced by (2S,6R)-4-((S)-11-chloro-3-(5-fluoropyridin-3-yl)-6-oxo- 10-(Trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl)-2,6 - tert-butyl dimethylpiperazine-1-carboxylate. The title compound was isolated in 72% yield as a yellow oil. MS (ESI) m/z: 866.1 [M+H]+. Step 3: (2S,6R)-4-((S)-11-(4-chlorothien-2-yl)-3-(5-fluoropyridin-3-yl)-6-oxo-10- (Trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl)-2,6-di Methylpiperazine-1-carboxylic acid tert-butyl ester

The title compound was prepared analogously to Example 23, step 2, wherein (2S,6R)-4-((S)-11-(4-chloro-5-(triisopropylsilyl)thiophen-2-yl)- 6-oxo-3-(pyrimidin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2, 3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester is replaced by (2S,6R)-4-((S)-11-( 4-Chloro-5-(triisopropylsilyl)thiophen-2-yl)-3-(5-fluoropyridin-3-yl)-6-oxo-10-(trifluoromethyl)-3 ,4-Dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tertiary butyl ester. The title compound was isolated in 73% yield as a colorless oil. MS (ESI) m/z: 710.0 [M+H]+. Step 4: (S)-11-(4-chlorothien-2-yl)-8-((3S,5R)-3,5-dimethylpiperazin-1-yl)-3-(5-fluoro Pyridin-3-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazoline-6 -ketone

The title compound was prepared analogously to Example 23, Step 3, wherein (2S,6R)-4-((S)-11-(4-chlorothien-2-yl)-6-oxo-3-(pyrimidine- 2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazoline-8 -yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester was replaced by (2S,6R)-4-((S)-11-(4-chlorothien-2-yl)-3 -(5-fluoropyridin-3-yl)-6-oxo-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2, 3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester. The title compound was isolated in 97% yield as a yellow oil. MS (ESI) m/z: 609.9 [M+H]+. Step 5: (S)-8-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-11-(4-chlorothien-2-yl)- 3-(5-fluoropyridin-3-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3,4-ij ]quinazolin-6-one

步驟1：3-氟-5-碘噻吩-2-甲酸甲酯

The title compound was prepared analogously to Example 23, step 4, wherein (S)-11-(4-chlorothien-2-yl)-8-((3S,5R)-3,5-dimethylpiperazine-1 -yl)-3-(pyrimidin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3, 4-ij] quinazolin-6-one was replaced by (S)-11-(4-chlorothiophen-2-yl)-8-((3S,5R)-3,5-dimethylpiperazine-1 -yl)-3-(5-fluoropyridin-3-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3 ,4-ij]quinazolin-6-one. The title compound was isolated in 69% yield as a white solid. MS (ESI) m/z: 664.0 [M+H]+. 1H NMR (400 MHz, CDCl3) δ 8.50 (s, 1H), 8.44 (d, J = 2.4 Hz, 1H), 8.12 (s, 1H), 7.54 - 7.45 (m, 1H), 7.43 - 7.31 (m, 1H), 7.08 - 6.85 (m, 1H), 6.69 - 6.59 (m, 1H), 6.43 (dd, J = 2.0, 16.8 Hz, 1H), 5.83 - 5.76 (m, 1H), 4.96 - 4.52 (m, 4H), 4.22 - 4.15 (m, 2H), 3.97 - 3.62 (m, 2H), 3.46 - 3.36 (m, 2H), 3.19 - 3.00 (m, 1H), 1.58 - 1.49 (m, 6H). Example 53 : (S)-8-((3S,5R)-4- acryloyl -3,5- dimethylpiperazin -1- yl )-11-(4- chlorothiophen -2- yl )- 3-(4- fluorothiophen -2- yl )-10-( trifluoromethyl )-3,4- dihydro -2H,6H-[1,4] thiazepine [2,3,4-ij ] quinazolin -6- one

Step 1: Methyl 3-fluoro-5-iodothiophene-2-carboxylate

A 1M solution of chloro-(2,2,6,6-tetramethyl-1-piperidinyl)magnesium-lithium chloride complex (18.7 mL) was added dropwise to methyl 3-fluorothiophene-2-carboxylate ( 19 mmol) in THF (30 mL) at -40°C. After 30 minutes, the mixture was cooled to -70 °C and iodine (19.7 mmol) in THF (10 mL) was added. The reaction was stirred at room temperature for one hour and quenched with water. The mixture was extracted twice with ethyl acetate, the combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to provide a residue which was analyzed by silica gel chromatography (4% in hexane ethyl acetate) purification. The title compound was isolated in 75% yield as a white solid. Step 2: 3-Fluoro-5-iodothiophene-2-carboxylic acid

Sodium hydroxide (42 mmol) was added to a solution of methyl 3-fluoro-5-iodothiophene-2-carboxylate (14.0 mmol) in THF (10 mL) and the mixture was stirred at room temperature for four hours. Most of the volatiles were removed under reduced pressure and the resulting aqueous solution was treated with 1M aqueous HCl (10 mL) to induce precipitation of the title compound, which was filtered and dried. The title compound was isolated in 79% yield as a white solid. Step 3: 4-Fluoro-2-iodothiophene

A solution of 3-fluoro-5-iodothiophene-2-carboxylic acid (11.0 mmol) in (methylsulfinyl)methane (30 mL) was treated with silver carbonate (1.10 mmol) and acetic acid (13.2 mmol) and heated at 120 The mixture was heated at °C for 3 hours. The mixture was cooled to room temperature and diluted with water, extracted three times with ethyl acetate and the combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford the title compound as a yellow oil in 99% yield. 1H NMR (400 MHz, CDCl3) δ ppm 6.72 - 6.74 (m, 1 H) 6.99 - 7.00 (m, 1 H). Step 4: tert-butyldimethyl((2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)propenyl)oxy ) silane

Under nitrogen, to tert-butyldimethyl(prop-2-yn-1-yloxy)silane (52.8 mmol), N,N'-bis(2-benzothiazolyl)-2,6- To a solution of pyridinedicarboxamide (58.1 mmol), copper chloride (5.28 mmol) and sodium tert-butoxide (7.93 mmol) in toluene (90 mL) was added tri-tert-butylphosphine (5.28 mmol). Methanol (4.28 mL) was added and the mixture was stirred at room temperature for 12 hours. The volatiles were removed under reduced pressure and the resulting residue was purified by silica gel chromatography (0-5% ethyl acetate in hexanes) to afford the title compound as a colorless oil in 76% yield. 1H NMR (400 MHz, CDCl3) δ ppm 0.93 (s, 9 H) 1.27 (s, 12 H) 4.29 – 4.30 (m, 2 H) 5.87 – 5.89 (m, 1 H) 5.97 (s, 1 H). Step 5: tert-butyl((2-(4-fluorothiophen-2-yl)propenyl)oxy)dimethylsilane

To 4-fluoro-2-iodothiophene (11.0 mmol) and tert-butyldimethyl ((2-(4,4,5,5-tetramethyl-1,3,2-dioxaborole To a solution of cyclo-2-yl)propenyl)oxy)silane (12.1 mmol) in dioxane (10 mL) and water (5 mL) was added potassium phosphate (32.9 mmol) and XPhos Pd G3 (0.22 mmol). The suspension was stirred at 60 °C for 2 hours and the volatiles were removed under reduced pressure to afford a residue which was purified by silica gel chromatography (4% ethyl acetate in hexane). The title compound was isolated in 54% yield as a yellow oil. Step 6: 3-((tertiarybutyldimethylsilyl)oxy)-2-(4-fluorothiophen-2-yl)propan-1-ol

The title compound was prepared analogously to Example 43, Step 2, wherein 2-(pyridin-3-yl)prop-2-en-1-ol was replaced by tert-butyl((2-(4-fluorothiophen-2-yl ) propenyl) oxy) dimethylsilane. The title compound was isolated in 35% yield as a yellow oil. 1H NMR (400 MHz, CDCl3) δ ppm 0.08 (s, 6 H) 0.91 (s, 9 H) 2.30 - 2.52 (m, 1 H) 3.20 (t, J=5.6 Hz, 1 H) 3.85 - 3.97 (m , 4 H) 6.55 (s, 1 H) 6.69 (s, 1 H). Step 7: S-(3-((tert-butyldimethylsilyl)oxy)-2-(4-fluorothien-2-yl)propyl)thioacetic acid

The title compound was prepared analogously to Example 43, Step 4, wherein 3-((tert-butyldiphenylsilyl)oxy)-2-(pyridin-3-yl)propan-1-ol was replaced by 3-( (tert-butyldimethylsilyl)oxy)-2-(4-fluorothien-2-yl)propan-1-ol. The title compound was isolated in 97% yield as a yellow oil. 1H NMR (400 MHz, methanol-d4) δ ppm 0.05 (s, 6 H) 0.91 (s, 9 H) 2.33 (s, 3 H) 3.12 - 3.15 (m, 2 H) 3.31 - 3.35 (m, 1 H) ) 3.73 - 3.77 (m, 1H) 3.84 - 3.86 (m, 1H) 6.53 - 6.54 (m, 1H) 6.67 - 6.68 (m, 1H). Step 8: (2S,6R)-4-(8-((3-((tert-butyldimethylsilyl)oxy)-2-(4-fluorothiophen-2-yl)propyl)sulfur Base)-7-chloro-2-oxo-6-(trifluoromethyl)-1,2-dihydroquinazolin-4-yl)-2,6-dimethylpiperazine-1-carboxylic acid tertiary butyl ester

The title compound was prepared analogously to Example 35, Step 7, wherein S-(3-hydroxy-2-(pyridin-4-yl)propyl)thioacetic acid was replaced by S-(3-((tertiary-butyldi (methylsilyl)oxy)-2-(4-fluorothien-2-yl)propyl)thioacetic acid. The title compound was isolated in 76% yield as a yellow solid. MS (ESI) m/z: 765.1 [M+H]+. Step 9: (2S,6R)-4-(7-Chloro-8-((2-(4-fluorothien-2-yl)-3-hydroxypropyl)thio)-2-oxo-6 -(Trifluoromethyl)-1,2-dihydroquinazolin-4-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester

The title compound was prepared analogously to Example 23, step 2, wherein (2S,6R)-4-((S)-11-(4-chloro-5-(triisopropylsilyl)thiophen-2-yl)- 6-oxo-3-(pyrimidin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2, 3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester is replaced by (2S,6R)-4-(8-((3-( (Third-butyldimethylsilyl)oxy)-2-(4-fluorothiophen-2-yl)propyl)thio)-7-chloro-2-oxo-6-(trifluoro Methyl)-1,2-dihydroquinazolin-4-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester. The title compound was isolated in 47% yield as a yellow oil. MS (ESI) m/z: 651.1 [M+H]+ Step 10: (2S,6R)-4-((S)-11-chloro-3-(4-fluorothiophen-2-yl)-6- Oxy-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl) -2,6-Dimethylpiperazine-1-carboxylic acid tert-butyl ester (int-5a) and (2S,6R)-4-((R)-11-chloro-3-(4-fluorothiophene-2 -yl)-6-oxo-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazole Lin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester (int-5b)

The title compound was prepared analogously to Example 29, Step 5, wherein (S)-7-chloro-8-((3-hydroxy-2-(pyrazin-2-yloxy)propyl)thio)-6- (Trifluoromethyl)quinazoline-2,4(1H,3H)-dione was replaced by (2S,6R)-4-(7-chloro-8-((2-(4-fluorothiophene-2- Base)-3-hydroxypropyl)thio)-2-oxo-6-(trifluoromethyl)-1,2-dihydroquinazolin-4-yl)-2,6-dimethyl Tributyl piperazine-1-carboxylate. The title compound was isolated as a 1:1 mixture of diastereomers. Int-5a and Int-5b were purified by SFC to obtain single diastereomers (column: Daicel Chiralpak IC (250mm*30mm, 10um); phase A: 55% methanol in acetonitrile, phase B : CO2) and characterized by SFC (column: Chirapak IC-3, 50x4.6 mm, I.D.=3 um, stabilized at 35°C with 100 bar back pressure. Mobile phase A: CO2 and mobile phase B: 40% methanol in acetonitrile with 0.05% diethylamine, flow rate = 3 mL/min. Detector: photodiode array).

Int-5a: SFC Rt= 1.18 min; MS (ESI) m/z: 633.2 [M+H]+. 1H NMR (400 MHz, CDCl3) δ ppm 1.51 (s, 9 H) 1.37 - 1.50 (m, 6 H) 3.18 - 3.36 (m, 3H) 3.68 - 3.83 (m, 1H) 3.98 - 4.18 (m, 3H) 4.31 - 4.45 (m, 2H) 4.53 - 4.72 (m, 1H) 4.83 (dd , J=13.2, 4.4 Hz, 1 H) 6.64 (s, 1 H) 6.89 (s, 1 H) 8.04 (s, 1 H).

Int-5b: SFC Rt= 2.23 min; MS (ESI) m/z: 633.2 [M+H]+. 1H NMR (400 MHz, CDCl3) δ ppm 1.51 (s, 9 H) 1.55 (s, 6 H) 3.16 - 3.34 (m, 3H) 3.68 - 3.80 (m, 1H) 3.97 - 4.17 (m, 3H) 4.31 - 4.44 (m, 2H) 4.53 - 4.71 (m, 1H) 4.83 (dd, J =13.6, 4.8 Hz, 1 H) 6.65 (s, 1 H) 6.89 (s, 1 H) 8.04 (s, 1 H). Step 11: (2S,6R)-4-((S)-11-(4-chloro-5-(triisopropylsilyl)thiophen-2-yl)-3-(4-fluorothiophene-2- Base)-6-oxo-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazoline -8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester

The title compound was prepared analogously to Example 23, Step 1, wherein (2S,6R)-4-((S)-11-chloro-6-oxo-3-(pyrimidin-2-yloxy)-10- (Trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl)-2,6-di Methylpiperazine-1-carboxylic acid tert-butyl ester was replaced by (2S,6R)-4-((S)-11-chloro-3-(4-fluorothiophen-2-yl)-6-oxo- 10-(Trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl)-2,6 - tert-butyl dimethylpiperazine-1-carboxylate. The title compound was isolated in 94% yield as a white solid. MS (ESI) m/z: 871.1 [M+H]+. Step 12: (2S,6R)-4-((S)-11-(4-chlorothiophen-2-yl)-3-(4-fluorothiophen-2-yl)-6-oxo-10- (Trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazolin-8-yl)-2,6-di Methylpiperazine-1-carboxylic acid tert-butyl ester

The title compound was prepared analogously to Example 23, step 2, wherein (2S,6R)-4-((S)-11-(4-chloro-5-(triisopropylsilyl)thiophen-2-yl)- 6-oxo-3-(pyrimidin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2, 3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester is replaced by (2S,6R)-4-((S)-11-( 4-Chloro-5-(triisopropylsilyl)thiophen-2-yl)-3-(4-fluorothiophen-2-yl)-6-oxo-10-(trifluoromethyl)-3 ,4-Dihydro-2H,6H-[1,4]thiazazo[2,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tertiary butyl ester. The title compound was isolated in 61% yield as a white solid. MS (ESI) m/z: 715.0 [M+H]+. Step 13: (S)-11-(4-Chlorothiophen-2-yl)-8-((3S,5R)-3,5-dimethylpiperazin-1-yl)-3-(4-fluoro Thiophen-2-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazoline-6 -ketone

The title compound was prepared analogously to Example 23, Step 3, wherein (2S,6R)-4-((S)-11-(4-chlorothien-2-yl)-6-oxo-3-(pyrimidine- 2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazeza[2,3,4-ij]quinazoline-8 -yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester is replaced by: (2S,6R)-4-((S)-11-(4-chlorothien-2-yl)- 3-(4-fluorothiophen-2-yl)-6-oxo-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2 ,3,4-ij]quinazolin-8-yl)-2,6-dimethylpiperazine-1-carboxylic acid tert-butyl ester. The title compound was isolated in 87% yield as a yellow solid. MS (ESI) m/z: 615.0 [M+H]+. Step 14: (S)-8-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-11-(4-chlorothien-2-yl)- 3-(4-fluorothiophen-2-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3,4-ij ]quinazolin-6-one

The title compound was prepared analogously to Example 23, step 4, wherein (S)-11-(4-chlorothien-2-yl)-8-((3S,5R)-3,5-dimethylpiperazine-1 -yl)-3-(pyrimidin-2-yloxy)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3, 4-ij] quinazolin-6-one was replaced by (S)-11-(4-chlorothiophen-2-yl)-8-((3S,5R)-3,5-dimethylpiperazine-1 -yl)-3-(4-fluorothiophen-2-yl)-10-(trifluoromethyl)-3,4-dihydro-2H,6H-[1,4]thiazepine[2,3 ,4-ij]quinazolin-6-one. The title compound was isolated in 42% yield as a white solid. MS (ESI) m/z: 669.0 [M+H]+. 1H NMR (400 MHz, CDCl3) δ ppm 1.51 - 1.61 (m, 6 H) 2.99 - 3.21 (m, 1 H) 3.29 - 3.46 (m, 2 H) 3.55 - 3.74 (m, 1 H) 3.86 - 4.05 (m, 1 H) 4.17 (t, J=11.2 Hz, 2 H) 4.43 - 5.00 (m, 4 H) 5.76 - 5.83 (m, 1 H) ) 6.43 (dd, J=16.8, 2.0 Hz, 1 H) 6.54 - 6.73 (m, 2 H) 6.82 (s, 1 H) 6.86 - 7.10 (m, 1 H) 7.37 (s, 1 H) 8.09 (s , 1H).

A. 檢定及活性資料KRAS G12C共價結合檢定執行如下： KRAS G12C 共價加合物形成 (CAF) 檢定 Compounds of the application synthesized according to any of the above procedures include:

A. Assay and activity data The KRAS G12C covalent binding assay is performed as follows: KRAS G12C covalent adduct formation (CAF) assay

This example provides a protocol for evaluating covalent adduct formation (CAF) between compounds of formula (I) and KRAS.

In vitro covalent adduct formation assay: Covalent adduct formation (CAF) reaction between Cys12 of KRAS 4B G12C protein and compounds disclosed herein was performed in vitro using liquid chromatography-mass spectrometry (LC-MS) Measure.

Recombinant human KRAS 4B protein containing the G12C mutation was used in compound screening experiments. This protein comprises a total of 188 amino acids including an N-terminal 6-histidine tag, followed by a tobacco etch virus (TEV) tag, followed by residues 1-169 of the native KRAS 4B sequence. The exact mass of the protein was 21,310 Da as determined by mass spectrometry. The complete amino acid sequence is shown below: MAHHHHHHAG GAENLYFQSM TEYKLVVVGA CGVGKSALTI QLIQNHFVDE YDPTIEDSYR KQVVIDGETC LLDILDTAGQ EEYSAMRDQY MRTGEGFLCV FAINNTKSFE DIHHYREQIK RVKDSEDVPM VLVGNKCDLP SRTVDTKQAQ DLARSYGIPF IETSAKTRQG VDDAFYTLVR EIRKHKEK (SEQ ID NO: 1)

In an alternative screen, the assay can be performed using the KRAS 4b G12C protein, which has 170 amino acids, a mass of 19,336 Da, and the amino acid sequence SMTEYKLVVVGA CGVGKSALTI QLIQNHFVDE YDPTIEDSYR KQVVIDGETC LDILDTAGQ EEYSAMRDQY MRTGEGFLCV FAINNTKSFE DIHHY REQIK RVKDSEDVPM VLVGNKCDLP SRTVDTKQAQ DLARSYGIPF IETSAKTRQG VDDAFYTLVR EIRKHKEK (SEQ ID NO: 2).

Recombinant proteins were expressed in E. coli BL21 cells and purified using affinity chromatography over Ni-NTA columns. The protein base was nucleotide exchanged to >95% GDP, concentrated to 4 mg/mL, and stored at -80°C in storage buffer (50 mM HEPES pH 7.4, 50 mM NaCl, 5 mM MgCl2, 1 mM DTT )middle. Pure KRAS 4B G12C protein was diluted to a concentration of 5 µM in Tris-buffered saline (pH 7.4). Compounds were dissolved in DMSO and added to diluted proteins to make 10 µM concentrations. The total DMSO concentration in the reaction was 4%. Reactions were mixed by pipetting and incubated for one hour at 22°C. Over time, reaction aliquots were obtained and diluted 2:1 in 0.1% formic acid. The intact mass of protein samples was measured by LC-MS using a QExactive+ mass spectrometer (Thermo Scientific). An amount of 500 ng of total protein was injected onto a C8 reverse-phase column, eluted with a seven-minute gradient of 30%-90% acetonitrile/0.1% formic acid, and analyzed for intact mass by mass spectrometry. Identified adducts were confirmed to be within 1 Dalton of the expected mass, and the relative ratio of free:adducted protein was used to quantify the percentage of protein bound by the compound. CAF reactions were performed in duplicate with a typical variability of ±5%.

示例性式(I)化合物對KRAS G12C介導之磷酸-ERK1/2抑制之抑制 Examples E1-E4 were evaluated at 60 minutes in the CAF assay above.

Inhibition of KRAS G12C-Mediated Phospho-ERK1/2 Inhibition by Exemplary Compounds of Formula (I)

This example shows that exemplary compounds of the disclosure inhibit KRAS G12C as measured by induced inhibition of ERK phosphorylation.

KRAS G12C mutant cell line, NCI H358 (ATCC, CRL-5807), and Ras Initiative (RI) KRAS G12C were cultured according to published protocols and maintained at 37°C in 5% CO ₂ . Phospho-ERK HTRF assay was performed following the supplier's protocol (CisBio #64AERPEH). Spread NCI-H358 or RI KRAS G12C cells at a density of 50,000 cells/well in a 96-well plate (Corning #3903) in the corresponding medium (for NCI-H358, RPMI + 10% FBS + 1% Pen/ Strep, and for RI KRAS G12C in DMEM + 10% FBS + 1% Pen/Strep + 4ug/ml blasticidin) and maintained at 37°C in 5% CO ₂ . Cells were allowed to adhere overnight and treated the next day with a Tecan D300e Digital Dispenser (Tecan Group Ltd., Switzerland) using an 11-point dose response of exemplified compounds starting at 2,500 nM followed by serial 1:3 dilutions up to 4 hours or 16 Hour. Following compound treatment, cells were washed once with ice-cold PBS. Cells were lysed by adding 50 µl of lysis buffer (1x) supplemented with 1x Pierce Halt Protease and Phosphatase inhibitor and incubated with shaking at 4°C for 30 minutes. After lysis, 16 µL of cell lysate from the 96-well cell culture plate was transferred to a 384-well plate (Perkin Elmer #6007290). Premix Antibody Solution was prepared by mixing (vol/vol) Advanced Phospho-ERK1/2 d2 Antibody and Advanced Phospho-ERK1/2 Eu Cryptate Antibody. Add the premixed antibody solution (4 µL) to the detection plate containing the cell lysate. The detection plate was incubated overnight at 4°C, and the HTRF signal was read the next day by using a Spectramax M5 or Spectramax i3 microplate reader (Molecular Devices, San Jose, CA, USA), and the data were read according to the manufacturer's instructions. plan to deal with.

Additional compounds were characterized for p-ERK inhibition, as described below. The results are summarized in Table 3.

While the foregoing embodiments have been described in some detail by way of illustration and example for purposes of clarity of understanding, those skilled in the art will recognize that certain changes and modifications may be practiced within the scope of the appended claims. In addition, each reference provided herein is incorporated by reference in its entirety to the same extent as if each reference was individually incorporated by reference. In the event of a conflict between the present application and the references provided herein, the present application shall control.

Claims (22)

A compound of formula (Ia) or a pharmaceutically acceptable salt thereof,

(Ia) wherein Z is S or O; m is 1 or 2; p is 1 or 2; L ¹ is

wherein k is an integer from 0 to 4; and ^each R is independently selected from methyl, and cyanomethyl, C ₂ -C ₄ alkyl, cyano, cycloalkyl, halo, haloalkyl, trifluoromethane group, and alkoxy group; or any two R ¹ combined to form a fused ring, bridge or spiro ring structure, optionally including a heteroatom selected from S, SO ₂ , O or N in the bridge or spiro ring, and wherein The bridge or spiro ring structure is optionally substituted by a side oxygen group; ^each R is independently selected from the group consisting of: alkyl, N-alkylamino, N,N-dialkylamino, alkylamide Alkyl, arylamidoalkyl, -OCH ₂ CONRR', wherein R and R' are independently selected from hydrogen, alkyl, and cycloalkyl, alkylsulfonamidoalkyl, arylsulfonamido Alkyl, N-Alkylaminoalkyl, N,N-Dialkylaminoalkyl, Alkoxy, Alkoxyalkyl, Cycloalkyl, Alkylcycloalkyl, Hydroxyalkyl, Halogen, Haloalkyl, aryl, aryloxy, aralkyl, heteroaryl, heteroarylalkyl, heterocyclyl, heterocyclylalkyl, and heteroaryloxy, any of which is optionally modified Substitution; or when m is 2, two R ² combine to form a spiro 3 to 6 membered ring optionally containing 1 to 3 heteroatoms selected from N, O, or S; R ³ , R ⁴ , R ⁵ , and R ⁶ are independently selected from halogen, hydrogen, hydroxyl, alkoxy, alkyl, cycloalkyl, amino, N-alkylamino, C-amide (-CONRR'), N-amide (-NHCOR), urea (-NHCONHR), ether (-OR), sulfonamide (-NHSO ₂ R or -SO ₂ NHR), and CF ₃ ; wherein each R and R' are independently hydrogen, alkyl, or cycloalkyl; or any two adjacent R ³ , R ⁴ , R ⁵ , or R ⁶ form an optionally substituted fused 5 member comprising 0 to 3 heteroatoms selected from N, O, or S, or 6-membered ring; the restriction is that one of R ³ , R ⁴ , R ⁵ , or R ⁶ is a bond connected to 2-quinazolone; and R ⁷ is alkyl, cyano, cycloalkyl, halogen , haloalkyl, trifluoromethyl, and alkoxy. The compound of formula (IIa) or a pharmaceutically acceptable salt thereof as claimed in item 1,

(IIa) wherein Ar is C ^- linked aryl, heteroaryl, heterocycle, or carbocycle; n is an integer from 0 to 3; and ^each R is independently selected from alkyl, amine, cyanohalogen, tris Fluoromethyl, heterocyclyl, or two R ¹⁰ combined to form a bicyclic fused heterocycle. The compound of formula (IIb) or a pharmaceutically acceptable salt thereof as claimed in item 1,

(IIb) wherein Ar is N ^- linked heteroaryl or heterocycle; n is an integer from 0 to 3; and ^each R is independently selected from alkyl, amino, cyanohalogen, trifluoromethyl, heterocyclyl , or two R ¹⁰ combine to form a bicyclic fused heterocycle. The compound of formula (IIc) or a pharmaceutically acceptable salt thereof as claimed in item 1,

(IIc) wherein Ar is aryl or heteroaryl; n is an integer from 0 to 3; and ^{each R} is independently selected from alkyl, amine, cyanohalogen, trifluoromethyl, heterocyclyl, or Two R ¹⁰ combine to form a bicyclic fused heterocycle. The compound as claimed in items 2 to 4, wherein n is 1. Compounds as claimed in items 2 to 4, wherein n is 2. The compound as claimed in items 2 to 4, wherein n is 3. The compound according to any one of claims 1 to 7, wherein p is 1. The compound according to any one of claims 1 to 7, wherein p is 2. The compound as any one of claims 1 to 9, wherein ^L is selected from:

,

,

,

,

,

,

,

,

,in

Linkage is via either of the two nitrogen atoms of L ¹ . A compound as claimed in any one of claims 1 to 10, wherein R ² is selected from methoxy, amino, MeOCH ₂ -, EtOCH ₂ -, MeO(CH ₂ ) ₂ NH-,

,

,

,

,

,

,

,

,

,

,and

, C -linked aryl or heteroaryl, N -linked heteroaryl or heterocyclyl, wherein R and R' are independently selected from hydrogen, alkyl, and cycloalkyl. The compound according to any one of claims 1 to 11, wherein R ³ , R ⁴ , R ⁵ , and R ⁶ are defined as fused thiophenes selected from the following:

,

,

,and

; Wherein each W, X, Y, and Z are independently selected from C=O, NH, O, S, CH, CQ, wherein Q is amino, halogen, methyl, -O-alkyl, -O-ring Alkyl, or trifluoromethyl. The compound according to any one of claims 1 to 11, wherein R ³ , R ⁴ , R ⁵ , and R ⁶ are defined as thiophenes selected from the following:

,

,

,

, wherein X is hydrogen, chlorine, methyl, or CF ₃ ,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,or

. The compound of claim 1, wherein the compound or a pharmaceutically acceptable salt thereof is:

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,or

. A pharmaceutical composition, which comprises a pharmaceutically effective amount of the compound according to any one of claims 1 to 14 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. The pharmaceutical composition according to claim 15, further comprising another therapeutic agent. A method of treating an individual with cancer characterized by the presence of a KRAS G12C mutation, the method comprising administering to the individual a therapeutically effective amount of a compound according to any one of claims 1 to 14, or a pharmaceutically acceptable amount thereof Accepted salts, or pharmaceutical compositions thereof. The method of claim 17, wherein the cancer is heart: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyosarcoma, fibroma, lipoma and teratoma; lung: bronchial carcinoma (squamous undifferentiated small cell, undifferentiated large cell, adenocarcinoma), alveolar (bronchiole) carcinoma, bronchial adenoma, sarcoma, lymphoma, enchondroma hamartoma, mesothelioma; gastrointestinal tract: esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumor, VIP tumor), small bowel (adenocarcinoma, lymphoma, carcinoid tumor, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large intestine (adenocarcinoma, tubular adenoma , villous adenoma, hamartoma, leiomyoma); urogenital tract: kidney (adenocarcinoma, Wilm's tumor (Wilms tumor), lymphoma, leukemia), bladder and urethra ( Squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testis (seminoma, teratoma, embryonal carcinoma, teratoma, choriocarcinoma, sarcoma, stromal cell carcinoma, fibroma, fibroadenoma, adenomatous tumor, lipoma); liver: hepatocellular carcinoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma; biliary tract: gallbladder carcinoma, ampullary carcinoma, cholangiocarcinoma; bone: osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulocyte sarcoma), Multiple myeloma, malignant giant cell tumor chordoma, osteochondroma (osteochondral exostosis), benign chondroma, chondroblastoma, chondromyxofibroma, osteoid osteoid, and giant cell tumor; nervous system: Skull (osteoma, hemangioma, granuloma, xanthoma, osteitis deformans), meninges (meningioma, meningiosarcoma, gliomatosis), brain (astrocytoma, medulloblastoma, glioma, Ependymoma, germ cell tumor (pineeal tumor), glioblastoma multiforme, oligodendroglioma, schwannoma, retinoblastoma, congenital tumor), neurofibroma of the spinal cord, meningioma , glioma, sarcoma); gynecology: uterine (endometrial carcinoma (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma), granulosa cell-oocystoma, Sertoli stromal cell tumor (Sertoli-Leydig cell tumor), dysgerminoma, malignant teratoma), vulva (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma , botryoid sarcoma (embryonal rhabdomyosarcoma), fallopian tube (carcinoma); hematology: blood (myeloid leukemia (acute and chronic), acute lymphoblastic leukemia, chronic lymphocytic leukemia, myeloproliferative disease, multiple myeloid myelodysplastic syndrome), Hodgkin's disease, non-Hodgkin's lymphoma (malignant lymphoma); skin: malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, nevus Dysplastic moles, lipomas, hemangiomas, dermatofibromas, keloids, psoriasis; or adrenal: neuroblastoma. The method according to claim 17, wherein the cancer is non-small cell lung cancer, small cell lung cancer, colorectal cancer, rectal cancer, or pancreatic cancer. Use of a compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in the manufacture of a medicament for treating a cancer in an individual characterized by the presence of a KRAS G12C mutation . The method of claim 20, wherein the cancer is heart: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyosarcoma, fibroma, lipoma and teratoma; lung: bronchial carcinoma (squamous undifferentiated small cell, undifferentiated large cell, adenocarcinoma), alveolar (bronchiole) carcinoma, bronchial adenoma, sarcoma, lymphoma, enchondroma hamartoma, mesothelioma; gastrointestinal tract: esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumor, VIP tumor), small bowel (adenocarcinoma, lymphoma, carcinoid tumor, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large intestine (adenocarcinoma, tubular adenoma, villous gland hamartoma, leiomyoma); urogenital tract: kidney (adenocarcinoma, Wilm's tumor (Wilms tumor), lymphoma, leukemia), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma , adenocarcinoma), prostate (adenocarcinoma, sarcoma), testis (seminoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, stromal cell carcinoma, fibroma, fibroadenoma, glandular tumor-like tumor, lipoma); liver: hepatocellular carcinoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma; biliary tract: gallbladder carcinoma, ampullary carcinoma, cholangiocarcinoma Bone: osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulocyte sarcoma), multiple myeloma, malignant giant cell tumor chordoma, Osteochondroma (exostoses of osteochondral), benign enchondroma, chondroblastoma, chondromyxofibroma, osteoid osteoid, and giant cell tumor; nervous system: skull (osteoma, hemangioma, granuloma, yellow osteitis deformans), meninges (meningioma, meningiosarcoma, gliomatosis), brain (astrocytoma, medulloblastoma, glioma, ependymoma, germ cell tumor (pineal tumor), glioblastoma multiforme, oligodendroglioma, schwannoma, retinoblastoma, congenital tumor), neurofibroma of the spinal cord, meningioma, glioma, sarcoma); gynecology: uterus ( Endometrial carcinoma (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma), granulosa cell-ootheca cell tumor, Sertoli stromal cell tumor, dysgerminoma, malignant teratoma), Vulva (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma (embryonal rhabdomyosarcoma), fallopian tube (carcinoma); hematology: Blood (myelogenous leukemia (acute and chronic), acute lymphoblastic leukemia, chronic lymphocytic leukemia, myeloproliferative disorders, multiple myeloma, myelodysplastic syndrome), Hodgkin's disease, non-Hodgkin's lymphoma (malignant lymphoma); skin: malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, nevus dysplastic nevus, lipoma, hemangioma, dermatofibroma, keloid, psoriasis; or adrenal: nerve Blastoma. The method according to claim 20, wherein the cancer is non-small cell lung cancer, small cell lung cancer, colorectal cancer, rectal cancer, or pancreatic cancer.