KRAS G12C Inhibitor and Method of Using the Same

Abstract

To provide KRAS G12C inhibitors useful for treating many disorders, including pancreatic, colonic and lung cancers, compositions of the same, and methods of using the same.SOLUTION: For example, the compounds of the formulas in the figure are presented.SELECTED DRAWING: None

Description

Technical Field

[0001] This specification provides KRAS G12C inhibitors, compositions thereof, and methods of using the same. These inhibitors are useful for treating a number of disorders including pancreatic cancer, colorectal cancer, and lung cancer.

Background Art

[0002] KRAS gene mutations are frequently found in pancreatic cancer, lung adenocarcinoma, colorectal cancer, gallbladder cancer, thyroid cancer, and cholangiocarcinoma. KRAS mutations are also observed in about 25% of NSCLC patients, and some studies have shown that KRAS mutations are poor prognostic factors for NSCLC patients. Recently, it has been found that V-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS) mutations confer resistance to therapies targeting epidermal growth factor receptor (EGFR) in colorectal cancer. Therefore, important information for prescribing TKI therapy can be obtained from the mutation status of KRAS. In summary, there is a need for new drug therapies for patients with pancreatic cancer, lung adenocarcinoma, or colorectal cancer, especially for patients diagnosed with such cancers characterized by KRAS mutations, including those who show progression after chemotherapy.

Summary of the Invention

[0003]

Chemical Formula

Chemical formula

Chemical formula

[0004] In another embodiment, the present specification provides a compound of formula (I)

Chemical formula

Chemical formula

[0005] Furthermore, formula (II)

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0006] In some embodiments, Q is C=O, and E 1 and E 2 are each CR 1 then (1) R 10 is C 1~3 Alkylene aryl, C 1~3 Alkylene heteroaryl, C0~3 Alkylene-C 3~8 Cycloalkyl, C 1~3 Alkylene-C 2~7 Heterocycloalkyl, or halo, or (2) R 13 is C 1~3 Haloalkyl or C 3~5 Cycloalkyl, whichever it is. In various embodiments, J is NR 10 and M is CR 13 In some embodiments, J is CR 10 and M is NR 13 In some embodiments, J is N and M is NR 13 In various embodiments, J is NR 10 and M is N.

[0007] Furthermore, formula (II)

Chemical formula

Chemical formula

Chemical formula

[0008] In some embodiments, Q is C=O, and E 1 and E 2 are each CR 1 When, (1) R 10 is C 1~3 alkylene aryl, C 1~3 alkylene heteroaryl, C 0~3 alkylene - C 3~8 cycloalkyl, C 1~3 alkylene - C 2~7 heterocycloalkyl, or halo, or (2) R 13 is C 1~3 haloalkyl or C 3~5 cycloalkyl. In various embodiments, J is NR 10 and M is CR 13 . In some embodiments, J is CR 10 and M is NR 13 . In some embodiments, J is N and M is NR 13 . In various embodiments, J is NR 10 and M is N.

[0009] Furthermore, formula (III) or (III’) [Chemical formula] [wherein, E 1 and E 2 are each independently N or CR 1 , and R 1 is independently H, hydroxy, C 1~6 alkyl, C 1~4 haloalkyl, C 1~4 alkoxy, NH - C 1~4 alkyl, N(C 1~4 alkyl)2, cyano, or halo, and R 2 is halo, C 1~6 alkyl, C 1~6 haloalkyl, OR’, N(R’)2, C 2~3 alkenyl, C 2~3 alkynyl, C 0~3 alkylene-C 3~14 cycloalkyl, C 0~3 alkylene-C 2~14 heterocycloalkyl, aryl, heteroaryl, C 0~3 alkylene-C 6~14 aryl, or C 0~3 alkylene-C 2~14 heteroaryl, and each R’ is independently H, C 1~6 alkyl, C 1~6 haloalkyl, C 3~14 cycloalkyl, C 2~14 heterocycloalkyl, C 2~3 alkenyl, C 2~3 alkynyl, aryl, or heteroaryl, or two R’ substituents are bonded to each other together with the nitrogen atom to which they are attached to form a 3- to 7-membered ring, R 3 is halo, C 1~3 alkyl, C 1~2 haloalkyl, C 1~3 alkoxy, C 3~4 cycloalkyl, C 2~14 heterocycloalkyl, C 2~3 alkenyl, C 2~3 alkynyl, aryl, or heteroaryl, R 4 is

Chemical formula

Chemical formula

[0010] Furthermore, formula (III) or (III’)

Chemical formula

Chemical formula

[0011] In some embodiments, the compound has the structure of formula (III). In other embodiments, the compound has the structure of formula (III’).

[0012] The compounds of formula (II) or (III) disclosed herein may have one or more of the following characteristics. In some embodiments, Q is C=O. In some embodiments, Q is C=S. In some embodiments, Q is C=NR 8 wherein. In various embodiments, R 8 is C 1~2 alkyl. In some embodiments, Q is CR 8 R 9 wherein. In various embodiments, Q is C=CR8 R 9 is. In some embodiments, R 8 and R 9 are bonded to each other together with the carbon atom to which they are attached to form a 3- to 4-membered ring. In some embodiments, R 8 is C 1~2 alkyl and R 9 is H.

[0013] Formula (IV) or (IV’)

Chemical formula

Chemical formula

Chemical formula

[0014] In some embodiments, the compounds disclosed herein have the structure of formula (IV). In various embodiments, the compounds disclosed herein have the structure of formula (IV’). In some embodiments, E 1 and E 2 are each CR 1 and R 8 is hydroxy, halo, nitro, or C 3~6 is cycloalkyl.

[0015] In some embodiments, R 8 is methyl. Further, formula (IV) or (IV’)

Chemical Formula

Chemical formula

[0016] In some embodiments, the compounds disclosed herein have the structure of formula (IV). In various embodiments, the compounds disclosed herein have the structure of formula (IV’). In some embodiments, E 1 and E 2 are each CR 1 and R 8 is hydroxy, halo, nitro, or C 3~6 cycloalkyl.

[0017] In some embodiments, R 8 is methyl.

[0018] Furthermore, formula (V)

Chemical formula

Chemical formula

Chemical formula

[0019] Furthermore, formula (V)

Chemical formula

[0020] The compounds of formula (I), (II), (III), (III'), (IV), (IV'), or (V) disclosed herein may have one or more of the following characteristics. In some embodiments, E1 and E 2 each is CR 1 In other embodiments, E 1 is CR 1 In other embodiments, E 2 is N. In some embodiments, E 1 is N, and E 2 is CR 1 In various embodiments, E 1 and E 2 each is N.

[0021] The compounds of formula (II), (III), (III’), (IV), (IV’), or (V) disclosed herein may have one or more of the following characteristics. In various embodiments, R 10 is C 1~6 alkyl, aryl, heteroaryl, C 3~14 cycloalkyl, C 2~14 heterocycloalkyl, C 1~6 alkoxy, O-C 0~6 alkylene-C 6~14 aryl, O-C 0~6 alkylene-C 2~14 heteroaryl, O-C 0~6 alkylene-C 3~14 cycloalkyl, O-C 0~6 alkylene-C 2~14 heterocycloalkyl, N-C 1~8 alkyl, N(C 1~8 alkyl)2, NH-C 0~6 alkylene-C 6~14 aryl, NH-C 0~6 alkylene-C 2~14 heteroaryl, NH-C 0~6 alkylene-C 3~14 cycloalkyl, or NH-C 0~6 alkylene-C 2~14 heterocycloalkyl. In various embodiments, R 10 is C 1~8 alkyl. In some embodiments, R 10 is C 0~3 alkylene-C 6~14 aryl. In some embodiments, R10 is C 0~3 alkylene-C 2~14 is heteroaryl. In some embodiments, R 10 is C 0~3 alkylene-C 3~14 is cycloalkyl. In some embodiments, R 10 is C 0~3 alkylene-C 2~14 is heterocycloalkyl. In other embodiments, R 10 is C 0~6 is alkyleneamine. For example, R 10 is i-Pr, t-Bu, phenyl, benzyl, OCH3, Cl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,

Chemical Structure

[0022] In some embodiments, R 10 includes ortho-substituted aryl, ortho-substituted heteroaryl, or 2-substituted cyclohexyl. For example, R 10 is

Chemical Structure

[0023] The compounds of formula (I), (II), (III), (III’), (IV), (IV’), or (V) disclosed herein may have one or more of the following characteristics. In some embodiments, R 1 is H. In some embodiments, R 1 is F. In some embodiments, R 1 is methyl.

[0024] The compounds of formula (I), (II), (III), (III'), (IV), (IV'), or (V) disclosed herein may have one or more of the following characteristics. In various embodiments, R 2 is aryl. In some embodiments, R 2 is heteroaryl. In various embodiments, R 2 is phenyl, naphthyl, pyridyl, indazolyl, indolyl, azaindolyl, indolinyl, benzotriazolyl, benzoxadiazolyl, imidazolyl, cinnolinyl, imidazopyridyl, pyrazolopyridyl, quinolinyl, isoquinolinyl, quinazolinyl, quinazolinonyl, indolinonyl, isoindolinonyl, tetrahydronaphthyl, tetrahydroquinolinyl, or tetrahydroisoquinolinyl. For example, R 2 is Cl, Br, CF3, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, piperidine, pyrrolidine, azetidine, OCH3, OCH2CH3, phenyl,

Chem.

[0025] In various embodiments, R 2 is bromine,

Chem.

[0026] The compounds of formula (I), (II), (III), (III'), (IV), (IV'), or (V) disclosed herein may have one or more of the following characteristics. In various embodiments, R 3 is halo. In various embodiments, R 3 is Cl. In various embodiments, R 3 is F. In some embodiments, R 3 is C 1~2is alkyl. In some embodiments, R 3 is methyl. In some embodiments, R 3 is C 1~2 haloalkyl. In various embodiments, R 3 is CF3.

[0027] The compounds of formula (I), (II), (III), (III’), (IV), (IV’), or (V) disclosed herein may have one or more of the following characteristics. In some embodiments, R 4 is

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

[0028] In various embodiments, R 4’ is H, C 1~8 alkyl, C 2~8 alkynyl, C 1~6 alkylene-O-C 1~4 alkyl, C 1~6 alkylene-OH, C 1~6 haloalkyl, C 0~3 alkylene-C 3~8 cycloalkyl, C 0~3 alkylene-C 2~7 heterocycloalkyl, C 0~3 alkylene-C 6~14 is aryl, or

Chemical formula

[0029] In various embodiments,

Chemical formula

[0030] In some embodiments, ring A is

Chemical formula

[0031] In some embodiments, ring A contains piperidinyl, piperazinyl, pyrrolidinyl, or azetidinyl. In some embodiments, ring A contains piperidinyl.

[0032] In various embodiments, ring A is

Chemical formula

[0033] In various embodiments, ring A is

Chemical formula

[0034] The compounds of formula (I), (II), (III), (III’), (IV), (IV’), or (V) disclosed herein may have one or more of the following characteristics.

[0035] In some embodiments, L is a bond.

[0036] In some embodiments, L is C 1~2 alkylene.

[0037] In various embodiments, L is O. In some embodiments, L is S.

[0038] In various embodiments, L is NH.

[0039] In some embodiments, R 5 is H or halo.

[0040] In some embodiments, R 5 is H, Br, Cl, F, CN, CH3, CF3, CH2Br, CH2OH, CH2CH2OH, CH2OCH2 phenyl, cyclopropyl, phenyl, CH2 phenyl, CH2OCH3, CH2N(CH3)2, CH2N(CH2CH3)2, CH2CO2H, CH2CO2CH3, CH2NHC(O)CH3, CH2C(O)NHCH3, CH2OC(O)CH3, or

Chemical formula

[0041] In some embodiments, R 6 is C 1~6 alkyl, C 1~6 alkylene-O-C 1~6Alkyl, C 1~6 Alkylene-OH, C 1~3 Haloalkyl, C 1~6 Alkylene-amine, C 0~6 Alkylene-amide, C 0~1 AlkyleneC(O)OC 1~3 Alkyl, C 0~1 Alkylene-C 2~14 Heterocycloalkyl, C 0~1 Alkylene-C 3~14 Cycloalkyl, or C 0~3 Alkylene-C 6~14 is aryl.

[0042]

Chem.

[0043] In various embodiments, R 6 is phenyl, cyclopropyl, CH3, CF3, CH2CH3, CH2NH2, CH(CH3)NH2, CH(CH3)2NH2, CH2Cl, CH2Br, CH2OCH3, CH2O phenyl, CH2OH, CO2H, CO2CH2CH3, CH2CO2H, CH2CH2NH2, CH2CH2OH, CH2CH2N(CH3)2, CH2NHCH3, C(O)NHCH3, C(O)N(CH3)2, CH2C(O)NH phenyl, CH2CHF2, CH2F, CHF2, CH2NHC(O)CH3, CH2NHCH2CH2OH, CH2NHCH2CO2H, CH2NH(CH3)CH2CO2CH3, CH2NHCH2CH2OCH3, CH2NH(CH3)CH2CH2OCH3, CH2NH(CH3)CH2C(O)N(CH3)2, CH2NH(CH3)CH2C(O)NHCH3, CH2CH2CCH, CH2NMe2, CH2NH(CH3)CH2CH2OH, CH2NH(CH3)CH2CH2F, CH2N + (CH3)3, CH2NHCH2CHF2, CH2NHCH2CH3,

Chem.

[0044] In various embodiments, R 5 and R 6 together are

Chemical formula

[0045] In some embodiments, R 5 and R 6 each is H.

[0046] In some embodiments, R 7 is H.

[0047] In some embodiments, R 7 is methyl.

[0048] In various embodiments, R 7 and R 5 together are -CH2- or -C(O)CH2-.

[0049] The compounds disclosed herein can be in pharmaceutically acceptable salt forms. The provided compounds can be formulated into pharmaceutical preparations containing the compounds disclosed herein and pharmaceutically acceptable excipients.

[0050] Also provided is a method of inhibiting KRAS G12C in cells, which includes contacting such cells with a compound or composition disclosed herein. Further provided is a method of treating cancer in a subject, which includes administering to such subject a therapeutically effective amount of a compound or composition disclosed herein. In some embodiments, the cancer is lung cancer, pancreatic cancer, or colorectal cancer.

Mode for Carrying Out the Invention

[0051] Definitions Abbreviations: The following abbreviations may be used herein.

Table 1

[0052] In the description of the present invention (particularly in the following claims), the use of the terms "a", "an", and "the", and similar indicators, is to be construed as including both the singular and the plural, unless otherwise specified. The detailed description of numerical ranges herein is used as a shorthand for referring individually to each distinct value falling within that range, unless otherwise specified herein, and each distinct value is incorporated into the specification as if it were individually recited herein. The use of all examples, or exemplary language (e.g., "such as") provided herein is intended to clarify the application and is not intended to limit the scope of the invention, unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention.

[0053] As used herein, the term "alkyl" refers to straight-chain and branched C1-C8 hydrocarbon groups, including, but not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, sec-butyl, t-butyl, n-pentyl, 2-methylbutyl, 3-methylbutyl, 2,2-dimethylpropyl, n-hexyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, 3,3-dimethylbutyl, and 2-ethylbutyl. The term C m-n means that the alkyl group has "m" to "n" carbon atoms. The term "alkylene" refers to an alkyl group having a substituent. An alkyl (e.g., methyl) group, or an alkylene (e.g., -CH2-) group, is independently selected, for example, halo, trifluoromethyl, trifluoromethoxy, hydroxy, alkoxy, nitro, cyano, alkylamino, C 1~8 alkyl, C2~8 Alkenyl, C 2~8 Alkynyl, -NC, amino, -CO2H, -CO2C1-C8 alkyl, -OCOC1-C8 alkyl, C3-C 10 Cycloalkyl, C3-C 10 Heterocycloalkyl, C5-C 10 Aryl, and C5-C 10 One or more of heteroaryl, typically 1 to 3, may be substituted. The term "haloalkyl" specifically refers to an alkyl group in which at least one, for example, 1 to 6, or all of the hydrogens of the alkyl group are substituted with halo atoms.

[0054] The terms "alkenyl" and "alkynyl" each refer to an alkyl group further containing a double bond or a triple bond.

[0055] As used herein, the term "halo" refers to fluoro, chloro, bromo, and iodo. The term "alkoxy" is defined as -OR where R is alkyl.

[0056] As used herein, the term "amino" or "amine" interchangeably refers to an -NR2 group, where each R is, for example, H or a substituent. In some embodiments, the amino group is further substituted to form an ammonium ion, for example, NR3 +is formed. The ammonium moiety is specifically included in the definition of "amino" or "amine". The substituent can be, for example, an alkyl group, an alkoxy group, a cycloalkyl group, a heterocycloalkyl group, an amide group, or a carboxylate group. The R group may be further substituted, for example, with one or more, such as 1 to 4 groups selected from a halo group, a cyano group, an alkenyl group, an alkynyl group, an alkyl group, a cycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroaryl group, a urea group, a carbonyl group, a carboxylate group, an amine group, and an amide group. The "amide" group or "amido" group is interchangeable and is similar to an amine group or an amino group, but further refers to a group containing C(O), for example, -C(O)NR2. As part of the intended amino group or amide group (optionally having an alkylene group, for example, alkylene - amino, or alkylene - amide), CH2NH2, CH(CH3)NH2, CH(CH3)2NH2, CH2CH2NH2, CH2CH2N(CH3)2, CH2NHCH3, C(O)NHCH3, C(O)N(CH3)2, CH2C(O)NH phenyl, CH2NHC(O)CH3, CH2NHCH2CH2OH, CH2NHCH2CO2H, CH2NH(CH3)CH2CO2CH3, CH2NHCH2CH2OCH3, CH2NH(CH3)CH2CH2OCH3, CH2NH(CH3)CH2C(O)N(CH3)2, CH2NH(CH3)CH2C(O)NHCH3, CH2CH2CCH, CH2NMe2, CH2NH(CH3)CH2CH2OH, CH2NH(CH3)CH2CH2F, CH2N + (CH3)3, CH2NHCH2CHF2, CH2NHCH2CH3, [Chemical formula] are exemplified.

[0057] As used herein, the term "aryl" refers to a C 6~14 monocyclic or polycyclic aromatic group, preferably a C 6~10 monocyclic or bicyclic aromatic group, or a C 10~14Refers to a polycyclic aromatic group. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, fluorenyl, azulenyl, anthryl, phenanthryl, pyrenyl, biphenyl, and terphenyl. Aryl also includes C 10~14 It also refers to bicyclic and tricyclic carbocyclic rings, in which case one ring is an aromatic ring and the other ring is a saturated, partially unsaturated, or aromatic ring, for example, dihydronaphthyl, indenyl, indanyl, or tetrahydronaphthyl (tetralinyl). Unless otherwise specified, aryl groups are, independently of each other, for example, halo, C 1~8 alkyl, C 2~8 alkenyl, C 2~8 alkynyl, -CF3, -OCF3, -NO2, -CN, -NC, -OH, alkoxy, amino, -CO2H, -CO2C1-C8 alkyl, -OCOC1-C8 alkyl, C3-C 10 cycloalkyl, C3-C 10 heterocycloalkyl, C5-C 10 aryl, and C5-C 10 heteroaryl, and may be substituted or unsubstituted by one or more groups selected therefrom, particularly 1 to 4 groups.

[0058] As used herein, the term "cycloalkyl" refers to a monocyclic or polycyclic non-aromatic carbocyclic ring, in which case the polycyclic ring can be a fused ring, a bridged ring, or a spiro ring. The carbocyclic ring can have 3 to 10 carbon ring atoms. Intended carbocyclic rings include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and cyclononyl.

[0059] As used herein, the term "heterocycloalkyl" refers to a monocyclic or polycyclic (e.g., bicyclic) saturated or partially unsaturated ring system containing a total of 3 or more atoms (e.g., 3 to 12, 4 to 10, 4 to 8, or 5 to 7), with 1 to 5 of those atoms (e.g., 1, 2, 3, 4, or 5) independently selected from nitrogen, oxygen, and sulfur. Non-limiting examples of heterocycloalkyl groups include azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, dihydropyrrolyl, morpholinyl, thiomorpholinyl, dihydropyridinyl, oxacycloheptyl, dioxacycloheptyl, thiacycloheptyl, and diazacycloheptyl.

[0060] Unless otherwise specified, a cycloalkyl group or a heterocycloalkyl group may or may not be substituted with one or more, particularly 1 to 4, groups. Some intended substituents include halo, C 1~8 alkyl, C 2~8 alkenyl, C 2~8 alkynyl, -OCF3, -NO2, -CN, -NC, -OH, alkoxy, amino, -CO2H, -CO2C1-C8 alkyl, -OCOC1-C8 alkyl, C3-C 10 cycloalkyl, C3-C 10 heterocycloalkyl, C5-C 10 aryl, and C5-C 10 heteroaryl.

[0061] As used herein, the term "heteroaryl" refers to a monocyclic or polycyclic (e.g., bicyclic) ring system containing 1 to 3 aromatic rings and having 1 to 4 heteroatoms (e.g., 1, 2, 3, or 4) selected from nitrogen, oxygen, and sulfur in the aromatic rings. In certain embodiments, the heteroaryl group has 5 to 20, 5 to 15, 5 to 10, or 5 to 7 ring atoms. Heteroaryl also includes C 10~14Bicyclic and tricyclic rings are also meant, where one ring is an aromatic ring and the other ring is a saturated, partially unsaturated, or aromatic ring. Examples of heteroaryl groups include, but are not limited to, furanyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl, triazolyl, benzofuranyl, benzimidazolyl, benzisoxazolyl, benzopyranyl, benzothiadiazolyl, benzothiazolyl, benzothienyl, benzothiophenyl, benzotriazolyl, benzoxazolyl, furopyridyl, imidazopyridinyl, imidazothiazolyl, indolizinyl, indolyl, indazolyl, isobenzofuranyl, isobenzothienyl, isoindolyl, isoquinolinyl, isothiazolyl, naphthyridinyl, oxazolopyridinyl, phthalazinyl, pteridinyl, purinyl, pyridopyridyl, pyrrolopyridyl, quinolinyl, quinoxalinyl, ciazolinyl, thiadiazolopyrimidyl, and thienopyridyl. Unless otherwise specified, the heteroaryl group may or may not be substituted with one or more, particularly 1 to 4 or 1 or 2 substituents. Intended substituents include halo, C 1~8 alkyl, C 2~8 alkenyl, C 2~8 alkynyl, -OCF3, -NO2, -CN, -NC, -OH, alkoxy, amino, -CO2H, -CO2C1-C8 alkyl, -OCOC1-C8 alkyl, C3-C 10 cycloalkyl, C3-C 10 heterocycloalkyl, C5-C 10 aryl, and C5-C 10 heteroaryl.

[0062] As used herein, the term Boc refers to the structure

Chemical formula

[0063] As used herein, the term "Cbz" refers to the structure [Chem.] .

[0064] As used herein, the term "Bn" refers to the structure [Chem.] .

[0065] [Chem.] .

[0066] As used herein, the term "trityl" refers to the structure [Chem.] .

[0067] As used herein, the term "tosyl" refers to the structure [Chem.] .

[0068] As used herein, the term "Troc" refers to the structure [Chem.] .

[0069] As used herein, the term "Teoc" refers to the structure [Chem.] .

[0070] As used herein, the term "Alloc" refers to the structure [Chem.] .

[0071] As used herein, the term Fmoc refers to the structure [Chemical Formula] as shown.

[0072] The disclosed compounds This specification provides KRAS inhibitors having one of the structures of Formulas I-V, which are discussed in detail below.

[0073] The compounds disclosed herein include all pharmaceutically acceptable isotopically labeled compounds in which one or more atoms of the disclosed compounds are replaced with atoms having the same atomic number but different atomic mass or mass number than the atomic mass or mass number typically found in nature. Examples of isotopes that can be incorporated into the disclosed compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, chlorine, and iodine, such as 2 H, 3 H, 11 C, 13 C, 14 C, 13 N, 15 N, 15 O, 17 O, 18 O, 31 P, 32 P, 35 S, 18 F, 36 Cl, 123 I, and 125 I, and the like. These radiolabeled compounds can be useful, for example, in elucidating characteristics such as the site or mechanism of action, or the binding affinity for a pharmacologically important site of action, and thereby can assist in determining or measuring the effectiveness of a compound. Certain isotopically labeled compounds of the present disclosure, for example, those incorporating a radioactive isotope, are useful in the study of the tissue distribution of drugs and / or substrates. For such purposes, the radioactive isotopes tritium, i.e., 3 H, and carbon-14, i.e., 14C is particularly useful considering its ease of incorporation and its immediate detection means.

[0074] Deuterium, i.e., 2 Substitution with a heavier isotope such as H results in certain therapeutic advantages due to higher metabolic stability, such as an extended in vivo half-life or a reduced required dose, and is thus preferred in some situations.

[0075] 11 C, 18 F, 15 O and 13 Substitution with a positron-emitting isotope such as N can be useful for examining substrate receptor occupancy in positron emission tomography (PET) studies. The isotope-labeled compounds of structure (I) can generally be prepared by conventional techniques known to those skilled in the art or by a process similar to the processes described in the following "Preparation" and "Examples" using appropriate isotope-labeled reagents in place of the unlabeled reagents used heretofore.

[0076] The isotope-labeled compounds disclosed herein can generally be prepared by conventional techniques known to those skilled in the art or by a process similar to the processes described in each of the attached examples and schemes using appropriate isotope-labeled reagents in place of the unlabeled reagents used heretofore.

[0077] The specific compounds disclosed herein may exist as stereoisomers (i.e., isomers that differ only in the spatial arrangement of atoms), such as optical isomers and conformational isomers (or rotamers). The compounds disclosed herein include all stereoisomers, both pure individual stereoisomer preparations and enriched preparations thereof, as well as racemic mixtures of such stereoisomers and both individual diastereomers and enantiomers that can be separated according to methods known to those skilled in the art. Further, the compounds disclosed herein include all tautomeric forms of the compounds.

[0078] The specific compounds disclosed herein may exist as atropisomers, i.e., conformational stereoisomers that result from steric interactions with other parts of the molecule and in which rotation about a single bond within the molecule is hindered or significantly slowed. The compounds disclosed herein include all atropisomers, including both pure individual atropisomer preparations, each enriched preparation, or each unspecified mixture. When the rotational barrier about the single bond is sufficiently large and the interconversion between conformations is sufficiently slow, separation and isolation of the isomeric species may be possible. For example, but not limited to, the following R 10 group

Chem.

[0079] This disclosure relates to formula (I)

Chem.

Chemical formula

[0080] The compound of formula I is of the formula (I - A), (I - B), (I - C), or (I - D): [Chemical formula] can be in the form of.

[0081] The present disclosure also relates to formula (II)

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0082] In various embodiments, J is NR10 where M is CR 13 In some embodiments, J is CR 10 where M is NR 13 In some embodiments, J is CR 10 where M is N. In various embodiments, J is N and M is NR 13 In some embodiments, J is N and M is CR 13 Some specifically contemplated R 13 include methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, trifluoromethyl, CH2NH2, and cyclopropyl. In some embodiments, J is NR 10 where M is N. In some embodiments, Q is C=O and each of E 1 and E 2 is CR 1 where (1) R 10 is C 1~3 alkylene aryl, C 1~3 alkylene heteroaryl, C 0~3 alkylene-C 3~8 cycloalkyl, C 1~3 alkylene-C 2~7 heterocycloalkyl, or halo, or (2) R 13 is C 1~3 haloalkyl or C 3~5 cycloalkyl, either of which.

[0083] The compound of formula II can be in the form of formula (II-A), (II-B), (II-C), (II-D), (II-E), (II-F), (II-G), (II-H), (II-J), (II-K), (II-L), (II-M), (II-N), (II-O), (II-P), or (II-Q):

Chemical formula

[0084] This disclosure also relates to formula (III) or formula (III’):

Chemical formula

Chemical formula

[0085] The compound of formula III is of formula (III-A), (III-B), (III-C), or (III-D):

Chemical formula

[0086] The compound of formula III' is of formula (III-A'), (III-B'), (III-C'), or (III-D'):

Chemical formula

[0087] This disclosure also relates to formula (IV) or formula (IV'):

Chemical formula

Chemical formula

Chemical formula

[0088] This specification also discloses formula (V): [Chemical formula] [wherein, E 1 and E 2 are each independently CR 1 or N, R 1 is independently H, hydroxy, C 1~4 alkyl, C 1~4 haloalkyl, C 1~4 alkoxy, NH-C 1~4 alkyl, N(C 1~4 alkyl)2, cyano, or halo, R 2 is halo, C 1~6 alkyl, C 1~6 haloalkyl, OR’, N(R’)2, C 2~3 alkenyl, C 2~3 alkynyl, C 0~3 alkylene-C 3~8 cycloalkyl, C 0~3 alkylene-C 2~7 heterocycloalkyl, C 0~3 alkylenearyl, or C 0~3 alkyleneheteroaryl, and each R’ is independently H, C 1~6 alkyl, C 1~6 haloalkyl, C 3~4 cycloalkyl, C 2~3 alkenyl, C 2~3 alkynyl, aryl, or heteroaryl, or two R’ substituents are bonded to each other together with the nitrogen atom to which they are attached to form a 3- to 7-membered ring, R 3 is halo, C 1~3 alkyl, C1~2 haloalkyl, C 1~3 alkoxy, C 3~4 cycloalkyl, C 2~3 alkenyl, C 2~3 alkynyl, aryl, or heteroaryl, R 4 is

Chem.

Chem.

[0089] In the case of the compounds of formulas (II), (III), and (III’), in some embodiments, Q is C=O. In some embodiments, Q is C=S. In some embodiments, Q is C=NR 8 is. R 8 is, C 1~2 alkyl, for example, may be methyl.

[0090] Q is, CR 8 R 9 or C=CR 8 R 9 can be. R 8 and R 9 can be bonded to each other together with the carbon atom to which they are attached to form a 3- to 4-membered ring, for example, a cyclopropyl ring. In some embodiments, R 8 is C 1~2 alkyl (for example, methyl), and R 9 is H.

[0091] In the case of the compounds of formulas (II), (III), (III’), (IV), (IV’), and (V), in various embodiments, R 10 is C 1~4 alkyl, aryl, heteroaryl, C 3~6 cycloalkyl, C 3~6 heterocycloalkyl, C 1~4 alkoxy, or aryloxy. In various embodiments, R 10 is C 1~8 alkyl, C 1~5 alkyl, or C 1~3 alkyl. In various embodiments, R 10 is C0~3 Alkylene aryl, C 0~1 Alkylene aryl, or phenyl. In various embodiments, R 10 is C 0~3 Alkylene heteroaryl, or C 0~1 Alkylene heteroaryl, and such heteroaryl can be, for example, pyridyl. In various embodiments, R 10 is C 0~3 Alkylene-C 3~8 Cycloalkyl, C 0~1 Alkylene-C 3~8 Cycloalkyl, or C 3~8 Cycloalkyl, and such cycloalkyl can be, for example, cyclohexyl. In various embodiments, R 10 is C 0~3 Alkylene-C 3~8 Heterocycloalkyl or C 0~1 Alkylene-C 3~8 Heterocycloalkyl. In various embodiments, R 10 is C 0~6 Alkylene amine or C 0~3 Alkylene amine or amine. Some specifically contemplated R 10 include i-Pr, t-Bu, phenyl, benzyl, OCH3, Cl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,

Chem.

Chem.

Chem.

[0092] For all compounds: R 1 can be a fraction. For example, R 1 is H, C 1~2 Alkyl (e.g., methyl), C 1~2 It can be haloalkyl (e.g., CF), or halo (e.g., F). Some specifically contemplated R 1 includes H, F, Me, Cl, and CF3.

[0093] R 2 is C 1~3 Alkyl, C 1~3 Haloalkyl, C 1~3 Alkoxy, C 0~1 Alkylene-C 3~8 Cycloalkyl, C 3~6 Cycloalkyl, C 0~1 alkylene aryl (e.g., aryl), or C 0~1 Alkylene may be heteroaryl (e.g., heteroaryl). Some specifically contemplated R 2 Groups include phenyl, naphthyl, pyridyl, indazolyl, indolyl, azaindolyl, indolinyl, benzotriazolyl, benzoxadiazolyl, imidazolyl, cinnolinyl, imidazopyridyl, pyrazolopyridyl, quinolinyl, isoquinolinyl, quinazolinyl, quinazolinonyl, indolinonyl, isoindolinonyl, tetrahydronaphthyl, tetrahydroquinolinyl, or tetrahydroisoquinolinyl groups. 2 These include Cl, Br, CF3, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, piperidine, pyrrolidine, azetidine, OCH3, OCH2CH3, phenyl, [ka] TIFF0007717224000100.tif253169TIFF0007717224000101.tif190165. In some embodiments, R 2 teeth [ka] is as follows.

[0094] R 3 is a halo (e.g., Cl, F), C 1~2 alkyl (e.g., methyl), or C 1~2 haloalkyl (e.g., CF3). Some specifically contemplated R 3 include Cl, F, Me, CF3, OMe, Et, C=CH2, and cyclopropyl.

[0095] L is a bond, C 1~6 alkylene, -O-C 0~5 alkylene, -S-C 0~5 alkylene, or -NH-C 0~5 alkylene, and in the case of C 2~6 alkylene, -O-C 2~5 alkylene, -S-C 2~5 alkylene, and NH-C 2~5 alkylene, one carbon atom of the alkylene group can optionally be replaced by O, S, or NH. For example, L can be -CH2-NH- when the carbon of the C2 alkylene group is replaced by NH, or -O-CH2CH2-O- when the carbon of the O-C3 alkylene group is replaced by O. Other options for substituting C3, C4, C5, or C6 alkylene with O, S, or NH are specifically contemplated. In some embodiments, L is C 1~2 alkylene, O, S, or NH. In some embodiments, L is a bond.

[0096] Ring A is a monocyclic 4- to 7-membered ring or a bicyclic, bridged, fused, or spiro 6- to 11-membered ring. Some specifically contemplated rings include cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, pyrrolidinyl, piperidinyl, azepanyl, imidazolidinyl, hexahydropyrimidinyl, hexahydropyridazinyl, tetrahydrofuranyl, tetrahydrothiophenyl, azetidinyl, spiroheptyl, spirooctyl, spirononyl, spirodecyl, diazabicyclodecyl, diazabicyclononyl, diazabicyclooctyl, diazabicycloheptyl, hexahydropyrrolopyridyl, octahydropyrrolopyridyl, and octahydropyrrolopyrimidinyl. In various embodiments, ring A can include piperidinyl, piperazinyl, pyrrolidinyl, or azetidinyl. In some embodiments, ring A includes piperidinyl. Ring A can be further substituted with 1 to 3 substituents. Some non-limiting examples of substitutions for ring A include 1 to 3 substituents selected from alkyl, alkenyl, alkynyl, hydroxyalkyl, carboxylic acid or ester, haloalkyl, alkylamine, C(O)NH2, oxo, halo, cyano, and isocyano.

[0097] R 4 is

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0098] R 4 is [Chemistry] In the case of, more specifically [Chemistry] it may be. In such embodiments, ring A is, for example, [Chemistry] it may be.

[0099] R 5 and R 6 are substituents of the acrylamide moiety of the KRAS inhibitors disclosed herein. In some embodiments, each of R 5 and R 6 is H. Some specifically contemplated R 5 substituents include H, Br, Cl, F, CN, CH3, CF3, CH2Br, CH2OH, CH2CH2OH, CH2OCH2 phenyl, cyclopropyl, phenyl, CH2 phenyl, CH2OCH3, CH2N(CH3)2, CH2N(CH2CH3)2, CH2CO2H, CH2CO2CH3, CH2NHC(O)CH3, CH2C(O)NHCH3, CH2OC(O)CH3, or [Chemistry] is included.

[0100] Some specifically contemplated R 6The substituents include phenyl, cyclopropyl, CH3, CF3, CH2CH3, CH2NH2, CH(CH3)NH2, CH(CH3)2NH2, CH2Cl, CH2Br, CH2OCH3, CH2O phenyl, CH2OH, CO2H, CO2CH2CH3, CH2CO2H, CH2CH2NH2, CH2CH2OH, CH2CH2N(CH3)2, CH2NHCH3, C(O)NHCH3, C(O)N(CH3)2, CH2C(O)NH phenyl, CH2CHF2, CH2F, CHF2, CH2NHC(O)CH3, CH2NHCH2CH2OH, CH2NHCH2CO2H, CH2NH(CH3)CH2CO2CH3, CH2NHCH2CH2OCH3, CH2NH(CH3)CH2CH2OCH3, CH2NH(CH3)CH2C(O)N(CH3)2, CH2NH(CH3)CH2C(O)NHCH3, CH2CH2CCH, CH2NMe2, CH2NH(CH3)CH2CH2OH, CH2NH(CH3)CH2CH2F, CH2N + (CH3)3, CH2NHCH2CHF2, CH2NHCH2CH3, [Chemical formula] TIFF0007717224000112.tif136167 is included.

[0101] R 5 and R 6 can be bonded to each other together with the atoms to which they are bonded to form a 4- to 6-membered ring, for example, a 5-membered or 6-membered ring. Such rings include those in which R 5 and R 6 are together [Chemical formula] is included.

[0102] In most embodiments, R 7 is H. However, in some embodiments, R 7 is methyl. In other embodiments, R 7 and R 5 together are -CH2- or -C(O)CH2-.

[0103] Some specifically contemplated partial alternatives

Chem.

[0104] Some specifically contemplated Rs 4 The substituents include

Chem.

[0105] Some specifically contemplated Rs 4’ The substituents include

Chem.

[0106] In another embodiment, the present invention

Chem.

[0107] In another embodiment, the present invention

Chem.

[0108] In another embodiment, the present invention [Chemical Formula] discloses a compound having a structure selected from, or a stereoisomer thereof, an atropisomer thereof, a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable salt of the stereoisomer thereof, or a pharmaceutically acceptable salt of the atropisomer thereof.

[0109] In another embodiment, the present invention [Chemical Formula] discloses a compound having a structure selected from, or a stereoisomer thereof, an atropisomer thereof, a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable salt of the stereoisomer thereof, or a pharmaceutically acceptable salt of the atropisomer thereof.

[0110] In another embodiment, the present invention [Chemical Formula] discloses a compound having a structure selected from, or a stereoisomer thereof, an atropisomer thereof, a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable salt of the stereoisomer thereof, or a pharmaceutically acceptable salt of the atropisomer thereof.

[0111] In another embodiment, the present invention [Chemical Formula] discloses a compound having a structure selected from, or a stereoisomer thereof, an atropisomer thereof, a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable salt of the stereoisomer thereof, or a pharmaceutically acceptable salt of the atropisomer thereof.

[0112] In another embodiment, the present invention [Chemical Formula] A compound having a structure selected from, or a stereoisomer thereof, an atropisomer thereof, a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable salt of the stereoisomer thereof, or a pharmaceutically acceptable salt of the atropisomer thereof is disclosed.

[0113] In another embodiment, the present invention

Chemical formula

[0114] In another embodiment, the present invention

Chemical formula

[0115] In another embodiment, the present invention

Chemical formula

[0116] In another embodiment, the present invention

Chemical formula

[0117] In another embodiment, the present invention

Chemical formula

[0118] In another embodiment, the present invention

Chemical formula

[0119] In another embodiment, the present invention

Chemical formula

[0120] In another embodiment, the present invention

Chemical formula

[0121] In another embodiment, the present invention

Chemical formula

[0122] In another embodiment, the present invention

Chemical formula

[0123] In another embodiment, these compounds can be used as intermediates in the process for producing the compounds of the present application.

[0124] In another embodiment, these compounds can be in pharmaceutically acceptable salt form.

[0125] In another embodiment, these compounds can be included in a pharmaceutical formulation comprising any one or more of the compounds and a pharmaceutically acceptable excipient.

[0126] In another embodiment, these compounds can be used in a method for inhibiting KRAS G12C in cells, which includes contacting the cells with any one of the compounds or a pharmaceutical formulation.

[0127] In another embodiment, these compounds can be used in a method for treating cancer in a subject, which includes administering a therapeutically effective amount of any one of the compounds or a composition to the subject.

[0128] In another embodiment, the cancer is lung cancer, pancreatic cancer, or colorectal cancer.

[0129] In another embodiment, the cancer is lung cancer.

[0130] In another embodiment, the cancer is pancreatic cancer.

[0131] In another embodiment, the cancer is colorectal cancer.

[0132] In another embodiment, the method further comprises administering to a patient in need thereof a therapeutically effective amount of a further pharmaceutically active compound.

[0133] In another embodiment, the further pharmaceutically active compound is carfilzomib.

[0134] In another embodiment, the further pharmaceutically active compound is cytarabine.

[0135] In another embodiment, the present invention comprises using one or more of the compounds for treating cancer in a subject.

[0136] In another embodiment, the present invention comprises using one or more of the compounds in the preparation of a medicament for treating cancer.

[0137] In another embodiment, the cancer is a hematological malignancy.

[0138] In another embodiment, the present invention comprises using one or more of the cancer-treating compounds, wherein the cancer is a hematological malignancy.

[0139] Examples 1 to 11 below are presented using a classification system, where the first number refers to the method used for compound synthesis, the second number refers to an identification number, and the third number, if described, refers to the elution order of the compound in the chromatographic separation step. If there is no third number, the compound is either a single compound or a mixture of isomers. In the classification system used for Examples 12 to 53, the first number is the identification number, and the second number, if described, refers to the elution order of the compound in the chromatographic separation step. If there is no second number, the compound is either a single compound or a mixture of isomers. For formatting purposes, the sequential numbering of the examples is interrupted and specific example numbers are intentionally omitted. "-" means no change or that there is no entry in the relevant box. Specifically intended compounds include those described in Table 1 and Table 1(a). [Table 2] TIFF0007717224000141.tif249168TIFF0007717224000142.tif244166TIFF0007717224000143.tif208168TIFF0007717224000144.tif225166TIFF0007717224000145.tif254169TIFF0007717224000146.tif224167TIFF0007717224000147.tif219167TIFF0007717224000148.tif196167TIFF0007717224000149.tif254168TIFF0007717224000150.tif249168TIFF0007717224000151.tif245167TIFF0007717224000152.tif244168TIFF0007717224000153.tif233167TIFF0007717224000154.tif232166TIFF0007717224000155.tif72166 [Table 3] TIFF0007717224000157.tif212166

[0140] Synthesis of the disclosed compounds The compounds disclosed herein can be synthesized by a number of specific methods. The examples outlining specific synthetic routes, and the general schemes below, are intended to provide guidance to synthetic chemists of ordinary skill, who will readily understand that solvents, concentrations, reagents, protecting groups, the order of synthetic steps, time, temperature, etc. can be adequately varied within the skill and judgment of one of ordinary skill in the art as needed.

[0141]

Chemical formula

[0142] Intended halogenating agents include, but are not limited to, chlorine, bromine, N-chlorosuccinimide, and N-bromosuccinimide, and optionally, for example, in the presence of a catalyst such as iron or aluminum. Synthetic chemists with ordinary skill will readily understand that the use of other halogenating agents and catalysts is possible.

[0143] Intended amidating agents include, but are not limited to, N,N'-diisopropylcarbodiimide, N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide, benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate, O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, thionyl chloride, isobutyl chloroformate, diethyl cyanophosphonate, carbonyldiimidazole, and polyphosphonic anhydride. Synthetic chemists with ordinary skill will readily understand that the use of other amidating agents is possible.

[0144] Intended sulfurizing agents include, but are not limited to, sulfur, phosphorus pentasulfide, and Lawesson's reagent. Synthetic chemists with ordinary skill will readily understand that the use of other sulfurizing agents is possible.

[0145] Intended oxidizing agents include, but are not limited to, hydrogen peroxide, iodobenzene diacetate, t-butyl hydroperoxide, N-bromosuccinimide, and ammonium peroxodisulfate. Synthetic chemists with ordinary skill will readily understand that the use of other oxidizing agents is possible.

[0146] Intended activating agents include, but are not limited to, sodium nitrite and t-butyl nitrite. Synthetic chemists with ordinary skill will readily understand that the use of other activating agents is possible.

[0147] Intended cross-coupling reactions include, but are not limited to, Suzuki coupling, Negishi coupling, Hiyama coupling, Kumada coupling, and Stille coupling. A chemist with ordinary skill will readily understand that the coupling as shown in Method 1 can be carried out under a number of conditions.

[0148]

Chemical formula

[0149]

Chemical formula

[0150]

Chemical formula

[0151]

Chemical Structure

[0152]

Chemical Structure

[0153] Contemplated metallation agents include, but are not limited to, bis(pinacolato)diboron, magnesium, zinc, hexamethyldistannane, and n-butyllithium. A synthetic chemist of ordinary skill will readily recognize that other metallation agents and catalysts are possible.

[0154] [ka] Method 7 Synthesis: Method 7 provides an alternative method for forming compounds of formula (I) disclosed herein. R is first prepared by cross-coupling of an aromatic or heteroaromatic acid intermediate shown in Step 1 with an X halide. 2 Then, in step 2, the acid moiety is converted to the appropriate R 4 The acid derivative is reacted with a (protected) reagent. Then, in step 3, the carbonyl group of the acid derivative is converted to a thiocarbonyl group using a sulfurizing agent. Then, in step 4, the thioacid derivative is reacted with an oxidizing agent to generate the isothiazole intermediate. Finally, R 4 Deprotection and acylation of the R 2 The group is deprotected.

[0155] [ka] Method 8 Synthesis: Compounds of formula (II) disclosed herein can be synthesized as outlined in Method 8. In Step 1, an appropriate aromatic or heteroaromatic acid is reacted with an amidating agent to generate a primary amide intermediate. The amide is then reacted with an isocyanate-forming reagent and R 10React with a substituted amine to form a urea intermediate. Intended isocyanate-forming agents include oxalyl chloride, thionyl chloride, and phosphorus oxychloride. Thereafter, in Step 3, the urea intermediate is reacted with a cyclizing agent to form the illustrated quinazolinedione ring. Intended cyclizing agents include bases such as potassium hexamethyldisilazide, potassium tert-butoxide, sodium hydride, and phosphazene, but are not limited thereto. Thereafter, in Step 4, an appropriate R 2 (protected) reagent and the X-halide of the quinazolinedione intermediate are subjected to a cross-coupling reaction to introduce the R 2 moiety. Thereafter, in Step 5, the oxo group of the quinazolinedione is converted to a leaving group using an activating agent. Intended activating agents include thionyl chloride, triflic anhydride, phosphorus oxychloride, and phosphorus pentachloride, but are not limited thereto. Thereafter, as shown in Step 6, the leaving group is replaced with a protected R 4 group to produce a substituted quinazolinone. The subsequent deprotection-acylation-deprotection sequence shown in Steps 7-9 is the same as in Step 8 of Method 1.

[0156]

Chemical Structure

[0157]

Chemical Structure

[0158]

Chemical Structure

[0159] Pharmaceutical Compositions, Administration, and Routes of Administration This specification also provides pharmaceutical compositions comprising the compounds disclosed herein together with pharmaceutically acceptable excipients, such as diluents or carriers, etc. Compounds and pharmaceutical compositions suitable for use in the present invention include those in which the compound can be administered in an effective amount to achieve its intended purpose. The administration of the compound is detailed below.

[0160] Suitable pharmaceutical formulations can be determined by those skilled in the art according to the route of administration and the desired dosage. See, for example, Remington’s Pharmaceutical Sciences, 1435 - 712 (18th ed., Mack Publishing Co, Easton, Pennsylvania, 1990). The formulation can affect the physical state, stability, in vivo release rate, and in vivo clearance rate of the administered drug. Depending on the route of administration, a suitable dosage may be calculated according to body weight, body surface area, or organ size. Further refinement of the calculations necessary for determining an appropriate therapeutic dosage can be routinely performed by those skilled in the art without undue experimentation, especially in light of dosage information and the evaluation methods disclosed herein as well as pharmacokinetic data obtainable through animal or human clinical trials.

[0161] The terms "pharmaceutically acceptable" or "pharmacologically acceptable" refer to molecular species and compositions that do not produce adverse reactions, allergic reactions, or other harmful reactions when administered to animals or humans. As used herein, "pharmaceutically acceptable" includes all solvents, dispersion media, coating agents, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. The use of such excipients for pharmaceutically active substances is well known in the art. Any conventional media or agent is intended to be used in a therapeutic composition as long as it is not incompatible with the therapeutic composition. Auxiliary active compounds can also be incorporated into the composition. In an exemplary embodiment, the formulation may include corn syrup solids, high oleic safflower oil, coconut oil, soybean oil, L-leucine, tricalcium phosphate, L-tyrosine, L-proline, L-lysine acetate, DATEM (emulsifier), L-glutamine, L-valine, dipotassium phosphate, L-isoleucine, L-arginine, L-alanine, glycine, L-asparagine monohydrate, L-serine, potassium citrate, L-threonine, sodium citrate, magnesium chloride, L-histidine, L-methionine, ascorbic acid, calcium carbonate, L-glutamic acid, L-cystine dihydrochloride, L-tryptophan, L-aspartic acid, choline chloride, taurine, m-inositol, iron sulfate, ascorbyl palmitate, zinc sulfate, L-carnitine, alpha-tocopheryl acetate, sodium chloride, niacinamide, mixed tocopherols, calcium pantothenate, copper sulfate, thiamine chloride hydrochloride, vitamin A palmitate, manganese sulfate, riboflavin, pyridoxine hydrochloride, folic acid, beta-carotene, potassium iodide, phylloquinone, biotin, sodium selenate, chromium chloride, sodium molybdate, vitamin D3, and cyanocobalamin.

[0162] The compound may be present in a pharmaceutical composition as a pharmaceutically acceptable salt. As used herein, "pharmaceutically acceptable salts" include, for example, base addition salts and acid addition salts.

[0163] Pharmaceutically acceptable base addition salts may be formed using a metal or an amine, such as an alkali metal and an alkaline earth metal or an organic amine. Pharmaceutically acceptable salts of the compounds may also be prepared using pharmaceutically acceptable cations. Suitable pharmaceutically acceptable cations are well known to those skilled in the art and include cations of alkali, alkaline earth, ammonium and quaternary ammonium. Carbonates or bicarbonates are also possible. Examples of metals used as cations are sodium, potassium, magnesium, ammonium, calcium, or ferrous, etc. Examples of suitable amines include isopropylamine, trimethylamine, histidine, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, dicyclohexylamine, ethylenediamine, N-methylglucamine, and procaine.

[0164] Pharmaceutically acceptable acid addition salts include inorganic acid salts or organic acid salts. Examples of suitable acid salts include hydrochloride, formate, acetate, citrate, salicylate, nitrate, and phosphate. Other suitable pharmaceutically acceptable salts are well known to those skilled in the art and include, for example, in addition to formic acid, acetic acid, citric acid, oxalic acid, tartaric acid, or mandelic acid, hydrochloric acid, hydrobromic acid, sulfuric acid or phosphoric acid, and also organic carboxylic acids, sulfonic acids, acids with a sulfonic or phosphoric group or N-substituted sulfamic acids, such as acetic acid, trifluoroacetic acid (TFA), propionic acid, glycolic acid, succinic acid, maleic acid, hydroxymaleic acid, methylmaleic acid, fumaric acid, malic acid, tartaric acid, lactic acid, oxalic acid, gluconic acid, glucaric acid, glucuronic acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, salicylic acid, 4-aminosalicylic acid, 2-phenoxybenzoic acid, 2-acetoxybenzoic acid, embonic acid, nicotinic acid or isonicotinic acid, and also, in addition to the 20 alpha amino acids used in protein synthesis in nature, such as amino acids like glutamic acid or aspartic acid, phenylacetic acid, methanesulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, ethane-1,2-disulfonic acid, benzenesulfonic acid, 4-methylbenzenesulfonic acid, naphthalene-2-sulfonic acid, naphthalene-1,5-disulfonic acid, 2- or 3-phosphoglyceric acid, glucose-6-phosphate, N-cyclohexylsulfamic acid (accompanied by cyclamate formation), or other acidic organic compounds, ascorbic acid, etc. are also included.

[0165] The pharmaceutical compositions containing the compounds disclosed herein can be manufactured by conventional methods, for example, by each step such as conventional mixing, dissolving, granulating, sugar coating tablet manufacturing, micronization, emulsification, encapsulation, entrapping, or lyophilization. Appropriate formulations depend on the selected route of administration.

[0166] In the case of oral administration, suitable compositions can be easily formulated by combining the compounds disclosed herein with pharmaceutically acceptable excipients such as carriers well-known in the art. Such excipients and carriers enable the formulation of the compounds of the present application for oral ingestion by the patient to be treated in the form of tablets, pills, dragees, capsules, solutions, gels, syrups, slurries, suspensions, etc. Oral pharmaceutical preparations can be prepared by adding the compounds disclosed herein using solid excipients, optionally grinding the resulting mixture, adding suitable auxiliaries as necessary, and then processing the granule mixture to obtain tablets or dragee cores. Suitable excipients include, for example, fillers and cellulose preparations. Disintegrants can be added as necessary. A wide variety of formulations are well-known as pharmaceutically acceptable raw materials, and they can be, for example, binders for various formulations (e.g., natural polymers or synthetic polymers), lubricants, surfactants, sweeteners and flavoring agents, coating materials, preservatives, pigments, thickeners, adjuvants, antibacterial agents, antioxidants, and carriers.

[0167] When administering a therapeutically effective amount of the compounds disclosed herein orally, the composition is typically in solid form (e.g., tablets, capsules, pills, powders, or lozenges) or in liquid form (e.g., aqueous suspensions, solutions, elixirs, or syrups).

[0168] When administered in tablet form, the composition can further contain functional solids and / or solid carriers, such as gelatin or adjuvants. Tablets, capsules, and powders can contain from about 1% to about 95% of the compound, preferably from about 15% to about 90% of the compound.

[0169] When administered in liquid or suspension form, functional liquids and / or liquid carriers such as water, petroleum, or oils of animal or plant origin can be added. The composition in liquid form can further contain physiological saline, sugar alcohol solutions, dextrose or other sugar solutions, or glycols. When administered in liquid or suspension form, the composition can contain from about 0.5 to about 90% by weight of the disclosed compounds herein, preferably from about 1 to about 50% of the disclosed compounds herein. In one intended embodiment, the liquid carrier is non-aqueous or substantially non-aqueous. For administration in liquid form, the composition can be provided as an immediate-dissolving solid formulation that is dissolved or suspended immediately prior to administration.

[0170] When a therapeutically effective amount of the disclosed compounds herein is administered by intravenous injection, intradermal injection, or subcutaneous injection, the composition is in the form of a pyrogen-free parenterally acceptable aqueous solution. The preparation of such parenterally acceptable solutions should be considered with respect to pH, isotonicity, stability, etc., but these are within the skill of those in the art. Preferred compositions for intravenous injection, intradermal injection, or subcutaneous injection typically contain an isotonic vehicle in addition to the compounds disclosed herein. Such compositions can be prepared for administration as an aqueous solution of the free base or a pharmaceutically acceptable salt, suitably mixed with a surfactant such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof, as well as in oils. Under normal storage and use conditions, these preparations can optionally contain preservatives to prevent the growth of microorganisms.

[0171] Injectable compositions can include sterile aqueous solutions, suspensions, or dispersions, and sterile powders for the extemporaneous preparation of sterile injectable solutions, suspensions, or dispersions. In all embodiments, such forms must be in a sterile form and be fluids that can easily pass through a hypodermic needle. They must be stable under manufacturing and storage conditions and, optionally, be resistant to the contaminating action of microorganisms such as bacteria and fungi by including preservatives. The carrier can be, for example, a solvent or dispersion medium containing water, ethanol, polyols (such as glycerol, propylene glycol, and liquid polyethylene glycol, etc.), suitable mixtures thereof, and vegetable oils. In one intended embodiment, the carrier is non-aqueous or substantially non-aqueous. Suitable fluidity can be maintained, for example, by using coatings such as lecithin, by maintaining the required particle size of the compounds in the dispersion embodiments, and by using surfactants. Prevention of microbial action can be provided by various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, etc. In many embodiments, it would be preferable to include isotonic agents such as sugars or sodium chloride. Sustained absorption of injectable compositions can be brought about by using agents that delay absorption, such as aluminum monostearate and gelatin, in the composition.

[0172] Sterile injectable solutions are prepared by incorporating the active compound in the required amount, optionally with other various aforementioned ingredients, into a suitable solvent and then filtering and sterilizing. Generally, dispersions are prepared by incorporating various sterile active ingredients into a sterile vehicle containing a base dispersion medium and other required ingredients from the above. In the case of embodiments of sterile powders for the preparation of sterile injectable solutions, the preferred preparation methods are the vacuum drying method and the freeze-drying method, which result in a powder obtained by adding the active ingredient and any further desired ingredients obtained from the previous sterile filtered solution.

[0173] Sustained release or controlled release formulations may be prepared to achieve controlled release of the active compound upon contact with body fluids in the gastrointestinal tract and to substantially maintain and be effective in the plasma level of the active compound. For example, release can be controlled by one or more of dissolution, diffusion, and ion exchange. Further, by making a controlled release formulation, absorption through saturable or rate-limiting pathways in the gastrointestinal tract can be enhanced. For example, for this purpose, the compound may be embedded in a polymeric matrix of a biodegradable polymer, a water-soluble polymer or a mixture of both, and optionally a suitable surfactant. In this case, embedding means incorporating fine particles into the matrix of the polymer. The controlled release formulation can also be obtained by encapsulating dispersed fine particles or emulsified microdroplets by known dispersion or emulsion coating techniques.

[0174] For administration by inhalation, the compounds of the present invention are conveniently delivered in the form of an aerosol spray presented from a pressurized pack or a nebulizer using a suitable propellant. In the case of a pressurized aerosol embodiment, the dosage unit can be determined by providing a valve to deliver a metered amount. Capsules and cartridges (e.g., made of gelatin) for use in an inhaler or inhalation device can be formulated to contain a powder mixture of the compound and a suitable powder base such as lactose or starch.

[0175] The compounds disclosed herein can be formulated for parenteral administration by injection (e.g., bolus injection or continuous infusion). Injectable formulations can be presented in unit dosage forms (e.g., in ampoules or multi-dose containers) with the addition of preservatives. The composition can take the form of a suspension, solution, or emulsion in an oily or aqueous vehicle and can contain substances for formulation such as suspending agents, stabilizers, and / or dispersing agents.

[0176] Pharmaceutical preparations for parenteral administration include aqueous solutions of the compound in water-soluble form. Further, suspensions of the compound can be prepared as suitable oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils or synthetic fatty acid esters. Aqueous injection suspensions can contain substances that increase the viscosity of the suspension. Optionally, suitable stabilizers or agents that increase the solubility of the compound can be included in the suspension to enable the preparation of highly concentrated solutions. Alternatively, the composition contained can be in powder form for constitution prior to use with a suitable vehicle (e.g., pyrogen-free sterile water).

[0177] The compounds disclosed herein can also be formulated into rectal compositions such as suppositories or retention enemas (e.g., containing conventional suppository bases). In addition to the formulations described above, the compounds can be formulated as depot preparations. Such long-acting formulations can be administered by implantation (e.g., subcutaneously or intramuscularly) or intramuscular injection. Thus, for example, the compound can be formulated using a suitable polymeric substance or a hydrophobic substance (e.g., as an emulsion in an acceptable oil) or an ion exchange resin, or as a slightly insoluble derivative, e.g., as a slightly insoluble salt.

[0178] In particular, the compounds disclosed herein can be administered orally, buccally, or sublingually in the form of tablets containing excipients such as starch or lactose, or in capsules or ovules (mixtures or single agents with excipients), or in the form of elixirs or suspensions containing flavoring or coloring agents. Such liquid preparations can be prepared using pharmaceutically acceptable additives such as suspending agents. The compound can also be administered parenterally, e.g., by intravenous, intramuscular, subcutaneous, or intracoronary injection. In the case of parenteral administration, it is best to use the compound in the form of a sterile aqueous solution that can contain other substances, e.g., salts, or sugar alcohols such as mannitol or glucose, to make the solution isotonic with the blood.

[0179] For animals, the disclosed compounds of the present specification are administered as a formulation that is preferably acceptable in accordance with normal veterinary practice. A veterinarian can readily determine the optimal dosage regimen and route of administration for an individual animal.

[0180] In some embodiments, all the components necessary for the treatment of KRAS-related disorders, in which the compounds disclosed herein are used alone or in combination with another agent or an intervention conventionally practiced for the treatment of such diseases, may be packaged in a kit. Specifically, the present invention provides a kit for use in a therapeutic intervention for a disease comprising a pharmaceutical package, said kit comprising a compound disclosed herein and a buffer and other components for preparing a deliverable form of said pharmaceutical, and / or a device for delivering such a pharmaceutical, and / or any agent for use in combination therapy using a compound disclosed herein, and / or instructions regarding the treatment of the disease, which are included with the pharmaceutical. The instructions may be fixed to any tangible medium, such as a printed matter, or a computer-readable magnetic or optical medium, or an instruction to refer to a data source of a remote computer, such as a World Wide Web page accessible via the Internet.

[0181] "Therapeutically effective amount" means an amount effective for treating or preventing the onset of, or reducing existing symptoms in, a subject being treated. Determination of an effective amount is well within the ability of one of ordinary skill in the art, particularly in light of the detailed disclosure provided herein. Generally, "therapeutically effective dosage" refers to the amount of a compound that results in the achievement of the desired effect. For example, in a preferred embodiment, a therapeutically effective amount of the disclosed compound of the present specification results in at least a 5%, at least a 10%, at least a 15%, at least a 20%, at least a 25%, at least a 30%, at least a 35%, at least a 40%, at least a 45%, at least a 50%, at least a 55%, at least a 60%, at least a 65%, at least a 70%, at least a 75%, at least an 80%, at least an 85%, or at least a 90% decrease in KRAS activity compared to a control.

[0182] The amount of the compound administered may depend on the subject being treated, the subject's age, health, sex, and weight, the type of concomitant treatment (if any), the severity of the affliction, the nature of the desired effect, the method and frequency of treatment, and the judgment of the prescribing physician. The frequency of administration may also depend on the pharmacodynamic effect on the arterial oxygen pressure. However, the most preferred dosage can be adjusted to suit the individual subject, which can be understood and determined by those skilled in the art without undue experimentation. In this case, typically, an adjustment of the standard dosage (for example, a reduction in dosage if the patient has a low body weight) is made.

[0183] Although individual needs vary, determining the optimal range of the effective amount of a compound is within the skill of those in the art. For example, when administered to humans in the curative or prophylactic treatment of the conditions and disorders specified herein, typical dosages of the compounds of the invention are from about 0.05 mg / kg / day to about 50 mg / kg / day, for example, at least 0.05 mg / kg, at least 0.08 mg / kg, at least 0.1 mg / kg, at least 0.2 mg / kg, at least 0.3 mg / kg, at least 0.4 mg / kg, or at least 0.5 mg / kg, and preferably 50 mg / kg or less, 40 mg / kg or less, 30 mg / kg or less, 20 mg / kg or less, or 10 mg / kg or less, which can be, for example, from about 2.5 mg / day (0.5 mg / kg × 5 kg) to about 5000 mg / day (50 mg / kg × 100 kg). For example, the dosage of the compound can be from about 0.1 mg / kg / day to about 50 mg / kg / day, from about 0.05 mg / kg / day to about 10 mg / kg / day, from about 0.05 mg / kg / day to about 5 mg / kg / day, from about 0.05 mg / kg / day to about 3 mg / kg / day, from about 0.07 mg / kg / day to about 3 mg / kg / day, from about 0.09 mg / kg / day to about 3 mg / kg / day, from about 0.05 mg / kg / day to about 0.1 mg / kg / day, from about 0.1 mg / kg / day to about 1 mg / kg / day, from about 1 mg / kg / day to about 10 mg / kg / day, from about 1 mg / kg / day to about 5 mg / kg / day, from about 1 mg / kg / day to about 3 mg / kg / day, from about 3 mg / day to about 500 mg / day, from about 5 mg / day to about 250 mg / day, from about 10 mg / day to about 100 mg / day, from about 3 mg / day to about 10 mg / day, or from about 100 mg / day to about 250 mg / day. Such dosages may be administered in a single dose or divided into multiple doses.

[0184] Methods of using KRAS G12C inhibitors The present disclosure provides a method of inhibiting RAS-mediated cell signaling, which comprises contacting cells with an effective amount of one or more of the disclosed compounds herein. The inhibition of RAS-mediated signaling can be evaluated and demonstrated by a variety of methods known in the art. Non-limiting examples include: (a) showing a decrease in the GTPase activity of RAS; (b) showing a decrease in GTP-binding affinity or an increase in GDP-binding affinity; (c) showing an increase in the Koff of GTP or a decrease in the Koff of GDP; (d) showing a decrease in the levels of signaling molecules downstream of the RAS pathway, such as a decrease in the levels of pMEK, pERK, or pAKT; and / or (e) showing a decrease in the binding of RAS complexes to downstream signaling molecules including, but not limited to, Raf. Kits and commercially available assays can be utilized for one or more of the above determinations.

[0185] The present disclosure also provides a method of treating a disease state including, but not limited to, a condition (such as cancer) in which a mutation in G12C KRAS, HRAS, or NRAS is involved, using a compound or pharmaceutical composition of the present disclosure.

[0186] In some embodiments, a method for cancer treatment is provided, and such method comprises administering to a subject in need thereof any of the above-described pharmaceutical compositions comprising an effective amount of the disclosed compounds herein. In some embodiments, the cancer is induced by a G12C mutation in KRAS, HRAS, or NRAS. In various embodiments, the cancer is pancreatic cancer, colorectal cancer, or lung cancer. In some embodiments, the cancer is gallbladder cancer, thyroid cancer, and cholangiocarcinoma.

[0187] In some embodiments, the present disclosure provides a method of treating a disorder in a subject in need of treatment for the disorder, wherein the method comprises determining whether such subject has a G12C mutation in KRAS, HRAS, or NRAS, and if the subject is determined to have a G12C mutation in KRAS, HRAS, or NRAS, administering to the subject a therapeutically effective dose of at least one of the disclosed compounds herein or a pharmaceutically acceptable salt thereof.

[0188] The disclosed compounds inhibit anchorage-independent cell growth and therefore have the ability to inhibit tumor metastasis. Accordingly, in another embodiment, the present disclosure provides a method of inhibiting tumor metastasis, such method comprising administering an effective amount of a compound disclosed herein.

[0189] G12C mutations in KRAS, HRAS, or NRAS have also been identified in hematological malignancies (e.g., cancers affecting the blood, bone marrow, and / or lymph nodes). Accordingly, certain embodiments are directed to administering the disclosed compounds (e.g., in the form of a pharmaceutical composition) to a patient in need of treatment for a hematological malignancy. Such malignancies include, but are not limited to, leukemias and lymphomas. For example, the disclosed compounds can be used to treat diseases such as acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), chronic myelogenous leukemia (CML), acute monocytic leukemia (AMoL), and / or other leukemias. In other embodiments, the compounds are useful in treating lymphomas, such as all subtypes of Hodgkin's lymphoma or non-Hodgkin's lymphoma. In various embodiments, the compounds are useful in the treatment of plasma cell malignancies, such as multiple myeloma, mantle cell lymphoma, and Waldenstrom's macroglobulinemia.

[0190] Determining whether a tumor or cancer contains a G12C KRAS, HRAS, or NRAS mutation can be performed by evaluating the nucleotide sequence encoding the KRAS, HRAS, or NRAS protein, evaluating the amino acid sequence of the KRAS, HRAS, or NRAS protein, or evaluating the characteristics of a putative KRAS, HRAS, or NRAS mutant protein. The sequence of wild-type human KRAS, HRAS, or NRAS is known in the art (e.g., Accession No. NP203524).

[0191] Methods for detecting mutations in KRAS, HRAS, or NRAS nucleotide sequences are known to those skilled in the art. These methods include, but are not limited to, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), polymerase chain reaction-single-strand conformational polymorphism (PCR-SSCP), real-time PCR, PCR sequencing, mutant allele-specific PCR amplification (MASA), direct sequencing, primer extension reaction, electrophoresis, oligonucleotide ligation assay, hybridization, TaqMan assay, SNP genotyping assay, high-resolution melting curve analysis, and microarray analysis. In some embodiments, a sample is evaluated for G12C KRAS, HRAS, or NRAS mutations by real-time PCR. In real-time PCR, a fluorescent probe specific for the G12C mutation of KRAS, HRAS, or NRAS is used. If the mutation is present, the probe binds and fluorescence is detected. In some embodiments, direct sequencing is used to identify G12C mutations in KRAS, HRAS, or NRAS in a specific region (e.g., exon 2 and / or exon 3) of the KRAS, HRAS, or NRAS gene. With this technique, all possible mutations in the sequenced region are identified.

[0192] Methods for detecting mutations in KRAS, HRAS, or NRAS proteins are known to those skilled in the art. These methods include, but are not limited to, detection of KRAS, HRAS, or NRAS mutants using a binding agent specific for the mutant protein (e.g., an antibody), protein electrophoresis and Western blot, and direct peptide sequencing.

[0193] A wide variety of samples can be used in the methods for determining whether a tumor or cancer contains a G12C KRAS, HRAS, or NRAS mutation. In some embodiments, the sample is obtained from a subject with a tumor or cancer. In some embodiments, the sample is a fresh tumor / cancer sample. In some embodiments, the sample is a frozen tumor / cancer sample. In some embodiments, the sample is a formalin-fixed, paraffin-embedded sample. In some embodiments, the sample is a circulating tumor cell (CTC) sample. In some embodiments, the sample is processed into a cell lysate. In some embodiments, the sample is processed into DNA or RNA.

[0194] The disclosure also relates to a method of treating a hyperproliferative disorder in a mammal, the method comprising administering to the mammal a therapeutically effective amount of a disclosed compound of the present specification, or a pharmaceutically acceptable salt thereof.In some embodiments, the method relates to the treatment of subjects suffering from cancer such as acute myeloid leukemia, adolescent cancer, pediatric adrenocortical carcinoma, AIDS-related cancers (e.g., lymphoma and Kaposi sarcoma), anal cancer, appendiceal cancer, astrocytoma, atypical teratoid rhabdoid tumor, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brainstem glioma, brain tumor, breast cancer, bronchial tumor, Burkitt lymphoma, carcinoid tumor, atypical teratoid rhabdoid tumor, fetal tumor, germ cell tumor, primary lymphoma, cervical cancer, pediatric cancer, chordoma, cardiac tumor, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), chronic myeloproliferative disorder, colon cancer, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, ductal carcinoma in situ (DCIS), fetal tumor, CNS cancer, endometrial cancer, epithelioma, esophageal cancer, nasal neuroblastoma, Ewing sarcoma, extracranial germ cell tumor, extragonadal germ cell tumor, eye cancer, bone malignant fibrous histiocytoma, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), germ cell tumor, gestational trophoblastic tumor, hairy cell leukemia, head and neck cancer, heart cancer, liver cancer, Hodgkin lymphoma, hypopharyngeal cancer, intraocular melanoma, islet cell tumor, pancreatic neuroendocrine tumor, kidney cancer, laryngeal cancer, lip and oral cavity cancer, liver cancer, lobular carcinoma in situ (LCIS), lung cancer, lymphoma, metastatic squamous neck cancer of unknown primary, midline cancer, oral cancer, multiple endocrine neoplasia syndrome, multiple myeloma / plasma cell neoplasm, mycosis fungoides, myelodysplastic syndrome, myelodysplastic / myeloproliferative neoplasm, multiple myeloma, Merkel cell carcinoma, malignant mesothelioma, bone malignant fibrous histiocytoma and osteosarcoma, nasal and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin lymphoma, non-small cell lung cancer (NSCLC), oral cancer, lip and oral cavity cancer, oropharyngeal cancer, ovarian cancer, pancreatic cancer, papillomatosis, paraganglioma, paranasal sinus and nasal cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pleuropulmonary blastoma, primary central nervous system (CNS) lymphoma, prostate cancer, rectal cancer, transitional cell carcinoma, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, skin cancer, gastric cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, T-cell lymphoma, testicular cancer, pharyngeal cancer, thymoma and thymic carcinoma, thyroid cancer, renal pelvis and ureteral transitional cell carcinoma, trophoblastic tumor, pediatric rare cancer, urethral cancer, uterine sarcoma, vaginal cancer, vulvar cancer, or virus-induced cancer.In some embodiments, the method relates to the treatment of non-cancerous hyperproliferative disorders such as benign skin hyperplasia (e.g., psoriasis), restenosis, or prostate (e.g., benign prostatic hyperplasia (BPH)).

[0195] In some embodiments, the method of treatment is directed to the treatment of lung cancer, and such method comprises administering to a subject in need thereof an effective amount of any of the above compounds (or a pharmaceutical composition comprising the same). In certain embodiments, the lung cancer is non-small cell lung cancer (NSCLC), such as adenocarcinoma, squamous cell carcinoma of the lung, or large cell carcinoma of the lung. In some embodiments, the lung cancer is small cell lung cancer. Other lung cancers treatable with the disclosed compounds include, but are not limited to, adenoid cystic tumors, carcinoid tumors, and undifferentiated cancers.

[0196] The present disclosure further provides a method of modulating the activity of a G12C mutant KRAS, HRAS, or NRAS protein by contacting the protein with an effective amount of the disclosed compound. The modulation can be inhibition or activation of protein activity. In some embodiments, the present disclosure provides a method of inhibiting protein activity by contacting a G12C mutant KRAS, HRAS, or NRAS protein with an effective amount of the disclosed compound solution. In some embodiments, the present disclosure provides a method of inhibiting the activity of a G12C mutant KRAS, HRAS, or NRAS protein by contacting a cell, tissue, or organ that expresses the protein of interest. In some embodiments, the present disclosure provides a method of inhibiting protein activity in a subject, including but not limited to rodents and mammals (e.g., humans), by administering an effective amount of the disclosed compound to the subject. In some embodiments, the modulation rate exceeds 25%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%. In some embodiments, the inhibition rate exceeds 25%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%.

[0197] In some embodiments, the present disclosure provides a method of inhibiting KRAS, HRAS, or NRAS G12C activity in a cell by contacting the cell with a disclosed compound in an amount sufficient to inhibit KRAS, HRAS, or NRAS G12C activity in the cell. In some embodiments, the present disclosure provides a method of inhibiting KRAS, HRAS, or NRAS G12C activity in a tissue by contacting the tissue with a disclosed compound in an amount sufficient to inhibit KRAS, HRAS, or NRAS G12C activity in the tissue. In some embodiments, the present disclosure provides a method of inhibiting KRAS, HRAS, or NRAS G12C activity in an organism by contacting the organism with a disclosed compound in an amount sufficient to inhibit KRAS, HRAS, or NRAS G12C activity in the organism. In some embodiments, the present disclosure provides a method of inhibiting KRAS, HRAS, or NRAS G12C activity in an animal by contacting the animal with a disclosed compound in an amount sufficient to inhibit KRAS, HRAS, or NRAS G12C activity in the animal. In some embodiments, the present disclosure provides a method of inhibiting KRAS, HRAS, or NRAS G12C activity in a mammal by contacting the mammal with a disclosed compound in an amount sufficient to inhibit KRAS, HRAS, or NRAS G12C activity in the mammal. In some embodiments, the present disclosure provides a method of inhibiting KRAS, HRAS, or NRAS G12C activity in a human by contacting the human with a disclosed compound in an amount sufficient to inhibit KRAS, HRAS, or NRAS G12C activity in the human. The present disclosure provides a method of treating a disease in a subject in need of treatment for the disease induced by KRAS, HRAS, or NRAS G12C activity.

[0198] Combination therapy: The present disclosure also provides methods of combination therapy, in which agents for which other pathway modulation is known, or other components of the same pathway, or overlapping sets of target enzymes, are used in combination with a compound of the present disclosure or a pharmaceutically acceptable salt thereof. In one aspect, such treatments include, but are not limited to, combining one or more disclosed compounds with chemotherapeutic agents, therapeutic antibodies, and radiation therapy to obtain a synergistic or additive therapeutic effect.

[0199] Currently, many chemotherapeutic agents are known in the art and can be used in combination with the disclosed compounds. In some embodiments, the chemotherapeutic agent is selected from the group consisting of a mitotic inhibitor, an alkylating agent, an antimetabolite, an intercalating antibiotic, a growth factor inhibitor, a cell cycle inhibitor, an enzyme, a topoisomerase inhibitor, a biological response modifier, an antihormone, an angiogenesis inhibitor, and an antiandrogen. Non-limiting examples include chemotherapeutic agents, cytotoxic agents, and non-peptide small molecules, such as Gleevec® (imatinib mesylate), Kyprolis® (carfilzomib), Velcade® (bortezomib), Casodex (bicalutamide), Iressa® (gefitinib), and Adriamycin, etc., and are hosts for chemotherapeutic agents. Non-limiting examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclophosphamide (CYTOXAN™); alkyl sulfonic acids such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carbocon, meturedopa, and uredopa; ethyleneimine and methylmelamine including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolmelamine; nitrogen mustards such as chlorambucil, chloronaphazine, colophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, noburemabicin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosoureas such as carmustine, chloroozotocin, fotemustine, lomustine, nimustine, ranimustine;Antibiotics such as aclacinomycin, actinomycin, aurodox, azaserine, bleomycin, cactinomycin, calicheamicin, carabicin, calminomycin, cardifilin, Casodex (trademark), chromomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin, idarubicin, marcellomycin, mitomycin, mycophenolic acid, nogalamycin, olivomycin, peplomycin, potfiromycin, puromycin, keramycin, rhodomycin, streptonigrin, streptozocin, tubercidin, ubenimex, dinostatin, zorubicin; Antimetabolites such as methotrexate and 5-fluorouracil (5-FU); Folic acid analogs such as denopterin, methotrexate, pteropterin, trimetrexate; Purine analogs such as fludarabine, 6-mercaptopurine, thiampurine, thioguanine; Pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, didoxuridine, doxifluridine, enocitabine, floxuridine, etc.; Androgens such as calusterone, drostanolone propionate, epithiostanol, mepitiostane, testolactone; Anti-adrenal agents such as aminoglutethimide, mitotane, trilostane; Folic acid supplements such as folinic acid; Aceglutamide; Aldophosphamide glycoside; Aminolevulinic acid; Amsacrine; Bestrabucil; Bisantrene; Edatraxate; Defofamine; Dexamethasone; Diacodone; Elformithine; Elliptinium acetate; Etoglucid; Gallium nitrate; Hydroxyurea; Lentinan; Lonidamine; Mitoguazone; Mitoxantrone; Mopidamol; Nitracrine; Pentostatin; Phenamet; Pirarubicin; Podophyllinic acid; 2-ethylhydrazide; Procarbazine; PSK; Razoxane; Schizophyllan; Spirogermanium; Tenuaazonic acid; Triazocine; 2,2’,2’’-trichloroethylamine; Urethane; Vinblastine; Dacarbazine; Mannomustine; Mitobronitol; Mitolactol; Pipobroman; Gacitabine; Arabinoside (“Ara-C”); Cyclophosphamide; Thiotepa;Taxanes, such as paclitaxel and docetaxel; retinoic acid; esperamicin; capecitabine; and pharmaceutically acceptable salts, acids or derivatives of any of the foregoing.

[0200] Suitable chemotherapeutic cell conditioners include antihormonal agents that act to regulate or inhibit hormonal action in tumors, such as antiestrogens including tamoxifen, (Nolvadex(trademark)), raloxifene, aromatase inhibitor 4(5)-imidazole, 4-hydroxytamoxifen, trioxifene, keoxifene, LY117018, onapristone, and toremifene (Fareston), and antiandrogens such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine; novantrone; teniposide; daunomycin; aminopterin; Xeloda; ibandronate; camptothecin-11 (CPT-11); topoisomerase inhibitor RFS2000; difluoromethylornithine (DMFO) are also included.

[0201] If desired, the compounds or pharmaceutical compositions of the present disclosure can generally be used in combination with commonly prescribed anti-cancer agents, including Herceptin®, Avastin®, Arimidex®, Rituxan®, Taxol®, Arimidex®, Taxotere®, ABVD, Avicine, Abagovomab, Acridine carboxamide, Adecatumumab, 17-N-allylamino-17-demethoxygeldanamycin, Alpharadin, Albosidib, 3-aminopyridine-2-carboxaldehyde thiosemicarbazone, Amonafide, Anthracenedione, Anti-CD22 immunotoxin, Anti-cancer drug, Anti-tumor herb, Apaziquone, Atiprimod, Azathioprine, Belotecan, Bendamustine, BIBW2992, Biliquidar, Brostallicin, Bryostatin, Buthionine sulfoximine, CBV (chemotherapy), Caliculin, Cell cycle non-specific anti-cancer drug, Dichloroacetic acid, Discodermolide, Elsamitrucin, Enocitabine, Epothilone, Eribulin, Everolimus, Exatecan, Exisulind, Ferginol, Holothurin, Phosphoestrol, ICE chemotherapy regimen, IT-101, Imexon, Imiquimod, Indolocarbazole, Irofulven, Lanidomide, Larotaxel, Lenalidomide, Lukantone, Lurtotecan, Mafosfamide, Mitozolomide, Nafoxidine, Nedaplatin, Olaparib, Ortataxel, PAC-1, Porfiromycin, Pixantrone, Proteasome inhibitor, Rebecamycin, Resiquimod, Rubitecan, SN-38, Salinosporamide A, Sapacitabine, Stanford V, Swainsonine, Talaporfin, Talidomide, Tegafur-uracil, Temodal, Tesetaxel, Triplatin tetranitrate, Tris(2-chloroethyl)amine, Troxacitabine, Uramustine, Vazimezan, Vinflunine, ZD6126 or Zosuquidar, etc.

[0202] The present disclosure further relates to methods of using the compounds or pharmaceutical compositions provided herein in combination with radiation therapy for inhibiting abnormal cell proliferation or treating hyperproliferative disorders in mammals. Techniques for performing radiation therapy are known in the art and these techniques can be used in the combination therapies described herein. Administration of the disclosed compounds in such combination therapies can be determined as described herein.

[0203] Radiation therapy can be performed by one of several methods, or a combination of several methods, including but not limited to external beam radiotherapy, brachytherapy, interstitial irradiation, stereotactic radiosurgery, total body irradiation, radiosurgery, and permanent or temporary implant brachytherapy. As used herein, the term "brachytherapy" refers to radiation therapy delivered by inserting a spatially confined radioactive material into a tumor site or other hyperproliferative tissue disease site or in the vicinity thereof in the body. Such term shall include, but not be limited to, exposure to radioactive isotopes (e.g., At-211, I-131, I-125, Y-90, Re-186, Re-188, Sm-153, Bi-212, P-32, and radioactive isotopes of Lu). Suitable sources for use as cell modulators in the present disclosure include both solids and liquids. By way of non-limiting example, the source can be a radionuclide, such as I-125, I-131, Yb-169, Ir-192 as a solid source, I-125 as a solid source, or other radionuclides that emit photons, beta particles, gamma rays, or other therapeutic light rays. The radioactive material can be a fluid made from any solution of the radionuclide(s), e.g., a solution of I-125 or I-131, or a radioactive fluid made using a suitable fluid slurry containing small particles of a solid radionuclide such as Au-198, Y-90. Further, the radionuclide(s) can be realized in a gel or radioactive microspheres.

[0204] The compounds or pharmaceutical compositions of the present disclosure can be used in combination in an amount with one or more substances selected from anti-angiogenic agents, signal transduction inhibitors, anti-proliferative agents, glycolysis inhibitors, or autophagy inhibitors.

[0205] Anti-angiogenic agents such as MMP-2 (matrix metalloproteinase 2) inhibitors, MMP-9 (matrix metalloproteinase 9) inhibitors, and COX-11 (cyclooxygenase 11) inhibitors can be used in combination with the disclosed compounds and the pharmaceutical compositions described herein. Examples of anti-angiogenic agents include, for example, rapamycin, temsirolimus (CCI-779), everolimus (RAD001), sorafenib, sunitinib, and bevacizumab. Examples of useful COX-II inhibitors include celecoxib, valdecoxib, and rofecoxib. Examples of useful matrix metalloproteinase inhibitors are described in WO96 / 33172, WO96 / 27583, European Patent Publication No. EP0818442, European Patent Publication No. EP1004578, WO98 / 07697, WO98 / 03516, WO98 / 34918, WO98 / 34915, WO98 / 33768, WO98 / 30566, European Patent Publication No. 606046, European Patent Publication No. 931788, WO90 / 05719, WO99 / 52910, WO99 / 52889, WO99 / 29667, WO1999007675, European Patent Publication No. EP1786785, European Patent Publication No. EP1181017, US Publication No. US20090012085, US Publication No. US5863949, US Publication No. US5861510, and European Patent Publication No. EP0780386, all of which are hereby incorporated by reference in their entirety. Preferred MMP-2 and MMP-9 inhibitors have little or no MMP-1 inhibitory activity. More preferred are those that selectively inhibit MMP-2 and / or AMP-9 against other matrix metalloproteinases (i.e., MAP-1, MMP-3, MMP-4, MMP-5, MMP-6, MMP-7, MMP-8, MMP-10, MMP-11, MMP-12, and MMP-13). Some specific examples of MMP inhibitors useful in the present disclosure include AG-3340, RO32-3555, and RS13-0830.

[0206] The compound of the present application may be used in combination therapy with other anti-tumor drugs. For example, acemannan, aclarubicin, aldesleukin, alemtuzumab, alitretinoin, altretamine, amifostine, aminolevulinic acid, amrubicin, amsacrine, anagrelide, anastrozole, ANCER, ansetim, ARGLABIN, arsenic trioxide, BAM002 (Novelos), bexarotene, bicalutamide, broxuridine, capecitabine, sermolukin, cetrorelix, cladribine, clotrimazole, cytarabine ocfosfate, DA3030 (Dong-A), daclizumab, denileukin diftitox, deslorelin, dexrazoxane, dirazep, docetaxel, docosanol, doxercalciferol, doxifluridine, doxorubicin, bromocriptine, carmustine, cytarabine, fluorouracil, HIT diclofenac, interferon alpha, daunorubicin, doxorubicin, tretinoin, edelfosine, edrecolomab, eflornithine, emitefur, epirubicin, epoetin beta, etoposide phosphate, exemestane, exisulind, fadrozole, filgrastim, finasteride, fludarabine phosphate, formestane, fotemustine, gallium nitrate, gemcitabine, gemtuzumab ozogamicin, gimeracil / oteracil / tegafur combination drug, glycopyrronium, goserelin, heptaplatin, human chorionic gonadotropin, human fetal alpha-fetoprotein, ibandronic acid, idarubicin, (imiquimod, interferon alpha, interferon alpha, natural, interferon alpha-2, interferon alpha-2a, interferon alpha-2b, interferon alpha-N1, interferon alpha- n3, interferon alpha con-1, interferon alpha, natural, interferon beta, interferon beta-1a, interferon beta-1b, interferon gamma, natural interferon gamma-1a, interferon gamma-1b, interleukin-1 beta, yobenguan, irinotecan, ilsogladine, lanreotide, LC9018 (Yakult), leflunomide, lenograstim, lentinan sulfate, letrozole, leukocyte alpha interferon, leuprorelin, levamisole + fluorouracil, rialozole, lobaplatin, lonidamine, lovastatin, masoprocol, melarsoprol, metoclopramide, mifepristone, miltefosine, milimostim, mismatched double-stranded RNA, mitoguazone, mitolactol, mitoxantrone, molgramostim, nafarelin, naloxone + pentazocine, nartograstim, nedaplatin, nilutamide, noscapine, novel erythropoietic stimulating protein, NSC631570 octreotide, oprelvekin, osaterone, oxaliplatin, paclitaxel, pamidronic acid, pegaspargase, peginterferon alpha-2b, pentosan polysulfate sodium, pentostatin, picibanil, pirarubicin, rabbit anti-thymocyte polyclonal antibody, polyethylene glycol interferon alpha-2a, porfimer sodium, raloxifene, raltitrexed, rasburicase, rhenium Re186 etidronate, RII retinamide, rituximab, romurtide, samarium (153Sm) lexidronam, sargramostim, sizofiran, sobuzoxan, sonermin, strontium chloride 89, suramin, tasonermin, tazarotene, tegafur, temoporfin, temozolomide, teniposide, tetrachlorodecaoxide, thalidomide, simalufasin, thyrotropin alpha, topotecan, toremifene, tositumomab iodine 131, trastuzumab, treosulfan, tretinoin, trilostane, trimethoprim, tryptorelin, tumor necrosis factor alpha, natural, ubenimex, bladder cancer vaccine, Maruyama vaccine, melanoma lysate vaccine, valrubicin, verteporfin, vinorelbine, VIRULIZIN, dinostatin stimalamer,or zoledronic acid; abarelix; AE941 (Aeterna), ambamustine, antisense oligonucleotide, bcl-2 (Genta), APC8015 (Dendreon), cetuximab, decitabine, dexaminoglutethimide, diacozone, EL532 (Elan), EM800 (Endorecherche), eniluracil, etanidazole, fenretinide, filgrastim SD01 (Amgen), fulvestrant, gallocitabine, gastrin 17 immunogen, HLA-B7 gene therapy (Vical), granulocyte macrophage colony stimulating factor, histamine dihydrochloride, ibritumomab tiuxetan, iromostat, IM862 (Cytran), interleukin-2, ibuprofen, LDI200 (Milkhaus), leridistim, lym-1-iodine 131 MAb (Techniclone), polymorphic epithelial mucin-yttrium 90 MAb (Antisoma), marimastat, menogaril, mitsumomab, motexafin gadolinium, MX6 (Galderma), nelarabine, nolatrexed, P30 protein, pegvisomant, pemetrexed, porfiromycin, prinomastat, RL0903 (Shire), rubitecan, satraplatin, sodium phenylacetate, strontium ranelate, SRL172 (SR Pharma), SU5416 (SUGEN), TA077 (Tanabe), tetrathiomolybdate, taliblastine, thrombopoietin, ethyl etiopurpurin tin, tirapazamine, cancer vaccine (Biomira), melanoma vaccine (New York University), melanoma vaccine (Sloan Kettering Institute), melanoma tumor lysate vaccine (New York Medical College), melanoma cell virus lysate vaccine (Royal Newcastle Hospital), or can be used in combination with valspodar, etc.

[0207] The compounds of the present invention may further be used in combination with a VEGFR inhibitor. Additionally, the compounds described in the following patents and patent applications can be used in combination therapies: US6,258,812, US2003 / 0105091, WO01 / 37820, US6,235,764, WO01 / 32651, US6,630,500, US6,515,004, US6,713,485, US5,521,184, US5,770,599, US5,747,498, WO02 / 68406, WO02 / 66470, WO02 / 55501, WO04 / 05279, WO04 / 07481, WO04 / 07458, WO04 / 09784, WO02 / 59110, WO99 / 45009, WO00 / 59509, WO99 / 61422, US5,990,141, WO00 / 12089, and WO00 / 02871.

[0208] In some embodiments, in combination, the composition of the present invention is combined with at least one anti-angiogenic agent. The agents include, but are not limited to, chemically synthesized compositions, antibodies, antigen-binding regions, radionuclides, and combinations and conjugates thereof prepared by in vitro synthesis. The agent can be an agonist, antagonist, allosteric modulator, toxin, or more generally, act to inhibit or stimulate its target (e.g., activation or inhibition of a receptor or enzyme), thereby promoting cell death or arresting cell growth.

[0209] Exemplary anti-angiogenic agents include Erbitux (trademark) (IMC-C225), KDR (kinase domain receptor) inhibitors (e.g., antibodies and antigen-binding regions that specifically bind to the kinase domain receptor), anti-VEGF agents (e.g., antibodies or antigen-binding regions that specifically bind to VEGF and / or soluble VEGF receptor or its ligand-binding region), such as AVASTIN (trademark) or VEGF-TRAP (trademark), and anti-VEGF receptor agents (e.g., antibodies or antigen-binding regions that specifically bind to it), EGFR inhibitors (e.g., antibodies or antigen-binding regions that specifically bind to it), such as Vectibix (panitumumab), IRESSA (trademark) (gefitinib), TARCEVA (trademark) (erlotinib), anti-Ang1 and anti-Ang2 agents (e.g., antibodies or antigen-binding regions that specifically bind to them or their receptors, such as Tie2 / Tek, etc.), and anti-Tie2 kinase inhibitors (e.g., antibodies or antigen-binding regions that specifically bind to it). The pharmaceutical composition of the present invention may include one or more agents (e.g., antibodies, antigen-binding regions, or soluble receptors) that specifically bind to a growth factor and inhibit its activity, such as an antagonist of hepatocyte growth factor (HGF, also known as scatter factor), and antibodies or antigen-binding regions that specifically bind to its receptor "c-met".

[0210] Other anti-angiogenic agents include Campath, IL-8, B-FGF, Tek antagonists (Ceretti et al., US Publication No. 2003 / 0162712; US Patent No. 6,413,932), anti-TWEAK agents (e.g., specifically binding antibodies or antigen-binding regions, or soluble TWEAK receptor antagonists; see Wiley, US Patent No. 6,727,225), ADAM disintegrin domains that antagonize the binding of integrin to its ligand (Fanslow et al., US Publication No. 2002 / 0042368), specifically binding anti-eph receptor and / or anti-ephrin antibodies or antigen-binding regions (US Patent Nos. 5,981,245; 5,728,813; 5,969,110; 6,596,852; 6,232,447; 6,057,124 and their patent family members), and anti-PDGF-BB antagonists (e.g., specifically binding antibodies or antigen-binding regions) and antibodies or antigen-binding regions that specifically bind to the PDGF-BB ligand, and PDGFR kinase inhibitors (e.g., specifically binding antibodies or antigen-binding regions thereto).

[0211] Additional anti-angiogenic / anti-tumor drugs include SD-7784 (Pfizer, USA); Sirenigitide (Merck KGaA, Germany, EPO770622); pegaptanib octasodium (Gilead Sciences, USA); alphastatin (BioActa, UK); M-PGA (Celgene, USA, US5712291); iromostat (Arriva, USA, US5892112); emaxinib (Pfizer, USA, US5792783); batranib (Novartis, Switzerland); 2-methoxyestradiol (EntreMed, USA); TLC ELL-12 (Elan, Ireland); anecortave acetate (Alcon, USA); alpha-D148Mab (Amgen, USA); CEP-7055 (Cephalon, USA); anti-Vn Mab (Crucell, Netherlands) DAC: anti-angiogenic drug (ConjuChem, Canada); angiostatin (InKine Pharmaceutical, USA); KM-2550 (Kyowa Hakko, Japan); SU-0879 (Pfizer, USA); CGP-79787 (Novartis, Switzerland, EP970070); ARGENT technology (Ariad, USA); YIGSR-stealth (Johnson & Johnson, USA); fibrinogen-E fragment (BioActa, UK); angiogenesis inhibitor (Trigen, UK); TBC-1635 (Encysive Pharmaceuticals, USA); SC-236 (Pfizer, USA); ABT-567 (Abbott, USA); metastatin (EntreMed, USA); angiogenesis inhibitor (Tripep, Sweden); maspin (Sosei, Japan); 2-methoxyestradiol (Oncology Sciences Corporation, USA); ER-68203-00 (IVAX, USA); benefin (Lane Labs, USA); Tz-93 (Tsumura, Japan); TAN-1120 (Takeda, Japan); FR-111142 (Fujisawa, Japan, JP02233610);Platelet Factor 4, (RepliGen, USA, EP407122); Vascular Endothelial Growth Factor Antagonist, (Borean, Denmark); Bevacizumab (pINN), (Genentech, USA); Angiogenesis Inhibitor, (SUGEN, USA); XL784, (Exelixis, USA); XL647, (Exelixis, USA); MAb, alpha5beta3 Integrin, Second Generation, (Applied Molecular Evolution, USA and MedImmune, USA); Gene Therapy, Retinopathy, (Oxford BioMedica, UK); Enzastaurin Hydrochloride (USAN), (Lilly, USA); CEP7055, (Cephalon, USA and Sanofi-Synthelabo, France); BC1, (Genoa Institute of Cancer Research, Italy); Angiogenesis Inhibitor, (Alchemia, Australia); VEGF Antagonist, (Regeneron, USA); rBPI21 and Anti-Angiogenic Agent Derived from BPI, (XOMA, USA); PI88, (Progen, Australia); Sirukumab (pINN), (Merck KGaA, German; Munich Technical University, Germany, Scripps Clinic and Research Foundation, USA); Cetuximab (INN), (Aventis, France); AVE8062, (Ajinomoto, Japan); AS1404, (Cancer Research Laboratory, New Zealand); SG292, (Telios, USA); Endostatin, (Boston Childrens Hospital, USA); ATN161, (Attenuon, USA); Angiostatin, (Boston Childrens Hospital, USA); 2-Methoxyestradiol, (Boston Childrens Hospital, USA); ZD6474, (AstraZeneca, UK); ZD6126, (Angiogene Pharmaceuticals, UK); PPI2458, (Praecis, USA); AZD9935, (AstraZeneca, UK);AZD2171, (AstraZeneca, UK); Batranib (pINN), (Novartis, Switzerland and Schering AG, Germany); Tissue Factor Pathway Inhibitor, (EntreMed, USA); Pegaptanib (Pinn), (Gilead Sciences, USA); Xanthriol, (Yonsei University, South Korea); Gene Vaccine, VEGF-2, (Scripps Clinic and Research Foundation, USA); SPV5.2, (Supratek, Canada); SDX103, (University of California at San Diego, USA); PX478, (ProlX, USA); Metastatin, (EntreMed, USA); Troponin I, (Harvard University, USA); SU6668, (SUGEN, USA); OXI4503, (OXiGENE, USA); o-Guanidine, (Dimensional Pharmaceuticals, USA); Motuporamine C, (British Columbia University, Canada); CDP791, (Celltech Group, UK); Actiprimod (pINN), (GlaxoSmithKline, UK); E7820, (Eisai, Japan); CYC381, (Harvard University, USA); AE941, (Aeterna, Canada); Vaccine, Angiogenesis, (EntreMed, USA); Urokinase Plasminogen Activation Inhibitor, (Dendreon, USA); Oglufanide (pINN), (Melmotte, USA); HIF-1 Alpha Inhibitor, (Xenova, UK); CEP5214, (Cephalon, USA); BAY RES2622, (Bayer, Germany); Angiocidin, (InKine, USA); A6, (Angstrom, USA); KR31372, (Korea Research Institute of Chemical Technology, South Korea); GW2286, (GlaxoSmithKline, UK); EHT0101, (ExonHit, France);CP868596, (Pfizer, USA); CP564959, (OSI, USA); CP547632, (Pfizer, USA); 786034, (GlaxoSmithKline, UK); KRN633, (Kirin Brewery, Japan); drug delivery system, intraocular, 2-methoxyestradiol, (EntreMed, USA); Anginex, (Maastricht University, Netherlands, and Minnesota University, USA); ABT510, (Abbott, USA); AAL993, (Novartis, Switzerland); VEGI, (ProteomTech, USA); tumor necrosis factor alpha inhibitor, (National Institute on Aging, USA); SU11248, (Pfizer, USA and SUGEN USA); ABT518, (Abbott, USA); YH16, (Yantai Rongchang, China); S-3APG, (Boston Childrens Hospital, USA and EntreMed, USA); MAb, KDR, (ImClone Systems, USA); MAb, alpha5beta1, (Protein Design, USA); KDR kinase inhibitor, (Celltech Group, UK, and Johnson&Johnson, USA); GFB116, (South Florida University, USA and Yale University, USA); CS706, (Sankyo, Japan); combretastatin A4 prodrug, (Arizona State University, USA); chondroitinase AC, (IBEX, Canada); BAY RES2690, (Bayer, Germany); AGM1470, (Harvard University, USA, Takeda, Japan, and TAP, USA); AG13925, (Agouron, USA); tetrathiomolybdate, (University of Michigan, USA); GCS100, (Wayne State University, USA) CV247, (Ivy Medical, UK);CKD732, (Chong Kun Dang, South Korea); MAb, vascular endothelial growth factor, (Xenova, UK); Iruxogratinib (INN), (Nippon Shinyaku, Japan), RG13577, (Aventis, France); WX360, (Wilex, Germany); Squaramine (pINN), (Genaera, USA), RPI4610, (Sirna, USA); cancer treatment, (Marinova, Australia); heparanase inhibitor, (InSight, Israel); KL3106, (Kolon, South Korea); Honokiol, (Emory University, USA); ZK CDK, (Schering AG, Germany); ZK Angio, (Schering AG, Germany); ZK229561, (Novartis, Switzerland, and Schering AG, Germany); XMP300, (XOMA, USA); VGA1102, (Taisho, Japan); VEGF receptor regulator, (Pharmacopeia, USA); VE-cadherin-2 antagonist, (ImClone Systems, USA); Basostatin, (National Institutes of Health, USA); vaccine, Flk-1, (ImClone Systems, USA); TZ93, (Tsumura, Japan); TAMSTATIN, (Beth Israel Hospital, USA); truncated soluble FLT1 (vascular endothelial growth factor receptor 1), (Merck&Co, USA); Tie-2 ligand, (Regeneron, USA); and thrombospondin 1 inhibitor, (Allegheny Health, Education and Research Foundation, USA) are included.;

[0212] Examples of autophagy inhibitors include, but are not limited to, chloroquine, 3-methyladenine, hydroxychloroquine (Plaquenil™), bafilomycin A1, 5-amino-4-imidazolecarboxamide riboside (AICAR), okadaic acid, autophagy-inhibiting algal toxins that inhibit type 2A or type 1 protein phosphatases, cAMP analogs, and drugs that increase cAMP levels such as adenosine, LY204002, N6-mercaptopurine riboside, and vinblastine. Furthermore, antisense or siRNA that inhibits protein expression, including but not limited to ATG5 (involved in autophagy), may be used.

[0213] Further pharmaceutically active compounds / drugs that can be used in cancer treatment and can be combined with one or more compounds of the present invention include epoetin alpha; darbepoetin alpha; panitumumab; pegfilgrastim; palifermin; filgrastim; denosumab; ancetastim; AMG102; AMG386; AMG479; AMG655; AMG745; AMG951, and AMG706, or pharmaceutically acceptable salts thereof.

[0214] In certain embodiments, the compositions provided herein are administered in combination with a chemotherapeutic agent. Suitable chemotherapeutic agents include natural product-derived substances such as vinca alkaloids (e.g., vinblastine, vincristine, and vinorelbine), paclitaxel, epipodophyllotoxins (e.g., etoposide and teniposide), antibiotics (e.g., dactinomycin (actinomycin D), daunorubicin, doxorubicin, and idarubicin), anthracyclines, mitoxantrone, bleomycin, plicamycin (mithramycin), mitomycin, etc., enzymes (e.g., L-asparaginase which metabolizes L-asparagine systemically and renders cells lacking their own asparagine synthesis ability in a depleted state), antiplatelet agents, anti-proliferative / anti-mitotic alkylating agents such as nitrogen mustards (e.g., mechlorethamine, cyclophosphamide analogs, melphalan, and chlorambucil), ethyleneimines and methylmelamines (e.g., hexamethylmelamine and thiotepa), etc., CDK inhibitors (e.g., seliciclib, UCN-01, P1446A-05, PD-0332991, dinaciclib, P27-00, AT-7519, RGB286638, and SCH727965), alkyl sulfonic acids (e.g., busulfan), nitrosoureas (e.g., carmustine (BCNU) analogs, and streptozocin), temozolomide-dacarbazine (DTIC), anti-proliferative / anti-mitotic antimetabolites such as folic acid analogs (e.g., methotrexate), pyrimidine analogs (e.g., fluorouracil, floxuridine, and cytarabine), purine analogs and related inhibitors (e.g., mercaptopurine, thioguanine, pentostatin and 2-chlorodeoxyadenosine), aromatase inhibitors (e.g., anastrozole, exemestane, and letrozole), and platinum coordination complexes (e.g., cisplatin and carboplatin), etc., procarbazine, hydroxyurea, mitotane, aminoglutethimide, histone deacetylase (HDAC) inhibitors (e.g., trichostatin, sodium butyrate, apicidin, suberoylanilide hydroxamic acid, vorinostat, LBH589, romidepsin, ACY-1215, and panobinostat), mTor inhibitors (e.g., temsirolimus, everolimus, ridaforolimus, and sirolimus),KSP (Eg5) inhibitors (e.g., Array520), DNA binders (e.g., zalypsis), PI3K delta inhibitors (e.g., GS-1101 and TGR-1202), PI3K delta and gamma inhibitors (e.g., CAL-130), multi-kinase inhibitors (e.g., TG02 and sorafenib), hormones (e.g., estrogen) and hormone agonists, e.g., luteinizing hormone releasing hormone (LHRH) agonists (e.g., goserelin, leuprolide and triptorelin), etc., BAFF neutralizing antibodies (e.g., LY2127399), IKK inhibitors, p38MAPK inhibitors, anti-IL-6 (e.g., CNTO328), telomerase inhibitors (e.g., GRN163L), aurora kinase inhibitors (e.g., MLN8237), cell surface monoclonal antibodies (e.g., anti-CD38 (HUMAX-CD38), anti-CS1 (e.g., elotuzumab)), HSP90 inhibitors (e.g., 17AAG and KOS953), P13K / Akt inhibitors (e.g., perifosine), Akt inhibitors (e.g., GSK-2141795), PKC inhibitors (e.g., enzastaurin), FTI (e.g., Zarnestra (trademark)), anti-CD138 (e.g., BT062), Torc1 / 2 specific kinase inhibitors (e.g., INK128), kinase inhibitors (e.g., GS-1101), ER / UPR targeting agents (e.g., MKC-3946), cFMS inhibitors (e.g., ARRY-382), JAK1 / 2 inhibitors (e.g., CYT387), PARP inhibitors (e.g., olaparib and veliparib (ABT-888)), BCL-2 antagonists may be included. Other chemotherapeutic agents may include mechlorethamine, camptothecin, ifosfamide, tamoxifen, raloxifene, gemcitabine, navelbine, sorafenib, or any analog or derived variant of the above chemotherapeutic agents.,

[0215] The compounds of the present invention may be used in combination with radiotherapy, hormone therapy, surgery and immunotherapy, and those therapies are well known to those skilled in the art.

[0216] In certain embodiments, the pharmaceutical compositions provided herein are administered in combination with a steroid. Suitable steroids include, but are not limited to, 21-acetoxypregnenolone, alclometasone, algestone, amcinonide, beclomethasone, betamethasone, budesonide, chloroprednisone, clobetasol, clocortolone, cloprednol, corticosterone, cortisone, cortivazol, deflazacort, desonide, desoximetasone, dexamethasone, diflorasone, diflucortolone, difuprednate, enoxolone, fluazacort, flucloronide, flumethasone, flunisolide, fluocinonide acetonide, fluocinonide, fluocortin butyl, fludrocortisone, fluorometholone, flupredolone acetate, fluprednisolone acetate, fluprednisolone, flurandrenolide, fluticasone propionate, formocortal, halcinonide, halobetasol propionate, halomethasone, hydrocortisone, loteprednol etabonate, madipredone, medrysone, meprednisone, methylprednisolone, mometasone furoate, paramethasone, prednicarbate, prednisone, prednival, prednylidene, rimexolone, tixocortol, triamcinolone, triamcinolone acetonide, triamcinolone benetonide, triamcinolone hexacetonide, and salts and / or derivatives thereof. In certain embodiments, the compounds of the invention can be used in combination with additional pharmaceutically active agents for treating nausea. Examples of agents that can be used for treating nausea include, but are not limited to, dronabinol, granisetron, metoclopramide, ondansetron, and prochlorperazine, or pharmaceutically acceptable salts thereof.

[0217] The compounds or pharmaceutical compositions of the present disclosure can also be used in combination with any amount of one or more substances selected from EGFR inhibitors, MEK inhibitors, PI3K inhibitors, AKT inhibitors, TOR inhibitors, and various agents such as anti-PD-1, anti-PDL-1, anti-CTLA4, anti-LAG1, and anti-OX40, GITR agonists, CAR-T cells, and BiTE immunotherapies.

[0218] EGFR inhibitors include, but are not limited to, small molecule antagonists, antibody inhibitors, or specific antisense nucleotides or siRNAs. Useful antibody inhibitors of EGFR include cetuximab (Erbitux), panitumumab (Vectibix), zalutumumab, nimotuzumab, and matuzumab. Small molecule antagonists of EGFR include gefitinib, erlotinib (Tarceva), and more recently lapatinib (Tykerb). See, for example, Yan L, et.al., Pharmacogenetics and Pharmacogenomics In Oncology Therapeutic Antibody Development, BioTechniques 2005;39(4):565-8, and Paez J G, et.al., EGFR Mutations In Lung Cancer Correlation With Clinical Response To Gefitinib Therapy, Science 2004;304(5676):1497-500.

[0219] Non-limiting examples of low molecular weight EGFR inhibitors include any EGFR inhibitor described in the following patent publications, as well as all pharmaceutically acceptable salts and solvates of said EGFR inhibitors: European Patent Application EP520722 (publication date December 30, 1992); European Patent Application EP566226 (publication date October 20, 1993); PCT International Publication WO96 / 33980 (publication date October 31, 1996); US Patent No. 5,747,498 (issue date May 5, 1998); PCT International Publication WO96 / 30347 (publication date October 3, 1996); European Patent Application EP787772 (publication date August 6, 1997); PCT International Publication WO97 / 30034 (publication date August 21, 1997); PCT International Publication WO97 / 30044 (publication date August 21, 1997); PCT International Publication WO97 / 38994 (publication date October 23, 1997); PCT International Publication WO97 / 49688 (publication date December 31, 1997); European Patent Application EP837063 (publication date April 22, 1998); PCT International Publication WO98 / 02434 (publication date January 22, 1998); PCT International Publication WO97 / 38983 (publication date October 23, 1997); PCT International Publication WO95 / 19774 (publication date July 27, 1995); PCT International Publication WO95 / 19970 (publication date July 27, 1995); PCT International Publication WO97 / 13771 (publication date April 17, 1997); PCT International Publication WO98 / 02437 (publication date January 22, 1998); PCT International Publication WO98 / 02438 (publication date January 22, 1998); PCT International Publication WO97 / 32881 (publication date September 12, 1997); German Application DE19629652 (publication date January 29, 1998); PCT International Publication WO98 / 33798 (publication date August 6, 1998); PCT International Publication WO97 / 32880 (publication date September 12, 1997); PCT International Publication WO97 / 32880 (publication date September 12, 1997); European Patent Application EP682027 (publication date November 15, 1995); PCT International Publication WO97 / 02266 (publication date January 23, 1997); PCT International Publication WO97 / 27199 (publication date July 31, 1997); PCT International Publication WO98 / 07726 (publication date February 26, 1998);PCT International Publication Gazette WO97 / 34895 (publication date: September 25, 1997); PCT International Publication Gazette WO96 / 31510' (publication date: October 10, 1996); PCT International Publication Gazette WO98 / 14449 (publication date: April 9, 1998); PCT International Publication Gazette WO98 / 14450 (publication date: April 9, 1998); PCT International Publication Gazette WO98 / 14451 (publication date: April 9, 1998); PCT International Publication Gazette WO95 / 09847 (publication date: April 13, 1995); PCT International Publication Gazette WO97 / 19065 (publication date: May 29, 1997); PCT International Publication Gazette WO98 / 17662 (publication date: April 30, 1998); U.S. Patent No. 5,789,427 (issue date: August 4, 1998); U.S. Patent No. 5,650,415 (issue date: July 22, 1997); U.S. Patent No. 5,656,643 (issue date: August 12, 1997); PCT International Publication Gazette WO99 / 35146 (publication date: July 15, 1999); PCT International Publication Gazette WO99 / 35132 (publication date: July 15, 1999); PCT International Publication Gazette WO99 / 07701 (publication date: February 18, 1999); and PCT International Publication Gazette WO92 / 20642 (publication date: November 26, 1992). Further non-limiting examples of small molecule EGFR inhibitors include any of the EGFR inhibitors described in Traxler, P., 1998, Exp. Opin. Ther. Patents 8(12):1599-1625.;

[0220] The anti-EGFR inhibitors include any anti-EGFR antibody or antibody fragment that can partially or completely block EGFR activation by its natural ligand. Non-limiting examples of anti-EGFR inhibitors include those described in Modjtahedi, H., et al., 1993, Br. J. Cancer 67:247-253; Teramoto, T., et al., 1996, Cancer 77:639-645; Goldstein et al., 1995, Clin. Cancer Res. 1:1311-1318; Huang, S.M., et al., 1999, Cancer Res. 15:59(8):1935-40; and Yang, X., et al., 1999, Cancer Res. 59:1236-1243. Thus, the EGFR inhibitor can be the monoclonal antibody Mab E7.6.3 (Yang, 1999, supra), or Mab C225 (ATCC accession number HB-8508), or an antibody or an antibody fragment having its binding specificity.

[0221] Examples of MEK inhibitors include, but are not limited to, CI-1040, AZD6244, PD318088, PD98059, PD334581, RDEA119, ARRY-142886, ARRY-438162, and PD-325901.

[0222] Examples of PI3K inhibitors include wortmannin, 17-hydroxy wortmannin analogs described in WO06 / 044453, 4-[2-(1H-indazol-4-yl)-6-[[4-(methylsulfonyl)piperazin-1-yl]methyl]thieno[3,2-d]pyrimidin-4-yl]morpholine (also known as GDC0941 and described in PCT publications WO09 / 036,082 and WO09 / 055,730), 2-methyl-2-[4-[3-methyl-2-oxo-8-(quinolin-3-yl)-2,3-dihydroimidazo[4,5-c]quinolin-1-yl]phenyl]propionitrile (also known as BEZ235 or NVP-BEZ235 and described in PCT publication number WO06 / 122806), (S)-1-(4-((2-(2-aminopyrimidin-5-yl)-7-methyl-4-morpholinothieno[3,2-d]pyrimidin-6-yl)methyl)piperazin-1-yl)-2-hydroxypropan-1-one (described in PCT publication number WO2008 / 070740), LY294002 (2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one, available from Axon Medchem), PI103 hydrochloride (3-[4-(4-morpholinylpyrido-[3′,2′:4,5]furo[3,2-d]pyrimidin-2-yl]phenol hydrochloride, available from Axon Medchem), PIK75 (N′-[(1E)-(6-bromoimidazo[1,2-a]pyridin-3-yl)methylene]-N,2-dimethyl-5-nitrobenzenesulfono-hydrazide hydrochloride, available from Axon Medchem), PIK90 (N-(7,8-dimethoxy-2,3-dihydro-imidazo[1,2-c]quinazolin-5-yl)-nicotinamide, available from Axon Medchem), GDC-0941 bismesylate (2-(1H-indazol-4-yl)-6-(4-methanesulfonyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine bismesylate, available from Axon Medchem), AS-252424 (5-[1-[5-(4-fluoro-2-hydroxy-phenyl)-furan-2-yl]-meth-(Z)-ylidene]-thiazolidine-2,4-Dion, available from Axon Medchem), and TGX-221 (7-methyl-2-(4-morpholinyl)-9-[1-(phenylamino)ethyl]-4H-pyrido-[1,2-a]pyrimidin-4-one, available from Axon Medchem), XL-765, and XL-147, but not limited thereto. Other PI3K inhibitors include demethoxybiridin, perifosine, CAL101, PX-866, BEZ235, SF1126, INK1117, IPI-145, BKM120, XL147, XL765, palomid 529, GSK1059615, ZSTK474, PWT33597, IC87114, TG100-115, CAL263, PI-103, GNE-477, CUDC-907, and AEZS-136.,

[0223] Examples of AKT inhibitors include Akt-1-1 (inhibits Akt1) (Barnett et al. (2005) Biochem. J., 385 (Pt. 2), 399-408); Akt-1-1,2 (inhibits Ak1 and Ak2) (Barnett et al. (2005) Biochem. J. 385 (Pt. 2), 399-408); API-59CJ-Ome (e.g., Jin et al. (2004) Br. J. Cancer 91, 1808-12); 1-H-imidazo[4,5-c]pyridinyl compounds (e.g., WO05011700); indole-3-carbinol and its derivatives (e.g., U.S. Patent No. 6,656,963; Sarkar and Li (2004) J Nutr. 134 (12 Suppl), 3493S-3498S); perifosine (e.g., interferes with Akt membrane localization; Dasmahapatra et al. (2004) Clin. Cancer Res. 10 (15), 5242-52, 2004); phosphatidylinositol ether lipid analogs (e.g., Gills and Dennis (2004) Expert. Opin. Investig. Drugs 13, 787-97); and triciribine (TCN or API-2 or NCI identifier: NSC154020; Yang et al. (2004) Cancer Res. 64, 4394-9), but not limited thereto.

[0224] Examples of TOR inhibitors include, but are not limited to, AP-23573, CCI-779, everolimus, RAD-001, rapamycin, temsirolimus, ATP-competitive TORC1 / TORC2 inhibitors such as PI-103, PP242, PP30, and Torin1. FKBP12 enhancers; other TOR inhibitors included in rapamycin and its derivatives include CCI-779 (temsirolimus), RAD001 (everolimus; WO9409010), and AP23573; rapalogs such as those disclosed in WO98 / 02441 and WO01 / 14387, such as AP23573, AP23464, or AP23841; 40-(2-hydroxyethyl)rapamycin, 40-[3-hydroxy(hydroxymethyl)methylpropanoate]-rapamycin (also called CC1779), 40-epi-(tetrazolito)-rapamycin (also called ABT578), 32-deoxorapamycin, 16-pentynilyloxy-32(S)-dihydrorapamycin, and other derivatives disclosed in WO05005434; derivatives disclosed in U.S. Patent No. 5,258,389, WO94 / 090101, WO92 / 05179, U.S. Patent No. 5,118,677, U.S. Patent No. 5,118,678, U.S. Patent No. 5,100,883, U.S. Patent No. 5,151,413, U.S. Patent No. 5,120,842, WO93 / 111130, WO94 / 02136, WO94 / 02485, WO95 / 14023, WO94 / 02136, WO95 / 16691, WO96 / 41807, WO96 / 41807, and U.S. Patent No. 5,256,790; phosphorus-containing rapamycin derivatives (e.g., WO05016252); 4H-1-benzopyran-4-one derivatives (e.g., U.S. Provisional Patent Application No. 60 / 528,340).

[0225] Immunotherapies include, but are not limited to, anti-PD-1 agents, anti-PDL-1 agents, anti-CTLA-4 agents, anti-LAG1 agents, and anti-OX40 agents. Exemplary anti-PD-1 antibodies and methods of use thereof are described in Goldberg et al., Blood 110(1):186-192(2007), Thompson et al., Clin. Cancer Res. 13(6):1757-1761(2007), and Korman et al. International Application No. PCT / JP2006 / 309606 (Publication No. WO2006 / 121168A1), each of which is hereby expressly incorporated by reference herein and includes Yervoy (trademark) (ipilimumab) or tremelimumab (anti-CTLA-4), galiximab (anti-B7.1), BMS-936558 (anti-PD-1), MK-3475 (anti-PD-1), AMP224 (anti-B7DC), BMS-936559 (anti-B7-H1), MPDL3280A (anti-B7-H1), MEDI-570 (anti-ICOS), AMG557 (anti-B7H2), MGA271 (anti-B7H3), IMP321 (anti-LAG-3), BMS-663513 (anti-CD137), PF-05082566 (anti-CD137), CDX-1127 (anti-CD27), anti-OX40 (Providence Health Services), huMAbOX40L (anti-OX40L), atacicept (anti-TACI), CP-870893 (anti-CD40), lucatumumab (anti-CD40), dacetuzumab (anti-CD40), muromonab-CD3 (anti-CD3), ipilimumab (anti-CTLA-4). Immunotherapies also include genetically engineered T cells (e.g., CAR-T cells) and bispecific antibodies (e.g., BiTE).

[0226] Examples of GITR agonists include GITR fusion proteins and anti-GITR antibodies (e.g., bivalent anti-GITR antibodies), such as the GITR fusion proteins described in U.S. Patent No. 6,111,090 box.c, European Patent No. 090505B1, U.S. Patent No. 8,586,023, PCT Publication Nos. WO2010 / 003118 and 2011 / 090754, or anti-GITR antibodies, such as those described in U.S. Patent No. 7,025,962, European Patent No. 1947183B1, U.S. Patent No. 7,812,135, U.S. Patent No. 8,388,967, U.S. Patent No. 8,591,886, European Patent No. EP1866339, PCT Publication Nos. WO2011 / 028683, PCT Publication No. WO2013 / 039954, PCT Publication No. WO2005 / 007190, PCT Publication No. WO2007 / 133822, PCT Publication No. WO2005 / 055808, PCT Publication No. WO99 / 40196, PCT Publication No. WO2001 / 03720, PCT Publication No. WO99 / 20758, PCT Publication No. WO2006 / 083289, PCT Publication No. WO2005 / 115451, U.S. Patent No. 7,618,632, and PCT Publication No. WO2011 / 051726, but are not limited thereto.

[0227] In some embodiments, a compound having the structure

Chemical Formula

[0228] In some embodiments, a structure

Chemical formula

[0229] In some embodiments, a structure

Chemical formula

[0230] In some embodiments, a structure

Chemical formula

[0231] In some embodiments, the structure

Chemical formula

[0232] In some embodiments, the structure

Chemical formula

[0233] In some embodiments, the structure

Chemical Structure

[0234] In some embodiments, the structure

Chemical Structure

[0235] In some embodiments, the structure

Chemical Structure

[0236] In some embodiments, the structure

Chemical Structure

[0237] In some embodiments, the structure

Chemical formula

[0238] In some embodiments, the structure

Chemical formula

[0239] In some embodiments, the structure

Chemical formula

[0240] In some embodiments, the structure

Chemical formula

[0241] In some embodiments, the structure

Chemical Structure

[0242] The compounds described herein can be used in combination with the agents disclosed herein or other suitable agents depending on the condition to be treated. Thus, in some embodiments, one or more compounds of the disclosure will be co-administered with other agents as described above. When the compounds described herein are used in combination therapy, they are administered simultaneously or separately with the second agent. Such co-administration can include simultaneous administration of the two agents in the same dosage form, simultaneous administration in individual dosage forms, and separate administration. That is, the compounds described herein and any of the above agents can be formulated together in the same dosage form for simultaneous administration. Alternatively, the disclosed compounds and any of the above agents can be co-administered, where both agents are included in separate formulations. In another alternative, any of the above agents can be administered after administration of the compounds of the disclosure, or vice versa. In some embodiments of the separate administration protocol, the disclosed compounds and any of the above agents are administered at intervals of minutes, or hours, or days.

[0243] In one aspect of the invention, treatment of a disease / condition using a combination of pharmaceutically active compounds that can be administered separately is contemplated, and the invention further relates to packaging separate pharmaceutical compositions in kit form. The kit includes two separate pharmaceutical compositions, namely a compound of the invention and a second pharmaceutical compound. The kit includes a container for holding the separate compositions, such as a divided bottle or a divided foil packet. Further examples of containers include syringes, boxes, and bags. In some embodiments, the kit includes instructions for use of the separate components. The kit form is particularly convenient when it is preferred to administer the separate components in different dosage forms (e.g., oral and parenteral dosage forms), at different dosing intervals, or when the prescribing medical professional desires a titration of the individual components of the combination.

Examples

[0244] Method 1 Example 1-1: 1-(4-(6-(2-Bromo-5-hydroxyphenyl)-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazin-1-yl)prop-2-en-1-one

Chemical Structure

[0245] Step 2: 2-Amino-4-bromo-5-chloro-3-fluorobenzamide (Intermediate B). A mixed N,N-dimethylformamide solution (30 mL) of 2-amino-4-bromo-5-chloro-3-fluorobenzoic acid (Intermediate A, 3.96 g, 14.7 mmol) and TBTU (4.97 g, 15.5 mmol, Advanced ChemTech, Louisville, KY, USA) was sequentially added with ammonium chloride (1.10 g, 20.6 mmol) and diisopropylethylamine (5.13 mL, 29.5 mmol). The resulting mixture was stirred at room temperature for 30 minutes. Then, the reaction mixture was added to a saturated aqueous sodium bicarbonate solution and stirred for 15 minutes. The resulting precipitate was collected by filtration, washed with water, and dried under vacuum to obtain 2-amino-4-bromo-5-chloro-3-fluorobenzamide. 1 H NMR (400 MHz, DMSO-d6) δ 8.03 (1H, br s), 7.72 (1H, d, J = 2.0 Hz), 7.47 (1H, br s), 6.86 (2H, s). 19 F NMR (376 MHz, DMSO-d6) δ -120.79 (1F, s). m / z (ESI, +ve) 268.9 (M+H) + 。

[0246] Step 3: 2-Amino-4-bromo-5-chloro-3-fluorobenzothioamide Lawesson's reagent (2.81 g, 6.95 mmol) was added to a THF solution (77 mL) of 2-amino-4-bromo-5-chloro-3-fluorobenzamide (Intermediate B, 3.10 g, 11.59 mmol). The resulting mixture was stirred at room temperature for 1 hour. Then, the reaction mixture was diluted with EtOAc (75 mL) and washed successively with 2M aqueous HCl solution (50 mL), saturated aqueous sodium bicarbonate solution (50 mL), and brine (50 mL). Then, the organic extract was dried over Na2SO4, collected by filtration, and concentrated under reduced pressure. The residue was purified by chromatography (silica gel, DCM containing 0 - 3% MeOH) to obtain 2-amino-4-bromo-5-chloro-3-fluorobenzothioamide: 11H NMR (400 MHz, DMSO-d6) δ 9.93 - 10.15 (1H, m), 9.63 (1H, br s), 7.28 (1H, d, J = 1.96 Hz), 6.34 (2H, s). 19 19F NMR (376 MHz, DMSO-d6) δ -119.52 (1 F, s). m / z (ESI, +ve) 284.8 (M+H) + .

[0247] Step 4: 6-Bromo-5-chloro-7-fluorobenzo[c]isothiazol-3-amine Hydrogen peroxide (in water, 30 wt%, 2.93 mL, 28.7 mmol) was added dropwise to a solution of 2-amino-4-bromo-5-chloro-3-fluorobenzothioamide (2.71 g, 9.55 mmol) in ice-cooled pyridine (32 mL). Subsequently, the resulting mixture was warmed to room temperature and stirred for 24 h. Water (50 mL) was added, and the precipitated solid was collected by filtration, washed with water, and dried in vacuo to give 6-bromo-5-chloro-7-fluorobenzo[c]isothiazol-3-amine: 1 1H NMR (400 MHz, DMSO-d6) δ 8.12 - 8.26 (2H, m), 7.95 - 8.06 (1H, m). 19 19F NMR (376 MHz, DMSO-d6) δ -114.32 (1 F, s). m / z (ESI, +ve) 283.0 (M+H) + .

[0248] Step 5: 6-Bromo-3,5-dichloro-7-fluorobenzo[c]isothiazole (Intermediate C) To an ice-cooled mixture of 6-bromo-5-chloro-7-fluorobenzo[c]isothiazol-3-amine (2.47 g, 8.78 mmol), water (12 mL), and concentrated hydrochloric acid (37 wt%, 12 mL, 395 mmol) was slowly added an aqueous solution (2.0 mL) of sodium nitrite (0.788 g, 11.4 mmol). The resulting mixture was stirred at 0 °C for 2.5 h, and then a mixture of copper(I) chloride (1.39 g, 14.1 mmol) dissolved in concentrated hydrochloric acid (37 wt%, 12 mL, 395 mmol) was added at 0 °C. Subsequently, the reaction mixture was warmed to room temperature and stirred for 20 h. The reaction mixture was diluted with water (50 mL), and the precipitated solid was collected by filtration and dried under vacuum. The recovered material was taken up in DCM:MeOH (3:1) (200 mL) and washed successively with water (200 mL) and brine (100 mL). Then, the organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by chromatography (silica gel, heptane containing 0 - 20% EtOAc) to give 6-bromo-3,5-dichloro-7-fluorobenzo[c]isothiazole: 1 H NMR (400 MHz, DMSO-d6) δ 7.99 (1H, d, J = 1.57 Hz). 19 F NMR (376 MHz, DMSO-d6) δ -111.48 (1 F, s). m / z (ESI, +ve) 425.0 (M+H) + 。

[0249] Step 6: 4-(6-Bromo-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazine-1-carboxylic acid tert-butyl (Intermediate D) A mixed N,N-dimethylformamide solution (2.0 mL) of 6-bromo-3,5-dichloro-7-fluorobenzo[c]isothiazole (Intermediate C, 150 mg, 0.497 mmol) and 1-Boc-piperazine (204 mg, 1.09 mmol) was stirred at room temperature for 20 h. Then, the reaction mixture was adsorbed onto silica gel and purified by chromatography (silica gel, heptane containing 0 - 20% EtOAc) to give 4-(6-bromo-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazine-1-carboxylic acid tert-butyl.1 1H NMR (400 MHz, chloroform-d) δ 7.60 (1H, d, J = 1.56 Hz), 3.68 - 3.79 (4H, m), 3.40 - 3.51 (4H, m), 1.26 (9H, s). m / z (ESI, +ve) 451.8 (M+H) + 。

[0250] Step 7: 4-(6-(2-Bromo-5-methoxyphenyl)-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazine-1-carboxylic acid tert-butyl A mixture of tert-butyl 4-(6-bromo-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazine-1-carboxylate (Intermediate D, 111 mg, 0.247 mmol), 2-bromo-5-methoxybenzeneboronic acid (0.114 mL, 0.494 mmol), sodium carbonate (0.041 mL, 0.988 mmol), and tetrakis(triphenylphosphine)palladium (14.3 mg, 0.012 mmol) in a mixture of 1,4-dioxane solution (1.6 mL) and water (0.4 mL) was heated at 90 °C for 21 h. Then, the reaction mixture was concentrated under reduced pressure, adsorbed on silica gel, and purified by column chromatography (silica gel, heptane containing 0 - 20% EtOAc / EtOH(3:1)) to give tert-butyl 4-(6-(2-bromo-5-methoxyphenyl)-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazine-1-carboxylate: m / z (ESI, +ve) 558.1 (M+H) + 。

[0251] Step 8: 1-(4-(6-(2-Bromo-5-hydroxyphenyl)-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazin-1-yl)prop-2-en-1-one Hydrogen chloride (4M, 1,4-dioxane solution, 2.0 mL, 8.0 mmol) was added to a mixture of tert-butyl 4-(6-(2-bromo-5-methoxyphenyl)-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazine-1-carboxylate (107 mg, 0.192 mmol) and methanol (2.0 mL), and the resulting mixture was stirred at room temperature for 1 hour. Subsequently, the reaction mixture was concentrated under reduced pressure to obtain 6-(2-bromo-5-methoxyphenyl)-5-chloro-7-fluoro-3-(piperazin-1-yl)benzo[c]isothiazole: m / z (ESI, +ve) 458.0 (M+1) + 。

[0252] To this substance (88 mg) was added a dichloromethane solution (2 mL) of N,N-diisopropylethylamine (0.101 mL, 0.578 mmol), and the resulting mixture was cooled to 0 °C. Acryloyl chloride (0.26 M DCM solution, 0.75 mL, 0.19 mmol) was added, and the resulting mixture was stirred at 0 °C for 10 minutes. The reaction mixture was concentrated under reduced pressure to obtain 1-(4-(6-(2-bromo-5-methoxyphenyl)-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazin-1-yl)prop-2-en-1-one: m / z (ESI, +ve) 512.0 (M+H) + 。

[0253] For the compound without the methyl ether protecting group, the crude material was purified at this stage. For the compound with the methyl ether protecting group, it was used in the next transformation without purification of the crude material.

[0254] The obtained 1-(4-(6-(2-bromo-5-methoxyphenyl)-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazin-1-yl)prop-2-en-1-one was taken up in 1,2-dichloroethane (2.0 mL) and cooled to 0 °C. Boron tribromide solution (1.0 M hexane solution, 0.97 mL, 0.97 mmol) was added, and the resulting mixture was stirred at 0 °C for 1 hour. Subsequently, the reaction mixture was added to saturated aqueous sodium hydrogen carbonate solution (2.0 mL), and extracted with DCM / MeOH (2:1) (10 mL). The organic extract was dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by chromatography (silica gel, DCM containing 0 - 3% MeOH) to obtain 1-(4-(6-(2-bromo-5-hydroxyphenyl)-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazin-1-yl)prop-2-en-1-one: 1 H NMR (400 MHz, DMSO-d6) δ 9.99 (br s, 1 H), 8.04 (s, 1 H), 7.55 (d, J = 8.7 Hz, 1 H), 6.81 - 6.94 (m, 2 H), 6.79 (d, J = 2.9 Hz, 1 H), 6.19 (dd, J = 16.7, 2.2 Hz, 1 H), 5.77 (dd, J = 10.5, 2.2 Hz, 1 H), 3.87 (br d, J = 19.5 Hz, 4 H), 3.63 (br t, J = 5.1 Hz, 4 H). 19 F NMR (376 MHz, DMSO-d6) δ -124.16 (1F, s). m / z (ESI, +ve) 498.0 (M+H) + 。

Table 4

[0255] Method 2 Example 2-1: 1-(4-(5-chloro-6-(3-hydroxy-1-naphthalenyl)[1,2]thiazolo[3,4-b]pyridin-3-yl)-1-piperazinyl)-2-propen-1-one

Chemical formula

[0256] Step 2: 4-(2-Amino-6-bromo-5-chloronicotinoyl)piperazine-1-carboxylic acid tert-butyl. To a DMF solution (14 mL) of 2-amino-6-bromo-5-chloronicotinic acid (1.12 g, 4.5 mmol) was added TBTU (1.93 g, 6.0 mmol). After 5 minutes, the reaction mixture was treated successively with 1-Boc-piperazine (912 mg, 4.9 mmol) and DIPEA (2.33 mL, 13.4 mmol). The resulting solution was stirred at room temperature for 25 h, saturated aqueous NaHCO3 (75 mL) was added, and the resulting mixture was extracted with DCM. The organic layer was separated, washed successively with water (2×), dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by chromatography (silica gel, DCM containing 0 - 7% MeOH) to give tert-butyl 4-(2-amino-6-bromo-5-chloronicotinoyl)piperazine-1-carboxylate: 1 H NMR (400 MHz, DMSO-d6) δ 7.58 (s, 1H), 6.66 (s, 2H), 3.33 (s, 8H), 1.40 (s, 9H). m / z (ESI, +ve) 419.0 (M+H) + 。

[0257] Step 3: tert-Butyl 4-(2-amino-6-bromo-5-chloropyridine-3-carbonothioyl)piperazine-1-carboxylate. Lawesson's reagent (353 mg, 0.87 mmol) was added to a THF solution (7.5 mL) of tert-butyl 4-(2-amino-6-bromo-5-chloronicotinoyl)piperazine-1-carboxylate (610 mg, 1.45 mmol), and the resulting solution was stirred at 50 °C for 2.5 h. The reaction mixture was then cooled to room temperature and treated successively with water (10 mL) and 1N aqueous HCl (4 mL). The resulting mixture was extracted with EtOAc (2×), the combined extracts were dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by chromatography (silica gel, DCM containing 0 - 6% MeOH) to give tert-butyl 4-(2-amino-6-bromo-5-chloropyridine-3-carbonothioyl)piperazine-1-carboxylate: 11H NMR (400 MHz, DMSO-d6) δ 7.47 (s, 1H), 6.58 (br. s, 2H), 4.30 (ddd, J = 13.3, 6.3, 3.3 Hz, 1H), 4.01 - 4.13 (m, 2H), 3.68 - 3.77 (m, 1H), 3.51 - 3.59 (m, 1H), 3.40 - 3.50 (m, 3H), 1.41 (s, 9H). m / z (ESI, +ve) 434.9 (M+H) + 。

[0258] Step 4: tert-Butyl 4-(5,6-dichloro-iso-thiazolo[3,4-b]pyridin-3-yl)piperazine-1-carboxylate. NCS (116 mg, 0.87 mmol) was added to a solution of tert-butyl 4-(2-amino-6-bromo-5-chloropyridine-3-carbonothioyl)piperazine-1-carboxylate (343 mg, 0.79 mmol) in THF (8 mL), and the resulting solution was stirred at room temperature for 20 minutes. Then, a mixture of water (10 mL) and 1 M aqueous sodium sulfite solution (5 mL) was added, and the resulting mixture was extracted with EtOAc (2 times). The combined extracts were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by chromatography (silica gel, DCM containing 0 - 4% MeOH) to obtain tert-butyl 4-(5,6-dichloro-iso-thiazolo[3,4-b]pyridin-3-yl)piperazine-1-carboxylate: 1 1H NMR (400 MHz, chloroform-d) δ 8.10 (s, 1H), 3.69 - 3.80 (m, 4H), 3.50 - 3.57 (m, 4H), 1.51 (s, 9H). m / z (ESI, +ve) 389.0 (M+H) + 。

[0259] Step 5: tert-Butyl 4-(5-chloro-6-(3-methoxynaphthalen-1-yl)iso-thiazolo[3,4-b]pyridin-3-yl)piperazine-1-carboxylate. A mixture of tert-butyl 4-(5,6-dichloroiso-thiazolo[3,4-b]pyridin-3-yl)piperazine-1-carboxylate (154 mg, 0.36 mmol), (3-methoxynaphthalen-1-yl)boronic acid (287 mg, 1.42 mmol), and cesium carbonate (463 mg, 1.42 mmol) was dissolved in a mixture of 1,4-dioxane solution (8 mL) and water (2 mL). Argon was bubbled through the mixture, and then tetrakis(triphenylphosphine)palladium (41 mg, 0.04 mmol) was added. The reaction mixture was bubbled with argon again and then heated at 100 °C for 25 h in a sealed tube. After cooling to room temperature, the reaction mixture was diluted with brine (40 mL) and extracted with EtOAc (twice). The combined extracts were dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by chromatography (silica gel, DCM containing 0-3.5% MeOH) to give tert-butyl 4-(5-chloro-6-(3-methoxynaphthalen-1-yl)iso-thiazolo[3,4-b]pyridin-3-yl)piperazine-1-carboxylate: m / z (ESI, +ve) 511.1 (M+H) + .

[0260] Step 6: 5-Chloro-6-(3-methoxynaphthalen-1-yl)-3-(piperazin-1-yl)iso-thiazolo[3,4-b]pyridine. Trifluoroacetic acid (560 μL, 7.6 mmol) was added to a DCM solution (6 mL) of tert-butyl 4-(5-chloro-6-(3-methoxynaphthalen-1-yl)iso-thiazolo[3,4-b]pyridin-3-yl)piperazine-1-carboxylate (155 mg, 0.30 mmol). The resulting solution was stirred at room temperature for 2.3 h and then concentrated under reduced pressure. The residue was purified by chromatography (silica gel, DCM containing 0-25% MeOH) to give 5-chloro-6-(3-methoxynaphthalen-1-yl)-3-(piperazin-1-yl)iso-thiazolo[3,4-b]pyridine as the TFA salt: 11H NMR (400 MHz, DMSO-d6) δ 8.78 (s, 1H), 7.94 (d, J = 8.2 Hz, 1H), 7.46 - 7.53 (m, 2H), 7.31 (d, J = 3.7 Hz, 2H), 7.19 (d, J = 2.4 Hz, 1H), 3.95 (s, 3H), 3.76 - 3.83 (m, 4H), 3.35 - 3.43 (m, 4H). m / z (ESI, +ve) 411.0 (M+H) + .

[0261] Step 7: 1-(4-(5-Chloro-6-(3-methoxy-1-naphthalenyl)[1,2]thiazolo[3,4-b]pyridin-3-yl)-1-piperazinyl)-2-propen-1-one. To a slurry of ice-cooled 5-chloro-6-(3-methoxynaphthalen-1-yl)-3-(piperazin-1-yl)isothiazolo[3,4-b]pyridine (TFA salt; 100 mg, 0.19 mmol) in DCM (5 mL) were sequentially added DIPEA (100 μL, 0.57 mmol) and acryloyl chloride (23 μL, 0.29 mmol). The resulting solution was stirred at 0 °C for 70 minutes, and saturated aqueous NaHCO3 (15 mL) was added. The resulting mixture was extracted with DCM (3 times), and the combined extracts were dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by chromatography (silica gel, DCM containing 0 - 7% MeOH) to give 1-(4-(5-chloro-6-(3-methoxy-1-naphthalenyl)[1,2]thiazolo[3,4-b]pyridin-3-yl)-1-piperazinyl)-2-propen-1-one: 1 1H NMR (400 MHz, DMSO-d6) δ 8.73 (s, 1H), 7.93 (d, J = 8.2 Hz, 1H), 7.45 - 7.54 (m, 2H), 7.25 - 7.39 (m, 2H), 7.19 (d, J = 2.5 Hz, 1H), 6.86 (dd, J = 16.7, 10.3 Hz, 1H), 6.19 (dd, J = 16.7, 2.3 Hz, 1H), 5.77 (dd, J = 10.5, 2.3 Hz, 1H), 3.94 (s, 3H), 3.81 - 3.94 (m, 4H), 3.69 - 3.76 (m, 4H). m / z (ESI, +ve) 465.0 (M+H)+ 。

[0262] Step 8: 1-(4-(5-Chloro-6-(3-hydroxy-1-naphthalenyl)[1,2]thiazolo[3,4-b]pyridin-3-yl)-1-piperazinyl)-2-propen-1-one. Boron tribromide (1.0 M hexane solution, 400 μL, 0.40 mmol) was added dropwise to a solution of 1-(4-(5-chloro-6-(3-methoxynaphthalen-1-yl)isothiazolo[3,4-b]pyridin-3-yl)piperazin-1-yl)prop-2-en-1-one (37.3 mg, 0.08 mmol) in ice-cooled 1,2-dichloroethane (4 mL). The resulting mixture was stirred at 0 °C for 2.3 h. Then, saturated aqueous NaHCO3 (5 mL) was added, and the resulting mixture was extracted with DCM:MeOH (4:1) (2 times). The combined extracts were dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by chromatography (silica gel, DCM containing 0 - 6% MeOH) to give 1-(4-(5-chloro-6-(3-hydroxy-1-naphthalenyl)[1,2]thiazolo[3,4-b]pyridin-3-yl)-1-piperazinyl)-2-propen-1-one: 1 H NMR (400 MHz, DMSO-d6) δ 9.97 (br. s, 1H), 8.72 (s, 1H), 7.79 (d, J = 8.6 Hz, 1H), 7.42 (t, J = 7.1 Hz, 1H), 7.17 - 7.28 (m, 3H), 7.09 (d, J = 2.1 Hz, 1H), 6.86 (dd, J = 16.7, 10.5 Hz, 1H), 6.19 (dd, J = 16.7, 2.3 Hz, 1H), 5.74 - 5.79 (m, 1H), 3.81 - 3.95 (m, 4H), 3.68 - 3.76 (m, 4H). m / z (ESI, +ve) 451.0 (M+H) + 。

Table 5

[0263] Method 3 Example 3-1: 1-(4-(5-chloro-7-fluoro-6-(3-hydroxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)piperazin-1-yl)prop-2-en-1-one

Chemical formula

[0264] Step 2: 1-(4-(5-chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)piperazin-1-yl)prop-2-en-1-one (Intermediate E). A mixture of 1-(4-(6-bromo-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazin-1-yl)prop-2-en-1-one (Intermediate D, 79 mg, 0.20 mmol), (3-methoxynaphthalen-1-yl)boronic acid (47.3 mg, 0.234 mmol), tetrakis(triphenylphosphine)palladium (22.5 mg, 0.020 mmol), and sodium carbonate (83 mg, 0.78 mmol) in a mixture of water (0.500 mL) and 1,4-dioxane (2.0 mL) was heated at 100 °C for 16 h. The reaction mixture was then adsorbed onto silica gel and purified by chromatography (silica gel, DCM containing 0~3% MeOH) to give 1-(4-(5-chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)piperazin-1-yl)prop-2-en-1-one: m / z (ESI, +ve) 482.0 (M+H) + .

[0265] Step 3: 1-(4-(5-chloro-7-fluoro-6-(3-hydroxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)piperazin-1-yl)prop-2-en-1-one. Boron tribromide (1.0 M hexane solution, 0.664 mL, 0.664 mmol) was added to a solution of 1-(4-(5-chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)piperazin-1-yl)prop-2-en-1-one (64 mg, 0.13 mmol) in ice-cooled 1,2-dichloroethane (2.0 mL), and the resulting mixture was stirred at 0 °C for 1 hour. Then, the reaction mixture was added to saturated aqueous sodium hydrogen carbonate (2.0 mL), and the resulting mixture was extracted with DCM:MeOH (2:1) (10 mL). The organic extract was dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by chromatography (silica gel, DCM containing 0 - 3% MeOH) to give 1-(4-(5-chloro-7-fluoro-6-(3-hydroxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)piperazin-1-yl)prop-2-en-1-one: 1 H NMR (400 MHz, DMSO-d6) δ 9.90 - 10.04 (1H, m), 8.10 (1H, s), 7.80 (1H, d, J = 8.41 Hz), 7.43 (1H, ddd, J = 1.96, 6.11, 8.17 Hz), 7.16 - 7.31 (3H, m), 7.07 (1H, d, J = 2.35 Hz), 6.87 (1H, dd, J = 10.47, 16.73 Hz), 6.19 (1H, dd, J = 2.25, 16.73 Hz), 5.77 (1H, dd, J = 2.25, 10.47 Hz), 3.88 (4H, br d, J = 19.56 Hz), 3.61 - 3.72 (4H, m). 19 F NMR (376 MHz, DMSO-d6) δ -123.78 (s, 1F). m / z (ESI, +ve) 468.0 (M+H) + 。

[0266] Alternative synthesis method of Intermediate E

Chemical Structure

Table 6

[0267] Method 4 Example 4-1: 1-(6-(5-Chloro-7-fluoro-6-(3-hydroxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)-2,6-diazaspiro[3.3]heptan-2-yl)prop-2-en-1-one.

Chem.

[0268] Step 2: 1-(6-(6-bromo-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)-2,6-diazaspiro[3.3]heptan-2-yl)prop-2-en-1-one. A hydrogen chloride solution (4 M, 1,4-dioxane solution, 5.0 mL, 20 mmol) was added to a methanol solution (10 mL) of tert-butyl 6-(6-bromo-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylate (249 mg, 0.538 mmol), and the resulting mixture was stirred at room temperature for 2 hours. Thereafter, the reaction mixture was concentrated under reduced pressure to obtain 6-bromo-5-chloro-7-fluoro-3-(2,6-diazaspiro[3.3]heptan-2-yl)benzo[c]isothiazole: m / z (ESI, +ve) 363.8 (M+1) + .

[0269] To this substance was added a dichloromethane solution (3.0 mL) of N,N-diisopropylethylamine (0.281 mL, 1.61 mmol), and the resulting mixture was cooled to 0 °C. Thereafter, acryloyl chloride (0.2 M DCM solution, 2.69 mL, 0.538 mmol) was added, and the resulting mixture was stirred at 0 °C for 10 minutes. Thereafter, the reaction mixture was concentrated under reduced pressure, and the residue was purified by chromatography (silica gel, DCM containing 0 - 10% EtOAc / EtOH (3:1)) to obtain 1-(6-(6-bromo-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)-2,6-diazaspiro[3.3]heptan-2-yl)prop-2-en-1-one: 1 H NMR (400 MHz, DMSO-d6) δ 7.65 (1 H, d, J = 1.4 Hz), 6.25 - 6.36 (1 H, m), 6.10 (1 H, dd, J = 17.0, 2.3 Hz), 5.64 - 5.72 (1 H, m), 4.58 (4 H, s), 4.47 (2 H, s), 4.18 (2 H, s). 19 F NMR (376 MHz, DMSO-d6) δ -113.54 (1F, s). m / z (ESI, +ve) 418.0 (M+H) + .

[0270] Step 3: 1-(6-(5-Chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)-2,6-diazaspiro[3.3]heptan-2-yl)prop-2-en-1-one. A mixture of 1-(6-(6-Bromo-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)-2,6-diazaspiro[3.3]heptan-2-yl)prop-2-en-1-one (102 mg, 0.245 mmol), (3-Methoxynaphthalen-1-yl)boronic acid (59.3 mg, 0.294 mmol), tetrakis(triphenylphosphine)palladium (28.3 mg, 0.024 mmol), and sodium carbonate (104 mg, 0.979 mmol) in a mixture of water (0.5 mL) and 1,4-dioxane (2.0 mL) was heated at 100 °C for 1 hour. Then, the reaction mixture was adsorbed on silica gel and purified by chromatography (silica gel, DCM containing 0 - 5% MeOH). The purified substance was sonicated in MeOH, the suspended solid was filtered off and collected, washed with MeOH, and then dried under vacuum to obtain 1-(6-(5-Chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)-2,6-diazaspiro[3.3]heptan-2-yl)prop-2-en-1-one: 1 H NMR (400 MHz, DMSO-d6) δ 7.93 (1 H, d, J = 8.4 Hz), 7.67 (1 H, s), 7.45 - 7.57 (2 H, m), 7.23 - 7.36 (2 H, m), 7.16 (1 H, d, J = 2.5 Hz), 6.27 - 6.39 (1 H, m), 6.11 (1 H, dd, J = 17.0, 2.2 Hz), 5.65 - 5.76 (1 H, m), 4.58 - 4.67 (4 H, m), 4.50 (2 H, s), 4.22 (2 H, s), 3.93 (3 H, s). 19 F NMR (376 MHz, DMSO-d6) δ -123.88 (1F, s). m / z (ESI, +ve) 494.0 (M+H) + .

[0271] Step 4: 1-(6-(5-chloro-7-fluoro-6-(3-hydroxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)-2,6-diazaspiro[3.3]heptan-2-yl)prop-2-en-1-one. Boron tribromide (1.0 M hexane solution, 0.638 mL, 0.638 mmol) was added to a solution of 1-(6-(5-chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)-2,6-diazaspiro[3.3]heptan-2-yl)prop-2-en-1-one (63 mg, 0.128 mmol) in ice-cooled 1,2-dichloroethane (2.0 mL). The resulting mixture was stirred at 0 °C for 2 hours. Then, the reaction mixture was added to saturated aqueous sodium hydrogen carbonate (2.0 mL), and the resulting mixture was extracted with DCM:MeOH (2:1) (10 mL). The organic extract was dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by chromatography (silica gel, MeOH (containing 2 M ammonia) 0 - 2% in DCM) to give 1-(6-(5-chloro-7-fluoro-6-(3-hydroxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)-2,6-diazaspiro[3.3]heptan-2-yl)prop-2-en-1-one: 1 H NMR (400 MHz, DMSO-d6) δ 9.82 - 10.04 (1 H, m), 7.79 (1 H, d, J = 8.2 Hz), 7.66 (1 H, s), 7.43 (1 H, dt, J = 8.3, 4.0 Hz), 7.26 (1 H, d, J = 2.3 Hz), 7.22 (2 H, d, J = 3.7 Hz), 7.05 (1 H, d, J = 2.3 Hz), 6.26 - 6.38 (1 H, m), 6.12 (1 H, dd, J = 16.8, 2.2 Hz), 5.66 - 5.72 (1 H, m), 4.58 - 4.67 (4 H, m), 4.50 (2 H, s), 4.22 (2 H, s). 19 F NMR (376 MHz, DMSO-d6) δ -123.98 (1F, s). m / z (ESI, +ve) 480.0 (M+H) + .

Table 7

[0272] Method 5 Example 5-1: N-(1-(5-chloro-7-fluoro-6-(3-hydroxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-3-azetidinyl)-N-methyl-2-propenamide [Chemical formula] Step 1: 2-amino-5-chloro-3-fluoro-4-(3-methoxynaphthalen-1-yl)benzamide. (3-Methoxynaphthalen-1-yl)boronic acid (2.04 g, 10.1 mmol), 2-amino-4-bromo-5-chloro-3-fluorobenzamide (Intermediate B (1.93 g, 7.20 mmol), tetrakis(triphenylphosphine)palladium (0.832 g, 0.720 mmol), an aqueous solution of sodium carbonate (1.2 mL, 28.8 mmol) dissolved in water (9.6 mL), and a 1,4-dioxane solution (38.4 mL) were heated at 90 °C for 2 days. Then, the reaction mixture was filtered through a Celite pad and washed with EtOAc. The filtrate was diluted with saturated aqueous NaHCO3 (50 mL) and extracted with EtOAc (50 mL, 3 times). The organic extract was washed with brine (30 mL) and dried over Na2SO4. Then, the solution was filtered and the filtrate was concentrated under reduced pressure. The residue was suspended in MeOH (5 mL), and the suspended solid was collected by filtration, washed with MeOH, and then dried to obtain 2-amino-5-chloro-3-fluoro-4-(3-methoxynaphthalen-1-yl)benzamide. The concentrated filtrate was purified by chromatography (silica gel, heptane containing 0% - 100% EtOAc-EtOH (3:1)) to obtain additional 2-amino-5-chloro-3-fluoro-4-(3-methoxynaphthalen-1-yl)benzamide. 11H NMR (400 MHz, DMSO-d6) δ 8.01 - 8.17 (m, 1H), 7.92 (d, J = 8.2 Hz, 1H), 7.75 (s, 1H), 7.43 - 7.55 (m, 3H), 7.23 - 7.34 (m, 2H), 7.10 (d, J = 2.5 Hz, 1H), 6.73 (s, 2H), 3.93 (s, 3H). m / z (ESI, +ve) 345.0 (M+H) + .

[0273] Step 2: 2-Amino-5-chloro-3-fluoro-4-(3-methoxynaphthalen-1-yl)benzothioamide. To a solution of 2-amino-5-chloro-3-fluoro-4-(3-methoxynaphthalen-1-yl)benzamide (2.11 g, 6.12 mmol) in tetrahydrofuran (41 mL) was added Lawesson's reagent (1.49 mL, 3.67 mmol), and the resulting mixture was stirred at room temperature for 1 hour. Then, the reaction mixture was diluted with EtOAc (60 mL) and washed successively with 2M HCl (60 mL), saturated aqueous NaHCO3 (60 mL), and brine (60 mL). The organic extract was dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was sonicated in DCM (5 mL), and the resulting precipitate was collected by filtration, washed with DCM, and dried under vacuum to give 2-amino-5-chloro-3-fluoro-4-(3-methoxynaphthalen-1-yl)benzothioamide. The filtrate was purified by chromatography (silica gel, heptane containing 0% - 100% EtOAc-EtOH (3:1)) to give additional 2-amino-5-chloro-3-fluoro-4-(3-methoxynaphthalen-1-yl)benzothioamide: m / z (ESI, +ve) 361.0 (M+H) + .

[0274] Step 3: 5-Chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazol-3-amine. A hydrogen peroxide solution (30% aqueous solution, 2.2 mL, 21.3 mmol) was slowly added to a ice-cooled pyridine solution (18 mL) of 2-amino-5-chloro-3-fluoro-4-(3-methoxynaphthalen-1-yl)benzothioamide (1.92 g, 5.33 mmol). The resulting mixture was warmed to room temperature and stirred at room temperature for 18 h. Then, the reaction mixture was diluted with water (60 mL), and the resulting precipitate was collected by filtration, washed successively with water and MeOH, and then dried in vacuo to give 5-chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazol-3-amine: 1 H NMR (400 MHz, DMSO-d6) δ 8.14 (s, 2H), 7.99 - 8.03 (m, 1H), 7.93 (d, J = 8.3 Hz, 1H), 7.48 - 7.55 (m, 1H), 7.47 (d, J = 2.3 Hz, 1H), 7.31 (d, J = 3.9 Hz, 2H), 7.16 (d, J = 2.5 Hz, 1H), 3.94 (s, 3H). 19 F NMR (376 MHz, DMSO-d6) δ -124.71 (s, 1F). m / z (ESI, +ve) 359.0 (M+H) + 。

[0275] Step 4: 3,5-Dichloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazole. 5-Chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazol-3-amine (1.55 g, 4.31 mmol) was added portionwise to a suspension of copper(II) chloride (0.870 g, 6.47 mmol) and tert-butyl nitrite (0.77 mL, 6.47 mmol) in acetonitrile (43 mL) at 65 °C over 15 minutes. The resulting mixture was stirred at 65 °C for 30 minutes, then cooled to room temperature and diluted with ice water (50 mL). The precipitated solid was filtered off, washed with water and dried in vacuo. The residue was sonicated in DCM (10 mL), the suspended solid was filtered off, washed with DCM and dried in vacuo to recover unreacted 5-chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazol-3-amine. The filtrate was concentrated under reduced pressure to afford 3,5-dichloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazole. 1 H NMR (400 MHz, DMSO-d6) δ 7.98 (s, 1H), 7.96 (d, J = 8.2 Hz, 1H), 7.49 - 7.56 (m, 2H), 7.28 - 7.36 (m, 2H), 7.24 - 7.28 (m, 1H), 3.95 (s, 3H). 19 F NMR (376 MHz, DMSO-d6) δ -122.17 (s, 1F). m / z (ESI, +ve) 378.0 (M+H) + 。

[0276] Step 5: (1-(5-Chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)azetidin-3-yl)(methyl)carbamic acid tert-butyl. A mixed DMF solution (1.3 mL) of 3,5-dichloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazole (100 mg, 0.264 mmol), DIPEA (0.14 mL, 0.793 mmol), and 3-Boc-3-methylaminoazetidine (0.098 mL, 0.529 mmol, Beta Pharma Scientific, Inc.) was stirred at room temperature for 18 h. Then, ice water (3 mL) was added and the resulting mixture was stirred for 15 min. Thereafter, the precipitated solid was filtered off, collected, washed with water, and dried in vacuo to give tert-butyl (1-(5-chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)azetidin-3-yl)(methyl)carbamate: m / z (ESI, +ve) 528.0 (M+H) + 。

[0277] Step 6: N-(1-(5-chloro-7-fluoro-6-(3-hydroxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-3-azetidinyl)-N-methyl-2-propenamide. The title compound was prepared in 3 steps from tert-butyl (1-(5-chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)azetidin-3-yl)(methyl)carbamate (131.1 mg, 0.248 mmol) according to the procedure reported in Method 1, Step 8: 1 H NMR (400 MHz, DMSO-d6) δ 9.89 - 10.10 (m, 1H), 7.79 (d, J = 8.4 Hz, 1H), 7.73 (s, 1H), 7.43 (ddd, J = 8.2, 5.1, 2.9 Hz, 1H), 7.20 - 7.30 (m, 3H), 7.05 (d, J = 2.2 Hz, 1H), 6.81 (dd, J = 16.7, 10.5 Hz, 1H), 6.10 - 6.23 (m, 1H), 5.69 - 5.81 (m, 1H), 5.37 - 5.59 (m, 1H), 4.63 - 4.74 (m, 3H), 4.53 - 4.61 (m, 1H), 3.14 - 3.23 (m, 3H). 1919F NMR (376 MHz, DMSO-d6) δ -124.10 (s, 1F). m / z (ESI, +ve) 468.0 (M+H) + .

Table 8

[0278] Method 6 Example 6-1: 1-(4-(6-(6-Amino-3-chloro-2-pyridinyl)-5-chloro-7-fluoro-2,1-benzothiazol-3-yl)-1-piperazinyl)-2-propen-1-one

Chemical

[0279] Step 2: 4-(6-(6-Amino-3-chloropyridin-2-yl)-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazine-1-carboxylic acid tert-butyl. A mixture of 4-(5-Chloro-7-fluoro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[c]isothiazol-3-yl)piperazine-1-carboxylic acid tert-butyl (99.5 mg, 0.200 mmol), SPhos Pd G3 (17.3 mg, 0.020 mmol), 6-Bromo-5-chloropyridin-2-amine (Combi-blocks Inc., San Diego, CA, USA, 124 mg, 0.6 mmol), and sodium carbonate (85 mg, 0.80 mmol) was dissolved in an aqueous solution (0.25 mL), and the mixture with 1,2-DCE (0.75 mL) was heated at 50 °C for 2 hours. The reaction mixture was concentrated under reduced pressure and purified by chromatography (silica gel, heptane containing 0% - 100% EtOAc-EtOH (3:1)), and 4-(6-(6-Amino-3-chloropyridin-2-yl)-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazine-1-carboxylic acid tert-butyl was obtained: m / z (ESI, +ve) 498.0 (M+H) + .

[0280] Step 3: 1-(4-(6-(6-Amino-3-chloro-2-pyridinyl)-5-chloro-7-fluoro-2,1-benzothiazol-3-yl)-1-piperazinyl)-2-propen-1-one. The title compound was prepared in two steps from tert-butyl 4-(6-(6-amino-3-chloropyridin-2-yl)-5-chloro-7-fluorobenzo[c]isothiazol-3-yl)piperazine-1-carboxylate (31.6 mg, 0.063 mmol) according to the procedure reported in Method 1, Step 8: 1 H NMR (400 MHz, DMSO-d6) δ 7.97 - 8.10 (m, 1H), 7.60 (d, J = 8.9 Hz, 1H), 6.86 (dd, J = 16.6, 10.6 Hz, 1H), 6.57 (d, J = 8.9 Hz, 1H), 6.38 (s, 2H), 6.19 (dd, J = 16.8, 2.3 Hz, 1H), 5.71 - 5.84 (m, 1H), 3.86 (br d, J = 19.9 Hz, 4H), 3.63 (br d, J = 1.0 Hz, 4H). 19 F NMR (376 MHz, DMSO-d6) δ -126.04 (s, 1F). m / z (ESI, +ve) 452.0 (M+H) + .

Table 9

[0281] Method 7 Example 7-1: 1-((3R)-4-(5-chloro-7-fluoro-6-(3-hydroxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-3-(difluoromethyl)-1-piperazinyl)-2-propen-1-one | 1-((3S)-4-(5-chloro-7-fluoro-6-(3-hydroxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-3-(difluoromethyl)-1-piperazinyl)-2-propen-1-one

Chem.

[0282] Step 2: 4-(2-Amino-5-chloro-3-fluoro-4-(3-methoxynaphthalen-1-yl)benzoyl)-3-(difluoromethyl)piperazine-1-carboxylic acid tert-butyl. A DMF solution (4 mL) of a mixture of 2-amino-5-chloro-3-fluoro-4-(3-methoxynaphthalen-1-yl)benzoic acid (0.150 g, 0.434 mmol), TBTU (0.188 g, 0.586 mmol), 3-(difluoromethyl)piperazine-1-carboxylic acid tert-butyl (0.123 g, 0.521 mmol), and DIPEA (0.23 mL, 1.302 mmol) was stirred at room temperature for 3 hours. Then, the reaction mixture was washed with saturated aqueous NaHCO3 and the aqueous washings were extracted with EtOAc. The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by chromatography (silica gel, EtOAc 0-40% / heptane) to give 4-(2-amino-5-chloro-3-fluoro-4-(3-methoxynaphthalen-1-yl)benzoyl)-3-(difluoromethyl)piperazine-1-carboxylic acid tert-butyl: m / z (ESI, +ve) 586 (M+Na) + .

[0283] Step 3: 4-(2-Amino-5-chloro-3-fluoro-4-(3-methoxynaphthalen-1-yl)phenylcarbonothioyl)-3-(difluoromethyl)piperazine-1-carboxylic acid tert-butyl. Lawson's reagent (0.041 mL, 0.10 mmol) was added to a THF solution (4 mL) of tert-butyl 4-(2-amino-5-chloro-3-fluoro-4-(3-methoxynaphthalen-1-yl)benzoyl)-3-(difluoromethyl)piperazine-1-carboxylate (0.095 g, 0.168 mmol), and the resulting mixture was stirred at 50 °C for 18 h. Thereafter, the reaction mixture was concentrated under reduced pressure and purified by column chromatography (silica gel, 0 - 30% EtOAc / heptane) to obtain tert-butyl 4-(2-amino-5-chloro-3-fluoro-4-(3-methoxynaphthalen-1-yl)phenylcarbonothioyl)-3-(difluoromethyl)piperazine-1-carboxylate: m / z (ESI, +ve) 602.2 (M+Na) + .

[0284] Step 4: tert-Butyl 4-(5-chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)-3-(difluoromethyl)piperazine-1-carboxylate. NBS (0.022 g, 0.17 mmol) was added to a THF solution (7 mL) of tert-butyl 4-(2-amino-5-chloro-3-fluoro-4-(3-methoxynaphthalen-1-yl)phenylcarbonothioyl)-3-(difluoromethyl)piperazine-1-carboxylate, and the resulting mixture was stirred at room temperature for 15 min. The reaction mixture was diluted with water and washed with 10% sodium thiosulfate. The aqueous washings were extracted with EtOAc, and the combined organic layers were concentrated under reduced pressure to obtain tert-butyl 4-(5-chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)-3-(difluoromethyl)piperazine-1-carboxylate: m / z (ESI, +ve) 578.2 (M+H) + .

[0285] Step 5: 1-((3R)-4-(5-chloro-7-fluoro-6-(3-hydroxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-3-(difluoromethyl)-1-piperazinyl)-2-propen-1-one | 1-((3S)-4-(5-chloro-7-fluoro-6-(3-hydroxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-3-(difluoromethyl)-1-piperazinyl)-2-propen-1-one. Method 1: Prepared using a procedure similar to the procedure described in Step 8: 1 H NMR (400 MHz, DMSO-d6) δ 10.13 (br. s., 1 H) 8.12 (d, J = 2.2 Hz, 1 H) 7.80 (d, J = 8.2 Hz, 1 H) 7.43 (br t, J = 7.0 Hz, 1 H) 7.20 - 7.30 (m, 3 H) 7.08 (dd, J=5.8, 2.2 Hz, 1 H) 6.78 - 6.91 (m, 1 H) 6.27 - 6.70 (m, 1 H) 6.20 (dd, J = 16.6, 2.0 Hz, 1 H) 5.76 - 5.84 (m, 1 H) 4.73 - 4.87 (m, 1 H) 4.19 - 4.72 (m, 2 H) 3.55 - 3.90 (m, 3 H) 3.36 - 3.47 (m, 1 H). m / z (ESI, +ve) 518.0 (M+H) + .

Table 10

[0286] Method 8 Example 8-1: 6-chloro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-(2-propanyl)phenyl)-4-(4-(2-propenoyl)-1-piperazinyl)-2(1H)-quinazolinone

Chem.

[0287] Step 2: 4-Bromo-5-chloro-2-fluoro-N-((2-isopropylphenyl)carbamoyl)benzamide. A mixed DCE solution (100 mL) of 4-bromo-5-chloro-2-fluorobenzamide (5.90 g, 23.4 mmol) and oxalyl chloride (1 M, DCM solution; 12.9 mL, 25.7 mmol) was stirred at 80 °C for 1 hour using a reflux condenser. Thereafter, the reaction mixture was cooled to room temperature and 2-isopropylaniline (6.62 mL, 46.7 mmol) was added. The resulting mixture was stirred at room temperature for 15 minutes and then cooled to 0 °C. The precipitated solid was removed by filtration, and the collected filtrate was concentrated under reduced pressure to obtain 4-bromo-5-chloro-2-fluoro-N-((2-isopropylphenyl)carbamoyl)benzamide: 1 H NMR (400 MHz, DMSO-d6) δ 11.06 (br. s., 1H) 10.31 (s, 1H) 7.97 - 8.05 (m, 2H) 7.82 (d, J = 7.2 Hz, 1H) 7.32 - 7.38 (m, 1H) 7.14 - 7.25 (m, 2H) 3.11 (spt, J = 6.8 Hz, 1H) 1.24 (d, J = 6.8 Hz, 6H). 19 F NMR (376 MHz, DMSO-d6) δ -113.6 (s, 1 F). m / z (ESI, +ve) 412.7 and 414.6 (M+H) + .

[0288] Step 3: 7-Bromo-6-chloro-1-(2-isopropylphenyl)quinazoline-2,4(1H,3H)-dione (Intermediate F). KHMDS (1M THF solution, 8.30 mL, 8.30 mmol) was added to a mixed THF solution (19 mL) of 4-bromo-5-chloro-2-fluoro-N-((2-isopropylphenyl)carbamoyl)benzamide (1.56 g, 3.77 mmol) at -20 °C, and the resulting mixture was warmed to room temperature over 1 hour. Then, the reaction mixture was diluted with EtOAc (150 mL) and washed with saturated aqueous ammonium chloride solution (100 mL, 2 times). The organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was suspended in DCM (5 mL), sonicated, collected by filtration, and dried in vacuo to give 7-bromo-6-chloro-1-(2-isopropylphenyl)quinazoline-2,4(1H,3H)-dione: 1 1H NMR (400 MHz, CDCl3) δ 9.43 (br. s., 1H) 8.29 (s, 1H) 7.55 - 7.59 (m, 2H) 7.39 - 7.44 (m, 1H) 7.16 (d, J = 7.8 Hz, 1H) 6.75 (s, 1H) 2.59 - 2.77 (m, 1H) 1.17 - 1.24 (m, 3H) 1.11 (d, J = 6.8 Hz, 3H). m / z (ESI, +ve) 392.9 and 395.0 (M+H) + 。

[0289] Step 4: 6-Chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropylphenyl)quinazoline-2,4(1H,3H)-dione. A DME solution (30 mL) of a mixture of 7-bromo-6-chloro-1-(2-isopropylphenyl)quinazoline-2,4(1H,3H)-dione (Intermediate F, 1.17 g, 2.96 mmol), (2-fluoro-6-methoxyphenyl)boronic acid (2.02 g, 11.9 mmol), SPhos Pd G3 (0.128 g, 0.148 mmol), and potassium carbonate (2 M, aqueous solution, 4.45 mL, 8.90 mmol) was stirred at 85 °C for 16 h. Then, the reaction mixture was diluted with EtOAc (150 mL) and washed with saturated aqueous NaHCO3 (100 mL, 3 times). The organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by chromatography (silica gel, heptane containing 0 - 50% EtOAc) to give 6-chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropylphenyl)quinazoline-2,4(1H,3H)-dione: 1 H NMR (400 MHz, DMSO-d6) δ 11.90 (d, J = 1.2 Hz, 1H) 8.11 (d, J = 3.3 Hz, 1H) 7.53 - 7.59 (m, 1 H) 7.48 (tt, J = 7.0, 2.2 Hz, 1H) 7.38 - 7.44 (m, 1H) 7.32 - 7.37 (m, 2H) 6.93 (dd, J = 8.4, 4.3 Hz, 1H) 6.86 (t, J = 8.7 Hz, 1H) 6.15 (s, 1H) 3.66 (d, J = 30 Hz, 3H) 2.73 (dq, J = 14.2, 7.0 Hz, 1H) 1.11 (t, J = 7.1 Hz, 3H) 1.03 (dd, J = 12.7, 6.8 Hz, 3H). 19 F NMR (376 MHz, DMSO-d6) δ -113.8 (s, 1F) -115.2 (s, 1F). m / z (ESI, +ve) 439.1 (M+H) + .

[0290] Step 5: 4,6-Dichloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropylphenyl)quinazolin-2(1H)-one. A solution of 6-chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropylphenyl)quinazoline-2,4(1H,3H)-dione (0.395 g, 0.900 mmol) and Et3N (0.753 mL, 5.40 mmol) in acetonitrile (9 mL) was added phosphorus oxychloride (0.503 mL, 5.40 mmol), and the resulting solution was stirred at 80 °C for 1.5 h. The reaction mixture was concentrated under reduced pressure to give 4,6-dichloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropylphenyl)quinazolin-2(1H)-one: m / z (ESI, +ve) 457.1 (M+H) + .

[0291] Alternative procedure for Step 5 (used as noted in the table below): Phosphorus oxychloride (6.0 eq) was added to a stirred mixture of the product obtained in Step 4 (1.0 eq), triethylamine (18.0 eq), and 1H-benzotriazole (12 eq) in acetonitrile solution (0.07 M), and the resulting reaction mixture was stirred at 80 °C for 3.5 h. Then, the reaction mixture was slowly poured into rapidly stirred water (100 mL) at 10 °C. The aqueous suspension was stirred for 15 min and then extracted with EtOAc (100 mL). The organic layer was washed with brine (100 mL), dried over MgSO4, filtered, and concentrated under reduced pressure to give the benzotriazole adduct intermediate, which was used directly in Step 6.

[0292] Step 6: tert-Butyl 4-(6-chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropylphenyl)-2-oxo-1,2-dihydroquinazolin-4-yl)piperazine-1-carboxylate. 4,6-Dichloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropylphenyl)quinazolin-2(1H)-one (obtained in Method 8, Step 5), tert-butyl piperazine-1-carboxylate (0.335 g, 1.80 mmol), and Et3N (0.753 mL, 5.40 mmol) in DCE (9 mL) were stirred at 60 °C for 20 minutes. The reaction mixture was diluted with EtOAc (100 mL) and washed with saturated aqueous NaHCO3 (75 mL, 3 times). The organic layer was dried over Na2SO4 and concentrated under reduced pressure. The residue was purified by chromatography (silica gel, heptane containing 0 - 60% EtOAc - EtOH (3:1)) to give tert-butyl 4-(6-chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropylphenyl)-2-oxo-1,2-dihydroquinazolin-4-yl)piperazine-1-carboxylate: m / z (ESI, +ve) 607.3 (M+H) + .

[0293] Note: When (S)-1-(3-methylpiperazin-1-yl)prop-2-en-1-one 2,2,2-trifluoroacetate was used, it was synthesized as follows:

[0294] (S)-1-(3-methylpiperazin-1-yl)prop-2-en-1-one 2,2,2-trifluoroacetate

Chemical formula

[0295] Step 6-a: (S)-tert-butyl 4-acryloyl-2-methylpiperazine-1-carboxylate. Acryloyl chloride (1.34 mL, 16.5 mmol) was added to a solution of (S)-1-boc-2-methyl-piperazine (3.00 g, 15.0 mmol, Boc Sciences, Shirley, NY) in THF (30.0 mL) at -10 °C, and the resulting mixture was stirred at -10 °C for 5 minutes. Then, triethylamine (6.26 mL, 44.9 mmol) was slowly added, and the resulting mixture was stirred at -10 °C for 15 minutes and then warmed to room temperature. The reaction mixture was partitioned between EtOAc and saturated aqueous NaHCO3. The aqueous layer was extracted with EtOAc (3 times), and the combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by chromatography (silica gel, heptane containing 0 - 100% EtOAc) to give (S)-tert-butyl 4-acryloyl-2-methylpiperazine-1-carboxylate: 1 H NMR (400 MHz, DMSO-d6) δ 6.72 - 6.85 (m, 1H) 6.10 - 6.18 (m, 1H) 5.68 - 5.76 (m, 1H) 4.08 - 4.32 (m, 2H) 3.68 - 4.03 (m, 2H) 2.86 - 3.14 (m, 2H) 2.66 - 2.80 (m, 1H) 1.38 - 1.43 (s, 9H) 0.96 - 1.04 (m, 3H). m / z (ESI, +ve) 277.3 (M+Na) + 。

[0296] Step 6-b: (S)-1-(3-methylpiperazin-1-yl)prop-2-en-1-one 2,2,2-trifluoroacetate. A mixed DCM solution (16 mL) of (S)-tert-butyl 4-acryloyl-2-methylpiperazine-1-carboxylate (3.21 g, 12.62 mmol) and TFA (4.7 mL, 63.1 mmol) was stirred at room temperature for 24 hours. Then, the reaction mixture was concentrated under reduced pressure to give (S)-1-(3-methylpiperazin-1-yl)prop-2-en-1-one 2,2,2-trifluoroacetate: 11H NMR (400 MHz, DMSO-d6) δ 8.70 - 8.99 (m, 1H) 6.74 - 6.91 (m, 1H) 6.12 - 6.26 (m, 1H) 5.70 - 5.84 (m, 1H) 4.25 - 4.44 (m, 1H) 4.07 - 4.25 (m, 1H) 3.49 - 3.53 (m, 1H) 3.22 - 3.32 (m, 2H) 2.92 - 3.08 (m, 2H) 1.14 - 1.29 (m, 3H). m / z (ESI, +ve) 155.1 (M+H) + 。

[0297] Step 7: 6-Chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropylphenyl)-4-(piperazin-1-yl)quinazolin-2(1H)-one. A solution of tert-butyl 4-(6-chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropylphenyl)-2-oxo-1,2-dihydroquinazolin-4-yl)piperazine-1-carboxylate (0.594 g, 0.978 mmol) in TFA (4 mL) was stirred at room temperature for 30 minutes. The reaction mixture was concentrated under reduced pressure to give 6-chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropylphenyl)-4-(piperazin-1-yl)quinazolin-2(1H)-one: m / z (ESI, +ve) 507.2 (M+H) + 。

[0298] Step 8: 4-(4-Acryloylpiperazin-1-yl)-6-chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropylphenyl)quinazolin-2(1H)-one. At 0 °C, acryloyl chloride (0.079 mL, 0.98 mmol) was added to a solution of 6-chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropylphenyl)-4-(piperazin-1-yl)quinazolin-2(1H)-one and DIPEA (0.85 mL, 4.9 mmol) in ice-cooled DCM (10 mL). The resulting mixture was stirred at 0 °C for 30 minutes. Subsequently, the reaction mixture was diluted with EtOAc (100 mL) and washed with saturated aqueous NaHCO3 (75 mL, 3 times). The organic layer was dried over Na2SO4, decanted, and concentrated under reduced pressure. The residue was purified by chromatography (silica gel, heptane containing 0 - 100% EtOAc - EtOH (3:1)) to give 4-(4-acryloylpiperazin-1-yl)-6-chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropylphenyl)quinazolin-2(1H)-one: 1 H NMR (400 MHz, CDCl3) δ 7.86 (d, J = 1.2 Hz, 1H) 7.41 - 7.54 (m, 2H) 7.29 - 7.37 (m, 2H) 7.14 (dt, J = 7.8, 1.7 Hz, 1H) 6.70 - 6.79 (m, 2H) 6.58 - 6.68 (m, 1H) 6.50 (d, J = 7.4 Hz, 1H) 6.39 (dd, J = 16.8, 1.8 Hz, 1H) 5.75 - 5.84 (m, 1H) 3.79 - 4.06 (m, 8H) 3.75 (s, 2H) 3.66 (s, 1H) 2.69 (tt, J = 13.4, 6.8 Hz, 1H) 1.20 - 1.24 (m, 3H) 1.07 (dd, J = 6.8, 3.9 Hz, 3H). 19 F NMR (377 MHz, CDCl3) δ -113.05 (s, 1F) -113.55 (s, 1F). m / z (ESI, +ve) 561.2 (M+H) + .

[0299] Step 9: 6-chloro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-(2-propanyl)phenyl)-4-(4-(2-propenoyl)-1-piperazinyl)-2(1H)-quinazolinone. BBr3 (1 M, DCE solution, 3.3 mL, 3.3 mmol) was added to a ice-cooled DCE solution (1.7 mL) of 4-(4-acryloylpiperazin-1-yl)-6-chloro-7-(2-fluoro-6-methoxyphenyl)-1-(2-isopropylphenyl)quinazolin-2(1H)-one (0.372 g, 0.663 mmol). The resulting mixture was stirred at 0 °C for 20 minutes, then warmed to room temperature and stirred at room temperature for 2 hours. Saturated aqueous NaHCO3 was added to the reaction mixture, followed by EtOAc (150 mL). The organic layer was separated and washed with saturated aqueous NaHCO3 (100 mL, 3 times). Then, the organic layer was dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by chromatography (silica gel, heptane containing 0 - 100% EtOAc-EtOH (3:1)) to obtain 6-chloro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-(2-propanyl)phenyl)-4-(4-(2-propenoyl)-1-piperazinyl)-2(1H)-quinazolinone: 1 H NMR (400 MHz, DMSO-d6) δ 10.06 (br. d., J = 15.1 Hz, 1H) 8.03 (d, J = 1.2 Hz, 1H) 7.51 - 7.56 (m, 1H) 7.45 (t, J = 7.6 Hz, 1H) 7.33 (tdd, J = 7.5, 7.5, 3.8, 1.4 Hz, 1H) 7.14 - 7.25 (m, 2H) 6.84 (dd, J = 16.8, 10.4 Hz, 1H) 6.62 - 6.74 (m, 2H) 6.14 - 6.26 (m, 2H) 5.71 - 5.78 (m, 1H) 3.71 - 3.99 (m, 8H) 2.52 - 2.59 (m, 1H) 1.02 - 1.12 (m, 6H). 19 F NMR (377 MHz, DMSO-d6) δ -113.6 (s, 1F) -114.8 (s, 1F). m / z (ESI, +ve) 547.1 (M+H) + 。

Table 11

[0300] Method 9 Example 9-1: 6-Chloro-7-(2,3-dichloro-5-hydroxyphenyl)-4-((2S)-2-methyl-4-(2-propenoyl)-1-piperazinyl)-1-(2-(2-propanyl)phenyl)-2(1H)-quinazolinone

Chemical Structure

[0301] Step 2: (S)-4-(4-Acryloyl-2-methylpiperazin-1-yl)-7-bromo-6-chloro-1-(2-isopropylphenyl)quinazolin-2(1H)-one. A DMF solution (2.3 mL) of a mixture of 7-bromo-4,6-dichloro-1-(2-isopropylphenyl)quinazolin-2(1H)-one (492 mg, 1.19 mmol), (S)-4-N-boc-2-methylpiperazine (478 mg, 2.39 mmol), and DIPEA (0.623 mL, 3.58 mmol) was stirred at room temperature for 10 minutes. Then, ice water (10 mL) was added, and the resulting mixture was stirred for 15 minutes. The precipitated solid was filtered off, washed with water, and dried under vacuum to obtain (S)-tert-butyl 4-(7-bromo-6-chloro-1-(2-isopropylphenyl)-2-oxo-1,2-dihydroquinazolin-4-yl)-3-methylpiperazine-1-carboxylate: m / z (ESI, +ve) 577.1 (M+H) + .

[0302] TFA (2.0 mL, 26.8 mmol) was added to a DCM solution (2.0 mL) of (S)-tert-butyl 4-(7-bromo-6-chloro-1-(2-isopropylphenyl)-2-oxo-1,2-dihydroquinazolin-4-yl)-3-methylpiperazine-1-carboxylate (297 mg, 0.516 mmol), and the resulting mixture was stirred at room temperature for 15 minutes. The resulting mixture was concentrated under reduced pressure to obtain (S)-7-bromo-6-chloro-1-(2-isopropylphenyl)-4-(2-methylpiperazin-1-yl)quinazolin-2(1H)-one: m / z (ESI, +ve) 477.0 (M+H) + .

[0303] Acryloyl chloride (0.258 M DCM solution, 4.0 mL, 1.031 mmol) was added to a ice-cooled mixed DCM solution (2.0 mL) of (S)-7-bromo-6-chloro-1-(2-isopropylphenyl)-4-(2-methylpiperazin-1-yl)quinazolin-2(1H)-one and DIPEA (0.269 mL, 1.547 mmol). The resulting mixture was stirred at 0 °C for 20 minutes. After concentration under reduced pressure, the residue was purified by chromatography (silica gel, heptane containing 0 - 100% EtOAc-EtOH (3:1)), and (S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-7-bromo-6-chloro-1-(2-isopropylphenyl)quinazolin-2(1H)-one was obtained: 1 H NMR (400 MHz, DMSO-d6) δ 7.91 - 8.08 (m, 1H), 7.49 - 7.67 (m, 2H), 7.41 (br d, J = 5.8 Hz, 1H), 7.21 (br s, 1H), 6.76 - 6.98 (m, 1H), 6.52 - 6.67 (m, 1H), 6.09 - 6.29 (m, 1H), 5.75 (br s, 1H), 4.61 - 4.96 (m, 1H), 4.23 - 4.48 (m, 1H), 3.93 - 4.21 (m, 2H), 3.50 - 3.77 (m, 1H), 3.33 - 3.49 (m, 1H), 3.23 - 3.28 (m, 1H), 2.94 - 3.24 (m, 1H), 1.27 (br d, J = 9.3 Hz, 6H), 1.09 (br s, 3H). m / z (ESI, +ve) 531.1 (M+H) + .

[0304] Step 3: (S)-4-(4-Acryloyl-2-methylpiperazin-1-yl)-6-chloro-7-(2,3-dichloro-5-methoxyphenyl)-1-(2-isopropylphenyl)quinazolin-2(1H)-one. (S)-4-(4-Acryloyl-2-methylpiperazin-1-yl)-7-bromo-6-chloro-1-(2-isopropylphenyl)quinazolin-2(1H)-one (120 mg, 0.226 mmol), 2-(2,3-dichloro-5-methoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (82 mg, 0.272 mmol), Na2CO3 (96 mg, 0.906 mmol), and Pd(PPh3)4 (26.2 mg, 0.023 mmol) were dissolved in a mixture of 1,4-dioxane solution (1.6 mL) and water (0.4 mL), and the mixture was heated at 90 °C for 17 h. Then, the reaction mixture was concentrated under reduced pressure and purified by chromatography (silica gel, heptane containing 0 - 100% EtOAc-EtOH (3:1)) to obtain (S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-6-chloro-7-(2,3-dichloro-5-methoxyphenyl)-1-(2-isopropylphenyl)quinazolin-2(1H)-one: m / z (ESI, +ve) 627.0 (M+H) + .

[0305] Step 4: 6-Chloro-7-(2,3-dichloro-5-hydroxyphenyl)-4-((2S)-2-methyl-4-(2-propenoyl)-1-piperazinyl)-1-(2-(2-propanyl)phenyl)-2(1H)-quinazolinone. BBr3 (1 M hexane solution, 0.32 mL, 0.320 mmol) was added to an ice-cooled mixture of (S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-6-chloro-7-(2,3-dichloro-5-methoxyphenyl)-1-(2-isopropylphenyl)quinazolin-2(1H)-one (40 mg, 0.064 mmol) and DCE (1.0 mL), and the resulting mixture was stirred at 0 °C for 30 minutes. Saturated aqueous NaHCO3 (2.0 mL) was added, and the resulting mixture was extracted with DCM / MeOH (2:1) (5 mL). The organic extract was dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by chromatography (silica gel, DCM containing 0 - 10% MeOH) to give 6-chloro-7-(2,3-dichloro-5-hydroxyphenyl)-4-((2S)-2-methyl-4-(2-propenoyl)-1-piperazinyl)-1-(2-(2-propanyl)phenyl)-2(1H)-quinazolinone: 1 H NMR (400 MHz, DMSO-d6) δ 10.42 (br d, J = 17.0 Hz, 1H), 7.86 - 8.11 (m, 1H), 7.50 - 7.63 (m, 1H), 7.47 (br t, J = 6.0 Hz, 1H), 7.36 (t, J = 7.5 Hz, 1H), 7.15 - 7.26 (m, 1H), 7.05 (d, J = 2.3 Hz, 1H), 6.78 - 6.96 (m, 1H), 6.44 - 6.58 (m, 1H), 6.11 - 6.29 (m, 2H), 5.71 - 5.82 (m, 1H), 4.68 - 4.98 (m, 1H), 3.96 - 4.52 (m, 3H), 3.52 - 3.85 (m, 2H), 3.34 - 3.51 (m, 1H), 2.95 - 3.26 (m, 1H), 1.27 - 1.41 (m, 3H), 0.95 - 1.13 (m, 6H). m / z (ESI, +ve) 611.0 (M+H) + . [Table 12] TIFF0007717224000223.tif254166TIFF0007717224000224.tif253166TIFF0007717224000225.tif253167TIFF0007717224000226.tif246167TIFF0007717224000227.tif158167

[0306] Method 10 Example 10-1: 1-(4-(7-chloro-6-(2-fluoro-6-hydroxyphenyl)-4-(2-methylphenyl)-1-phthalazinyl)-1-piperazinyl)-2-propen-1-one.

Chemical formula

[0307] Step 2: 6-chloro-7-(2-fluoro-6-hydroxyphenyl)-2,3-dihydrophthalazine-1,4-dione. A DME solution (60 mL) of a mixture of 6,7-dichloro-2,3-dihydrophthalazine-1,4-dione (intermediate G, 3.80 g, 16.45 mmol), 2-fluoro-6-hydroxyphenylboronic acid (10.26 g, 65.8 mmol, Combi-blocks Inc., San Diego, CA, USA), SPhos Pd G3 (1.423 g, 1.645 mmol), and 2M aqueous Na2CO3 solution (32.9 mL, 65.8 mmol) was stirred at 80 °C for 16 h. The reaction mixture was cooled to room temperature and diluted with water (200 mL) and EtOAc (300 mL). The aqueous layer was separated, acidified with 5N HCl, and extracted with EtOAc (300 mL). The combined organic layers were washed with brine (200 mL), dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was suspended in DCM (50 mL), collected by filtration, and 6-chloro-7-(2-fluoro-6-hydroxyphenyl)-2,3-dihydrophthalazine-1,4-dione was obtained: m / z (ESI, +ve) 307.0 (M+H) + .

[0308] Step 3: 6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-2,3-dihydrophthalazine-1,4-dione. tert-Butyl(chloro)diphenylsilane (2.67 mL, 10.25 mmol) was added to an ice-cooled mixed acetonitrile solution (40 mL) of 6-chloro-7-(2-fluoro-6-hydroxyphenyl)-2,3-dihydrophthalazine-1,4-dione (2.62 g, 8.54 mmol), TEA (4.75 mL, 34.2 mmol). The resulting mixture was stirred at 0 °C for 15 minutes, then warmed to room temperature and stirred for 1.5 hours. Additional tert-butyl(chloro)diphenylsilane (2.67 mL, 10.25 mmol) was added and the resulting mixture was stirred at room temperature for 16 hours. Subsequently, the reaction mixture was diluted with water (300 mL), acidified with 5N HCl and extracted with EtOAc (300 mL). The organic layer was separated, washed successively with brine (250 mL), dried over MgSO4, filtered and concentrated under reduced pressure. The residue was taken up in DCM (200 mL), TFA (20 mL) was added and the resulting mixture was stirred at room temperature for 45 minutes. Thereafter, the reaction mixture was diluted with saturated aqueous NaHCO3 (200 mL) and extracted with DCM (250 mL, 2 times). The combined organic extracts were dried over MgSO4, filtered and concentrated under reduced pressure to afford 6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-2,3-dihydrophthalazine-1,4-dione: m / z (ESI, +ve) 545.2 (M+H) + .

[0309] Step 4: 6-(2-((tert-Butyldiphenylsilyl)oxy)-6-fluorophenyl)-1,4,7-trichlorophthalazine. Pyridine (1.45 mL, 17.1 mmol) was added to a mixture of 6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-2,3-dihydrophthalazine-1,4-dione (4.66 g, 8.55 mmol) and phosphorus oxychloride (6.39 mL, 68.4 mmol), and the resulting mixture was heated at 100 °C for 1.5 h. Then, the reaction mixture was cooled to room temperature and slowly poured into stirred water (300 mL) while maintaining the internal temperature below 10 °C. After stirring for 15 min, the resulting mixture was extracted with EtOAc (400 mL), and the organic extract was washed successively with brine (250 mL), dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by chromatography (silica gel, heptane containing 0-25% EtOAc) to give 6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-1,4,7-trichlorophthalazine: m / z (ESI, +ve) 581.1 (M+H) + .

[0310] Step 5: tert-Butyl 4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-4,7-dichlorophthalazin-1-yl)piperazine-1-carboxylate (Intermediate H). 1-Boc-piperazine (5.00 g, 26.9 mmol) was added to a mixed DCM solution (35 mL) of 6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-1,4,7-trichlorophthalazine (5.21 g, 8.95 mmol) and triethylamine (3.77 mL, 26.9 mmol), and the resulting mixture was stirred at room temperature for 19 hours. Then, the reaction mixture was partitioned between DCM (300 mL) and saturated aqueous NaHCO3 (200 mL). The organic layer was separated, dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by chromatography (silica gel, heptane containing 0 - 50% EtOAc) to obtain a mixture of tert-butyl 4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-4,7-dichlorophthalazin-1-yl)piperazine-1-carboxylate and tert-butyl 4-(7-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-4,6-dichlorophthalazin-1-yl)piperazine-1-carboxylate. The individual positional isomers were isolated by chiral SFC purification (OJ-H column (30 × 250 mm, 5 μm), 15% (MeOH containing 20 mM NH3) in supercritical CO2) to obtain tert-butyl 4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-4,7-dichlorophthalazin-1-yl)piperazine-1-carboxylate as the second eluted isomer: 1 H NMR (400 MHz, CDCl3) δ 8.27 (s, 1H) 8.17 (s, 1H) 7.56 - 7.61 (m, 4H) 7.40 - 7.46 (m, 2H) 7.31 - 7.37 (m, 4H) 6.99 - 7.07 (m, 1H) 6.77 (t, J = 8.61 Hz, 1H) 6.42 (d, J = 8.22 Hz, 1H) 3.72 - 3.77 (m, 4H) 3.53 - 3.59 (m, 4H) 1.51 (s, 9H) 0.66 (s, 9H). m / z (ESI, +ve) 731.2 (M+H) + .

[0311] Step 6: 6-(2-((tert-Butyldiphenylsilyl)oxy)-6-fluorophenyl)-4,7-dichloro-1-(piperazin-1-yl)phthalazine. Trifluoroacetic acid (2 mL, 26.8 mmol) was added to a stirred solution of tert-butyl 4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-4,7-dichlorophthalazin-1-yl)piperazine-1-carboxylate (Intermediate H, 1.21 g, 1.654 mmol) in DCM (10 mL), and the resulting mixture was stirred at room temperature for 1.5 h. The reaction mixture was then diluted with saturated aqueous NaHCO3 (75 mL) and extracted with DCM (100 mL, 2 times). The combined organic extracts were dried over MgSO4, filtered and concentrated in vacuo to give 6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-4,7-dichloro-1-(piperazin-1-yl)phthalazine: m / z (ESI, +ve) 631.3 (M+H) + .

[0312] Step 7: 1-(4-(6-(2-((tert-Butyldiphenylsilyl)oxy)-6-fluorophenyl)-4,7-dichlorophthalazin-1-yl)piperazin-1-yl)prop-2-en-1-one. Acryloyl chloride (0.148 mL, 1.81 mmol) was added to a mixed solution of 6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-4,7-dichloro-1-(piperazin-1-yl)phthalazine (1.04 g, 1.647 mmol) and triethylamine (0.694 mL, 4.94 mmol) in DCM (10 mL), and the resulting mixture was stirred at room temperature for 45 min. Saturated aqueous NaHCO3 (75 mL) was added and the resulting mixture was extracted with DCM (100 mL, 3 times). The combined organic extracts were dried over MgSO4, filtered and concentrated in vacuo to give 1-(4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-4,7-dichlorophthalazin-1-yl)piperazin-1-yl)prop-2-en-1-one: m / z (ESI, +ve) 685.1 (M+H) + .

[0313] Step 8: 1-(4-(4,7-Dichloro-6-(2-fluoro-6-hydroxyphenyl)phthalazin-1-yl)piperazin-1-yl)prop-2-en-1-one (Intermediate I). TBAF (1 M THF solution, 3.3 mL, 3.30 mmol) was added to a THF solution (10 mL) of 1-(4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-4,7-dichlorophthalazin-1-yl)piperazin-1-yl)prop-2-en-1-one (1.13 g, 1.648 mmol), and the resulting mixture was stirred at room temperature for 15 minutes. The reaction mixture was concentrated under reduced pressure, and the residue was purified by column chromatography (silica gel, heptane containing 0 - 100% EtOAc) to obtain 1-(4-(4,7-dichloro-6-(2-fluoro-6-hydroxyphenyl)phthalazin-1-yl)piperazin-1-yl)prop-2-en-1-one: 1 H NMR (400 MHz, DMSO-d6) δ 10.26 (br s, 1H) 8.31 (s, 1H) 8.14 (s, 1H) 7.31 - 7.40 (m, 1H) 6.78 - 6.92 (m, 3H) 6.17 (dd, J = 16.63, 2.35 Hz, 1H) 5.74 (dd, J = 10.37, 2.35 Hz, 1H) 3.79 - 3.92 (m, 4H) 3.46 - 3.55 (m, 4H). m / z (ESI, +ve) 447.0 (M+H) + 。

[0314] Step 9: 1-(4-(7-Chloro-6-(2-fluoro-6-hydroxyphenyl)-4-(o-tolyl)phthalazin-1-yl)piperazin-1-yl)prop-2-en-1-one. A mixture of 1-(4-(4,7-dichloro-6-(2-fluoro-6-hydroxyphenyl)phthalazin-1-yl)piperazin-1-yl)prop-2-en-1-one (Intermediate I, 25 mg, 0.056 mmol), 2-tolylboronic acid (30.4 mg, 0.224 mmol, Frontier Scientific Inc., Logan UT, USA), Pd(PPh3)4 (6.46 mg, 5.59 μmol, Strem Chemicals Inc., NewburyPort, MA, USA), and an aqueous solution of 2M Na2CO3 (0.084 mL, 0.168 mmol) in 1,4-dioxane (0.3 mL) was stirred at 40 °C for 18 h. Then, the reaction mixture was diluted with EtOAc (20 mL) and washed with water (15 mL). The organic layer was separated, washed successively with brine (15 mL), dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by chromatography (silica gel, heptane containing 0 - 100% EtOAc) to give 1-(4-(7-chloro-6-(2-fluoro-6-hydroxyphenyl)-4-(o-tolyl)phthalazin-1-yl)piperazin-1-yl)prop-2-en-1-one: 1 H NMR (400 MHz, DMSO-d6) δ 10.15 (br s, 1H) 8.33 (s, 1H) 7.36 - 7.45 (m, 2H) 7.24 - 7.36 (m, 4H) 6.90 (dd, J = 16.63, 10.37 Hz, 1H) 6.70 - 6.80 (m, 2H) 6.18 (dd, J = 16.73, 2.25 Hz, 1H) 5.75 (dd, J = 10.56, 2.15 Hz, 1H) 3.83 - 3.97 (m, 4H) 3.47 - 3.62 (m, 4H) 1.98 - 2.06 (m, 3H). m / z (ESI, +ve) 503.1 (M+H) + .

Table 13

[0315] Method 11 Example 11-1: 6-Chloro-7-(5-methyl-1H-indazol-4-yl)-1-(2-(2-propanyl)phenyl)-4-(4-(2-propenoyl)-1-piperazinyl)-2(1H)-quinazolinone

Chemical formula

[0316] Step 2: 4-(7-Bromo-6-chloro-1-(2-isopropylphenyl)-2-oxo-1,2-dihydroquinazolin-4-yl)piperazine-1-carboxylic acid tert-butyl. tert-Butyl piperazine-1-carboxylate (268 mg, 1.438 mmol) was added to a stirred mixture of crude 4-(1H-benzo[d][1,2,3]triazol-1-yl)-7-bromo-6-chloro-1-(2-isopropylphenyl)quinazolin-2(1H)-one (647 mg, 1.308 mmol), triethylamine (3.68 mL, 26.2 mmol) in dimethyl sulfoxide (6 mL). The reaction mixture was stirred at 80 °C for 30 minutes. The reaction mixture was diluted with EtOAc (100 mL) and washed with water (75 mL). The organic layer was separated, washed with brine (75 mL), dried over MgSO4, filtered and concentrated in vacuo. The residue was purified by chromatography (silica gel, heptane containing 0 - 100% EtOAc) to afford tert-butyl 4-(7-bromo-6-chloro-1-(2-isopropylphenyl)-2-oxo-1,2-dihydroquinazolin-4-yl)piperazine-1-carboxylate. 1 H NMR (400 MHz, chloroform-d) δ 7.79 (1 H, s) 7.49 - 7.59 (2 H, m) 7.36 - 7.42 (1 H, m) 7.11 (1 H, d, J = 7.63 Hz) 6.80 (1 H, s) 3.79 - 3.92 (4 H, m) 3.62 - 3.73 (4 H, m) 2.60 (1 H, spt, J = 6.80 Hz) 1.49 - 1.54 (9 H, m) 1.22 (3 H, d, J = 6.85 Hz) 1.08 (3 H, d, J = 6.85 Hz). m / z (ESI) M+H: 561.0.

[0317] Step 3: tert-Butyl 4-(6-chloro-1-(2-isopropylphenyl)-7-(5-methyl-1H-indazol-4-yl)-2-oxo-1,2-dihydroquinazolin-4-yl)piperazine-1-carboxylate. Under an argon atmosphere in a sealed vial, tert-butyl 4-(7-bromo-6-chloro-1-(2-isopropylphenyl)-2-oxo-1,2-dihydroquinazolin-4-yl)piperazine-1-carboxylate (115 mg, 0.205 mmol), 4-borono-5-methyl-1H-indazole (0.144 mL, 0.819 mmol, Ark Pharm Inc., Arlington Heights, IL, USA), Sphos Pd G3 (0.016 mL, 0.020 mmol), and sodium carbonate (2 M aqueous solution, 0.409 mL, 0.819 mmol) were mixed in 1,2-dimethoxyethane (1 mL). The reaction mixture was stirred at 100 °C for 24 hours. The reaction mixture was cooled to room temperature and diluted with EtOAc (50 mL) and water (40 mL). The organic layer was separated, washed with brine (40 mL), dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by chromatography (silica gel, heptane containing 0 - 50% (EtOAc / EtOH 3:1)) to obtain tert-butyl 4-(6-chloro-1-(2-isopropylphenyl)-7-(5-methyl-1H-indazol-4-yl)-2-oxo-1,2-dihydroquinazolin-4-yl)piperazine-1-carboxylate. m / z (ESI) M+H: 613.2.

[0318] Step 4: 6-Chloro-1-(2-isopropylphenyl)-7-(5-methyl-1H-indazol-4-yl)-4-(piperazin-1-yl)quinazolin-2(1H)-one. Trifluoroacetic acid (0.5 mL, 6.71 mmol) was added to a stirred dichloromethane solution (1 mL) of tert-butyl 4-(6-chloro-1-(2-isopropylphenyl)-7-(5-methyl-1H-indazol-4-yl)-2-oxo-1,2-dihydroquinazolin-4-yl)piperazine-1-carboxylate (78 mg, 0.127 mmol). The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain crude 6-chloro-1-(2-isopropylphenyl)-7-(5-methyl-1H-indazol-4-yl)-4-(piperazin-1-yl)quinazolin-2(1H)-one. m / z (ESI) M+H: 513.2.

[0319] Step 5: 6-Chloro-7-(5-methyl-1H-indazol-4-yl)-1-(2-(propan-2-yl)phenyl)-4-(4-(prop-2-enoyl)-1-piperazinyl)-2(1H)-quinazolinone. Acryloyl chloride (10.33 μl, 0.127 mmol) was added to a stirred mixture of 6-chloro-1-(2-isopropylphenyl)-7-(5-methyl-1H-indazol-4-yl)-4-(piperazin-1-yl)quinazolin-2(1H)-one (65 mg, 0.127 mmol), triethylamine (0.178 mL, 1.267 mmol) in dichloromethane (2 mL) at 0 °C. The reaction mixture was stirred at 0 °C for 20 minutes. Further acryloyl chloride (5.17 μl, 0.064 mmol) was added and the reaction mixture was stirred at 0 °C for a further 20 minutes. The reaction mixture was diluted with DCM (25 mL) and the reaction was quenched with saturated aqueous sodium bicarbonate (20 mL). The organic layer was separated, dried over MgSO4, filtered and concentrated in vacuo. The residue was purified by chromatography (silica gel, heptane containing (EtOAc / EtOH 3:1) 0 - 80%) to give an impure product. The impure product was further purified by chromatography (silica gel, heptane containing acetone 0 - 100%) to give the separated diastereomers. The first eluting diastereomer was 6-chloro-7-(5-methyl-1H-indazol-4-yl)-1-(2-(propan-2-yl)phenyl)-4-(4-(prop-2-enoyl)-1-piperazinyl)-2(1H)-quinazolinone (Example 11-1-1). 11H NMR (400 MHz, chloroform-d) δ 10.28 (1 H, br s) 7.94 (1 H, s) 7.35 - 7.49 (4 H, m) 7.25 - 7.31 (2 H, m) 7.11 (1 H, d, J = 7.67 Hz) 6.64 (1 H, dd, J = 16.79, 10.57 Hz) 6.54 (1 H, s) 6.41 (1 H, dd, J = 16.79, 1.87 Hz) 5.81 (1 H, dd, J = 10.57, 1.66 Hz) 3.83 - 4.07 (8 H, m) 2.74 (1 H, spt, J = 6.84 Hz) 2.13 (3 H, s) 1.23 (3 H, d, J = 6.84 Hz) 1.04 (3 H, d, J = 6.84 Hz). m / z (ESI) M+H: 567.2. The second eluted diastereomer was further purified by column chromatography (silica gel, heptane containing 0 - 80% (EtOAc / EtOH 3:1)) to obtain 6-chloro-7-(5-methyl-1H-indazol-4-yl)-1-(2-(2-propanyl)phenyl)-4-(4-(2-propenoyl)-1-piperazinyl)-2(1H)-quinazolinone (Example 11-1-2). 1 1H NMR (400 MHz, chloroform-d) δ 10.37 (1 H, br s) 7.94 (1 H, s) 7.34 - 7.50 (4 H, m) 7.21 - 7.31 (2 H, m) 7.13 (1 H, d, J = 7.67 Hz) 6.64 (1 H, dd, J = 16.90, 10.68 Hz) 6.55 (1 H, s) 6.41 (1 H, dd, J = 16.79, 1.66 Hz) 5.81 (1 H, dd, J = 10.47, 1.55 Hz) 3.83 - 4.08 (8 H, m) 2.70 (1 H, spt, J = 6.84 Hz) 2.13 (3 H, s) 1.22 (3 H, d, J = 6.84 Hz) 1.03 (3 H, d, J = 6.84 Hz). m / z (ESI) M+H: 567.2. The above Method 11, Steps 1 to 5 are as follows:

Table 14

[0320] Item 2 - Individual Examples Example 12 1-(4-(7-chloro-4-cyclopropyl-6-(2-fluoro-6-hydroxyphenyl)-1-phthalazinyl)-1-piperazinyl)-2-propen-1-one [Chemical Structure] Step 1: tert-Butyl 4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-cyclopropylphthalazin-1-yl)piperazine-1-carboxylate. To a 20 mL vial charged with tert-butyl 4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-4,7-dichlorophthalazin-1-yl)piperazine-1-carboxylate (Intermediate H, 0.060 g, 0.082 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) dichloromethane complex (0.033 g, 0.041 mmol) and 2-methyltetrahydrofuran (2.0 mL) were added. The resulting mixture was sealed and stirred at room temperature for 10 minutes, then cyclopropylzinc bromide (0.5 M THF solution, 0.820 mL, 0.410 mmol; Rieke Metals, Lincoln, NE, USA) was added via syringe. The reaction mixture was heated at 80 °C for 3 hours, then cooled to room temperature and partitioned between EtOAc (30 mL) and water (10 mL). The aqueous layer was extracted once more with EtOAc (20 mL). The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure. The crude product was purified by column chromatography (silica gel 24 g, heptane containing 0 - 30% acetone) to give tert-butyl 4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-cyclopropylphthalazin-1-yl)piperazine-1-carboxylate. 11H NMR (chloroform-d) δ: 8.31 - 8.38 (m, 1H), 8.15 - 8.23 (m, 1H), 7.55 - 7.64 (m, 4H), 7.39 - 7.47 (m, 2H), 7.29 - 7.38 (m, 4H), 6.99 - 7.09 (m, 1H), 6.74 - 6.85 (m, 1H), 6.36 - 6.47 (m, 1H), 3.68 - 3.79 (m, 4H), 3.37 - 3.51 (m, 4H), 2.37 - 2.48 (m, 1H), 1.48 - 1.54 (m, 9H), 1.37 - 1.45 (m, 1H), 1.30 - 1.33 (m, 1H), 1.00 - 1.15 (m, 2H), 0.61 - 0.71 (m, 9H). m / z (ESI) M+H: 737.4.

[0321] Step 2: 6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-cyclopropyl-1-(piperazin-1-yl)phthalazine. Trifluoroacetic acid (0.316 mL, 4.10 mmol) was added to a DCM solution (0.7 mL) of tert-butyl 4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-cyclopropylphthalazin-1-yl)piperazine-1-carboxylate. The resulting mixture was sealed and stirred at room temperature for 30 minutes. The reaction mixture was diluted with DCM (10 mL) and made basic using saturated aqueous NaHCO3 (5 mL). The aqueous layer was extracted once more with DCM (10 mL). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure to obtain 6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-cyclopropyl-1-(piperazin-1-yl)phthalazine. 11H NMR (chloroform-d) δ: 8.30 - 8.36 (m, 1H), 8.18 - 8.24 (m, 1H), 7.55 - 7.64 (m, 4H), 7.40 - 7.46 (m, 2H), 7.33 (q, J = 7.1 Hz, 4H), 6.97 - 7.09 (m, 1H), 6.74 - 6.83 (m, 1H), 6.36 - 6.46 (m, 1H), 3.45 - 3.55 (m, 4H), 3.16 - 3.26 (m, 4H), 2.35 - 2.49 (m, 1H), 1.37 - 1.46 (m, 1H), 1.30 - 1.33 (m, 1H), 1.06 - 1.12 (m, 2H), 0.61 - 0.70 (m, 9H). m / z (ESI) M+H: 637.2.

[0322] Step 3: 1-(4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-cyclopropylphthalazin-1-yl)piperazin-1-yl)prop-2-en-1-one. To a 20 mL vial containing 6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-cyclopropyl-1-(piperazin-1-yl)phthalazine (0.023 g, 0.036 mmol), triethylamine (16 μl, 0.114 mmol) and dichloromethane (1.0 mL) were added. The resulting mixture was sealed and stirred at room temperature for 10 minutes, then acryloyl chloride (4.0 μl, 0.049 mmol) was added via syringe. The reaction mixture was sealed and stirring was continued at room temperature for 20 minutes. The reaction was quenched with saturated aqueous NaHCO3 (3 mL) and diluted with DCM (10 mL). The aqueous layer was extracted once more with DCM (5 mL). The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure to obtain 1-(4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-cyclopropylphthalazin-1-yl)piperazin-1-yl)prop-2-en-1-one. 11H NMR (chloroform-d) δ: 8.32 - 8.38 (m, 1H), 8.16 - 8.24 (m, 1H), 7.55 - 7.65 (m, 4H), 7.40 - 7.48 (m, 2H), 7.31 - 7.38 (m, 4H), 6.98 - 7.10 (m, 1H), 6.75 - 6.84 (m, 1H), 6.60 - 6.72 (m, 1H), 6.41 - 6.47 (m, 1H), 6.31 - 6.40 (m, 1H), 5.72 - 5.82 (m, 1H), 3.79 - 4.08 (m, 4H), 3.44 - 3.62 (m, 4H), 2.38 - 2.49 (m, 1H), 1.40 - 1.45 (m, 1H), 1.33 - 1.37 (m, 1H), 1.04 - 1.13 (m, 2H), 0.62 - 0.68 (m, 9H). m / z (ESI) M+H: 691.2.

[0323] Step 4: 1-(4-(7-Chloro-4-cyclopropyl-6-(2-fluorophenol)-1-phthalazinyl)-1-piperazinyl)-2-propen-1-one. To a 20 mL vial charged with 1-(4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-cyclopropylphthalazin-1-yl)piperazin-1-yl)prop-2-en-1-one (0.022 g, 0.032 mmol) was added tetrahydrofuran (2.0 mL), followed by tetrabutylammonium fluoride (1.0 M THF solution, 0.070 mL, 0.070 mmol). The vial was sealed and stirred at room temperature for 30 minutes. The reaction mixture was concentrated under reduced pressure. The crude product was purified by column chromatography (silica 24 g, DCM containing 0 - 5% MeOH) to obtain 1-(4-(7-Chloro-4-cyclopropyl-6-(2-fluorophenol)-1-phthalazinyl)-1-piperazinyl)-2-propen-1-one. 11H NMR (chloroform-d) δ: 8.30 - 8.37 (m, 1H), 8.11 - 8.18 (m, 1H), 7.29 - 7.38 (m, 1H), 6.96 - 7.18 (m, 1H), 6.88 - 6.94 (m, 1H), 6.76 - 6.85 (m, 1H), 6.59 - 6.72 (m, 1H), 6.31 - 6.42 (m, 1H), 5.73 - 5.84 (m, 1H), 3.73 - 4.05 (m, 4H), 3.35 - 3.62 (m, 4H), 2.40 - 2.52 (m, 1H), 1.35 - 1.42 (m, 1H), 1.29 - 1.34 (m, 1H), 1.03 - 1.14 (m, 2H). m / z (ESI) M+H: 453.2.

[0324] Example 13 1-(4-(4-Anilino-7-chloro-6-(2-fluoro-6-hydroxyphenyl)-1-phthalazinyl)-1-piperazinyl)-2-propen-1-one

Chemical formula

[0325] Step 2: 7-(2-((tert-Butyldiphenylsilyl)oxy)-6-fluorophenyl)-6-chloro-N-phenyl-4-(piperazin-1-yl)phthalazin-1-amine. Example 12. Similar to Step 2, reaction of 4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-(phenylamino)phthalazin-1-yl)piperazine-1-carboxylic acid tert-butyl gave 7-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-6-chloro-N-phenyl-4-(piperazin-1-yl)phthalazin-1-amine. 1 H NMR (chloroform-d) δ: 8.19 - 8.26 (m, 1H), 7.75 - 7.80 (m, 1H), 7.60 - 7.68 (m, 5H), 7.49 - 7.55 (m, 2H), 7.39 - 7.46 (m, 3H), 7.32 - 7.37 (m, 5H), 7.02 - 7.11 (m, 2H), 6.75 - 6.84 (m, 1H), 6.59 - 6.67 (m, 1H), 6.43 - 6.53 (m, 1H), 3.35 - 3.47 (m, 4H), 3.16 - 3.27 (m, 4H), 0.70 - 0.76 (m, 9H). m / z (ESI) M+H: 688.2.

[0326] Step 3: 1-(4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-(phenylamino)phthalazin-1-yl)piperazin-1-yl)prop-2-en-1-one. Example 12. Similar to Step 3, reaction of 7-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-6-chloro-N-phenyl-4-(piperazin-1-yl)phthalazin-1-amine gave 1-(4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-(phenylamino)phthalazin-1-yl)piperazin-1-yl)prop-2-en-1-one. 11H NMR (chloroform-d) δ: 8.16 - 8.24 (m, 1H), 7.77 - 7.84 (m, 1H), 7.62 - 7.67 (m, 4H), 7.52 - 7.55 (m, 1H), 7.41 - 7.46 (m, 3H), 7.32 - 7.38 (m, 6H), 7.02 - 7.11 (m, 2H), 6.77 - 6.84 (m, 1H), 6.65 - 6.71 (m, 1H), 6.46 - 6.51 (m, 1H), 6.30 - 6.39 (m, 2H), 5.73 - 5.81 (m, 1H), 3.86 - 4.05 (m, 4H), 3.37 - 3.53 (m, 4H), 0.69 - 0.75 (m, 9H). m / z (ESI) M+H: 742.3.

[0327] Step 4: 1-(4-(4-Anilino-7-chloro-6-(2-fluoro-6-hydroxyphenyl)-1-phthalazinyl)-1-piperazinyl)-2-propen-1-one. Example 12, similar to Step 4, 1-(4-(4-anilino-7-chloro-6-(2-fluoro-6-hydroxyphenyl)-1-phthalazinyl)-1-piperazinyl)-2-propen-1-one was produced by the reaction of 1-(4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-(phenylamino)phthalazin-1-yl)piperazin-1-yl)prop-2-en-1-one. 1 1H NMR (chloroform-d) δ: 7.96 - 8.09 (m, 2H), 7.46 - 7.57 (m, 2H), 7.37 - 7.44 (m, 1H), 7.29 - 7.33 (m, 1H), 7.20 - 7.26 (m, 1H), 6.96 - 7.07 (m, 1H), 6.81 - 6.87 (m, 1H), 6.70 - 6.77 (m, 1H), 6.54 - 6.67 (m, 1H), 6.29 - 6.41 (m, 1H), 5.68 - 5.82 (m, 1H), 3.74 - 3.96 (m, 4H), 3.12 - 3.43 (m, 4H). m / z (ESI) M+H: 504.2.

[0328] Example 14 1-(4-(7-Chloro-4-cyclopentyl-6-(2-fluoro-6-hydroxyphenyl)-1-phthalazinyl)-1-piperazinyl)-2-propen-1-one

Chem.

[0329] Step 2: 6-(2-((tert-Butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-cyclopentyl-1-(piperazin-1-yl)phthalazine. Example 12. Similar to Step 2, 6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-cyclopentyl-1-(piperazin-1-yl)phthalazine was produced by the reaction of 4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-cyclopentylphthalazin-1-yl)piperazine-1-carboxylic acid tert-butyl. 1 H NMR (chloroform-d) δ: 8.17 - 8.21 (m, 1H), 8.12 - 8.16 (m, 1H), 7.61 - 7.66 (m, 2H), 7.51 - 7.56 (m, 2H), 7.40 - 7.46 (m, 2H), 7.34 - 7.38 (m, 2H), 7.29 - 7.33 (m, 2H), 6.99 - 7.08 (m, 1H), 6.74 - 6.82 (m, 1H), 6.37 - 6.45 (m, 1H), 3.58 - 3.67 (m, 4H), 3.27 - 3.36 (m, 4H), 2.18 - 2.22 (m, 1H), 2.08 - 2.12 (m, 2H), 1.86 - 1.91 (m, 3H), 1.69 - 1.77 (m, 3H), 0.59 - 0.67 (m, 9H). m / z (ESI) M+H: 665.2.

[0330] Step 3: 1-(4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-cyclopentylphthalazin-1-yl)piperazin-1-yl)prop-2-en-1-one. Example 12. Similar to Step 3, 1-(4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-cyclopentylphthalazin-1-yl)piperazin-1-yl)prop-2-en-1-one was produced by the reaction of 6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-cyclopentyl-1-(piperazin-1-yl)phthalazine. 11H NMR (chloroform-d) δ: 8.18 - 8.25 (m, 1H), 8.13 - 8.17 (m, 1H), 7.61 - 7.67 (m, 2H), 7.50 - 7.57 (m, 2H), 7.39 - 7.48 (m, 2H), 7.28 - 7.37 (m, 4H), 6.99 - 7.10 (m, 1H), 6.75 - 6.83 (m, 1H), 6.62 - 6.71 (m, 1H), 6.33 - 6.43 (m, 2H), 5.73 - 5.81 (m, 1H), 3.84 - 4.07 (m, 4H), 3.71 - 3.82 (m, 1H), 3.49 - 3.65 (m, 4H), 1.80 - 1.96 (m, 4H), 1.67 - 1.77 (m, 4H), 0.62 - 0.67 (m, 9H). m / z (ESI) M+H: 719.2.

[0331] Step 4: 1-(4-(7-chloro-4-cyclopentyl-6-(2-fluoro-6-hydroxyphenyl)-1-phthalazinyl)-1-piperazinyl)-2-propen-1-one. Example 12, similar to Step 4, 1-(4-(7-chloro-4-cyclopentyl-6-(2-fluoro-6-hydroxyphenyl)-1-phthalazinyl)-1-piperazinyl)-2-propen-1-one was produced by the reaction of 1-(4-(6-(2-((tert-butyldiphenylsilyl)oxy)-6-fluorophenyl)-7-chloro-4-cyclopentylphthalazin-1-yl)piperazin-1-yl)prop-2-en-1-one. 1 1H NMR (chloroform-d) δ: 8.10 - 8.22 (m, 2H), 7.29 - 7.38 (m, 1H), 6.86 - 6.93 (m, 1H), 6.77 - 6.85 (m, 1H), 6.61 - 6.72 (m, 1H), 6.33 - 6.44 (m, 1H), 5.74 - 5.85 (m, 1H), 3.82 - 4.05 (m, 4H), 3.75 - 3.82 (m, 1H), 3.40 - 3.63 (m, 4H), 2.06 - 2.24 (m, 4H), 1.81 - 1.96 (m, 2H), 1.67 - 1.79 (m, 2H). m / z (ESI) M+H: 481.2.

[0332] Example 15 1-(4-(7-chloro-6-(2-fluoro-6-hydroxyphenyl)-4-(piperidin-1-yl)-1-phthalazinyl)-1-piperazinyl)-2-propen-1-one

Chem.

[0333] Step 2: 2-(7-chloro-1-(piperazin-1-yl)-4-(piperidin-1-yl)phthalazin-6-yl)-3-fluorophenol. Example 12. Similar to Step 2, 2-(7-chloro-1-(piperazin-1-yl)-4-(piperidin-1-yl)phthalazin-6-yl)-3-fluorophenol was produced by the reaction of tert-butyl 4-(7-chloro-6-(2-fluoro-6-hydroxyphenyl)-4-(piperidin-1-yl)phthalazin-1-yl)piperazine-1-carboxylate. 1 H NMR (chloroform-d) δ: 8.09 - 8.13 (m, 1H), 7.95 - 8.03 (m, 1H), 7.28 - 7.38 (m, 1H), 6.83 - 6.89 (m, 1H), 6.75 - 6.82 (m, 1H), 3.39 - 3.48 (m, 4H), 3.31 - 3.38 (m, 4H), 3.12 - 3.21 (m, 4H), 1.75 - 1.80 (m, 4H), 1.64 - 1.69 (m, 2H). m / z (ESI) M+H: 442.2.

[0334] Step 3: 1-(4-(7-chloro-6-(2-fluoro-6-hydroxyphenyl)-4-(1-piperidinyl)-1-phthalazinyl)-1-piperazinyl)-2-propen-1-one. Example 12. Similar to Step 3, 1-(4-(7-chloro-6-(2-fluoro-6-hydroxyphenyl)-4-(1-piperidinyl)-1-phthalazinyl)-1-piperazinyl)-2-propen-1-one was produced by the reaction of 2-(7-chloro-1-(piperazin-1-yl)-4-(piperidin-1-yl)phthalazin-6-yl)-3-fluorophenol. 1 H NMR (chloroform-d) δ: 8.08 - 8.15 (m, 1H), 7.98 - 8.05 (m, 1H), 7.29 - 7.39 (m, 1H), 6.86 - 6.94 (m, 1H), 6.76 - 6.85 (m, 1H), 6.59 - 6.70 (m, 1H), 6.30 - 6.43 (m, 1H), 5.72 - 5.84 (m, 1H), 3.77 - 4.05 (m, 4H), 3.40 - 3.56 (m, 4H), 3.32 - 3.38 (m, 4H), 1.73 - 1.85 (m, 4H), 1.64 - 1.70 (m, 2H). m / z (ESI) M+H: 496.2.

[0335] Example 16 1-(4-(7-chloro-6-(2-fluoro-6-hydroxyphenyl)-4-phenoxy-1-phthalazinyl)-1-piperazinyl)-2-propen-1-one

Chemical formula

[0336] Step 2: 2-(7-chloro-4-phenoxy-1-(piperazin-1-yl)phthalazin-6-yl)-3-fluorophenol Example 12. Similar to Step 2, 2-(7-chloro-4-phenoxy-1-(piperazin-1-yl)phthalazin-6-yl)-3-fluorophenol was produced by the reaction of 4-(7-chloro-6-(2-fluoro-6-hydroxyphenyl)-4-phenoxyphtalazin-1-yl)piperazine-1-carboxylate. 1 1H NMR (chloroform-d) δ: 8.37 - 8.42 (m, 1H), 8.14 - 8.19 (m, 1H), 7.37 - 7.45 (m, 2H), 7.29 - 7.34 (m, 1H), 7.19 - 7.25 (m, 2H), 6.89 - 6.98 (m, 1H), 6.76 - 6.87 (m, 4H), 3.36 - 3.45 (m, 4H), 3.13 - 3.22 (m, 4H). m / z (ESI) M+H: 451.2.

[0337] Step 3: 1-(4-(7-chloro-6-(2-fluoro-6-hydroxyphenyl)-4-phenoxy-1-phthalazinyl)-1-piperazinyl)-2-propen-1-one. Example 12. Similar to Step 3, 1-(4-(7-chloro-6-(2-fluoro-6-hydroxyphenyl)-4-phenoxy-1-phthalazinyl)-1-piperazinyl)-2-propen-1-one was produced by the reaction of 2-(7-chloro-4-phenoxy-1-(piperazin-1-yl)phthalazin-6-yl)-3-fluorophenol. 1 1H NMR (chloroform-d) δ: 8.41 - 8.45 (m, 1H), 8.17 - 8.20 (m, 1H), 7.40 - 7.45 (m, 2H), 7.28 - 7.37 (m, 2H), 7.20 - 7.26 (m, 1H), 6.78 - 6.87 (m, 2H), 6.59 - 6.70 (m, 1H), 6.31 - 6.41 (m, 1H), 5.97 - 6.06 (m, 1H), 5.74 - 5.81 (m, 1H), 3.76 - 4.03 (m, 4H), 3.38 - 3.53 (m, 4H). m / z (ESI) M+H: 505.2.

[0338] Examples 17-1 and 17-2 (2E)-1-(4-(5-chloro-7-fluoro-6-(3-methoxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-4-(dimethylamino)-2-buten-1-one (Example 17-1) and (2E)-1-(4-(5-chloro-7-fluoro-6-(3-hydroxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-4-(dimethylamino)-2-buten-1-one (Example 17-2)

Chemical Structure

[0339] Step 2: 5-Chloro-6-(3-methoxynaphthalen-1-yl)-3-(piperazin-1-yl)benzo[c]isothiazole. To a solution of tert-butyl 4-(5-chloro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)piperazine-1-carboxylate (327 mg, 0.56 mmol) in DCM (6 mL) was added trifluoroacetic acid (1.04 mL, 13.9 mmol) via syringe. The resulting yellow solution was stirred at room temperature for 4 h and then concentrated. The residue was purified by silica gel chromatography (eluent: DCM containing 0 - 25% MeOH) to afford the mono-TFA salt of 5-chloro-6-(3-methoxynaphthalen-1-yl)-3-(piperazin-1-yl)benzo[c]isothiazole. m / z (ESI) M+H: 428.0.

[0340] Step 3: (2E)-1-(4-(5-chloro-7-fluoro-6-(3-methoxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-4-(dimethylamino)-2-buten-1-one. To a solution of 5-chloro-6-(3-methoxynaphthalen-1-yl)-3-(piperazin-1-yl)benzo[c]isothiazole (74 mg of mono-TFA salt, 0.14 mmol) and trans-4-dimethylaminocrotonic acid hydrochloride (38 mg, 0.23 mmol) in DMA (2 mL) was added thionyl chloride (41 μL, 0.69 mmol) via syringe. The resulting brown solution was stirred at room temperature for 2.5 h. The reaction of the reaction mixture was quenched with water (50 mL) and extracted with DCM / MeOH (8:1). The organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: DCM containing 0 - 15% MeOH) to afford (2E)-1-(4-(5-chloro-7-fluoro-6-(3-methoxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-4-(dimethylamino)-2-buten-1-one. 11H NMR (400 MHz, DMSO-d6) δ 8.10 (s, 1H), 7.94 (d, J = 8.4 Hz, 1H), 7.46 - 7.55 (m, 2H), 7.27 - 7.35 (m, 2H), 7.19 (d, J = 2.5 Hz, 1H), 6.61 - 6.72 (m, 2H), 3.94 (s, 3H), 3.80 - 3.93 (m, 4H), 3.62 - 3.68 (m, 4H), 3.07 (d, J = 4.3 Hz, 2H), 2.18 (s, 6H). m / z (ESI) M+H: 539.2.

[0341] Step 4: (2E)-1-(4-(5-chloro-7-fluoro-6-(3-hydroxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-4-(dimethylamino)-2-buten-1-one. At 0 °C, boron tribromide (1.0 M hexane solution, 218 μL, 0.22 mmol) was added dropwise via syringe to a solution of (2E)-1-(4-(5-chloro-7-fluoro-6-(3-methoxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-4-(dimethylamino)-2-buten-1-one (23.5 mg, 0.044 mmol) in 1,2-dichloroethane (4 mL). The resulting yellow slurry was stirred at 0 °C for 2.75 h, and then the reaction was quenched with saturated aqueous NaHCO3 (4 mL). The mixture was extracted twice with a 4:1 mixture of DCM / MeOH. The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: DCM containing 0 - 18% MeOH) to give (2E)-1-(4-(5-chloro-7-fluoro-6-(3-hydroxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-4-(dimethylamino)-2-buten-1-one. 11H NMR (400 MHz, DMSO-d6) δ 9.97 (s, 1H), 8.10 (s, 1H), 7.80 (d, J = 8.4 Hz, 1H), 7.40 - 7.46 (m, 1H), 7.19 - 7.30 (m, 3H), 7.07 (d, J = 2.4 Hz, 1H), 6.62 - 6.71 (m, 2H), 3.80 - 3.93 (m, 4H), 3.62 - 3.69 (m, 4H), 3.07 (d, J = 4.1 Hz, 2H), 2.17 (s, 6H). m / z (ESI) M+H: 525.0.

[0342] Examples 18-1 to 18-3 1-(4-(5-Chloro-7-fluoro-6-(3-methoxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-2-(hydroxymethyl)-2-propen-1-one. (Example 18-1) and 2-(bromomethyl)-1-(4-(5-chloro-7-fluoro-6-(3-hydroxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-2-propen-1-one (Example 18-2) and 1-(4-(5-chloro-7-fluoro-6-(3-hydroxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-2-(hydroxymethyl)-2-propen-1-one (Example 18-3)

Chemical Structure

[0343] Step 2: 2-(Bromomethyl)-1-(4-(5-chloro-7-fluoro-6-(3-hydroxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-2-propene-1-one and 1-(4-(5-chloro-7-fluoro-6-(3-hydroxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-2-(hydroxymethyl)-2-propene-1-one. At 0 °C, boron tribromide solution (1.0 M hexane solution, 167 μL, 0.17 mmol) was added dropwise via syringe to a solution of 1-(4-(5-chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-2-(hydroxymethyl)-2-propen-1-one (17.1 mg, 0.033 mmol) in 1,2-dichloroethane (4 mL). The resulting slurry was stirred at 0 °C for 40 minutes and then the reaction was quenched with saturated aqueous NaHCO3 (5 mL). The mixture was extracted twice with a 4:1 mixture of DCM / MeOH. The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: DCM containing 0 - 7% MeOH) to give two products.

[0344] First elution peak: 2-(Bromomethyl)-1-(4-(5-chloro-7-fluoro-6-(3-hydroxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)piperazin-1-yl)prop-2-en-1-one. 1 H NMR (400 MHz, DMSO-d6) δ 9.96 (br. s, 1H), 8.13 (s, 1H), 7.80 (d, J = 8.2 Hz, 1H), 7.40 - 7.47 (m, 1H), 7.19 - 7.29 (m, 3H), 7.07 (d, J = 2.4 Hz, 1H), 5.78 (s, 1H), 5.41 (s, 1H), 4.38 (s, 2H), 3.84 - 3.93 (m, 4H), 3.62 - 3.72 (m, 4H). m / z (ESI) M+H: 560.0

[0345] Second elution peak: 1-(4-(5-chloro-7-fluoro-6-(3-hydroxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)piperazin-1-yl)-2-(hydroxymethyl)prop-2-en-1-one. 11H NMR (400 MHz, DMSO-d6) δ 9.98 (br. s, 1H), 8.11 (s, 1H), 7.79 (d, J = 8.2 Hz, 1H), 7.37 - 7.48 (m, 1H), 7.17 - 7.28 (m, 3H), 7.07 (d, J = 2.4 Hz, 1H), 5.43 (br. s, 1H), 5.20 (br. s, 1H), 5.07 - 5.14 (m, 1H), 4.12 (br. s, 2H), 3.78 - 3.86 (m, 4H), 3.57 - 3.66 (m, 4H). m / z (ESI) M+H: 498.0

[0346] Examples 19-1 to 19-3 1-(4-(5-Chloro-7-fluoro-6-(5-methoxy-1-methyl-1H-indazol-7-yl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-2-propen-1-one (Example 19-1) and 1-(4-(5-chloro-7-fluoro-6-(5-hydroxy-1-methyl-1H-indazol-7-yl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-2-propen-1-one (Example 19-2) and 1-(4-(5-chloro-7-fluoro-6-(5-hydroxy-2-methyl-2H-indazol-7-yl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-2-propen-1-one (Example 19-3) [Chemical Structure] TIFF0007717224000244.tif105167 Step 1: 4-(5-Chloro-7-fluoro-6-(5-methoxy-1H-indazol-7-yl)benzo[c]isothiazol-3-yl)piperazine-1-carboxylic acid tert-butyl. Intermediate D (232 mg, 0.51 mmol), 5-methoxy-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole (535 mg, 1.95 mmol, see the following synthesis), and cesium carbonate (636 mg, 1.95 mmol) were dissolved in a mixture of 1,4-dioxane (8 mL) and water (2 mL), and the resulting slurry was degassed under an argon stream. Tetrakis(triphenylphosphine)palladium (59 mg, 0.05 mmol) was added, and the mixture was degassed again under an argon stream. The reaction mixture was sealed and heated at 100 °C for 18 h. The reaction was cooled to room temperature and partitioned between brine (40 mL) and EtOAc. The aqueous layer was extracted twice with EtOAc, and the combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: DCM 0 - 4.5% / MeOH) to give tert-butyl 4-(5-chloro-7-fluoro-6-(5-methoxy-1H-indazol-7-yl)benzo[c]isothiazol-3-yl)piperazine-1-carboxylate. LCMS-ESI (positive ion) m / z: 518.2 (M+H) + 。 1 H NMR (400 MHz, DMSO-d6) δ 12.90 (br. s, 1H), 8.06 (s, 1H), 8.03 (s, 1H), 7.31 (d, J = 1.4 Hz, 1H), 6.99 (d, J = 2.2 Hz, 1H), 3.83 (s, 3H), 3.61 - 3.69 (m, 4H), 3.54 - 3.60 (m, 4H), 1.45 (s, 9H).

[0347]

Chemical Structure

[0348] Step 2: tert-Butyl 4-(5-chloro-7-fluoro-6-(5-methoxy-1-methyl-1H-indazol-7-yl)benzo[c]isothiazol-3-yl)piperazine-1-carboxylate and tert-Butyl 4-(5-chloro-7-fluoro-6-(5-methoxy-2-methyl-2H-indazol-7-yl)benzo[c]isothiazol-3-yl)piperazine-1-carboxylate. A solution of tert-butyl 4-(5-chloro-7-fluoro-6-(5-methoxy-1H-indazol-7-yl)benzo[c]isothiazol-3-yl)piperazine-1-carboxylate (115 mg, 0.22 mmol) in THF (5 mL) was added with sodium hydride (60% mineral oil dispersion, 44.5 mg, 1.1 mmol). After 10 minutes, iodomethane (69 μL, 1.1 mmol) was added and the reaction mixture was stirred at room temperature for an additional 15 minutes, then partitioned between saturated aqueous ammonium chloride solution (10 mL) and DCM. The aqueous layer was extracted twice with DCM, and the combined organic layers were dried over anhydrous sodium sulfate and concentrated to give a mixture of tert-butyl 4-(5-chloro-7-fluoro-6-(5-methoxy-1-methyl-1H-indazol-7-yl)benzo[c]isothiazol-3-yl)piperazine-1-carboxylate and tert-butyl 4-(5-chloro-7-fluoro-6-(5-methoxy-2-methyl-2H-indazol-7-yl)benzo[c]isothiazol-3-yl)piperazine-1-carboxylate. The crude mixture was used in the subsequent step without purification. LCMS-ESI (positive ion) m / z: 532.0 (M+H) + 。

[0349] Step 3: 5-Chloro-7-fluoro-6-(5-methoxy-1-methyl-1H-indazol-7-yl)-3-(piperazin-1-yl)benzo[c]isothiazole (Intermediate J) and 5-chloro-7-fluoro-6-(5-methoxy-2-methyl-2H-indazol-7-yl)-3-(piperazin-1-yl)benzo[c]isothiazole (Intermediate K). A solution of a mixture of tert-butyl 4-(5-chloro-7-fluoro-6-(5-methoxy-1-methyl-1H-indazol-7-yl)benzo[c]isothiazol-3-yl)piperazine-1-carboxylate and tert-butyl 4-(5-chloro-7-fluoro-6-(5-methoxy-2-methyl-2H-indazol-7-yl)benzo[c]isothiazol-3-yl)piperazine-1-carboxylate (143 mg) in DCM (6 mL) was added trifluoroacetic acid (484 μL, 6.5 mmol) with a syringe. The resulting solution was stirred at room temperature for 25 minutes and then concentrated. The residue was purified by silica gel chromatography (eluent: DCM 0 - 25% / MeOH).

[0350] First elution peak: Mono-TFA salt of 5-chloro-7-fluoro-6-(5-methoxy-1-methyl-1H-indazol-7-yl)-3-(piperazin-1-yl)benzo[c]isothiazole (Intermediate J). LCMS-ESI (positive ion) m / z: 432.0 (M+H) + 。 1 H NMR (400 MHz, DMSO-d6) δ 8.16 (s, 1H), 8.03 (s, 1H), 7.35 (d, J = 2.4 Hz, 1H), 6.99 (d, J = 2.4 Hz, 1H), 3.84 (s, 3H), 3.67 - 3.76 (m, 4H), 3.56 (s, 3H), 3.36 - 3.42 (m, 4H).

[0351] Second elution peak: Mono-TFA salt of 5-chloro-7-fluoro-6-(5-methoxy-2-methyl-2H-indazol-7-yl)-3-(piperazin-1-yl)benzo[c]isothiazole (Intermediate K). LCMS-ESI (positive ion) m / z: 432.0 (M+H) + 。

[0352] Step 4: 1-(4-(5-Chloro-7-fluoro-6-(5-methoxy-1-methyl-1H-indazol-7-yl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-2-propen-1-one. To a slurry of the ice-cooled DCM solution (5 mL) of intermediate J (108 mg, 0.20 mmol) in its mono-TFA salt, DIPEA (104 μL, 0.60 mmol) was then added dropwise with a syringe followed by acryloyl chloride (24 μL, 0.30 mmol). The resulting solution was stirred at 0 °C for 3 hours, and then the reaction was quenched with saturated aqueous NaHCO3 (15 mL) and extracted twice with DCM. The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: DCM 0 - 7% / MeOH) to obtain 1-(4-(5-chloro-7-fluoro-6-(5-methoxy-1-methyl-1H-indazol-7-yl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-2-propen-1-one. LCMS-ESI (cation) m / z: 486.0 (M+H) + 。 1 H NMR (400 MHz, DMSO-d6) δ 8.13 (s, 1H), 8.02 (s, 1H), 7.33 (d, J = 2.2 Hz, 1H), 6.99 (d, J = 2.4 Hz, 1H), 6.85 (dd, J = 16.6, 10.6 Hz, 1H), 6.18 (dd, J = 16.7, 2.3 Hz, 1H), 5.76 (dd, J = 10.5, 2.3 Hz, 1H), 3.85 - 3.95 (m, 4H), 3.84 (s, 3H), 3.62 - 3.72 (m, 4H), 3.56 (s, 3H).

[0353] Step 5: 1-(4-(5-chloro-7-fluoro-6-(5-hydroxy-1-methyl-1H-indazol-7-yl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-2-propen-1-one. To a solution of 1-(4-(5-chloro-7-fluoro-6-(5-methoxy-1-methyl-1H-indazol-7-yl)benzo[c]isothiazol-3-yl)piperazin-1-yl)prop-2-en-1-one (72.5 mg, 0.15 mmol) in ice-cooled 1,2-dichloroethane (5 mL) was added dropwise boron tribromide solution (1.0 M hexane solution, 746 μL, 0.75 mmol) via syringe. The resulting slurry was stirred at 0 °C for 3.75 h, then the reaction was quenched with saturated aqueous NaHCO3 (5 mL), and extracted twice with a 4:1 mixture of DCM / MeOH. The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: DCM 0 - 6% / MeOH) to give 1-(4-(5-chloro-7-fluoro-6-(5-hydroxy-1-methyl-1H-indazol-7-yl)benzo[c]isothiazol-3-yl)piperazin-1-yl)prop-2-en-1-one. LCMS-ESI (cation) m / z: 472.0 (M+H) + 。 1 H NMR (400 MHz, DMSO-d6) δ 9.40 (s, 1H), 8.12 (s, 1H), 7.92 (s, 1H), 7.12 (d, J = 2.2 Hz, 1H), 6.81 - 6.91 (m, 2H), 6.18 (dd, J = 16.7, 2.5 Hz, 1H), 5.76 (dd, J = 10.4, 2.4 Hz, 1H), 3.81 - 3.94 (m, 4H), 3.62 - 3.70 (m, 4H), 3.52 (s, 3H).

[0354] For the synthesis of 1-(4-(5-chloro-7-fluoro-6-(5-hydroxy-2-methyl-2H-indazol-7-yl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-2-propen-1-one.

[0355] Steps 4 and 5 were carried out as described above using the intermediate K of Step 3, and 1-(4-(5-chloro-7-fluoro-6-(5-hydroxy-2-methyl-2H-indazol-7-yl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-2-propen-1-one was generated. LCMS-ESI (positive ion) m / z: 472.0 (M+H) + 。 1 H NMR (400 MHz, DMSO-d6) δ 9.28 (s, 1H), 8.11 (s, 1H), 8.01 (s, 1H), 6.95 (d, J = 2.0 Hz, 1H), 6.77 - 6.90 (m, 2H), 6.18 (dd, J = 16.7, 2.5 Hz, 1H), 5.76 (dd, J = 10.4, 2.2 Hz, 1H), 4.03 (s, 3H), 3.80 - 3.94 (m, 4H), 3.58 - 3.66 (m, 4H).

[0356] Example 20 1-(4-(5-chloro-7-fluoro-6-(3-hydroxynaphthalen-1-yl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-4-hydroxy-2-methylidene-1-butanone

Chemical Structure

[0357] Step 2: 1-(4-(5-chloro-7-fluoro-6-(3-hydroxy-1-naphthalenyl)-2,1-benzothiazol-3-yl)-1-piperazinyl)-4-hydroxy-2-methylidene-1-butanone. To a solution of 4-((tert-butyldiphenylsilyl)oxy)-1-(4-(5-chloro-7-fluoro-6-(3-methoxynaphthalen-1-yl)benzo[c]isothiazol-3-yl)piperazin-1-yl)-2-methylenebutan-1-one (85 mg, 0.11 mmol) and DCM (2 mL) was added dropwise a 2 M solution of BBr3 (0.28 mL, 0.56 mmol) in DCM at 0 °C. The reaction of the reaction mixture ...

Claims

1. 【Fig. 1】 A compound, or a pharmaceutically acceptable salt thereof.

2. A pharmaceutical composition comprising the compound according to claim 1 and a pharmaceutically acceptable excipient.

3. The pharmaceutical composition according to claim 2 for use in the treatment of cancer.

4. The pharmaceutical composition according to claim 3, wherein the cancer is non-small cell lung cancer, small intestine cancer, appendiceal cancer, colorectal cancer, endometrial cancer, pancreatic cancer, skin cancer, gastric cancer, nasal cancer, or cholangiocarcinoma.

5. The pharmaceutical composition according to claim 4, wherein the cancer is non-small cell lung cancer.

6. The pharmaceutical composition according to claim 4, wherein the cancer is colorectal cancer.

7. The pharmaceutical composition according to claim 4, wherein the cancer is pancreatic cancer.

8. The pharmaceutical composition according to any one of claims 3 to 7, wherein the cancer is mediated by a KRAS G12C mutation.

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