Cyclic Complex Compounds, Compositions Thereof, and Treatment Methods Using the Same

Abstract

In this specification, compounds having the following structure: 【Chemical 1】 TIFF2025524597000214.tif48165 (wherein the substituents are as defined in this specification), compositions comprising an effective amount of the compound, and methods of modulating the activity of KRAS G12D are provided.

Description

Technical Field

[0001] This specification provides a heterocyclic compound useful for treating cancer, a pharmaceutical composition containing the compound, and a method of using the compound for treating a cancer or a medical condition treatable or preventable by inhibition of KRAS G12D activity, the method comprising administering an effective amount of the compound to a subject in need thereof.

Background Art

[0002] Ras is a protein family that associates with the cell membrane via its C-terminal membrane targeting region and is well known as a molecular switch in the intracellular signaling network (Cox AD, Der CJ. Ras history: The saga continues. Small GTPases. 2010;1(1):2-27). Ras proteins bind to either GTP or GDP and switch between "on" and "off" states. When a Ras protein is bound to GDP, the Ras protein is in an off (or inactive) state. Also, when switched on by specific growth-promoting stimuli such as growth factors, the Ras protein is induced to exchange the GDP bound to it for GTP and changes to an on (or active) state (Malumbres M, Barbacid M. RAS oncogenes: The first 30 years. Nat Rev Cancer. 2003;3(6):459-465). By switching to the active state, the Ras protein can interact with various downstream proteins and activate the associated signaling pathways (Berndt N, Hamilton AD, Sebti SM. Targeting protein prenylation for cancer therapy. Nat Rev Cancer. 2011;11(11):775-791). The Ras superfamily contains various subfamilies including Ras, Ral, Rap, Rheb, Rad, Rit, and Miro (Wennerberg K, Rossman KL, Der CJ. The Ras superfamily at a glance. J Cell Sci. 2005;118(Pt 5):843-846). Since HRas, NRas, and KRas are the most common cancer genes in human cancers, these proteins are the most well-studied in the Ras family (O’Bryan JP. Pharmacological targeting of RAS: Recent success with direct inhibitors. Pharmacol Res. 2019;139:503-511).

[0003] KRas is one of the most frequently mutated genes in human cancers. Based on data from the Catalogue of Somatic Mutations (COSMIC) database, KRas mutations can be found in approximately 20% of human cancers, including pancreatic cancer, colorectal cancer, lung cancer, skin cancer, etc. (O’Bryan JP. Pharmacological targeting of RAS: Recent success with direct inhibitors. Pharmacol Res. 2019;139:503-511). The most common KRas mutations are found at positions G12 and G13, where the GTP hydrolysis activity of KRas, which is stimulated by the GTPase-activating protein (GAP), is blocked (Wang W, Fang G, Rudolph J. Ras inhibition via direct Ras binding--is there a path forward?. Bioorg Med Chem Lett. 2012;22(18):5766-5776). As a result, the KRas protein is overactivated, and ultimately, cell growth is uncontrolled, leading to cancer.

[0004] Among various cancers, pancreatic cancer is considered the most KRas-dependent cancer type. KRas mutations are observed in 94.1% of pancreatic ductal adenocarcinoma (PDAC). Two of the KRas G12D mutation (41%) and G12V mutation (34%) are the most dominant mutations in all KRas mutant PDAC (Waters AM, Der CJ. KRAS: The Critical Driver and Therapeutic Target for Pancreatic Cancer. Cold Spring Harb Perspect Med. 2018;8(9):a031435). In vivo data generated by mouse models have shown that the progression and maintenance of pancreatic cancer greatly rely on the constitutive activation of KRas downstream signaling (Siveke JT, Schmid RM. Chromosomal instability in mouse metastatic pancreatic cancer--it’s Kras and Tp53 after all. Cancer Cell. 2005;7(5):405-407). This indicates that the mutant KRas protein is a very attractive drug target for pancreatic cancer and other cancers with KRas mutations. Since the Kras protein is generally considered an intractable drug target, there are no therapeutic drugs clinically available that can selectively target the Kras protein with the G12D mutation.

[0005] Therefore, the KRas G12D mutation is a very attractive target for pancreatic cancer and other cancers with this mutation. Thus, small molecule therapeutic drugs that can selectively bind to Kras G12D and inhibit its function should be highly useful.

[0006] Any citation or identification of a reference in this section of the present application should not be construed as an admission that the reference is prior art to the present application.

Summary of the Invention

[0007] In this specification, a compound having the following formula (I): [Chem.] Also provided are its pharmaceutically acceptable salts, tautomers, stereoisomers, enantiomers, atropisomers, isotopic molecular species, and prodrugs, wherein the substituents are as defined herein. In one embodiment, the compound is not any one of the compounds of Table 1.

[0008] In one embodiment, the compound is selected from Table 2.

[0009] In one embodiment, provided herein is a method of inhibiting the activity of an intracellular KRAS mutant protein, comprising contacting the cell with an effective amount of a compound provided herein, or a pharmaceutically acceptable salt, tautomer, isotopic molecular species, stereoisomer, or prodrug thereof, optionally, wherein the KRAS mutant protein is a KRAS G12D mutant protein.

[0010] In one embodiment, provided herein is a method for treating or preventing cancer, comprising administering to a subject in need of treatment or prevention of cancer an effective amount of a compound provided herein, or a pharmaceutically acceptable salt, tautomer, isotopic molecular species, stereoisomer, or prodrug thereof, optionally, wherein the cancer is mediated by a KRAS mutation, preferably a KRAS G12D mutation.

BRIEF DESCRIPTION OF THE INVENTION

[0011] Definitions As used herein, the "KRAS gene" refers to a gene selected from the group consisting of: DIRAS1, DIRAS2, DIRAS3, ERAS, GEM, HRAS, KRAS, MRAS, NKIRAS1, NKIRAS2, NRAS, RALA, RALB, RAP1A, RAP1B, RAP2A, RAP2B, RAP2C, RASD1, RASD2, RASL10A, RASL10B, RASL11A, RASL11B, RASL12, REM1, REM2, RERG, RERGL, RRAD, RRAS, RRAS2, and variants thereof.

[0012] As used herein, the "KRAS protein" refers to a protein or an isoform thereof expressed by the KRAS gene (Scolnick EM, Papageoege AG, Shih TY (1979), "Guanine nucleotide-binding activity for src protein of rat-derived murine sarcoma viruses," Proc Natl Acad Sci USA. 76(5):5355 - 5559, Kranenburg O (November 2005) "The KRAS oncogene: past, present, and future," Biochimica et Biophysica Acta (BBA)-Reviews on Cancer, 1756(2):81 - 2).

[0013] As used herein, the "G12D mutation" refers to a mutation from glycine to aspartic acid at the 12th amino acid residue located in the G domain of the KRAS protein.

[0014] As used herein, "KRAS G12D" or "G12D" refers to a KRAS protein having the G12D mutation.

[0015] As used herein, and as used in the specification and the appended claims, the indefinite articles "a" and "an" and the definite article "the" include the plural referents as well as the singular referents unless the context clearly dictates otherwise.

[0016] As used herein, and unless otherwise specified, the terms "about" and "approximately" when used in connection with the dosage, amount, or weight percentage of the components of a composition or dosage form mean a dosage, amount, or weight percentage that is recognized by one of ordinary skill in the art to provide a pharmacologically equivalent effect to that obtained from the specified dosage, amount, or weight percentage. In certain embodiments, the terms "about" and "approximately" when used in this context contemplate a dosage, amount, or weight percentage that is within 30%, 20%, 15%, 10%, or 5% of the specified dosage, amount, or weight percentage.

[0017] As used herein, and unless otherwise specified, the terms "about" and "approximately" are used in connection with a numerical value or values provided to characterize a particular solid form, e.g., a particular temperature or temperature range (such as those describing a melting, dehydration, desolvation, or glass transition temperature), a mass change (such as a mass change as a function of temperature or humidity), a solvent or water content (e.g., from the perspective of mass or percentage), or a peak position (such as in an analysis by IR or Raman spectroscopy or XRPD), to indicate that the value or values of the range may deviate to the extent reasonably considered by one of ordinary skill in the art, yet still describe the solid form. Techniques for characterizing crystalline forms and amorphous solids include, but are not limited to, thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), X-ray powder diffraction (XRPD), single crystal X-ray diffraction, vibrational spectroscopy, e.g., infrared (IR) and Raman spectroscopy, solid and solution nuclear magnetic resonance (NMR) spectroscopy, optical microscopy, hot stage optical microscopy, scanning electron microscopy (SEM), electron crystallography and quantitative analysis, particle size analysis (PSA), surface area analysis, solubility studies, and dissolution studies. In certain embodiments, the terms "about" and "approximately", when used in this context, indicate that the numerical value or values of the range may vary within 30%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1.5%, 1%, 0.5%, or 0.25% of the recited value or values of the range. For example, in some embodiments, the value of an XRPD peak position may vary by up to ±0.2° 2θ (or ±0.2 degrees 2θ), yet still describe a particular XRPD peak.

[0018] An "alkyl" group is a saturated, partially saturated, or unsaturated straight-chain or branched-chain acyclic hydrocarbon having from 1 to 10 carbon atoms, typically from 1 to 8 carbons, and in some embodiments from 1 to 6, 1 to 4, or 2 to 6 carbon atoms. Representative alkyl groups include -methyl, -ethyl, -n-propyl, -n-butyl, -n-pentyl, and -n-hexyl, and saturated branched-chain alkyls include -isopropyl, -sec-butyl, -isobutyl, -tert-butyl, -isopentyl, -neopentyl, tert-pentyl, -2-methylpentyl, -3-methylpentyl, -4-methylpentyl, -2,3-dimethylbutyl, and the like. Examples of unsaturated alkyl groups include, among others, vinyl, allyl, -CH=CH(CH3), -CH=C(CH3)2, -C(CH3)=CH2, -C(CH3)=CH(CH3), -C(CH2CH3)=CH2, -C≡CH, -C≡C(CH3), -C≡C(CH2CH3), -CH2C≡CH, -CH2C≡C(CH3), and -CH2C≡C(CH7CH3), but are not limited thereto. The alkyl group may be substituted or unsubstituted. When the alkyl groups described herein are said to be "substituted", they are those found in the exemplary compounds and embodiments disclosed herein, as well as halogen (chloro, iodo, bromo, or fluoro), alkyl, hydroxyl, alkoxy, alkoxyalkyl, amino, alkylamino, carboxy, nitro, cyano, thiol, thioether, imine, imide, amidine, guanidine, enamine, aminocarbonyl, acylamino, phosphonato, phosphine, thiocarbonyl, sulfonyl, sulfone, sulfonamide, ketone, aldehyde, ester, urea, urethane, oxime, hydroxylamine, alkoxyamine, aralkoxyamine, N-oxide, hydrazine, hydrazide, hydrazone, azide, isocyanate, isothiocyanate, cyanate, thiocyanate, B(OH)2, or may be substituted with any substituent or plurality of substituents such as O(alkyl)aminocarbonyl.

[0019] The "cycloalkyl" group is a saturated, partially saturated, or unsaturated cyclic alkyl group having 3 to 10 carbon atoms, with a single cyclic ring or multiple fused or bridged rings, optionally substituted with 1 to 3 alkyl groups. In some embodiments, the cycloalkyl group has 3 to 8 ring members, while in other embodiments, the number of ring carbon atoms ranges from 3 to 5, 3 to 6, or 3 to 7. Cycloalkyl containing more than one ring may be fused, spiro, or bridged, or a combination thereof. Examples of such cycloalkyl groups include, for example, monocyclic structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, 1-methylcyclopropyl, 2-methylcyclopentyl, 2-methylcyclooctyl, etc., or polycyclic or bridged ring structures such as 1-bicyclo[1.1.1]pentyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.2]octyl, adamantyl, etc. Examples of unsaturated cycloalkyl groups include, among others, cyclohexenyl, cyclopentenyl, cyclohexadienyl, butadienyl, pentadienyl, hexadienyl. The cycloalkyl group may be substituted or unsubstituted. Examples of such substituted cycloalkyl groups include, for example, cyclohexanol, etc.

[0020] The "aryl" group is an aromatic carbocyclic group having 6 to 14 carbon atoms, with a single ring (e.g., phenyl) or multiple fused rings (e.g., naphthyl or anthryl). In some embodiments, the aryl group has 6 to 14 carbons in the ring portion of the group, while in other embodiments, it contains 6 to 12 or even 6 to 10 carbon atoms. Specific aryls include phenyl, biphenyl, naphthyl, etc. The aryl group may be substituted or unsubstituted. The term "aryl group" also includes groups containing fused rings, such as fused aromatic aliphatic ring systems (e.g., indanyl, tetrahydronaphthyl, etc.).

[0021] "Heterocyclyl" is an aromatic (also called heteroaryl) or non-aromatic cycloalkyl in which 1 to 4 of the ring carbon atoms are independently replaced by heteroatoms from the group consisting of O, S, and N. In some embodiments, the heterocyclyl group contains 3 to 10 ring members, while other such groups have 3 to 5, 3 to 6, or 3 to 8 ring members. Heterocyclyl can also be attached to other groups at any ring atom (i.e., any carbon atom or heteroatom of the heterocyclic ring). The heterocyclyl group may be substituted or unsubstituted. Examples of heterocyclyl groups include, but are not limited to, bicyclic rings, tricyclic rings, and tetracyclic rings, as well as bridged ring systems or spirocyclic ring systems, and multiple fused rings. Heterocyclyl groups include unsaturated, partially saturated, and saturated ring systems, such as imidazolyl, imidazolinyl, and imidazolidinyl (e.g., imidazolidin-4-one or imidazolidine-2,4-dionyl) groups. The term heterocyclyl includes, for example, fused ring species that include fused aromatic and non-aromatic groups such as 1- and 2-aminotetralin, benzotriazolyl (e.g., 1H-benzo[d][1,2,3]triazolyl), benzimidazolyl (e.g., 1H-benzo[d]imidazolyl), 2,3-dihydrobenzo[1,4]dioxinyl, and benzo[1,3]dioxolyl. The term also includes bridged polycyclic ring systems containing heteroatoms, such as, but not limited to, quinuclidinyl. Representative examples of heterocyclyl groups include aziridinyl, azetidinyl, azepanyl, oxetanyl, pyrrolidyl, imidazolidinyl (e.g., imidazolidin-4-one or imidazolidine-2,(4 - Dionyl), pyrazolidinyl, thiazolidinyl, tetrahydrothiophenyl, tetrahydrofuranyl, dioxolyl, furanyl, thiophenyl, pyrrolyl, pyrrolinyl, imidazolyl, imidazolinyl, pyrazolyl, pyrazolinyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, benzisoxazolyl (e.g., benzo[d]isoxazolyl), thiazolyl, thiazolinyl, isothiazolyl, thiadiazolyl, oxadiazolyl, piperidyl, piperazinyl (e.g., piperazin - 2 - onyl), morpholinyl, thiomorpholinyl, tetrahydropyranyl (e.g., tetrahydro - 2H - pyranyl), tetrahydrothiopyranyl, oxathianyl, dioxyl, dithianyl, pyranyl, pyridyl, pyrimidyl, pyridazinyl, pyrazinyl, triazinyl, dihydropyridyl, dihydrodithiinyl, dihydrodithionyl, 1,4 - dioxaspiro[4.5]decanyl, 2 - oxo - 1 - oxa - 3,8 - diazaspiro[4.5]decane, 1 - oxo - 2,8 - diazaspiro[4.5]decane, 3 - oxo - 2,8 - diazaspiro[4.5]decane, 3 - oxo - 1 - oxa - 4,9 - diazaspiro[5.5]undecane, 2 - oxo - 1 - oxa - 3,9 - diazaspiro[5.5]undecane, homopiperazinyl, quinuclidinyl, indolyl (e.g., indolyl - 2 - onyl or isoindolin - 1 - onyl), indolinyl, isoindolyl, isoindolinyl, azaindolyl (pyrrolopyridyl or 1H - pyrrolo[2,3 - b]pyridyl), indazolyl, indolizinyl, benzotriazolyl (e.g., 1H - benzo[d][1,2,3]triazolyl), benzimidazolyl (e.g., 1H - benzo[d]imidazolyl or 1H - benzo[d]imidazol - 2(3H) - onyl), benzofuranyl, benzothiophenyl, benzothiazolyl, benzoxadiazolyl, benzoxazinyl, benzodithiinyl, benzoxathiinyl, benzothiazinyl, benzoxazolyl (i.e., benzo[d]oxazolyl), benzothiazolyl, benzothiadiazolyl, benzo[1,3]dioxolyl, pyrazolopyridyl (e.g., 1H - pyrazolo[3,4-b]pyridyl, 1H-pyrazolo[4,3-b]pyridyl), imidazopyridyl (e.g., azabenzimidazolyl or 1H-imidazo[4,5-b]pyridyl), triazolopyridyl, isoxazolopyridyl, purinyl, xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl (e.g., 3,4-dihydroisoquinolin-1(2H)-onyl), quinolidinyl, quinoxalinyl, quinazolinyl, cinnolinyl, phthalazinyl, naphthyridinyl, pteridinyl, thianaphthalenyl, dihydrobenzothiazinyl, dihydrobenzofuranyl, dihydroindolyl, dihydrobenzodioxinyl, tetrahydroindolyl, tetrahydroindazolyl, tetrahydrobenzimidazolyl, tetrahydrobenzotriazolyl, tetrahydropyrrolopyridyl, tetrahydropyrazolopyridyl, tetrahydroimidazopyridyl, tetrahydrotriazolopyridyl, tetrahydropyrimidin-2(1H)-one, and tetrahydroquinolinyl groups, but are not limited thereto. Representative non-aromatic heterocyclyl groups do not include fused ring species containing a fused aromatic group. Examples of non-aromatic heterocyclyl groups include aziridinyl, azetidinyl, azepanyl, pyrrolidinyl, imidazolidinyl (e.g., imidazolidin-4-onyl or imidazolidin-2,4-dionyl), pyrazolidinyl, thiazolidinyl, tetrahydrothiophenyl, tetrahydrofuranyl, piperidinyl, piperazinyl (e.g., piperazin-2-onyl), morpholinyl, thiomorpholinyl, tetrahydropyranyl (e.g., tetrahydro-2H-pyranyl), tetrahydrothiopyranyl, oxathianyl, dithianyl, 1,4-dioxaspiro[4.5]decanyl, homopiperazinyl, quinuclidyl, or tetrahydropyrimidin-2(1H)-one. Representative substituted heterocyclyl groups may be mono-substituted or multi-substituted, for example, with various substituents such as those described below, and may be 2-, 3-, 4-, 5-, or 6-substituted, or may be a di-substituted pyridyl or morpholinyl group, but are not limited thereto.,

[0022] A "heteroaryl" group is an aryl ring system having a heteroaromatic ring system with 1 to 4 heteroatoms as ring atoms, and the remaining atoms of the ring system are carbon atoms. In some embodiments, the heteroaryl group contains 3 to 6 ring atoms in the ring portion of the group, and in other embodiments, 6 to 9, and even 6 to 10 atoms. Suitable heteroatoms include oxygen, sulfur, and nitrogen. In certain embodiments, the heteroaryl ring system is monocyclic or bicyclic. Non-limiting examples include pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, benzisoxazolyl (e.g., benzo[d]isoxazolyl), thiazolyl, pyrrolyl, pyridazinyl, pyrimidyl, pyrazinyl, thiophenyl, benzothiophenyl, furanyl, benzofuranyl, indolyl (e.g., indol-2-onyl or isoindolin-1-onyl), azaindolyl (pyrrolopyridyl or 1H-pyrrolo[2,3-b]pyridyl), indazolyl, benzimidazolyl (e.g., 1H-benzo[d]imidazolyl), imidazopyridyl (e.g., azabenzimidazolyl or 1H-imidazo[4,5-b]pyridyl), pyrazolopyridyl, triazolopyridyl, benzotriazolyl (e.g., 1H-benzo[d][1,2,3]triazolyl), benzoxazolyl (e.g., benzo[d]oxazolyl), benzothiazolyl, benzothiadiazolyl, isoxazolopyridyl, thianaphthalenyl, purinyl, xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl (e.g., 3,4-dihydroisoquinolin-1(2H)-onyl), tetrahydroquinolinyl, quinoxalinyl, and quinazolinyl groups, among others, but not limited thereto.

[0023] As used herein, a "spirocyclic ring" refers to two or more rings in which adjacent rings are joined through a single atom. The individual rings within a spirocyclic ring may be the same or different. The individual rings within a spirocyclic ring may be substituted or unsubstituted, and may have substituents different from those of other individual rings within a set of spirocyclic rings.

[0024] The "cycloalkylalkyl" group is a radical of the formula: -alkyl-cycloalkyl, where alkyl and cycloalkyl are as defined above. A substituted cycloalkylalkyl group may be substituted at the alkyl, cycloalkyl, or both the alkyl and cycloalkyl moieties of the group. Representative cycloalkylalkyl groups include, but are not limited to, methylcyclopropyl, methylcyclobutyl, methylcyclopentyl, methylcyclohexyl, ethylcyclopropyl, ethylcyclobutyl, ethylcyclopentyl, ethylcyclohexyl, propylcyclopentyl, propylcyclohexyl, and the like.

[0025] The "aralkyl" group is a radical of the formula: -alkyl-aryl, where alkyl and aryl are as defined above. A substituted aralkyl group may be substituted at the alkyl, aryl, or both the alkyl and aryl moieties of the group. Representative aralkyl groups include, but are not limited to, benzyl and phenethyl groups and fused (cycloalkylaryl)alkyl groups such as 4-ethyl-indanyl.

[0026] The "heterocyclylalkyl" group is a radical of the formula: -alkyl-heterocyclyl, where alkyl and heterocyclyl are as defined above. A substituted heterocyclylalkyl group may be substituted at the alkyl, heterocyclyl, or both the alkyl and heterocyclyl moieties of the group. Representative heterocyclylalkyl groups include, but are not limited to, 4-ethyl-morpholinyl, 4-propylmorpholinyl, furan-2-ylmethyl, furan-3-ylmethyl, pyridin-3-ylmethyl, tetrahydrofuran-2-ylethyl, and indol-2-ylpropyl.

[0027] "Halogen" is fluorine, chlorine, bromine, or iodine.

[0028] The "hydroxyalkyl" group is the above alkyl group substituted with one or more hydroxy groups.

[0029] An "alkoxy" or "alkoxyl" group is -O-(alkyl), where alkyl is as defined above.

[0030] An "alkoxyalkyl" group is -(alkyl)-O-(alkyl), where alkyl is as defined above.

[0031] An "amino" group is a radical of the formula: -NH2.

[0032] An "alkylamino" group is a radical of the formula: -NH-alkyl or -N(alkyl)2, where each alkyl is independently as defined above.

[0033] A "carboxy" group is a radical of the formula: -C(O)OH.

[0034] An "aminocarbonyl" group is a radical of the formula: -C(O)N(R # )2, -C(O)NH(R # ), or -C(O)NH2, where each R # is independently a substituted or unsubstituted alkyl, cycloalkyl, aryl, aralkyl, heterocyclyl, or heterocyclic group as defined herein.

[0035] An "acylamino" group is a radical of the formula: -NHC(O)(R # ) or -N(alkyl)C(O)(R # ), where each alkyl and R # is independently as defined above.

[0036] A "sulfonylamino" group is a radical of the formula: -NHSO2(R # ) or -N(alkyl)SO2(R # ), where each alkyl and R # is as defined above.

[0037] The "urea" group is a radical of the formula: -N(alkyl)C(O)N(R # )2, -N(alkyl)C(O)NH(R # ), -N(alkyl)C(O)NH2, -NHC(O)N(R # )2, -NHC(O)NH(R # ), or -NH(CO)NHR # , where each alkyl and R # is independently as defined above.

[0038] With the exception of alkyl groups, when the groups described herein are said to be "substituted", they may be substituted with any suitable substituent or substituents. Exemplary examples of substituents are those found in the exemplary compounds and embodiments disclosed herein, and halogen (chloro, iodo, bromo, or fluoro), alkyl, hydroxyl, alkoxy, alkoxyalkyl, amino, alkylamino, carboxy, nitro, cyano, thiol, thioether, imine, imide, amidine, guanidine, enamine, aminocarbonyl, acylamino, phosphonate, phosphine, thiocarbonyl, sulfonyl, sulfone, sulfonamide, ketone, aldehyde, ester, urea, urethane, oxime, hydroxylamino, alkoxyamine, aralkoxyamine, N-oxide, hydrazine, hydrazide, hydrazone, azide, isocyanate, isothiocyanate, cyanate, thiocyanate, oxo (=O), B(OH)2, O(alkyl)aminocarbonyl, cycloalkyl which may be monocyclic or fused or non-fused polycyclic (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl), or heterocyclyl which may be monocyclic or fused or non-fused polycyclic (e.g., pyrrolidyl, piperidyl, piperazinyl, morpholinyl, or thiazinyl), monocyclic or fused or non-fused polycyclic aryl or heteroaryl (e.g., phenyl, naphthyl, pyrrolyl, indolyl, furanyl, thiophenyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, triazolyl, tetrazolyl, pyrazolyl, pyridyl, quinolinyl, isoquinolinyl, acridinyl, pyrazinyl, pyridazinyl, pyrimidinyl, benzimidazolyl, benzothiophenyl, or benzofuranyl), aryloxy, aralkyloxy, heterocyclyloxy, and heterocyclylalkoxy.

[0039] As used herein, the term "pharmaceutically acceptable salt(s)" refers to salts prepared from pharmaceutically acceptable non-toxic acids or bases, including inorganic acids and bases as well as organic acids and bases. Suitable pharmaceutically acceptable base addition salts of the compounds of formula (I) include, but are not limited to, those well known in the art. For example, see Remington’s Pharmaceutical Sciences, 18 th eds., Mack Publishing, Easton PA (1990) or Remington: The Science and Practice of Pharmacy, 19 th eds., Mack Publishing, Easton PA (1995).

[0040] As used herein and unless otherwise indicated, the term "stereoisomer" or "stereoisomerically pure" means one stereoisomer of a compound that is substantially free of other stereoisomers of the compound. For example, a stereoisomerically pure compound having one chiral center is substantially free of the opposite enantiomer of the compound. A stereoisomerically pure compound having two chiral centers is substantially free of other diastereomers of the compound. A typical stereoisomerically pure compound contains more than about 80% by weight of one stereoisomer of the compound and less than about 20% by weight of the other stereoisomers of the compound, more than about 90% by weight of one stereoisomer of the compound and less than about 10% by weight of the other stereoisomers of the compound, more than about 95% by weight of one stereoisomer of the compound and less than about 5% by weight of the other stereoisomers of the compound, or more than about 97% by weight of one stereoisomer of the compound and less than about 3% by weight of the other stereoisomers of the compound. Compounds can have chiral centers and can exist as racemic compounds, individual enantiomers or diastereomers, and mixtures thereof. All such isomeric forms, including mixtures thereof, are included in the embodiments disclosed herein.

[0041] The use of such compounds in stereoisomerically pure form, as well as the use of mixtures of those forms, are encompassed by the embodiments disclosed herein. For example, mixtures containing equal or unequal amounts of enantiomers of a particular compound can be used in the methods and compositions disclosed herein. These isomers may be synthesized asymmetrically or resolved using standard techniques such as chiral columns or chiral resolving agents. See, for example, Jacques, J., et al., Enantiomers, Racemates and Resolutions (Wiley-Interscience, New York, 1981), Wilen, S.H., et al., Tetrahedron 33:2725 (1977), Eliel, E.L., Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962), and Wilen, S.H., Tables of Resolving Agents and Optical Resolutions p.268 (E.L. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, IN, 1972).

[0042] It should also be noted that a compound can include E and Z isomers, or mixtures thereof, as well as cis and trans isomers, or mixtures thereof. In certain embodiments, the compound is isolated as either the E or Z isomer. In other embodiments, the compound is a mixture of the E and Z isomers.

[0043] As used herein and unless otherwise indicated, "atropisomer" refers to a stereoisomer resulting from restricted rotation about a single bond axis having a high enough rotational barrier to permit isolation of the individual rotational isomers.

[0044] "Tautomers" refer to the isomeric forms of a compound that are in equilibrium with each other. The concentration of the isomeric forms depends on the environment in which the compound is found and can vary, for example, depending on whether the compound is a solid or is in an organic or aqueous solution. For example, in an aqueous solution, pyrazole can exhibit the following isomeric forms, which are called tautomers of each other: [Chemical formula]

[0045] As will be readily understood by those skilled in the art, a wide variety of functional groups and other structures can exhibit tautomerism, and all tautomers of the compounds of formula (I) are within the scope of the present invention.

[0046] It should also be noted that a compound can contain an unnatural proportion of atomic isotopes in one or more of its atoms. For example, a compound can be radiolabeled with a radioactive isotope such as tritium ( 3 H), iodine-125 ( 125 I), sulfur-35 ( 35 S), or carbon-14 ( 14 C), or can contain deuterium ( 2 H), carbon-13 ( 13 C), or nitrogen-15 ( 15Isotopically enriched such as (N) may be used. As used herein, an "isotopic molecular species" is a compound that is isotopically enriched. The term "isotopically enriched" refers to an atom having an isotopic composition other than the natural isotopic composition of that atom. "Isotopically enriched" may also refer to a compound containing at least one atom having an isotopic composition other than the natural isotopic composition of that atom. The term "isotopic composition" refers to the amount of each isotope present in a given atom. Radiolabeled compounds and isotopically enriched compounds are useful as therapeutic agents, such as cancer and anti-inflammatory agents, research reagents, such as binding assay reagents, and diagnostic agents, such as in vivo imaging agents. All isotopic variations of the compounds described herein are intended to be included within the scope of the embodiments provided herein, whether radioactive or not. In some embodiments, isotopic molecular species of the compounds are provided, for example, the isotopic molecular species is a compound enriched in deuterium, carbon-13, or nitrogen-15.

[0047] As used herein, "treating" means alleviating, in whole or in part, a disorder, disease, or condition, or one or more symptoms associated with the disorder, disease, or condition, or delaying or halting further progression or worsening of those symptoms, or alleviating or eradicating the cause(s) of the disorder, disease, or condition itself. In some embodiments, "treating" means alleviating, in whole or in part, a disorder, disease, or condition, or delaying or halting further progression or worsening of those symptoms. In another embodiment, "treating" means alleviating, in whole or in part, a disorder, disease, or condition, or a symptom associated with the condition, where the condition is treatable or preventable by inhibition of KRAS G12D.

[0048] As used herein, "preventing" means a method of wholly or partially delaying and / or preventing the onset, recurrence, or spread of a disorder, disease, or medical condition, a method of preventing a subject from suffering from a disorder, disease, or medical condition, or a method of reducing the risk that a subject will suffer from a disorder, disease, or medical condition. In one embodiment, the medical condition is a condition treatable or preventable by inhibition of KRAS G12D.

[0049] The term "effective amount" in relation to a compound means an amount capable of treating or preventing a disorder, disease, or medical condition, or a symptom thereof, disclosed herein.

[0050] The term "subject" includes animals including animals such as cows, monkeys, horses, sheep, pigs, chickens, turkeys, quails, cats, dogs, mice, rats, rabbits, or guinea pigs, and in one embodiment includes mammals, and in another embodiment includes humans, but is not limited thereto.

[0051] Compound As used herein, a compound having the following formula (I):

Chemical formula

Chem.

Chem.

Chem.

Chem.

[0052] In one embodiment, the compound is not any one of the compounds in Table 1.

[0053]

Table 1-1

Table 1-2

[0054] In one embodiment, L 1 is -O-.

[0055] In one embodiment, R 5 is -CF3, and R 11 is

Chem.

[0056] In one embodiment, the heterocyclic alkyl contains at least one oxygen as a ring member in the heterocyclic moiety.

[0057] In one embodiment, R 8 is -L 8a -R 8a wherein L 8a is -(CH2) n - and n is an integer of 1, 2, or 3, R 8a is oxetanyl, tetrahydrofuryl, tetrahydro-2H-pyranyl, dihydro-2H-pyranyl, oxabicyclo[2.1.1]hexyl, oxabicyclo[2.2.1]heptyl, oxaspiro[3.3]heptyl, oxabicyclo[3.2.1]octyl, oxabicyclo[2.2.2]octyl, oxaspiro[3.5]nonyl, or oxaspiro[3.4]octyl, R 8a is optionally substituted.

[0058] In some embodiments, R 8a is optionally substituted with halogen, cyano, hydroxy, alkoxy, or alkyl optionally substituted with halogen, cyano, hydroxy, alkoxy, cycloalkyl or cycloalkyloxy.

[0059] In one embodiment, R 8 is

Chemical formula

[0060] In one embodiment, the heterocyclic alkyl contains at least one nitrogen as a ring member in the heterocyclic moiety.

[0061] In one embodiment, R 8 is -L 8a -R 8a wherein L8a is -(CH2) n and n is an integer of 1, 2, or 3, R 8a is azetidyl, pyridyl, isoxazolyl, oxazolyl, dihydro-2H-pyranyl, tetrahydro-2H-pyranyl, pyrrolidinonyl, azaspiro[3.3]heptyl, azabicyclo[2.1.1]hexyl, pyrrolidyl, 1H-pyrazolyl, R 8a is optionally substituted.

[0062] In some embodiments, R 8a is optionally substituted with halogen, cyano, hydroxy, alkoxy, or alkyl optionally substituted with halogen, cyano, hydroxy, or alkoxy.

[0063] In one embodiment, R 8 is

Chemical formula

[0064] In one embodiment, L 1 is a bond, R 5 is -CF3, R 11 is

Chemical formula

[0065] In one embodiment, R 8 is -L 8a -R 8a and L 8a is -(CH2) n and n is an integer of 0, 1, 2, or 3, and R 8a is cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or azetidyl, R 8a is optionally substituted. Here, R 5 is H, -Cl, -CF3, or unsubstituted C 1-4 alkyl, R 8 is unsubstituted or substituted alkyl, unsubstituted or substituted cycloalkyl, unsubstituted or substituted cycloalkylalkyl, unsubstituted or substituted aryl, unsubstituted or substituted aralkyl, unsubstituted or substituted heterocyclyl, unsubstituted or substituted heterocyclylalkyl, unsubstituted or substituted heteroaryl, unsubstituted or substituted heteroaralkyl, unsubstituted or substituted spirocyclic ring, or unsubstituted or substituted -alkyl-spirocyclic ring, R 11 is 3,8-diazabicyclo[3.2.1]oct-3-yl,

Chemical Structure

Chemical Structure

Chemical Structure

Chemical Structure

[0066] In one embodiment, L 1 is -O-.

[0067] In one embodiment, R 5 is -CF3, R 11 is

Chemical formula

[0068] In one embodiment, the heterocyclylalkyl contains at least one oxygen as a ring member in the heterocyclyl moiety.

[0069] In one embodiment, R 8 is -L 8a -R 8a and L 8a is -(CH2) n -, n is an integer of 1, 2, or 3, R 8a is oxetanyl, tetrahydrofuryl, tetrahydro-2H-pyranyl, dihydro-2H-pyranyl, oxabicyclo[2.1.1]hexyl, oxabicyclo[2,2,1]heptyl, oxaspiro[3.3]heptyl, oxabicyclo[3.2.1]octyl, oxabicyclo[2.2.2]octyl, oxaspiro[3.5]nonyl, or oxaspiro[3.4]octyl, R 8a is optionally substituted.

[0070] In one embodiment, R 8 is

Chemical formula

[0071] In one embodiment, the heterocyclylalkyl contains at least one nitrogen as a ring member in the heterocyclyl moiety.

[0072] In one embodiment, R 8 is -L 8a -R 8a and L 8a is -(CH2) n - where n is an integer of 1, 2, or 3, R 8a is azetidyl, pyridyl, isoxazolyl, oxazolyl, dihydro-2H-pyranyl, tetrahydro-2H-pyranyl, pyrrolidinonyl, azaspiro[3.3]heptyl, azabicyclo[2.1.1]hexyl, pyrrolidyl, 1H-pyrazolyl, R 8a is optionally substituted.

[0073] In one embodiment, R 8 is

Chemical formula

[0074] In one embodiment, L 1 is a bond, R 5 is -CF3, R 11 is

Chemical formula

[0075] In one embodiment, R 8 is -L 8a -R 8a and L 8a is -(CH2) n - where n is an integer of 0, 1, 2, or 3, R 8a ​​is cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or azetidyl, R 8a is optionally substituted.

[0076] In one embodiment, R 8 is

Chemical formula

[0077] In one embodiment, R 5 is -CF3, L 1 is -O-, R 11 is 3,8-diazabicyclo[3.2.1]oct-8-yl,

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0078] In one embodiment, the compound is

Chemical formula

[0079] In one embodiment, R 5 is -Cl, L 1 is -O-, R 8 is

Chem.

[0080] In one embodiment, the compound is

Chem.

[0081] In one embodiment, R 5 is unsubstituted C 1-4 alkyl, R 11 is

Chem.

Chem.

[0082] In one embodiment, the compound is

Chem.

[0083] In one embodiment, R 5 is -CF3, R 11 is 3 - oxa - 7,9 - diazabicyclo[3.3.1]non - 9 - yl,

Chem.

Chem.

[0084] In one embodiment, the compound is

Chem.

[0085] Aspect 2: In one embodiment, the compound is selected from Table 2.

[0086] Aspect 3: In one embodiment, provided herein is a pharmaceutical composition comprising an effective amount of a compound provided herein, or a pharmaceutically acceptable salt, tautomer, isotopic molecular species, stereoisomer, or prodrug thereof, and a pharmaceutically acceptable carrier, excipient, or vehicle.

[0087] Aspect 4: In one embodiment, provided herein is a method of inhibiting the activity of an intracellular KRAS mutant protein, the method comprising contacting the cell with an effective amount of a compound provided herein, or a pharmaceutically acceptable salt, tautomer, isotopic molecular species, stereoisomer, or prodrug thereof, wherein optionally, the KRAS mutant protein is a KRAS G12D mutant protein.

[0088] Aspect 5: In one embodiment, provided herein is a method for treating or preventing cancer, the method comprising administering to a subject in need of treating or preventing cancer an effective amount of a compound provided herein, or a pharmaceutically acceptable salt, tautomer, isotopic molecular species, stereoisomer, or prodrug thereof, optionally, wherein the cancer is mediated by a KRAS mutation, preferably a KRAS G12D mutation.

[0089] Aspect 6: Provided herein is a method for modulating the activity of KRAS G12D, the method comprising contacting the cell with an effective amount of a compound provided herein, or a pharmaceutically acceptable salt, tautomer, isotopic molecular species, stereoisomer, or prodrug thereof.

[0090] Aspect 7: Provided herein is a method for treating or preventing cancer, the method comprising administering to a subject in need of treating or preventing cancer an effective amount of a compound provided herein.

[0091] Aspect 8: Provided herein is a kit for treating cancer, the kit comprising (a) a pharmaceutical composition comprising a compound provided herein, and (b) instructions for administering an effective amount of a pharmaceutical composition comprising a KRAS G12D inhibitor provided herein for treating cancer in an individual.

[0092] This embodiment can be more fully understood by reference to the forms and examples for carrying out the invention, which are intended as illustrations of non-limiting embodiments.

[0093] Method for manufacturing the compound The compound can be produced using conventional organic synthesis methods and commercially available starting materials. By way of example, and not limitation, the compound of formula (I) can be prepared in the examples described herein, and also as outlined in Schemes 1-3 shown below. It should be noted that those skilled in the art should know how to modify the procedures shown in the exemplary schemes and examples in order to reach the desired product. General protecting groups can be used to prevent specific functional groups from undergoing unwanted reactions. Exemplary protecting groups are described in “Protective Groups in Organic Synthesis”, 4 th Edition, P.G.M. Wuts; T.W. Greene, John Wiley, 2007, and the references cited therein.

[0094] Scheme 1

Chemical formula

[0095] As shown in Scheme 1, in some embodiments, a method for preparing the compound defined as formula (I) is provided herein. The compound 1-1 substituted with LG (leaving group), where LG is Cl, F, methylsulfonyl, or methylsulfinyl, is converted to compound 1-2 under substitution conditions (e.g., NaH, THF) or Buchwald coupling conditions (e.g., RuPhos Pd catalyst, Cs2CO3, 1,4-dioxane), and then compound 1-2 is deprotected (e.g., TFA and DCM for deprotecting the Boc group when PG1 and PG2 are Boc) to obtain the compound defined as formula (I).

[0096] Scheme 2

Chemical formula

[0097] As shown in Scheme 2, in some embodiments, provided herein is a method for preparing a compound defined as formula (I). Compound 2-1 substituted with halogen (X1, X2, X3 are halogen) is converted to compound 2-2 under basic conditions (e.g., NaH, THF), and then compound 2-2 is converted to compound 2-3 under substitution conditions (KF, DMSO when LG is F, or MeSNa, MeOH followed by m-CPBA oxidation when LG is methylsulfonyl or methylsulfinyl), and compound 2-3 further undergoes a metal-catalyzed cross-coupling reaction such as Suzuki, Negishi, or Stille coupling (e.g., Pd(dtbpf)Cl2, K3PO4, 1,4-dioxane, water for Suzuki coupling) to obtain compound 2-4, where M can be boronic acid, boronic acid ester, metal (such as Zn), tributyltin, etc., and compound 2-4 is converted to compound 2-5 under substitution conditions (e.g., NaH, THF) or Buchwald coupling conditions (e.g., RuPhos Pd catalyst, Cs2CO3, 1,4-dioxane), and then compound 2-5 is deprotected (e.g., TFA and DCM for deprotecting the Boc group when PG1 and PG2 are Boc) to obtain the compound defined as formula (I).

[0098] Scheme 3

Chemical Structure

[0099] As shown in Scheme 3, in some embodiments, provided herein is a method for preparing a compound defined as formula (I). The compound 3-1 substituted with halogen (X1, X2, X3 are halogens) is converted to compound 3-2 under basic conditions (e.g., NaH, THF), and then compound 3-2 is converted to compound 3-3 under substitution conditions (when LG is F, KF, DMSO, or when LG is methylsulfonyl or methylsulfinyl, MeSNa, MeOH, followed by m-CPBA oxidation), compound 3-3 is converted to compound 3-4 under substitution conditions (e.g., NaH, THF) or Buchwald coupling conditions (e.g., RuPhos Pd catalyst, Cs2CO3, 1,4-dioxane), and compound 3-4 undergoes a metal-catalyzed cross-coupling reaction such as Suzuki, Negishi, or Stille coupling (e.g., Pd(dtbpf)Cl2, K3PO4, 1,4-dioxane, water for Suzuki coupling) to obtain compound 3-5, where M can be boronic acid, boronic acid ester, metal (such as Zn), tributyltin, etc., and then compound 3-5 is deprotected (e.g., TFA and DCM for deprotecting the Boc group when PG1 and PG2 are Boc) to obtain the compound defined as formula (I).

[0100] This embodiment can be more fully understood by referring to the forms and examples for carrying out the invention, which are intended to be illustrative of non-limiting embodiments.

Examples

[0101] The following examples are purely for illustrative purposes and should not be construed as limiting in any way. Unless otherwise specified, the experimental methods in the examples described below are conventional methods. Unless otherwise specified, all reagents and materials are commercially available. All solvents and chemicals used are of analytical grade or chemical purity. All solvents are redistilled before use. All anhydrous solvents are prepared according to standard methods or reference methods. Silica gel (100 - 200 mesh) for column chromatography and silica gel (GF254) for thin layer chromatography (TLC) are commercially available from Tsingdao Haiyang Chemical Co., Ltd. or Yantai Chemical Co., Ltd. in China and, unless otherwise specified, are all eluted with petroleum ether (60 - 90 °C) / ethyl acetate (v / v) and visualized with iodine or a solution of molybdophosphoric acid in ethanol. Unless otherwise specified, all extraction solvents are dried over anhydrous Na2SO4.

[0102] Unless otherwise instructed, the following reactions are carried out under a positive pressure of nitrogen or argon or in an anhydrous solvent using a drying tube, and the reaction flask is fitted with a rubber septum for introducing the substrate and reagents via a syringe, and the glassware is oven-dried and / or heat-dried.

[0103] Unless otherwise instructed, column chromatography purification is carried out using a silica gel column or a Biotage system (manufacturer: Dyax Corporation) with a silica SepPak cartridge (Waters), or using a Teledyne Isco Combiflash purification system with a prepacked silica gel cartridge.

[0104] 1 1H NMR spectra were recorded on a Varian instrument at 400 MHz or 500 MHz using TMS (tetramethylsilane) as an internal standard. 11H-NMR spectra were obtained using tetramethylsilane (0.00 ppm) or residual solvent (CDCl3: 7.25 ppm, CD3OD: 3.31 ppm, D2O: 4.79 ppm, d6-DMSO: 2.50 ppm, d6-acetone: 2.05, (CD3)2CO: 2.05) as reference standards with CDCl3, CD2Cl2, CD3OD, D2O, d6-DMSO, d6-acetone or (CD3)2CO as solvents. When the multiplicity of the peak is reported, the following abbreviations are used: s (singlet), d (doublet), t (triplet), q (quartet), qn (quintet), sx (sextet), m (multiplet), br (broad), dd (doublet of doublets), dt (doublet of triplets). The given coupling constant is reported in Hertz (Hz).

[0105] LC / MS data were recorded using an Agilent 1100, 1200 high performance liquid chromatography-ion trap mass spectrometer (LC-MSD Trap) equipped with a diode array detector (DAD) detecting at 214 nm and 254 nm and an ion trap (ESI source). All compound names except for the reagents were generated by ChemDraw® 19.1.

[0106] In the following examples, the following abbreviations are used. [Table 2-1] [Table 2-2] [Table 2-3]

[0107] Synthesis of Compounds

[0108] Example 1: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-2-((1-(difluoromethyl)-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-8-fluoro-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chemical formula

[0109] Step 1: 1-(Iodomethyl)-4-(((4-methoxybenzyl)oxy)methyl)-2-oxabicyclo[2.1.1]hexane

Chemical formula

[0110] At room temperature, NaH (1.20 g, 30.0 mmol) was added to a solution of (1-(iodomethyl)-2-oxabicyclo[2.1.1]hexan-4-yl)methanol (2.54 g, 10.0 mmol) in THF (20 mL). The resulting mixture was stirred at room temperature for 1 hour. Then, 1-(chloromethyl)-4-methoxybenzene was added to the reaction, and the mixture was stirred at room temperature for 1 hour. After completion, the reaction mixture was quenched with water (30 mL), extracted with EtOAc (100 mL), and washed with saturated aqueous NaCl solution (50 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to obtain a residue. The residue was purified by silica gel column chromatography, eluting with 0 - 30% ethyl acetate in petroleum ether to obtain the title product (1.87 g, 5.0 mmol).

[0111] Step 2: (4-(((4-methoxybenzyl)oxy)methyl)-2-oxabicyclo[2.1.1]hexan-1-yl)methanol

Chemical formula

[0112] At room temperature, KOAc (2.45 g, 25 mmol) was added to a solution of 1-(iodomethyl)-4-(((4-methoxybenzyl)oxy)methyl)-2-oxabicyclo[2.1.1]hexane (1.87 g, 5.0 mmol) in DMSO (30 mL). The resulting mixture was stirred at 110 °C for 16 hours. After completion, the reaction mixture was diluted with EtOAc (200 mL) and washed with saturated aqueous NaCl solution (50 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to obtain a residue. The residue was added to sodium ethoxide (680 mg, 10 mmol) and EtOH (10 mL). Then, the reaction mixture was stirred at room temperature for 1 hour. After completion, the reaction mixture was directly concentrated to obtain a residue. The residue was purified by silica gel column chromatography, eluting with 0-80% ethyl acetate in petroleum ether to obtain the title product (528.2 mg, 2.0 mmol).

[0113] Step 3: 4-(((4-Methoxybenzyl)oxy)methyl)-2-oxabicyclo[2.1.1]hexane-1-carbaldehyde

Chemical formula

[0114] At room temperature, DMP (848 mg, 2.0 mmol) was added to a solution of (4-(((4-methoxybenzyl)oxy)methyl)-2-oxabicyclo[2.1.1]hexan-1-yl)methanol (528.2 mg, 2.0 mmol) in DCM (10 mL). The resulting mixture was stirred at room temperature for 1 hour. After completion, the reaction mixture was diluted with EtOAc (100 mL) and washed with saturated aqueous NaCl solution (50 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to obtain the crude title product (1.0 g).

[0115] Step 4: 1-(Difluoromethyl)-4-(((4-methoxybenzyl)oxy)methyl)-2-oxabicyclo[2.1.1]hexane

Chemical formula

[0116] At room temperature, DAST (483.6 mg, 3.0 mmol) was added to a solution of crude 4-(((4-methoxybenzyl)oxy)methyl)-2-oxabicyclo[2.1.1]hexane-1-carbaldehyde (1.0 g) in DCM (10 mL). The resulting mixture was stirred at room temperature for 16 h. After completion, the reaction mixture was concentrated directly to give a residue. The residue was purified by silica gel column chromatography, eluting with 0-20% ethyl acetate in petroleum ether to give the title product (284.1 mg, 1.0 mmol).

[0117] Step 5: (1-(Difluoromethyl)-2-oxabicyclo[2.1.1]hexan-4-yl)methanol

Chem.

[0118] At room temperature, DDQ (454.0 mg, 2.0 mmol) was added to a solution of 1-(difluoromethyl)-4-(((4-methoxybenzyl)oxy)methyl)-2-oxabicyclo[2.1.1]hexane (284.1 mg, 1.0 mmol) and water (1 mL) in DCM (10 mL). The resulting mixture was stirred at room temperature for 2 h. After completion, the reaction mixture was diluted with water (30 mL), extracted with DCM (100 mL), and washed with saturated aqueous NaCl solution (20 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to give a residue. The residue was purified by silica gel column chromatography, eluting with 0-60% ethyl acetate in petroleum ether to give the title product (60.0 mg, 0.36 mmol).

[0119] Step 6: tert-Butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-2-((1-(difluoromethyl)-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-8-fluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

Chem.

[0120] At room temperature, sodium hydride (26.84 mg, 0.66 mmol) was added to a solution of (1-(difluoromethyl)-2-oxabicyclo[2.1.1]hexan-4-yl)methanol (60.0 mg, 0.36 mmol) in THF (5 mL). The resulting mixture was stirred at room temperature for 1 hour. Then, tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-2,8-difluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (155 mg, 0.28 mmol) was added to the reaction mixture and stirred at room temperature for 1 hour. After completion, the reaction mixture was diluted with EtOAc (50 mL) and washed with saturated NaCl aqueous solution (15 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to obtain a residue. The residue was purified by Prep-TLC (petroleum ether:EtOAc = 1:3) to obtain the title product (123.0 mg, 0.14 mmol). MS (ESI, m / e) [M+1] + 879.3.

[0121] Step 7: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-2-((1-(difluoromethyl)-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-8-fluoro-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

[0122] At room temperature, TFA (5 mL) was added to a solution of tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-2-((1-(difluoromethyl)-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-8-fluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (123 mg, 0.14 mmol) in DCM (5 mL). The resulting mixture was stirred at room temperature for 3 hours. After completion, the reaction mixture was concentrated to give a residue. The residue was purified by HPLC to give the title product (80 mg). It was then further purified by chiral HPLC to give isomer 1 (23.0 mg) and isomer 2 (25.0 mg). Isomer 1: 1 1H NMR (500 MHz, CD3OD) δ 8.12 (s, 1H), 7.22 - 7.20 (m, 1H), 7.02 - 6.99 (m, 1H), 6.12 - 5.90 (m, 1H), 4.77 - 4.74 (m, 2H), 4.56 - 4.45 (m, 2H), 3.87 (s, 2H), 3.71 - 3.63 (m, 4H), 2.06 (s, 2H), 1.83 - 1.75 (m, 6H); MS (ESI, m / e) [M+1] + 679.4. Isomer 2: 1 1H NMR (500 MHz, CD3OD) δ 8.12 (s, 1H), 7.22 - 7.20 (m, 1H), 7.02 - 6.99 (m, 1H), 6.12 - 5.90 (m, 1H), 4.77 - 4.74 (m, 2H), 4.53 - 4.45 (m, 2H), 3.87 (s, 2H), 3.71 - 3.63 (m, 4H), 2.06 (s, 2H), 1.83 - 1.75 (m, 6H). MS (ESI, m / e) [M+1] + 679.4.

[0123] Example 2: 4-(2-((2-Oxabicyclo[2.1.1]hexan-4-yl)methoxy)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-6-chloro-8-fluoroquinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile [Chemical formula]

[0124] Step 1: tert-Butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-6-chloro-2,8-difluoroquinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate [Chemical formula]

[0125] Under N2 atmosphere, a mixture of tert-butyl 3-(7-bromo-6-chloro-2,8-difluoroquinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (244 mg, 0.5 mmol), tert-butyl (3-cyano-7-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[b]thiophen-2-yl)carbamate, DPEphosPdCl2 (72 mg, 0.1 mmol) and Cs2CO3 (490 mg, 1.5 mmol) in toluene (17.5 mL) was stirred at 95 °C for 4 h. After completion, the reaction mixture was purified by combi-flash (petroleum ether / EtOAc = 3:1) to give the title product (35 mg, 10% yield). MS (ESI, m / e) [M+1] + 701.6.

[0126] Step 2: tert-Butyl 3-(2-((2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-6-chloro-8-fluoroquinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate [Chemical formula]

[0127] At room temperature, sodium hydride (8.0 mg, 0.334 mmol) was added to a solution of (2-oxabicyclo[2.1.1]hexan-4-yl)methanol (28 mg, 0.25 mmol) in THF (5 mL). The resulting mixture was stirred at room temperature for 1 hour. Then, tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-6-chloro-2,8-difluoroquinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (117 mg, 0.167 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature for 1 hour. After completion, the reaction mixture was diluted with EtOAc (50 mL) and washed with saturated aqueous NaCl solution (15 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to obtain a residue. The residue was purified by Prep-TLC (petroleum ether:EtOAc = 1:1) to give the title product (100 mg, 75.8%). MS (ESI, m / e) [M+1] + 795.6.

[0128] Step 3: 4-(2-((2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-6-chloro-8-fluoroquinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

[0129] At room temperature, TFA (5 mL) was added to a solution of tert-butyl 3-(2-((2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-6-chloro-8-fluoroquinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (100 mg, 0.126 mmol) in DCM (5 mL). The resulting mixture was stirred at room temperature for 3 hours. After completion, the reaction mixture was concentrated to obtain a residue. The residue was purified by prep-HPLC to obtain the title product (60 mg), which was separated by chiral HPLC (Prep-HPLC apparatus: Prep-HPLC-Gilson, column: i-Cellulose-5, 21.2 mm × 250 mm, 5 μm, mobile phase A: n-hexane, mobile phase B: EtOH (0.2% 2M NH3 in MeOH), flow rate: 20 mL / min, temperature: 25 °C, gradient: 40% B for 12 minutes, retention time - isomer 1: 7.5 minutes, retention time - isomer 2: 9.5 minutes) to obtain two isomers. Isomer 1 (20.47 mg, 27.3%): 1 H NMR (500 MHz, CD3OD) δ 7.86 (s, 1H), 7.21 - 7.18 (m, 1H), 7.04 - 7.01 (m, 1H), 4.76 - 4.71 (m, 2H), 4.56 - 4.42 (m, 3H), 3.73 (s, 2H), 3.66 - 3.59 (m, 4H), 1.96 - 1.95 (m, 2H), 1.89 - 1.78 (m, 4H), 1.64 - 1.61 (m, 2H); MS (ESI, m / e) [M+1] + 595.4. Isomer 2 (18.65 mg, 24.9%): 11H NMR (500 MHz, CD3OD) δ 7.86 (s, 1H), 7.21 - 7.18 (m, 1H), 7.04 - 7.01 (m, 1H), 4.76 - 4.71 (m, 2H), 4.56 - 4.41 (m, 3H), 3.73 (s, 2H), 3.66 - 3.58 (m, 4H), 1.96 - 1.95 (m, 2H), 1.89 - 1.78 (m, 4H), 1.64 - 1.61 (m, 2H); MS (ESI, m / e) [M+1] + 595.4.

[0130] Example 3: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((3-methoxytetrahydrofuran-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chem.

[0131] Step 1: tert-Butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-2-((3-methoxytetrahydrofuran-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

Chem.

[0132] A solution of (3-methoxytetrahydrofuran-3-yl)methanol (120 mg, 0.91 mmol) in THF (20 mL) was cooled to 0 °C. Then, NaH (36 mg, 0.91 mmol) was added in one portion and the mixture was stirred at 0 °C for 30 minutes. Next, tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-2,8-difluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (240 mg, 0.32 mmol) was added to the mixture. The resulting mixture was stirred at 0 °C for 1 hour, then quenched with H2O (10 mL), extracted three times with EtOAc (20 mL), washed with brine (20 mL), dried over Na2SO4, and filtered. The filtrate was concentrated and purified by column chromatography to give the title product (144 mg, 52.17%). MS (ESI, m / e) [M+H] + 847.5.

[0133] Step 2: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((3-methoxytetrahydrofuran-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

[0134] A solution of tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-2-((3-methoxytetrahydrofuran-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (140 mg, 0.16 mmol) in DCM (10 mL) was treated with TFA (2 mL). The mixture was stirred at room temperature for 2 - 3 h and the resulting mixture was concentrated to dryness. The residue was then diluted with DCM / MeOH = 10 / 1 (5 mL) and subsequently 5 drops of 7M NH3 in MeOH were added. The resulting solution was concentrated to dryness and purified by column chromatography to afford the title product (85 mg, 77.3%). MS (ESI, m / e) [M+H] + 647.5. The product was separated by chiral HPLC (first time: Prep-HPLC apparatus: Prep-HPLC-Gilson, column: Amylose-1, 21.2 mm×250 mm, 5 μm, mobile phase A: n-hexane, mobile phase B: EtOH (0.2% 2M NH3 in MeOH), flow rate: 20 mL / min, temperature: 25 °C, gradient: 60% B for 18 min, retention time - mixture 1: 5.5 min, retention time - mixture 2: 14.0 min, second time (simply to separate mixture 2): Prep-HPLC apparatus: Prep-HPLC-Gilson, column: Cellulose-C, 20 mm×250 mm, 5 μm, mobile phase A: n-hexane, mobile phase B: EtOH (0.2% 2M NH3 in MeOH), flow rate: 20 mL / min, temperature: 25 °C, gradient: 20% B for 40 min, retention time - isomer 3: 28 min, retention time - isomer 4: 36 min) to afford three isomers. Mixture 1 (a mixture of two stereoisomers): 11H NMR (500 MHz, CD3OD) δ 8.12 (s, 1H), 7.22 - 7.19 (m, 1H), 7.02 - 6.99 (m, 1H), 4.75 - 4.70 (m, 1H), 4.57 - 4.55 (m, 2H), 4.46 - 4.44 (m, 1H), 3.98 - 3.83 (m, 4H), 3.70 - 3.64 (m, 4H), 3.36 (s, 3H), 2.21 - 2.17 (m, 1H), 2.07 - 2.02 (m, 1H), 1.82 (s, 4H); MS (ESI, m / e) [M+H] + 647.5. Isomer 3: 1 1H NMR (500 MHz, CD3OD) δ 8.12 (s, 1H), 7.22 - 7.19 (m, 1H), 7.02 - 6.99 (m, 1H), 4.75 - 4.45 (m, 4H), 3.96 - 3.64 (m, 8H), 3.36 (s, 3H), 2.21 - 2.17 (m, 1H), 2.07 - 2.02 (m, 1H), 1.83 (s, 4H); MS (ESI, m / e) [M+H] + 647.5. Isomer 4: 1 1H NMR (500 MHz, CD3OD) δ 8.12 (s, 1H), 7.22 - 7.19 (m, 1H), 7.02 - 6.99 (m, 1H), 4.72 - 4.45 (m, 4H), 3.97 - 3.64 (m, 8H), 3.36 (s, 3H), 2.21 - 2.18 (m, 1H), 2.08 - 2.02 (m, 1H), 1.82 (s, 4H); MS (ESI, m / e) [M+H] + 647.5.

[0135] Example 4: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((4-methoxytetrahydro-2H-pyran-4-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chemical Structure

[0136] Example 4 was prepared by replacing (1-(difluoromethyl)-2-oxabicyclo[2.1.1]hexan-4-yl)methanol with (4-methoxytetrahydro-2H-pyran-4-yl)methanol according to the same procedure as described in Example 1 (Steps 6 and 7) to obtain the title compound. It was separated by chiral HPLC (Prep-HPLC apparatus: Prep-HPLC-Gilson, column: i-Cellulose-5, 21.2 mm × 250 mm, 5 μm, mobile phase A: n-hexane, mobile phase B: EtOH (0.2% 2M NH3 in MeOH), flow rate: 18 mL / min, temperature: 25 °C, gradient: 25% B for 15 minutes, retention time - isomer 1: 10.5 minutes, retention time - isomer 2: 12.5 minutes) to obtain isomer 1 (11.2 mg) and isomer 2 (19.2 mg). Isomer 1: 1 H NMR (500 MHz, CD3OD) δ 8.11 (s, 1H), 7.22 - 7.20 (m, 1H), 7.02 - 6.99 (m, 1H), 4.57 - 4.45 (m, 4H), 3.74 - 3.64 (m, 8H), 3.33 (s, 3H), 1.89 - 1.75 (m, 8H); MS(ESI, m / e) [M + H] + 661.5. Isomer 2: 1 H NMR(500 MHz, CD3OD) δ 8.11 (s, 1H), 7.22 - 7.20 (m, 1H), 7.02 - 7.01 (m, 1H), 4.50 - 4.45 (m, 4H), 3.74 - 3.64 (m, 8H), 3.33 (s, 3H), 1.89 - 1.75 (m, 8H); MS(ESI, m / e) [M + H] + 661.4.

[0137] Example 5: 4-(2-((2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-4-(3,8-diazabicyclo[3.2.1]octan-8-yl)-6-chloro-8-fluorobenzoxazol-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chemical formula

[0138] Example 5 was prepared by replacing tert-butyl 8-(7-bromo-2,8-difluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate with tert-butyl 8-(7-bromo-6-chloro-2,8-difluoroquinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate according to the same procedure as described in Example 9 (Step 1, Step 2 and Step 3) to obtain the title product. It was separated by chiral HPLC (Prep-HPLC apparatus: Prep-HPLC-Gilson, column: IE, 20 mm × 250 mm, 5 μm, mobile phase A: n-hexane, mobile phase B: EtOH (0.2% 2M NH3 in MeOH), flow rate: 18 mL / min, temperature: 25 °C, gradient: 60% B for 13 minutes, retention time - isomer 1: 7.5 minutes, retention time - isomer 2: 10 minutes) to obtain isomer 1 (1.03 mg) and isomer 2 (1.31 mg). Isomer 1: 1 H NMR (500 MHz, CD3OD) δ 7.89 (s, 1H), 7.22 - 7.19 (m, 1H), 7.05 - 7.01 (m, 1H), 4.94 - 4.72 (m, 2H), 4.75 - 4.71 (m, 2H), 4.57 - 4.56 (m, 1H), 3.73 (s, 2H), 3.26 - 3.23 (m, 2H), 2.93 - 2.90 (m, 2H), 2.08 - 2.05 (m, 4H), 1.95 - 1.94 (m, 2H), 1.61 - 1.60 (m, 2H); MS(ESI, m / e) [M+H] + 595.4. Isomer 2: 11H NMR (500 MHz, CD3OD) δ 7.89 (s, 1H), 7.22 - 7.19 (m, 1H), 7.05 - 7.01 (m, 1H), 4.94 - 4.72 (m, 2H), 4.75 - 4.71 (m, 2H), 4.57 - 4.56 (m, 1H), 3.73 (s, 2H), 3.25 - 3.23 (m, 2H), 2.92 - 2.90 (m, 2H), 2.07 - 2.05 (m, 4H), 1.95 - 1.94 (m, 2H), 1.61 - 1.60 (m, 2H); MS (ESI, m / e) [M+H]+ 595.3.

[0139] Example 6: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((3-hydroxyoxolan-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chemical formula

[0140] Step 1: tert-Butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-2-((3-hydroxyoxolan-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

Chemical formula

[0141] At room temperature, NaH (4 mg, 0.9 mmol) was added to a solution of tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-2,8-difluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (30 mg, 0.041 mmol) and 3-(hydroxymethyl)tetrahydrofuran-3-ol (10 mg, 0.082 mmol) in THF (10 mL). The resulting mixture was stirred at room temperature for 2 h. The reaction mixture was evaporated in vacuo. The residue was purified by pre-TLC (eluting with DCM / MeOH = 20 / 1) to give the title product (12 mg, 35.2% yield). MS (ESI, m / e) [M+1]+833.6.

[0142] Step 2: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((3-hydroxytetrahydrofuran-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

[0143] At room temperature, TFA (1 mL) was added to a solution of tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-2-((3-hydroxytetrahydrofuran-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (12 mg, 0.014 mmol) in DCM (2 mL). The reaction mixture was stirred at room temperature for 1 h. The solvent was concentrated and purified by pre-HPLC to give the title product (2 mg, 27% yield, formate salt). 11H NMR (500 MHz, CD3OD) δ 8.48 (s, 0.5H), 8.12 (s, 1H), 7.23 - 7.20 (m, 1H), 7.03 - 7.00 (m, 1H), 4.66 - 4.55 (m, 1H), 4.52 - 4.49 (m, 3H), 4.02 - 3.75 (m, 8H), 2.19 - 2.16 (m, 1H), 2.14 - 2.01 (m, 5H). MS (ESI, m / e) [M+1] + 633.5.

[0144] Example 7: 4-(2-((2-Oxabicyclo[2.2.1]heptan-4-yl)methoxy)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chemical Structure

[0145] Example 7 was prepared by replacing (3-methoxytetrahydrofuran-3-yl)methanol with (5-oxabicyclo[2.1.1]hexan-1-yl)methanol according to the same procedure as described in Example 3 to obtain the title product (2.09 mg). 1 1H NMR (500 MHz, CD3OD) δ 7.32 (s, 1H), 6.42 (s, 1H), 6.24 - 6.20 (m, 1H), 3.88 - 3.59 (m, 5H), 3.04 - 2.76 (m, 6H), 1.07 - 0.85 (m, 9H), 0.84 - 0.82 (m, 1H). MS (ESI, m / e) [M+H] + 643.4.

[0146] Example 8: Methyl 3-(((7-(2-amino-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-6-(trifluoromethyl)quinazolin-2-yl)oxy)methyl)-3-methylazetidine-1-carboxylate

Chemical Structure

[0147] Step 1: Methyl 3-(hydroxymethyl)-3-methylazetidine-1-carboxylate

Chemical Structure

[0148] At 0 °C, methyl chloroformate (124 mg, 1.31 mmol) was added to a solution of (3-methylazetidin-3-yl)methanol hydrochloride (200 mg, 1.46 mmol) and DIPEA (565 mg, 4.38 mmol) in DCM (5 mL). The reaction mixture was stirred at room temperature for 1 h. The solvent was added to H2O (20 mL). The organic layer was concentrated to give the crude compound (232 mg, 100% yield) as a colorless oil. MS (ESI, m / e) [M+1] + 160.1.

[0149] Example 8 was prepared by replacing 3-(hydroxymethyl)tetrahydrofuran-3-ol with methyl 3-(hydroxymethyl)-3-methylazetidine-1-carboxylate according to the same procedure as described in Example 6 to give the title product (4 mg, 12% yield, formate). 11H NMR (500 MHz, CD3OD) δ 8.48 (s, 0.5H), 8.12 (s, 1H), 7.23 - 7.20 (m, 1H), 7.03 - 7.00 (m, 1H), 4.65 - 4.60 (m, 1H), 4.56 - 4.53 (m, 1H), 4.50 - 4.48 (m, 2H), 4.10 - 3.80 (m, 4H), 3.73 - 3.44 (m, 7H), 2.02 - 1.96 (m, 4H), 1.42 (s, 3H). MS (ESI, m / e) [M+1] + 674.5.

[0150] Example 9: 4-(2-((2-Oxabicyclo[2.1.1]hexan-4-yl)methoxy)-4-(3,8-diazabicyclo[3.2.1]octan-8-yl)-8-fluoro-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chem.

[0151] Step 1: tert-Butyl 8-(7-bromo-2-chloro-8-fluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate

Chem.

[0152] At room temperature, tert-butyl 3,8-diazabicyclo[3.2.1]octane-3-carboxylate (5.86 g, 27.6 mmol) was added to a solution of 7-bromo-2,4-dichloro-8-fluoro-6-(trifluoromethyl)quinazoline (10.0 g, 27.6 mmol) and DIPEA (7.10 g, 55.2 mmol) in DCM (200 mL). The resulting mixture was stirred at room temperature for 2 hours. After completion, the reaction mixture was diluted with water (100 mL), extracted with DCM (500 mL), and washed with saturated NaCl (100 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to obtain a residue. The residue was purified by silica gel column chromatography, eluting with 0-40% ethyl acetate in petroleum ether to obtain the title product (10.0 g, 18.6 mmol).

[0153] Step 2: tert-Butyl 8-(7-bromo-2,8-difluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate

Chemical Structure

[0154] At room temperature, KF (5.8 g, 100 mmol) was added to a solution of tert-butyl 8-(7-bromo-2-chloro-8-fluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate (10.0 g, 18.6 mmol) in DMSO (300 mL). The resulting mixture was stirred at 100 °C for 16 hours. After completion, the reaction mixture was diluted with EtOAc (500 mL) and washed with saturated aqueous NaCl (100 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to obtain a residue. The residue was purified by silica gel column chromatography, eluting with 0-80% ethyl acetate in petroleum ether to obtain the title product (6.5 g, 12.4 mmol).

[0155] Step 3: 8-(2-((2-Oxabicyclo[2.1.1]hexan-4-yl)methoxy)-7-bromo-8-fluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylic acid tert-butyl

Chem.

[0156] At room temperature, sodium hydride (80.0 mg, 2.0 mmol) was added to a solution of (2-oxabicyclo[2.1.1]hexan-4-yl)methanol (114.1 mg, 1.0 mmol) in THF (5 mL). The resulting mixture was stirred at room temperature for 1 hour. Then, 8-(7-bromo-2,8-difluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylic acid tert-butyl (552 mg, 1.0 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature for 1 hour. After completion, the reaction mixture was quenched with water (10 mL), extracted with EtOAc (100 mL), and washed with saturated aqueous NaCl solution (15 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to obtain a residue. The residue was purified by silica gel column chromatography, eluting with 0-100% ethyl acetate in petroleum ether, to give the title product (492.8 mg, 0.8 mmol). MS (ESI, m / e) [M+1] + 617.1.

[0157] Step 4: 8-(2-((2-Oxabicyclo[2.1.1]hexan-4-yl)methoxy)-7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylic acid tert-butyl

Chem.

[0158] At room temperature, Cs2CO3 (325.8 mg, 1.0 mmol) was added to a solution of tert-butyl 8-(2-((2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-7-bromo-8-fluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate (308.5 mg, 0.5 mmol), tert-butyl (3-cyano-7-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[b]thiophen-2-yl)carbamate (418 mg, 1.0 mmol), and DPEPhosPdCl2 (71.6 mg, 0.1 mmol) in toluene (15 mL). The resulting mixture was stirred at 100 °C for 16 h. After completion, the reaction mixture was diluted with EtOAc (50 mL) and washed with saturated aqueous NaCl solution (25 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to obtain a residue. The residue was purified by Prep-TLC (pure EtOAc) to give the title product (162.6 mg, 0.20 mmol). MS (ESI, m / e) [M+1] + 829.4.

[0159] Step 5: 4-(2-((2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-4-(3,8-diazabicyclo[3.2.1]octan-8-yl)-8-fluoro-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

[0160] At room temperature, TFA (5 mL) was added to a solution of tert-butyl 8-(2-((2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate (162.6 mg, 0.20 mmol) in DCM (5 mL). The resulting mixture was stirred at room temperature for 3 hours. After completion, the reaction mixture was concentrated to obtain a residue. The residue was purified by HPLC to obtain the title product (80 mg), which was further purified by chiral HPLC (Prep-HPLC apparatus: Prep-HPLC-Gilson, column: IE, 20 mm × 250 mm, 5 μm, mobile phase A: n-hexane, mobile phase B: EtOH (0.2% 2M NH3 in MeOH), flow rate: 18 mL / min, temperature: 25 °C, gradient: 40% B for 16 minutes, retention time - isomer 1: 10 minutes, retention time - isomer 2: 14.5 minutes) to obtain isomer 1 (40.5 mg) and isomer 2 (19.0 mg). Isomer 1: 1 H NMR (500 MHz, CD3OD) δ 8.14 (s, 1H), 7.23 - 7.20 (m, 1H), 7.02 - 6.99 (m, 1H), 4.97 - 4.75 (m, 2H), 4.76 - 4.75 (m, 2H), 4.57 - 4.56 (m, 1H), 3.73 (s, 2H), 3.25 - 3.23 (m, 2H), 2.95 - 2.92 (m, 2H), 2.09 - 2.08 (m, 4H), 1.96 - 1.95 (m, 2H), 1.61 - 1.60 (m, 2H); MS (ESI, m / e) [M+1] + 629.4. Isomer 2: 11H NMR (500 MHz, CD3OD) δ 8.14 (s, 1H), 7.23 - 7.20 (m, 1H), 7.02 - 6.99 (m, 1H), 4.97 - 4.75 (m, 2H), 4.76 - 4.75 (m, 2H), 4.57 - 4.56 (m, 1H), 3.73 (s, 2H), 3.25 - 3.23 (m, 2H), 2.95 - 2.92 (m, 2H), 2.09 - 2.08 (m, 4H), 1.96 - 1.95 (m, 2H), 1.61 - 1.60 (m, 2H); MS (ESI, m / e) [M+1] + 629.3.

[0161] Example 10: 3 - ((((7 - (2 - Amino - 3 - cyano - 7 - fluorobenzo[b]thiophen - 4 - yl) - 4 - (3,8 - diazabicyclo[3.2.1]octan - 3 - yl) - 8 - fluoro - 6 - (trifluoromethyl)quinazolin - 2 - yl)oxy)methyl)tetrahydrofuran - 3 - carbonitrile

Chem.

[0162] Example 10 was prepared by replacing (1 - (difluoromethyl) - 2 - oxabicyclo[2.1.1]hexan - 4 - yl)methanol with 3 - (hydroxymethyl)tetrahydrofuran - 3 - carbonitrile according to the same procedure as described in Example 1 (Steps 6 and 7) to obtain the title compound. It was separated by chiral HPLC (Prep - HPLC apparatus: Prep - HPLC - Gilson, column: IE, 20 mm × 250 mm, 5 μm, mobile phase A: n - hexane, mobile phase B: EtOH (0.2% 2M NH3 in MeOH), flow rate: 18 mL / min, temperature: 25 °C, gradient: 40% B for 18 minutes, retention time - isomer 1: 9 minutes, retention time - isomer 2: 11 minutes, retention time - isomer 3: 13 minutes, retention time - isomer 4: 17 minutes) to obtain isomer 1 (30.5 mg), isomer 2 (27.2 mg), isomer 3 (26.5 mg) and isomer 4 (23.2 mg). Isomer 1: 11H NMR (500 MHz, CD3OD) δ 8.14 (s, 1H), 7.22 - 7.20 (m, 1H), 7.02 - 6.99 (m, 1H), 4.65 - 4.48 (m, 4H), 4.10 - 4.01 (m, 4H), 3.72 - 3.66 (m, 4H), 2.48 - 2.25 (m, 2H), 1.95 - 1.69 (m, 4H); MS (ESI, m / e) [M + H] + 642.3. Isomer 2: 1 1H NMR (500 MHz, CD3OD) δ 8.14 (s, 1H), 7.22 - 7.20 (m, 1H), 7.02 - 7.01 (m, 1H), 4.67 - 4.48 (m, 4H), 4.10 - 3.94 (m, 4H), 3.72 - 3.67 (m, 4H), 2.53 - 2.43 (m, 1H), 2.36 - 2.26 (m, 1H), 1.93 - 1.74 (m, 4H); MS (ESI, m / e) [M + H] + 642.3. Isomer 3: 1 1H NMR (500 MHz, CD3OD) δ 8.14 (s, 1H), 7.22 - 7.20 (m, 1H), 7.02 - 6.99 (m, 1H), 4.67 - 4.49 (m, 4H), 4.10 - 3.94 (m, 4H), 3.72 - 3.59 (m, 4H), 2.51 - 2.43 (m, 1H), 2.37 - 2.24 (m, 1H), 1.93 - 1.74 (m, 4H); MS (ESI, m / e) [M + H] + 642.3. Isomer 4: 1 1H NMR (500 MHz, CD3OD) δ 8.14 (s, 1H), 7.22 - 7.20 (m, 1H), 7.02 - 6.99 (m, 1H), 4.65 - 4.48 (m, 4H), 4.10 - 4.01 (m, 4H), 3.72 - 3.55 (m, 4H), 2.53 - 2.39 (m, 1H), 2.39 - 2.24 (m, 1H), 1.90 - 1.74 (m, 4H); MS (ESI, m / e) [M + H] + 642.3.

[0163] Example 11: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((1-(methoxymethyl)-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chemical formula

[0164] Step 1: (1-(Methoxymethyl)-2-oxabicyclo[2.1.1]hexan-4-yl)methanol

Chemical formula

[0165] At room temperature, sodium methoxide (5.40 g, 100.0 mmol) was added to a solution of (1-(iodomethyl)-2-oxabicyclo[2.1.1]hexan-4-yl)methanol (2.54 g, 10.0 mmol) in MeOH (20 mL). The resulting mixture was stirred at 70 °C for 16 h. After completion, the reaction mixture was diluted with water (30 mL), extracted with EtOAc (100 mL), and washed with saturated NaCl aqueous solution (50 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to obtain a crude product (500 mg).

[0166] Example 11 was prepared by replacing (1-(difluoromethyl)-2-oxabicyclo[2.1.1]hexan-4-yl)methanol with (1-(methoxymethyl)-2-oxabicyclo[2.1.1]hexan-4-yl)methanol according to the same procedure as described in Example 1 (Steps 6 and 7) to obtain the title product. It was separated by chiral HPLC (Prep-HPLC apparatus: Prep-HPLC-Gilson, column: IE, 20 mm×250 mm, 5 μm, mobile phase A: n-hexane, mobile phase B: EtOH (0.2% 2M NH3 in MeOH), flow rate: 18 mL / min, temperature: 25 °C, gradient: 40% B for 22 minutes, retention time - isomer 1: 9 minutes, retention time - isomer 2: 19 minutes) to obtain isomer 1 (1.79 mg) and isomer 2 (1.59 mg). Isomer 1: 1 1H NMR (500 MHz, CD3OD) δ 8.11 (s, 1H), 7.22 - 7.20 (m, 1H), 7.02 - 6.99 (m, 1H), 4.75 - 4.72 (m, 2H), 4.58 - 4.46 (m, 2H), 3.81 (s, 2H), 3.71 - 3.63 (m, 6H), 3.38 (s, 3H), 1.89 - 1.83 (m, 6H), 1.68 - 1.67 (m, 2H); MS (ESI, m / e) [M+H] + 673.4. Isomer 2: 1 1H NMR (500 MHz, CD3OD) δ 8.11 (s, 1H), 7.22 - 7.20 (m, 1H), 7.02 - 6.99 (m, 1H), 4.75 - 4.72 (m, 2H), 4.58 - 4.46 (m, 2H), 3.81 (s, 2H), 3.71 - 3.63 (m, 6H), 3.38 (s, 3H), 1.89 - 1.83 (m, 6H), 1.68 - 1.67 (m, 2H); MS (ESI, m / e) [M+H] + 673.4.

[0167] Example 12: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-2-((1-ethyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-8-fluoro-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile [Chemical formula]

[0168] Example 12 was prepared by replacing (1-(difluoromethyl)-2-oxabicyclo[2.1.1]hexan-4-yl)methanol with (1-ethyl-2-oxabicyclo[2.1.1]hexan-4-yl)methanol according to the same procedure as described in Example 1 (Steps 6 and 7), to obtain the title product (0.7 mg). 1 H NMR (500 MHz, CD3OD) δ 8.12 (s, 1H), 7.21 - 7.20 (m, 1H), 7.04 - 7.00 (m, 1H), 4.80 - 4.49 (m, 4H), 4.18 - 4.01 (m, 2H), 3.93 - 3.76 (m, 3H), 3.70 - 3.64 (m, 1H), 2.18 - 1.97 (m, 5H), 1.96 - 1.87 (m, 1H), 1.85 - 1.72 (m, 4H), 1.61 - 1.55 (m, 3H). MS (ESI, m / e) [M+H] + 657.5.

[0169] Example 13: 4-(2-((1-acetyl-3-methylazetidin-3-yl)methoxy)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile [Chemical formula]

[0170] Step 1: 1-(3-(hydroxymethyl)-3-methylazetidin-1-yl)ethan-1-one [Chemical formula]

[0171] To a 25 mL round-bottom flask were added (3-methylazetidin-3-yl)methanol hydrochloride (530 mg, 4.0 mmol), DCM (20 mL), pyridine (530 mg, 6.7 mmol) and acetyl chloride (220 mg, 2.8 mmol). The reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated under vacuum. The residue was purified by flash (MeOH:DCM = 15 - 20%) to give the title product (300 mg, 52.1% yield). MS (ESI, m / e) [M+H] + 144.3.

[0172] Step 2: tert-Butyl 3-(2-((1-acetyl-3-methylazetidin-3-yl)methoxy)-7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

Chemical Structure

[0173] To a 20 mL sealed tube were added 1-(3-(hydroxymethyl)-3-methylazetidin-1-yl)ethan-1-one (40 mg, 0.055 mmol), tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-2,8-difluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (40 mg, 0.278 mmol), THF (3 mL) and NaH (5 mg, 0.125 mmol, 60%). The mixture was stirred at room temperature for 16 hours. The reaction was quenched with saturated aqueous NH4Cl solution (10 mL) and extracted with EtOAc (20 mL x 3). The combined organic phases were dried over anhydrous Na2SO4 and concentrated under vacuum. The residue was purified by flash (MeOH:DCM = 5 - 10%) to give the title product (36 mg, 76.8% yield). MS (ESI, m / e) [M+H] +858.3.

[0174] Step 3: 4-(2-((1-Acetyl-3-methylazetidin-3-yl)methoxy)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

[0175] To a 25 mL round-bottom flask were added 3-(2-((1-acetyl-3-methylazetidin-3-yl)methoxy)-7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl (36 mg, 0.042 mmol) and TFA (4 mL). The reaction mixture was stirred at room temperature for 16 h. The reaction mixture was concentrated under vacuum. The residue was dissolved in EtOAc (20 mL), and saturated aqueous Na2CO3 solution (10 mL) was added. The solution was stirred at room temperature for 10 min and extracted with EtOAc (20 mL × 2). The combined organic phases were washed with brine (20 mL), dried over Na2SO4, and concentrated under vacuum. The residue was purified by Prep-HPLC (column: XBridge Shield RP18 OBD column, mobile phase A: water (10 mmol / L NH4HCO3 + 0.1% NH3·H2O), mobile phase B: ACN, gradient: 25% B - 65% B for 9 min) to give the title product (4.2 mg, 15.2% yield). 1 H NMR (500 MHz, CD3OD) δ 8.12 (s, 1H), 7.23 - 7.20 (m, 1H), 7.03 - 7.00 (m, 1H), 4.69 - 4.44 (m, 4H), 4.27 - 4.16 (m, 1H), 4.10 - 3.75 (m, 6H), 3.69 - 3.68 (m, 1H), 2.03 (s, 4H), 1.88 (s, 3H), 1.45 (s, 3H). MS (ESI, m / e) [M + H] + 658.3.

[0176] Example 14: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-2-((3-(difluoromethyl)tetrahydrofuran-3-yl)methoxy)-8-fluoro-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chemical formula

[0177] Example 14 was prepared by replacing (3-methoxytetrahydrofuran-3-yl)methanol with (3-(difluoromethyl)tetrahydrofuran-3-yl)methanol according to the same procedure as described in Example 3 to obtain the title racemic product (84.39 mg), which was separated by chiral HPLC (Prep-HPLC apparatus: Prep-HPLC-Gilson, column: IE, 20 mm × 250 mm, 5 μm, mobile phase A: n-hexane, mobile phase B: EtOH (0.2% 2M NH3 in MeOH), flow rate: 18 mL / min, temperature: 25 °C, gradient: 30% B for 22 minutes, retention time - isomer 1: 7.5 minutes, retention time - isomer 2: 10 minutes, retention time - isomer 3: 17 minutes, retention time - isomer 4: 18 minutes) to obtain four isomers. Isomer 1: 1 H NMR (500 MHz, CD3OD) δ 8.13 (s, 1H), 7.23 - 7.20 (m, 1H), 7.03 - 6.99 (m, 1H), 6.30 - 6.08 (m, 1H), 4.59 - 4.67 (m, 4H), 3.95 - 3.85 (m, 4H), 3.74 - 3.65 (m, 4H), 2.37 - 2.09 (m, 1H), 2.00 - 1.92 (m, 1H), 1.89 - 1.73 (m, 4H); MS (ESI, m / e) [M+H] + 667.5. Isomer 2: 11H NMR (500 MHz, CD3OD) δ 8.13 (s, 1H), 7.23 - 7.20 (m, 1H), 7.03 - 6.99 (m, 1H), 6.30 - 6.08 (m, 1H), 4.69 - 4.40 (m, 4H), 4.01 - 3.81 (m, 4H), 3.74 - 3.65 (m, 4H), 2.22 - 2.17 (m, 1H), 1.97 - 1.93 (m, 1H), 1.91 - 1.73 (m, 4H); MS (ESI, m / e) [M+H] + 667.4. Isomer 3: 1 1H NMR (500 MHz, CD3OD) δ 8.13 (s, 1H), 7.23 - 7.20 (m, 1H), 7.03 - 6.99 (m, 1H), 6.30 - 6.08 (m, 1H), 4.64 - 4.40 (m, 4H), 4.03 - 3.79 (m, 4H), 3.72 - 3.64 (m, 4H), 2.21 - 2.17 (m, 1H), 2.05 - 1.93 (m, 1H), 1.89 - 1.71 (m, 4H); MS (ESI, m / e) [M+H] + 667.4. Isomer 4: 1 1H NMR (500 MHz, CD3OD) δ 8.13 (s, 1H), 7.23 - 7.20 (m, 1H), 7.02 - 6.99 (m, 1H), 6.30 - 6.08 (m, 1H), 4.66 - 4.39 (m, 4H), 4.04 - 3.81 (m, 4H), 3.69 - 3.63 (m, 4H), 2.29 - 2.12 (m, 1H), 2.04 - 1.93 (m, 1H), 1.82 - 1.78 (m, 4H); MS (ESI, m / e) [M+H] + 667.4.

[0178] Example 15: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chemical Structure

[0179] Step 1: tert-Butyl 3-(7-bromo-2-chloro-8-fluoroquinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

Chemistry

[0180] At room temperature, tert-butyl 3,8-diazabicyclo[3.2.1]octane-8-carboxylate (4.24 g, 20 mmol) was added to a solution of 7-bromo-2,4-dichloro-8-fluoroquinazoline (6.0 g, 20 mmol) and DIEA (5.16 g, 40 mmol) in DCM (100 mL). The resulting mixture was stirred at room temperature for 2 hours. After completion, the reaction mixture was diluted with DCM (200 mL) and washed with saturated water (150 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to obtain a residue. The residue was purified by silica gel (petroleum ether:EtOAc = 3:1) to obtain the title product (6.5 g, 69.1%). MS (ESI, m / e) [M+1] + 471.4, 473.4.

[0181] Step 2: tert-Butyl 3-(7-bromo-2,8-difluoroquinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

Chemistry

[0182] A solution of tert-butyl (1R,5S)-3-(7-bromo-2-chloro-8-fluoroquinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (4.73 g, 10 mmol) and KF (5.8 g, 100 mmol) in DMSO (40 mL) was stirred at 100 °C for 15 h. After completion, the reaction mixture was cooled to 0 °C, decanted into ice water, and filtered to give the title product (2.5 g, 55.1%). 1 H NMR (500 MHz, DMSO) δ 7.89 - 7.87 (m, 1H), 7.65 - 7.62 (m, 1H), 4.40 - 4.38 (m, 2H), 4.26 - 4.23 (m, 2H), 3.62 - 3.60 (m, 2H), 1.81 - 1.77 (m, 2H), 1.65 - 1.63 (m, 2H), 1.46 (s, 9H). MS (ESI, m / e) [M+1] + 455.3, 457.3

[0183] Step 3: tert-Butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-2,8-difluoroquinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate [Chemical formula]

[0184] At room temperature, to a solution of tert-butyl 3-(7-bromo-2,8-difluoroquinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (400 mg, 0.87 mmol), tert-butyl (3-cyano-7-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[b]thiophen-2-yl)carbamate (365 mg, 0.87 mmol) and Pd(dtbpf)Cl2 (228 mg, 0.35 mmol) in 1,4-dioxane (15 mL) and H2O (3 mL) was added K3PO4 (556 mg, 2.62 mmol). The reaction mixture was then stirred at 90 °C for about 4 h. After completion, the reaction mixture was diluted with EtOAc (50 mL) and washed with saturated aqueous NaCl solution. The organic layer was dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated to give the crude product. The residue was purified by silica gel column chromatography, eluting with 0 - 40% DCM in ethyl acetate to give the title product (350 mg, 0.53 mmol). MS (ESI, m / e) [M+1] + 667.2.

[0185] Step 4: tert-Butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-2-((1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

Chem.

[0186] A solution of (1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methanol (19 mg, 0.15 mmol) in THF (2 mL) at 0 °C was added sodium hydride (6 mg, 0.15 mmol). The reaction mixture was stirred at 0 °C for 0.5 h. Then, tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-2,8-difluoroquinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (50 mg, 0.075 mmol) was added to the reaction mixture and the mixture was stirred at room temperature for 2 h. After completion, the reaction mixture was quenched with water (10 mL) and extracted with EtOAc (30 mL x 3) and washed with saturated aqueous NaCl solution. The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to give a residue. The residue was purified by silica gel column chromatography, eluting with 0 - 3% MeOH in DCM to give the title product (45 mg, 0.058 mmol). MS (ESI, m / e) [M+1] + 775.3.

[0187] Step 5: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

[0188] At room temperature, TFA (2 mL) was added to a solution of tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-2-((1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (45 mg, 0.058 mmol) in DCM (2 mL). The resulting mixture was stirred at room temperature for 3 hours. After completion, the reaction mixture was concentrated to obtain a residue, which was dissolved in 3 mL of DCM and the pH was adjusted to approximately 10 using 7M NH3 solution in MeOH. The residue was purified by pre-HPLC to obtain the title product (12 mg, 0.021 mmol, formate). 1 H NMR (500 MHz, CD3OD) δ 8.53 (s, 0.5H), 7.81 - 7.80 (m, 1H), 7.26 - 7.23 (m, 2H), 7.04 - 7.00 (m, 1H), 4.73 - 4.48 (m, 4H), 3.92 (s, 2H), 3.80 (s, 2H), 3.72 - 3.63 (m, 2H), 2.11 - 1.91 (m, 4H), 1.81 (s, 2H), 1.64 - 1.62 (m, 2H), 1.43 (s, 3H). δ MS (ESI, m / e) [M+1] + 575.3.

[0189] Example 16: 4-(4-(3-oxa-7,9-diazabicyclo[3.3.1]nonan-9-yl)-8-fluoro-2-((1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chemical Structure

[0190] Step 1: tert-Butyl 9-(7-bromo-2-chloro-8-fluoro-6-(trifluoromethyl)quinazolin-4-yl)-3-oxa-7,9-diazabicyclo[3.3.1]nonane-7-carboxylate [Chemical formula]

[0191] A mixture of 7-bromo-2,4-dichloro-8-fluoro-6-(trifluoromethyl)quinazoline (3.0 g, 8.2 mmol) and DIPEA (1.71 g, 13.14 mmol) in DCM (50 mL) was cooled to 0 °C. Then, tert-butyl 3-oxa-7,9-diazabicyclo[3.3.1]nonane-7-carboxylate was added thereto. The mixture was stirred at 0 °C for 2 hours, diluted with DCM (100 mL), washed with brine (20 mL), the operation was repeated 3 times, and filtered. The filtrate was concentrated and purified by chromatography to obtain the title product (3.79 g, 83.1%). MS (ESI, m / e) [M+1] + 556.4.

[0192] Step 2: tert-Butyl 9-(7-bromo-2,8-difluoro-6-(trifluoromethyl)quinazolin-4-yl)-3-oxa-7,9-diazabicyclo[3.3.1]nonane-7-carboxylate [Chemical formula]

[0193] To a solution of tert-butyl 9-(7-bromo-2-chloro-8-fluoro-6-(trifluoromethyl)quinazolin-4-yl)-3-oxa-7,9-diazabicyclo[3.3.1]nonane-7-carboxylate (3.6 g, 6.5 mmol) in DMSO (50 mL) was added KF (2.28 g, 39.1 mmol). The mixture was stirred at 90 °C for 2 - 3 hours. The resulting mixture was cooled to room temperature, diluted with EtOAc (50 mL), washed with brine (20 mL), and the operation was repeated 3 times. The organic phase was filtered, the filtrate was concentrated, and purified by chromatography to obtain the title product (2.91 g, 83.1%). MS (ESI, m / e) [M+1] + 539.7.

[0194] Step 3: tert-Butyl 9-(7-bromo-8-fluoro-2-((1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-6-(trifluoromethyl)quinazolin-4-yl)-3-oxa-7,9-diazabicyclo[3.3.1]nonane-7-carboxylate

Chemical Structure

[0195] A solution of (1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methanol (95 mg, 0.74 mmol) in THF (30 mL) was cooled to 0 °C. To this solution was added NaH (29 mg, 0.74 mmol) in one portion, and the mixture was stirred at room temperature for 30 minutes. Then, tert-butyl 9-(7-bromo-2,8-difluoro-6-(trifluoromethyl)quinazolin-4-yl)-3-oxa-7,9-diazabicyclo[3.3.1]nonane-7-carboxylate (398 mg, 0.71 mmol) was added to the mixture. The resulting mixture was stirred at room temperature for 4 - 5 hours, then quenched with H2O (20 mL), extracted 3 times with EtOAc (30 mL), washed 3 times with brine (20 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated and purified by column chromatography to obtain the title product (327 mg, 71.08%). MS (ESI, m / e) [M+H]+ 647.9.

[0196] Step 4: tert-Butyl 9-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-2-((1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-6-(trifluoromethyl)quinazolin-4-yl)-3-oxa-7,9-diazabicyclo[3.3.1]nonane-7-carboxylate

Chem.

[0197] A mixture of tert-butyl 9-(7-bromo-8-fluoro-2-((1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-6-(trifluoromethyl)quinazolin-4-yl)-3-oxa-7,9-diazabicyclo[3.3.1]nonane-7-carboxylate (40 mg, 0.061 mmol), tert-butyl (3-cyano-7-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[b]thiophen-2-yl)carbamate (104 mg, 0.024 mmol), Cs2CO3 (60 mg, 0.18 mmol) and DPEphosPdCl2 (10 mg, 0.012) in toluene (10 mL) was degassed three times with N2 and stirred at 100 °C for 16 h. The resulting mixture was filtered and the filtrate was concentrated. The residue was purified by column chromatography to give the title product (15 mg, 28.8%). MS (ESI, m / e) [M+H] + 859.2.

[0198] Step 5: 4-(4-(3-Oxa-7,9-diazabicyclo[3.3.1]nonan-9-yl)-8-fluoro-2-((1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

[0199] To a solution of tert-butyl 9-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-2-((1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-6-(trifluoromethyl)quinazolin-4-yl)-3-oxa-7,9-diazabicyclo[3.3.1]nonane-7-carboxylate (50 mg) in DCM (10 mL) was added TFA (2 mL). The mixture was stirred at room temperature for 2 - 3 h. The resulting mixture was concentrated to dryness, then diluted with DCM / MeOH = 10 / 1 (5 mL), and 5 drops of 7M NH₃ solution in MeOH were added. Next, the new solution was concentrated to dryness and purified by column chromatography to give the title product (21 mg, 55.2%). 1 H NMR (500 MHz, CD₃OD) δ 7.96 (s, 1H), 7.22 - 7.20 (m, 1H), 7.03 - 6.99 (m, 1H), 4.74 - 4.72 (m, 2H), 4.46 (s, 2H), 4.31 - 4.10 (m, 4H), 3.79 (s, 2H), 3.53 - 3.37 (m, 4H), 1.81 - 1.80 (m, 2H), 1.69 - 1.53 (m, 2H), 1.42 (s, 3H). MS (ESI, m / e) [M+H] + 659.8.

[0200] Example 17: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-2-(3-(cyanomethyl)-3-methylazetidin-1-yl)-8-fluoro-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chemical Structure

[0201] Step 1: tert-Butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-7-fluorobenzo[b]thiophen-4-yl)-2,8-difluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

Chem.

[0202] At room temperature, into a 1 L round-bottom flask were added tert-butyl 3-(7-bromo-2,8-difluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (50 g, 0.097 mol), tert-butyl (4-(5,5-dimethyl-1,3,2-dioxaborolan-2-yl)-7-fluorobenzo[b]thiophen-2-yl)carbamate (47.2 g, 0.11 mol), Pd(dtbpf)Cl2 (6.2 g, 0.096 mol), K3PO4 (36.5 g, 0.17 mol), dioxane (500 mL), and H2O (50 mL). Under a N2 atmosphere, the resulting mixture was stirred at 90 °C for 2 hours. The resulting mixture was extracted with EtOAc (2 L). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with petroleum ether:EtOAc = 100:0 to 70:30 to obtain the title product (26 g, 38.2% yield). MS (ESI, m / e) [M+H] + = 710.10.

[0203] Step 2: tert-Butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-7-fluoro-3-iodobenzo[b]thiophen-4-yl)-2,8-difluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

Chem.

[0204] At room temperature, 3-(7-(2-((tert-butoxycarbonyl)amino)-7-fluorobenzo[b]thiophen-4-yl)-2,8-difluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl (39 g, 0.055 mol), CH3CN (400 mL), and NIS (16 g, 0.07 mol) were placed in a 1 L round-bottom flask. The resulting mixture was stirred at room temperature for 2 hours. The resulting mixture was concentrated under vacuum. The residue was purified by silica gel column chromatography eluting with petroleum ether:EtOAc = 100:0 to 70:30 to obtain the title product (20 g, 43.5% yield). MS (ESI, m / e) [M+H] + = 836.00.

[0205] Step 3: 3-(7-(2-Amino-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-2,8-difluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl

Chemical formula

[0206] Under an N2 atmosphere at room temperature, tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-7-fluoro-3-iodobenzo[b]thiophen-4-yl)-2,8-difluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (100 mg, 0.12 mmol), Zn(CN)2 (70 mg, 0.6 mmol), Pd(PPh3)4 (83 mg, 0.07 mmol) and DMF (10 mL) were added to a 20 mL microwave tube. The resulting mixture was stirred for 10 minutes by microwave at 150 °C. The resulting mixture was diluted with water (30 mL) and extracted with EtOAc (2 x 30 mL). The combined organic layers were washed with brine (3 x 30 mL) and dried over anhydrous Na2SO4. The organic layer was concentrated under reduced pressure. The residue was purified by TLC (petroleum ether:EtOAc = 1:1) to give the title product (25 mg, 32.9% yield). MS (ESI, m / e) [M+H]= 635.05.

[0207] Step 4: tert-Butyl 3-(7-(2-amino-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-2-(3-(cyanomethyl)-3-methylazetidin-1-yl)-8-fluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

Chemical formula

[0208] At room temperature, tert-butyl 3-(7-(2-amino-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-2,8-difluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (20 mg, 0.03 mmol), THF (1 mL), 2-(3-methylazetidin-3-yl)acetonitrile TFA salt (17 mg, 0.15 mmol), and DIEA (41 mg, 0.3 mmol) were added to a 10 mL vial. The mixture was stirred at room temperature for 2 h. The mixture was concentrated to give the title crude product (40 mg, crude). MS (ESI, m / e) [M+H] + 725.15.

[0209] Step 5: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-2-(3-(cyanomethyl)-3-methylazetidin-1-yl)-8-fluoro-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

[0210] At room temperature, tert-butyl 3-(7-(2-amino-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-2-(3-(cyanomethyl)-3-methylazetidin-1-yl)-8-fluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (20 mg, 0.03 mmol), DCM (1 mL), and TFA (0.2 mL) were added to a 10 mL vial. The mixture was stirred at room temperature for 2 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash under the following conditions (column: XBridge Shield RP18 OBD column, 30*150 mm, 5 μm, mobile phase A: water (10 mmol / L NH4HCO3 + 0.1% NH3 . H2O), mobile phase B: ACN, flow rate: 60 mL / min, gradient: 20% B to 50% B for 9 min, 50% B, wavelength: 254 nm) to give the title product (5.1 mg, 29.6% yield, TFA salt). 11H NMR (300 MHz, DMSO-d6) δ 9.14 - 9.13 (m, 1H), 8.94 (s, 1H), 8.06 (s, 2H), 7.90 (s, 1H), 7.22 - 7.08 (m, 2H), 4.50 - 4.09 (m, 6H), 3.99 - 3.89 (m, 4H), 2.96 (s, 2H), 1.94 (s, 4H), 1.41 (s, 3H). 19F NMR: δ -57.00, -74.23, -116.68, -124.64. MS (ESI, m / e) [M+H] + 625.10.

[0211] Example 18: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((3-methyloxetan-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chem.

[0212] Example 18 was prepared by replacing (3-methoxytetrahydrofuran-3-yl)methanol with (3-methyloxetan-3-yl)methanol according to the same procedure as described in Example 3 to obtain the title product (12.75 mg). 1 1H NMR (500 MHz, CD3OD) δ 8.12 (s, 1H), 7.33 - 7.13 (m, 1H), 7.02 - 6.99 (m, 1H), 4.68 - 4.46 (m, 8H), 3.73 - 3.65 (m, 4H), 1.85 (s, 4H), 1.46 (s, 3H). MS (ESI, m / e) [M+H] + 617.5.

[0213] Example 19: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((4-hydroxytetrahydro-2H-pyran-4-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chem.

[0214] Example 19 was prepared by replacing (1-(difluoromethyl)-2-oxabicyclo[2.1.1]hexan-4-yl)methanol with 4-(hydroxymethyl)tetrahydro-2H-pyran-4-ol according to the same procedure as described in Example 1 (Steps 6 and 7) to give the title product (6.4 mg). 1 H NMR (500 MHz, CD3OD) δ 8.12 (s, 1H), 7.23 - 7.22 (m, 1H), 7.04 - 6.99 (m, 1H), 4.85 - 4.62 (m, 2H), 4.37 - 4.36 (m, 2H), 4.24 - 4.22 (m, 2H), 3.93 - 3.75 (m, 6H), 2.18 - 2.13 (m, 4H), 1.87 - 1.63 (m, 4H). MS (ESI, m / e) [M+H] + 647.5.

[0215] Example 20: Methyl 3-(((7-(2-amino-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-\ 8-fluoro-6-(trifluoromethyl)quinazolin-2-yl)oxy)methyl)-3-methoxyazetidine-1-carboxylate

Chem.

[0216] Example 20 was prepared by replacing (3-methylazetidin-3-yl)methanol hydrochloride with (3-methoxyazetidin-3-yl)methanol hydrochloride according to the same procedure as described in Example 8 to obtain the title product (1 mg, 9% yield, formate). 1 H NMR (500 MHz, CD3OD) δ 8.51 (s, 1H), 8.17 (s, 1H), 7.29 - 7.21 (m, 1H), 7.03 - 7.00 (m, 1H), 4.81 - 4.76 (m, 2H), 4.72 - 4.61 (m, 2H), 4.12 - 4.00 (m, 5H), 3.87 - 3.80 (m, 2H), 3.75 - 3.68 (m, 3H), 3.51 - 3.41 (m, 4H), 2.05 (s, 4H). MS (ESI, m / e) [M+1] + 690.5.

[0217] Example 21: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((1-(hydroxymethyl)-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chem.

[0218] Step 1: Methyl (4-(hydroxymethyl)-2-oxabicyclo[2.1.1]hexan-1-yl)acetate

Chem.

[0219] At room temperature, potassium acetate (980.0 mg, 10.0 mmol) was added to a solution of (1-(iodomethyl)-2-oxabicyclo[2.1.1]hexan-4-yl)methanol (254 mg, 1.0 mmol) in DMSO (20 mL). The resulting mixture was stirred at 100 °C for 16 hours. After completion, the reaction mixture was diluted with water (30 mL), extracted with EtOAc (100 mL), and washed with saturated aqueous NaCl solution (50 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to obtain the crude product (50 mg).

[0220] Example 2% was prepared by replacing (1-(difluoromethyl)-2-oxabicyclo[2.1.1]hexan-4-yl)methanol with methyl ((4-(hydroxymethyl)-2-oxabicyclo[2.1.1]hexan-1-yl)acetate by the same procedure as described in Example 1 (Steps 6 and 7) to obtain the title product (0.8 mg, formate). 1 H NMR (500 MHz, CD3OD) δ 8.50 (s, 1H), 8.11 (s, 1H), 7.24 - 7.16 (m, 1H), 7.03 - 7.00 (m, 1H), 4.80 - 4.46 (m, 4H), 3.91 - 3.65 (m, 8H), 2.16 - 1.78 (m, 8H). MS (ESI, m / e) [M+H] + 659.4

[0221] Example 22: 4-(((7-(2-Amino-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-6-(trifluoromethyl)quinazolin-2-yl)oxy)methyl)tetrahydro-2H-pyran-4-carbonitrile

Chemical formula

[0222] Example 22 was prepared by replacing (1-(difluoromethyl)-2-oxabicyclo[2.1.1]hexan-4-yl)methanol with 4-(hydroxymethyl)tetrahydro-2H-pyran-4-carbonitrile by the same procedure as described in Example 1 (Steps 6 and 7) to obtain the title product. It was separated by chiral HPLC (Prep-HPLC apparatus: Prep-HPLC-Gilson, column: IE, 20 mm × 250 mm, 5 μm, mobile phase A: n-hexane, mobile phase B: EtOH (0.2% 2M NH3 in MeOH), flow rate: 18 mL / min, temperature: 25 °C, gradient: 50% B for 14 minutes, retention time - isomer 1: 7 minutes, retention time - isomer 2: 11.5 minutes) to obtain isomer 1 (21.2 mg) and isomer 2 (20.2 mg). Isomer 1: 1 H NMR (500 MHz, CD3OD) δ 8.13 (s, 1H), 7.22 - 7.20 (m, 1H), 7.02 - 6.99 (m, 1H), 4.57 - 4.45 (m, 4H), 4.01 - 3.99 (m, 2H), 3.74 - 3.64 (m, 6H), 2.06 - 2.03 (m, 2H), 1.87 - 1.82 (m, 6H); MS(ESI, m / e) [M + H] + 656.5. Isomer 2: 1 H NMR(500 MHz, CD3OD) δ 8.13 (s, 1H), 7.22 - 7.20 (m, 1H), 7.02 - 6.99 (m, 1H), 4.57 - 4.45 (m, 4H), 4.01 - 3.99 (m, 2H), 3.74 - 3.62 (m, 6H), 2.06 - 2.03 (m, 2H), 1.87 - 1.82 (m, 6H); MS(ESI, m / e) [M + H] + 656.5.

[0223] Example 23: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((tetrahydro-2H-pyran-4-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chemical formula

[0224] Step 1: tert-Butyl 3-(7-bromo-8-fluoro-2-(methylthio)-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

Chem.

[0225] A mixture of tert-butyl 3-(7-bromo-2,8-difluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (14.8 g, 28.3 mmol) and sodium methanethiolate (1.96 g, 28.4 mmol) in THF (500 mL) was stirred at room temperature for 15 h. The reaction mixture was diluted with EtOAc (50 mL) and washed with saturated NaCl aqueous solution (15 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to obtain a residue. The residue was purified by combi-flash (petroleum ether:EtOAc = 10:1) to obtain the title product (10.0 g, 64.1%). MS (ESI, m / e) [M+1] + 551.4.

[0226] Step 2: tert-Butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-2-(methylthio)-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

Chem.

[0227] Under a N2 atmosphere, a mixture of tert-butyl 3-(7-bromo-8-fluoro-2-(methylthio)-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (5.5 g mg, 10 mmol), tert-butyl (3-cyano-7-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[b]thiophen-2-yl)carbamate (8.36 g, 20 mmol), Pd(dtbpf)Cl2 (3.26 g, 5 mmol) and K3PO4 (6.36 g, 30 mmol) in dioxane / water (550 mL / 55 mL) was stirred at 80 °C for 0.5 h. After completion, the reaction mixture was diluted with EtOAc (50 mL) and washed with saturated NaCl aqueous solution (15 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to obtain a residue. The residue was purified by combi-flash (petroleum ether:EtOAc = 3:1) to obtain the title product (2.9 g, 38.1%). MS (ESI, m / e) [M+1] + 763.5.

[0228] Step 3: tert-Butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-2-(methylsulfonyl)-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

Chemical Structure

[0229] A mixture of tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-2-(methylthio)-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (1.24 g, 1.63 mmol) and m-CPBA (561 mg, 3.26 mmol) in DCM (10 mL) was stirred at room temperature for 3 h. The reaction mixture was diluted with EtOAc (50 mL) and washed with saturated aqueous NaHCO3 (15 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to give a residue. The residue was used directly in the next step without further purification. MS (ESI, m / e) [M+1] + 795.6.

[0230] Step 4: tert-Butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-2-((tetrahydro-2H-pyran-4-yl)methoxy)-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

Chemical formula

[0231] A solution of (tetrahydro-2H-pyran-4-yl)methanol (12 mg, 0.1 mmol) in THF (2 mL) at 0 °C was added to LiHMDS (0.1 mL, 0.1 mmol). The reaction mixture was stirred at room temperature for 0.5 h. Then, tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-2-(methylsulfonyl)-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (40 mg, 0.05 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature for 1 h. After completion, the reaction mixture was diluted with EtOAc (40 mL) and washed with saturated aqueous NaCl solution (15 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to obtain a residue. The residue was purified by Prep-TLC (petroleum ether:EtOAc = 2:1) to obtain the title product (35 mg, 85.4%). MS (ESI, m / e) [M+1] + 831.6.

[0232] Step 5: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((tetrahydro-2H-pyran-4-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

[0233] At room temperature, TFA (2 mL) was added to a solution of tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-2-((tetrahydro-2H-pyran-4-yl)methoxy)-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (35 mg, 0.042 mmol) in DCM (2 mL). The resulting mixture was stirred at room temperature for 3 h. After completion, the reaction mixture was concentrated to obtain a residue. The residue was purified by prep-HPLC to obtain the title product (9.79 mg, 36.9%). 11H NMR (500 MHz, CD3OD) δ 8.10 (s, 1H), 7.28 - 7.15 (m, 1H), 7.03 - 6.99 (m, 1H), 4.66 - 4.52 (m, 2H), 4.34 - 4.32 (m, 2H), 4.00 - 3.91 (m, 4H), 3.82 - 3.72 (m, 2H), 3.53 - 3.44 (m, 2H), 2.14 - 2.11 (m, 1H), 2.04 - 1.94 (m, 4H), 1.79 - 1.77 (m, 2H), 1.56 - 1.37 (m, 2H). MS (ESI, m / e) [M+1] + 631.5.

[0234] Example 24: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((1-(1-methyl-1H-imidazol-2-yl)azetidin-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chemical formula

[0235] Step 1: (1-(1-Methyl-1H-imidazol-2-yl)azetidin-3-yl)methanol

Chemical formula

[0236] A solution of azetidin-3-ylmethanol hydrochloride (500 mg, 4.06 mmol), 2-iodo-1-methyl-1H-imidazole (527 mg, 4.87 mmol), CuI (78 mg, 0.41 mmol), (R)-proline (94 mg, 10.82 mmol) and Cs2CO3 (2.65 g, 8.12 mmol) in DMF (30 mL) was stirred at 100 °C for 3 h. The solvent was concentrated and the residue was purified by column chromatography on silica gel (DCM:MeOH = 15:1) to give the title product (220 mg, 32%). MS (ESI, m / e) [M+1] + 168.4.

[0237] Example 24 was prepared by replacing methyl 3-(hydroxymethyl)-3-methylazetidine-1-carboxylate with (1-(1-methyl-1H-imidazol-2-yl)azetidin-3-yl)methanol according to the same procedure as described in Example 8 to give the title product (5 mg, 14% yield, formate). 1 H NMR (500 MHz, CD3OD) δ 8.49 (s, 1H), 8.12 (s, 1H), 7.20 - 7.22 (m, 1H), 7.06 - 7.00 (m, 1H), 6.74 (s, 1H), 6.69 (s, 1H), 4.52 - 4.73 (m, 4H), 4.32 - 4.26 (m, 2H), 4.12 - 4.05 (m, 2H), 3.94 - 3.89 (m, 2H), 3.81 - 3.72 (m, 2H), 3.44 (s, 3H), 3.30 - 3.15 (m, 1H), 1.95 (s, 4H). MS (ESI, m / e) [M+1] + 682.5.

[0238] Example 25: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((3-fluorotetrahydrofuran-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile [Chemical formula]

[0239] Step 1: tert-Butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-2-((3-fluorotetrahydrofuran-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

Chem.

[0240] At 0 °C, tert-Butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-2,8-difluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (50 mg, 0.07 mmol) was added portionwise to a stirred mixture of (3-fluorotetrahydrofuran-3-yl)methanol (8.2 mg, 0.07 mmol) and NaH (3.3 mg, 0.14 mmol) in THF (1 mL). The resulting mixture was stirred at room temperature for 1 h. The reaction was quenched with saturated aqueous NH4Cl. The resulting mixture was extracted with EtOAc (3 x 30 mL). The combined organic layers were washed with brine (50 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (petroleum ether / EtOAc 1:1) to give the title product (30 mg, 69.47% yield). MS (ESI, m / e) [M+H]+ 835.35.

[0241] Step 2: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((3-fluorotetrahydrofuran-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

[0242] A solution of tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-2-((3-fluorotetrahydrofuran-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (30 mg, 0.04 mmol) and TFA (0.5 mL) in DCM (1.5 mL) was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC (column: Xselect CSH C18 OBD column 30*150 mm 5 μm, mobile phase A: water (0.1% TFA), mobile phase B: ACN, flow rate: 60 mL / min, gradient: 15% B - 45% B for 8 min, 45% B, wavelength: 254 nm, RT1 (min): 7) to give the title product (8.1 mg, 35.52% yield, TFA salt / formate). 1 H NMR (300 MHz, CD3OD) δ 8.47 (s, 0.5H), 8.13 (s, 1H), 7.21 (s, 1H), 7.02 (s, 1H), 4.76 - 4.57 (m, 4H), 4.26 - 3.68 (m, 8H), 2.45 - 2.16 (m, 2H), 2.05 (s, 4H). MS (ESI, m / e) [M+H] + 635.40.

[0243] Example 26: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-6-(trifluoromethyl)-2-((3-(trifluoromethyl)tetrahydrofuran-3-yl)methoxy)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chemical Structure

[0244] Step 1: Diethyl 2-(2-(benzyloxy)ethyl)malonate

Chemical Structure

[0245] At 0 °C, NaH (8.93 g, 223.16 mmol, 60%) was added portionwise to a solution of ((2-bromoethoxy)methyl)benzene (40 g, 185.97 mmol) in THF (500 mL). After stirring for 5 minutes, diethyl malonate (60 g, 371.94 mmol) was added. The resulting mixture was stirred at 0 °C for 12 hours. The resulting mixture was quenched with saturated aqueous NH4Cl at 0 °C and extracted with EtOAc (3 x 200 mL). The combined organic layers were washed with brine (2 x 200 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by C18 column chromatography, eluting with H2O:CH3CN (100:0 - 20:80) to give the title product (44 g, 80.38% yield). MS (ESI, m / e) [M+H] + 295.10.

[0246] Step 2: Diethyl 2-(2-(benzyloxy)ethyl)-2-(trifluoromethyl)malonate

Chemical Structure

[0247] A solution of diethyl 2-(2-(benzyloxy)ethyl)malonate (40 g, 135.89 mmol) in DMF (500 mL) at 0 °C was added NaH (6.52 g, 163.07 mmol, 60%) portionwise in several times. After stirring at room temperature for 1 h, at -60 °C, 2,8-difluoro-5-(trifluoromethyl)-5H-dibenzo[b,d]thiophen-5-ium trifluoromethanesulfonate (87.24 g, 203.84 mmol) was added. The resulting mixture was stirred at room temperature for 12 h. The resulting mixture was quenched with saturated aqueous NH4Cl solution at 0 °C and extracted with EtOAc (3 x 200 mL). The combined organic layers were washed with brine (2 x 200 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with petroleum ether:EtOAc (100:0 - 80:20) to give the title product (10 g, 20.31% yield). MS (ESI, m / e) [M+H] + 363.10.

[0248] Step 3: 2-(2-(Benzyloxy)ethyl)-2-(trifluoromethyl)propane-1,3-diol

Chemical Structure

[0249] A solution of diethyl 2-(2-(benzyloxy)ethyl)-2-(trifluoromethyl)malonate (9.5 g, 26.22 mmol) in THF (100 mL) at 0 °C was added LiAlH4 (2.99 g, 78.65 mmol) portionwise in several times. After stirring at room temperature for 1 h, the resulting mixture was quenched with Na2SO4 . 10H2O at 0 °C. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with petroleum ether:EtOAc (100:0 - 60:40) to give the title product (2.1 g, 28.78% yield). MS (ESI, m / e) [M+H]+279.00.

[0250] Step 4: 4-(Benzyloxy)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-2-(trifluoromethyl)butan-1-ol

Chem.

[0251] At 0 °C, TBDPSCl (2.28 g, 8.30 mmol) was added portionwise to a solution of 2-(2-(benzyloxy)ethyl)-2-(trifluoromethyl)propane-1,3-diol (2.1 g, 7.55 mmol) and 1H-imidazole (1.54 g, 22.64 mmol) in DCM (50 mL). After stirring at room temperature for 3 h, the mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with petroleum ether:EtOAc (100:0 - 70:30) to give the title product (2.5 g, 64.12% yield). MS (ESI, m / e) [M+Na] + 539.25. + 539.25.

[0252] Step 5: 2-(((tert-butyldiphenylsilyl)oxy)methyl)-2-(trifluoromethyl)butane-1,4-diol

Chem.

[0253] At room temperature, Pd / C (1.2 g, 10%) was added to a solution of 4-(benzyloxy)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-2-(trifluoromethyl)butan-1-ol (1.2 g, 2.323 mmol) in EtOH (50 mL). After stirring at 80 °C for 12 h under an H2(g) atmosphere, the mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with petroleum ether:EtOAc (100:0 - 60:40) to give the title product (500 mg, 40.38% yield). MS (ESI, m / e) [M+H] + 427.15. + 427.15.

[0254] Step 6: tert-Butyldiphenyl((3-(trifluoromethyl)tetrahydrofuran-3-yl)methoxy)silane

Chem.

[0255] At 0 °C, n-BuLi (0.54 mL, 1.35 mmol, 2.5 M) was added to a solution of 2-(((tert-butyldiphenylsilyl)oxy)methyl)-2-(trifluoromethyl)butane-1,4-diol (480 mg, 1.13 mmol) and TsCl (300.3 mg, 1.58 mmol) in THF (50 mL). After stirring at room temperature for 1.5 h, n-BuLi (0.54 mL, 1.35 mmol, 2.5 M in hexane) was added at 0 °C. The resulting mixture was stirred at room temperature for 12 h. The resulting mixture was quenched with saturated aqueous NH4Cl at 0 °C and extracted with EtOAc (3 x 50 mL). The combined organic layers were washed with brine (2 x 20 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with petroleum ether:EtOAc (100:0 - 90:10) to give the title product (190 mg, 41.33% yield).

[0256] Step 7: (3-(Trifluoromethyl)tetrahydrofuran-3-yl)methanol

Chem.

[0257] A solution of tert-butyldiphenyl((3-(trifluoromethyl)tetrahydrofuran-3-yl)methoxy)silane (90 mg, 0.22 mmol) in THF (50 mL) at 0 °C was added dropwise with TBAF (0.55 mL, 0.55 mmol, 1 M in THF). After stirring at room temperature for 12 h, the mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with petroleum ether:EtOAc (100:0 - 60:40) to give the title product (15 mg, 40.02%).

[0258] Step 8: tert-Butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-6-(trifluoromethyl)-2-((3-(trifluoromethyl)tetrahydrofuran-3-yl)methoxy)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate [Chemical formula]

[0259] To a solution of tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-2,8-difluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (20 mg, 0.03 mmol) and (3-(trifluoromethyl)tetrahydrofuran-3-yl)methanol (9.26 mg, 0.06 mmol) in THF (2 mL) at 0 °C was added NaH (2.18 mg, 0.06 mmol, 60%). After stirring at room temperature for 2 h, the mixture was quenched with saturated aqueous NH4Cl at 0 °C and extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (2 x 20 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC with petroleum ether:EtOAc (4:1) to give the title product (13 mg, 53.97% yield). MS (ESI, m / e) [M+H]+ 885.25.

[0260] Step 9: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-6-(trifluoromethyl)-2-((3-(trifluoromethyl)tetrahydrofuran-3-yl)methoxy)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

[0261] To a stirred solution of tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-6-(trifluoromethyl)-2-((3-(trifluoromethyl)tetrahydrofuran-3-yl)methoxy)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (13 mg, 0.015 mmol) in DCM (0.6 mL) was added TFA (0.2 mL). The resulting mixture was stirred at room temperature for 1 h and concentrated under reduced pressure. The crude product was purified by Prep-HPLC (column: XBridge Shield RP18 OBD column, 30*150 mm, 5 μm, mobile phase A: water (0.05% TFA), mobile phase B: ACN, flow rate: 60 mL / min, gradient: 32% B - 62% B for 9 min, 62% B, wavelength: 220 nm, RT1 (min): 7) to give the title product (0.8 mg, 6.82% yield, TFA salt). 1 H NMR (300 MHz, CD3OD) δ 8.13(s, 1H), 7.24 -7.20 (m, 1H), 7.09- 6.98 (m, 1H),4.81 - 4.54 (m,4H), 4.23 (s, 2H),4.04 - 3.83 (m,6H), 2.39 - 2.05(m, 6H). MS (ESI,m / e) [M+H] + 685.20.

[0262] Example 27: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro- 2-((4-hydroxy-4-methyltetrahydrofuran-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzothiophene-3-carbonitrile

Chemical formula

[0263] Step 1: Methyl 4-hydroxytetrahydrofuran-3-carboxylate

Chemical formula

[0264] At 0 °C, NaBH4 (5.2 g, 83.26 mmol) was added portionwise to a solution of methyl 4-oxooxolane-3-carboxylate (10 g, 69.38 mmol) in MeOH (1 L). Under a nitrogen atmosphere, the resulting mixture was stirred at room temperature for 2 h. The mixture was quenched with saturated aqueous NH4Cl solution at 0 °C and extracted with DCM (5 × 500 mL). The resulting mixture was concentrated under vacuum. The crude product (5.7 g) was used directly in the next step without further purification.

[0265] Step 2: Methyl 4-((tert-butyldimethylsilyl)oxy)tetrahydrofuran-3-carboxylate

Chemical formula

[0266] In a nitrogen atmosphere, a solution of methyl 4-hydroxyoxolane-3-carboxylate (5.7 g, 39.00 mmol), TBSCl (7.64 g, 50.00 mmol) and imidazole (5.31 g, 78.00 mmol) in DMF (57 mL) was stirred at room temperature overnight. The reaction was quenched with water (30 mL) at room temperature. The aqueous layer was extracted with EtOAc (3 x 30 mL). The combined organic layers were washed with brine and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with petroleum ether:EtOAc (5%-10%) to give the title product (9.5 g, 94% yield) as a colorless oil.

[0267] Step 3: 4-(Hydroxymethyl)oxolan-3-ol

Chemical formula

[0268] LiAlH4 (2.4 g, 63.16 mmol) was suspended in THF (48 mL). At 0 °C, a solution of methyl 4-((tert-butyldimethylsilyl)oxy)oxolane-3-carboxylate (9.5 g, 36.48 mmol) in THF (57 mL) was slowly added dropwise and the mixture was stirred at 0 °C for 1 h. LiAlH4 (2.4 g, 63.16 mmol) was added and the mixture was stirred at room temperature for 2.5 h. The mixture was heated to reflux for 1 h. The reaction solution was cooled to room temperature. Then, water (2.9 mL), 15% aqueous sodium hydroxide solution (2.9 mL) and water (5.8 mL) were added at 0 °C to quench the reaction. The resulting mixture was stirred at room temperature for 15 min and dried over anhydrous MgSO4. The precipitated insoluble material was filtered through celite. The filtrate was concentrated. The crude product (4.5 g) was used directly in the next step without further purification.

[0269] Step 4: 4-(((tert-Butyldimethylsilyl)oxy)methyl)oxolan-3-ol

Chemical formula

[0270] A solution of 4-(hydroxymethyl)tetrahydrofuran-3-ol (2 g, 16.93 mmol), TBSCl (1.54 g, 10.20 mmol) and imidazole (2.31 g, 33.86 mmol) in DMF (100 mL) was stirred at room temperature overnight. The reaction was quenched with water (50 mL) at room temperature. The aqueous layer was extracted with EtOAc (3 x 50 mL). The combined organic layers were washed with brine and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with petroleum ether:EtOAc (5%-10%) to give the title product (1.7 g, 43% yield) as a colorless oil. MS (ESI, m / e) [M+H] + 233.15.

[0271] Step 5: 4-(((tert-butyldimethylsilyl)oxy)methyl)dihydrofuran-3(2H)-one

Chemical formula

[0272] A solution of 4-(((tert-butyldimethylsilyl)oxy)methyl)tetrahydrofuran-3-ol (1.7 g, 7.31 mmol) and DMP (6.21 g, 14.63 mmol) in DCM (16 mL) was stirred at 40 °C for 5 h. The resulting mixture was filtered and the filter cake was washed with DCM (3 x 10 mL). The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with petroleum ether:EtOAc (5%-20%) to give the title product (1.3 g, 77% yield) as a colorless oil. MS (ESI, m / e) [M+H] + 231.15.

[0273] Step 6: 4-(((tert-butyldimethylsilyl)oxy)methyl)-3-methyltetrahydrofuran-3-ol

Chemical formula

[0274] At -78 °C, MeMgBr (1.0 M solution, 0.87 mL) was added to a solution of 4 - ((((tert-butyldimethylsilyl)oxy)methyl)dihydrofuran-3(2H)-one (1.2 g, 5.21 mmol) in THF (10 mL). The mixture was stirred at room temperature for 1 hour and quenched with saturated aqueous NH4Cl. After removing all volatile substances under reduced pressure, the residue was diluted with DCM. The organic layer was washed twice with brine, dried over MgSO4, and filtered. The filtrate was concentrated to give a residue, which was purified by silica gel column chromatography, eluting with petroleum ether:EtOAc (5% - 20%) to give the title product (1 g, 78% yield) as a colorless oil. MS (ESI, m / e) [M+H] + 247.15.

[0275] Step 7: 4-(Hydroxymethyl)-3-methyltetrahydrofuran-3-ol

Chemical formula

[0276] Under a nitrogen atmosphere, a solution of 4 - ((((tert-butyldimethylsilyl)oxy)methyl)-3-methyltetrahydrofuran-3-ol (500 mg, 2.03 mmol) and TBAF (6 mL, 6.09 mmol, 1 M in THF) in THF (10 mL) was stirred at room temperature for 2 hours. The resulting mixture was diluted with water (5 mL). The aqueous layer was extracted with EtOAc (5 x 5 mL). The residue was purified by silica gel column chromatography, eluting with DCM / MeOH (0% - 5%) to give the title product (10 mg, 3.73% yield) as a colorless oil.

[0277] Step 8: tert-Butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-2-((4-hydroxy-4-methyltetrahydrofuran-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

Chem.

[0278] Under a nitrogen atmosphere at 0 °C, NaH (2.42 mg, 0.101 mmol) was added portionwise to a solution of 4-(hydroxymethyl)-3-methyltetrahydrofuran-3-ol (10 mg, 0.08 mmol) and tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-2,8-difluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (37.1 mg, 0.051 mmol) in THF (0.5 mL). The resulting mixture was stirred at room temperature for 2 h under a nitrogen atmosphere. The reaction was quenched with saturated aqueous NH4Cl solution (2 mL) at 0 °C. The aqueous layer was extracted with EtOAc (3 x 10 mL). The resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-TLC (petroleum ether:EtOAc = 1:2) to give the title product (13 mg, 30% yield) as a white solid. MS (ESI, m / e) [M+H] + 847.30.

[0279] Step 9: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((4-hydroxy-4-methyltetrahydrofuran-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

[0280] A solution of tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-2-((4-hydroxy-4-methyltetrahydrofuran-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (12 mg, 0.014 mmol) and TFA (0.2 mL) in DCM (1 mL) was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC under the following conditions (column: XBridge Shield RP18 OBD column, 30 * 150 mm, 5 μm, mobile phase A: water (0.05% TFA), mobile phase B: ACN, flow rate: 60 mL / min, gradient: 22% B - 52% B for 9 min) to obtain the title product (1 mg, 11% yield, TFA salt). 1 H NMR (300 MHz, CD3OD) δ 8.11 (s, 1H), 7.24 - 7.19 (m, 1H), 7.05 - 6.99 (m, 1H), 4.79 - 4.60 (m, 3H), 4.54 - 4.42 (m, 1H), 4.23 (s, 2H), 4.14 - 4.11 (m, 1H), 3.98 - 3.80 (m, 3H), 3.79 - 3.66 (m, 2H), 2.64 - 2.48 (m, 1H), 2.13 (s, 4H), 1.44 (s, 3H). MS (ESI, m / e) [M+H] + 647.25.

[0281] Example 28: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((3-methyloxetan-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chemical Structure

[0282] Example 28 was prepared by replacing (3-methoxytetrahydrofuran-3-yl)methanol with (3-methoxyoxetan-3-yl)methanol according to the same procedure as described in Example 3 to obtain the title product (2.65 mg). 1 H NMR (500 MHz, CD3OD) δ 8.13 (s, 1H), 7.22 - 7.20 (m, 1H), 7.02 - 6.99 (m, 1H), 4.84 - 4.82 (m, 2H), 4.74 - 4.72 (m, 2H), 4.64 - 4.62 (m, 2H), 4.59 - 4.56 (m, 1H), 4.47 - 4.45 (m, 1H), 3.73 - 3.70 (m, 1H), 3.66 - 3.64 (m, 3H), 3.40 (s, 3H), 1.87 - 1.77 (m, 4H). MS (ESI, m / e) [M+H] + 633.4.

[0283] Example 29: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)naphthalen-2-ol

Chem.

[0284] Step 1: tert-Butyl 3-(8-fluoro-7-(3-(methoxymethoxy)naphthalen-1-yl)-2-((1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

Chem.

[0285] A mixture of tert-butyl 3-(7-bromo-8-fluoro-2-((1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (50 mg, 0.08 mmol), 2-(3-(methoxymethoxy)naphthalen-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (49 mg, 0.16 mmol), DPEPhosPdCl2 (11 mg, 0.016 mmol) and Cs2CO3 (77 mg, 0.24 mmol) was added to toluene (5 mL). The solution was degassed three times with N2. The mixture was stirred at 100 °C for 3 hours. The cooled obtained mixture was filtered and the filtrate was concentrated. The residue was purified by silica chromatography column to give the title product (44 mg). MS (ESI, m / e) [M+H] + 739.5.

[0286] Step 2: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)naphthalen-2-ol

[0287] A solution of tert-butyl 3-(8-fluoro-7-(3-(methoxymethoxy)naphthalen-1-yl)-2-((1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (44 mg, 0.06 mmol) in DCM (10 mL) was added dropwise with TFA (2 mL). The mixture was stirred at room temperature for 2 - 3 h. The resulting mixture was concentrated to dryness and then diluted with DCM / MeOH = 10 / 1 (5 mL). Next, 5 drops of 7M NH3 / MeOH solution were added to the mixture. The solution was further concentrated to dryness and purified by chromatography to obtain the title product (2.64 mg). NMR (500 MHz, CD3OD) δ 8.21 (s, 1H), 7.74 - 7.72 (m, 1H), 7.41 - 7.37 (m, 1H), 7.25 - 7.24 (m, 1H), 7.17 - 7.16 (m, 2H), 7.04 - 7.02 (m, 1H), 4.75 - 4.70 (m, 2H), 4.57 - 4.53 (m, 2H), 3.79 (s, 2H), 3.72 - 3.69 (m, 4H), 1.87 - 1.81 (m, 6H), 1.62 - 1.61 (m, 2H), 1.42 (s, 3H). MS (ESI, m / e) [M+H] + 595.5.

[0288] Example 30: 4-(((7-(2-Amino-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-6-(trifluoromethyl)quinazolin-2-yl)oxy)methyl)-2-oxabicyclo[2.1.1]hexane-1-carbonitrile [Chemical Structure]

[0289] Step 1: 1-(Iodomethyl)-4-(((4-methoxybenzyl)oxy)methyl)-2-oxabicyclo[2.1.1]hexane [Chemistry]

[0290] At room temperature, NaH (1.20 g, 30.0 mmol) was added to a solution of (1-(iodomethyl)-2-oxabicyclo[2.1.1]hexan-4-yl)methanol (2.54 g, 10.0 mmol) in THF (20 mL). The resulting mixture was stirred at room temperature for 1 hour. Then, 1-(chloromethyl)-4-methoxybenzene was added to the reaction mixture and stirred at room temperature for 1 hour. After completion, the reaction mixture was quenched with water (30 mL), extracted with EtOAc (100 mL), and washed with saturated aqueous NaCl solution (50 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to obtain a residue. The residue was purified by silica gel column chromatography, eluting with 0 - 30% ethyl acetate in petroleum ether to give the title product (1.87 g, 5.0 mmol).

[0291] Step 2: (4-(((4-methoxybenzyl)oxy)methyl)-2-oxabicyclo[2.1.1]hexan-1-yl)methanol [Chemistry]

[0292] At room temperature, KOAc (2.45 g, 25 mmol) was added to a solution of 1-(iodomethyl)-4-(((4-methoxybenzyl)oxy)methyl)-2-oxabicyclo[2.1.1]hexane (1.87 g, 5.0 mmol) in DMSO (30 mL). The resulting mixture was stirred at 110 °C for 16 h. After completion, the reaction mixture was diluted with EtOAc (200 mL) and washed with saturated aqueous NaCl solution (50 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to obtain a residue. The residue was added to sodium ethoxide (680 mg, 10 mmol) and EtOH (10 mL). Then, the reaction mixture was stirred at room temperature for 1 h. After completion, the reaction mixture was concentrated directly to obtain a residue. The residue was purified by silica gel column chromatography, eluting with 0 - 80% ethyl acetate in petroleum ether to obtain the title product (528.2 mg, 2.0 mmol).

[0293] Step 3: 4-(((4-Methoxybenzyl)oxy)methyl)-2-oxabicyclo[2.1.1]hexane-1-carbaldehyde

Chemical formula

[0294] At room temperature, DMP (848 mg, 2.0 mmol) was added to a solution of (4-(((4-methoxybenzyl)oxy)methyl)-2-oxabicyclo[2.1.1]hexan-1-yl)methanol (528.2 mg, 2.0 mmol) in DCM (10 mL). The resulting mixture was stirred at room temperature for 1 h. After completion, the reaction mixture was diluted with EtOAc (100 mL) and washed with saturated aqueous NaCl solution (50 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to obtain the crude title product (1.0 g crude).

[0295] Step 4: 4-(((4-Methoxybenzyl)oxy)methyl)-2-oxabicyclo[2.1.1]hexane-1-carbonitrile

Chemical formula

[0296] At room temperature, hydroxylamine hydrochloride (87.0 mg, 1.26 mmol) was added to a solution of 4-(((4-methoxybenzyl)oxy)methyl)-2-oxabicyclo[2.1.1]hexane-1-carbaldehyde (100 mg crude) in pyridine (2 mL). The resulting mixture was stirred at room temperature for 16 h. After completion, the reaction mixture was concentrated directly to obtain the crude product (150 mg crude). Subsequently, triethylamine (0.15 ml, 1.1 mmol), methanesulfonyl chloride (0.06, 0.41 mmol) and DCM (5 mL) were added to the crude product. The resulting mixture was stirred at room temperature for 16 h. After completion, the residue was purified by silica gel chromatography, eluting with 0-60% ethyl acetate in petroleum ether to obtain the title product (60.0 mg, 0.23 mmol).

[0297] Step 5: 4-(Hydroxymethyl)-2-oxabicyclo[2.1.1]hexane-1-carbonitrile

Chem.

[0298] At room temperature, DDQ (90.8 mg, 0.4 mmol) was added to a solution of 4-(hydroxymethyl)-2-oxabicyclo[2.1.1]hexane-1-carbonitrile (60.0 mg, 0.23 mol) and water (1 mL) in DCM (5 mL). The resulting mixture was stirred at room temperature for 2 h. After completion, the reaction mixture was diluted with water (10 mL), extracted with DCM (10 mL), and washed with saturated aqueous NaCl solution (10 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to obtain a residue. The residue was purified by silica gel chromatography, eluting with 0-60% ethyl acetate in petroleum ether to obtain the title product (20.0 mg, 0.14 mmol).

[0299] Example 30 was prepared by replacing (1-(difluoromethyl)-2-oxabicyclo[2.1.1]hexan-4-yl)methanol with 4-(hydroxymethyl)-2-oxabicyclo[2.1.1]hexane-1-carbonitrile according to the same procedure as described in Example 1, to obtain the title product (4.4 mg). 1 H NMR (500 MHz, CD3OD) δ 8.12 (s, 1H), 7.22 - 7.20 (m, 1H), 7.02 - 6.99 (m, 1H), 4.85 - 4.55 (m, 3H), 3.97 (s, 2H), 3.85 - 3.75 (m, 5H), 2.39 (s, 2H), 2.02 - 1.99 (m, 6H). δ MS (ESI, m / e) [M+1] + 654.5.

[0300] Example 31: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-2-((1-(cyclopropanecarbonyl)-3-methylazetidin-3-yl)methoxy)-8-fluoro-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chem.

[0301] Example 31 was prepared by replacing acetyl chloride with cyclopropanecarbonyl chloride according to the same procedure as described in Example 13, to obtain the title product (4.4 mg). 11H NMR (500 MHz, CD3OD) δ 8.12 (s, 1H), 7.23 - 7.20 (m, 1H), 7.03 - 7.00 (m, 1H), 4.68 - 4.47 (m, 4H), 4.39 - 4.35 (m, 1H), 4.05 - 3.97 (m, 4H), 3.83 - 3.69 (m, 3H), 2.00 (s, 4H), 1.61 - 1.56 (m, 3H), 1.47 (s, 3H), 0.86 - 0.79 (m, 4H). MS (ESI, m / e) [M+1] + 684.5.

[0302] Example 32: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-2-((3-ethyloxetan-3-yl)methoxy)-8-fluoro-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chemical Structure

[0303] Step 1: tert-Butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-2-((3-ethyloxetan-3-yl)methoxy)-8-fluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

Chemical Structure

[0304] A solution of (3-ethyloxetan-3-yl)methanol (12 mg, 0.1 mmol) in THF (2 mL) at 0 °C was added to LiHMDS (0.1 mL, 0.1 mmol). The resulting mixture was stirred at room temperature for 0.5 h. Then, tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-2-(methylsulfonyl)-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (40 mg, 0.05 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature for 1 h. After completion, the reaction mixture was diluted with EtOAc (40 mL) and washed with saturated aqueous NaCl solution (15 mL×3). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to obtain a residue. The residue was purified by Prep-TLC (petroleum ether:EtOAc = 2:1) to obtain the title product (35 mg, 84.4%). MS (ESI, m / e) [M+1] + 831.5.

[0305] Step 2: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-2-((3-ethyloxetan-3-yl)methoxy)-8-fluoro-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

[0306] At room temperature, TFA (1 mL) was added to a solution of tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-2-((3-ethyloxetan-3-yl)methoxy)-8-fluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (70 mg, 0.0842 mmol) in DCM (10 mL). The resulting mixture was stirred at room temperature for 3 h. After completion, the reaction mixture was concentrated to obtain a residue. The residue was purified by prep-HPLC to obtain the title product (7.5 mg, 14.2%). 11H NMR (500 MHz, CD3OD) δ 8.12 (s, 1H), 7.23 - 7.20 (m, 1H), 7.04 - 6.96 (m, 1H), 4.64 - 4.60 (m, 4H), 4.60 - 4.54 (m, 1H), 4.49 - 4.46 (m, 3H), 3.69 - 3.59 (m, 4H), 1.93 - 1.84 (m, 6H), 1.01 - 0.98 (m, 3H). MS (ESI, m / e) [M+1] + 631.5.

[0307] Example 33: 4-(2-((2-Oxaspiro[3.3]heptan-6-yl)methoxy)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chem.

[0308] Example 33 was prepared by replacing (3-ethyloxetan-3-yl)methanol with (2-oxaspiro[3.3]heptan-6-yl)methanol according to the same procedure as described in Example 32 to obtain the title product (11.5 mg, 21.7%). 1 1H NMR (500 MHz, CD3OD) δ 8.10 (s, 1H), 7.22 - 7.19 (m, 1H), 7.02 - 6.98 (m, 1H), 4.72 (s, 2H), 4.66 (s, 2H), 4.55 - 4.53 (m, 1H), 4.45 - 4.42 (m, 1H), 4.37 - 4.36 (m, 2H), 3.71 - 3.62 (m, 4H), 2.64 - 2.58 (m, 1H), 2.45 - 2.40 (m, 2H), 2.19 - 2.15 (m, 2H), 1.89 - 1.76 (m, 4H). MS (ESI, m / e) [M+1] + 643.4.

[0309] Example 34: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-2-((3-(1,1-difluoroethyl)oxetan-3-yl)methoxy)-8-fluoro-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chem.

[0310] Example 34 was prepared by replacing (3-ethyloxetan-3-yl)methanol with (3-(1,1-difluoroethyl)oxetan-3-yl)methanol according to the same procedure as described in Example 32 to obtain the title product (9.5 mg, 17.7%). 1 H NMR (500 MHz, CD3OD) δ 8.13 (s, 1H), 7.23 - 7.20 (m, 1H), 7.03 - 6.99 (m, 1H), 4.80 (s, 2H), 4.78 - 4.77 (m, 2H), 4.71 - 4.70 (m, 2H), 4.59 - 4.56 (m, 1H), 4.48 - 4.46 (m, 1H), 3.73 - 3.65 (m, 4H), 1.83 - 1.71 (m, 7H). MS (ESI, m / e) [M+1] + 667.4.

[0311] Example 35: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-2-((4-(cyanomethyl)tetrahydro-2H-pyran-4-yl)methoxy)-8-fluoro-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chem.

[0312] Example 35 was prepared by replacing methyl tetrahydrofuran-3-carboxylate with ethyl tetrahydro-2H-pyran-4-carboxylate according to the same procedure as described in Example 37 to give the title product (4.5 mg, 19.56% yield). 1 H NMR (300 MHz, CD3OD) δ 8.13 (s, 1H), 7.26 - 7.19 (m, 1H), 7.06 - 6.97 (m, 1H), 4.60 - 4.46 (m, 4H), 3.87 - 3.73 (m, 4H), 3.72 - 3.57 (m, 4H), 2.84 (s, 2H), 1.95 - 1.76 (m, 6H), 1.75 - 1.66 (m, 2H). MS (ESI, m / e) [M+H] + 670.30.

[0313] Example 36: 4-(2-((1-Acetyl-3-fluoroazetidin-3-yl)methoxy)-4-(-3,8-diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chem.

[0314] Step 1: (3-Fluoroazetidin-3-yl)methanol trifluoroacetate

Chem.

[0315] At room temperature, TFA (5 ml) was added to a solution of tert-butyl 3-fluoro-3-(hydroxymethyl)azetidine-1-carboxylate (648 mg, 33 mmol) in DCM (10 ml). The reaction mixture was stirred at room temperature for 1 hour. The solvent was evaporated to dryness and used directly in the next step.

[0316] Example 36 was prepared by replacing (3-methylazetidin-3-yl)methanol hydrochloride with (3-fluoroazetidin-3-yl)methanol trifluoroacetate according to the same procedure as described in Example 8 to obtain the title product (10 mg, formate). 1 H NMR (500 MHz, CD3OD) δ 8.45 (s, 0.5H), 8.13 (s, 1H), 7.23 - 7.20 (m, 1H), 7.04 - 7.00 (m, 1H), 4.82 - 4.81 (m, 2H), 4.71 - 4.69 (m, 1H), 4.59 - 4.50 (m, 2H), 4.42 - 4.38 (m, 1H), 4.29 - 4.24 (m, 1H), 4.17 - 4.10 (m, 1H), 4.08 - 4.02 (m, 2H), 3.87 - 3.75 (m, 2H), 2.08 - 2.00 (m, 4H), 1.92 (s, 3H). MS (ESI, m / e) [M+1] + 662.5.

[0317] Example 37: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-2-((3-(cyanomethyl)tetrahydrofuran-3-yl)methoxy)-8-fluoro-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chem.

[0318] Step 1: Methyl 3-(cyanomethyl)tetrahydrofuran-3-carboxylate

Chem.

[0319] At -78 °C, HMPA (5.9 g, 32.92 mmol) was added to a solution of LDA (4.23 ml, 8.46 mmol) in THF (8 mL), turning the solution red. At -78 °C, a solution of methyl tetrahydrofuran-3-carboxylate (1 g, 7.7 mmol) in THF (2 mL) was added to the solution. The mixture was stirred at -78 °C for 20 minutes. Then, 2-bromoacetonitrile (1.85 g, 15.42 mmol) was added and the reaction was warmed to room temperature. The solution was stirred for an additional 16 hours. At room temperature, the reaction was quenched with saturated NH4Cl(aq.) (20 mL). The resulting mixture was extracted with EtOAc (100 mL x 3). The combined organic layers were washed with brine (100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with petroleum ether / EtOAc (100:0 - 85:15) to afford the title product (600 mg, 46% yield).

[0320] Step 2: 2-(3-(Hydroxymethyl)tetrahydrofuran-3-yl)acetonitrile

Chemical formula

[0321] At 0 °C, LiBH4 (74 mg, 3.40 mmol) was added portionwise to a stirred mixture of methyl 3-(cyanomethyl)tetrahydrofuran-3-carboxylate (250 mg, 1.48 mmol) in THF (3 mL). The resulting mixture was stirred at 80 °C for an additional 3 hours. At room temperature, the reaction was quenched with saturated aqueous NH4Cl solution. The resulting mixture was extracted with EtOAc (50 mL x 3). The combined organic layers were washed with brine (50 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with petroleum ether:EtOAc (100:0 - 68:32) to afford the title product (89 mg, 43% yield).

[0322] Step 3: tert-Butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-2-((3-(cyanomethyl)tetrahydrofuran-3-yl)methoxy)-8-fluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

Chem.

[0323] Under a nitrogen atmosphere at 0 °C, NaH (5.1 mg, 0.21 mmol, 60%) was added portionwise in several times to a stirred solution of 2-(3-(hydroxymethyl)tetrahydrofuran-3-yl)acetonitrile (15 mg, 0.11 mmol) and tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-2,8-difluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (78.0 mg, 0.11 mmol) in THF (2 mL). The resulting mixture was stirred at room temperature for an additional 1 hour. At room temperature, the reaction was quenched with saturated aqueous NH4Cl solution (20 mL). The resulting mixture was extracted with EtOAc (40 mL x 3). The combined organic layers were washed with brine (10 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (petroleum ether:EtOAc = 1:2) to give the title product (70 mg, 77% yield). MS (ESI, m / e) [M+H] + 856.40.

[0324] Step 4: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-2-((3-(cyanomethyl)tetrahydrofuran-3-yl)methoxy)-8-fluoro-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

[0325] At room temperature, tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-2-((3-(cyanomethyl)tetrahydrofuran-3-yl)methoxy)-8-fluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (65 mg, 0.076 mmol), DCM (1.5 mL) and TFA (0.5 mL) were added to an 8 mL vial. The resulting mixture was stirred at room temperature for an additional 2 hours. The resulting mixture was concentrated under reduced pressure. The mixture was basified to pH 9 with saturated aqueous NaHCO3. The resulting mixture was extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (10 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLC (column: Xselect CSH C18 OBD column 30 * 150 mm 5 μm, mobile phase A: water (0.1% formic acid), mobile phase B: CH3CN, flow rate: 60 mL / min, gradient: 15% B - 45% B for 8 minutes) to obtain the title product (10.2 mg, 20.48% yield, formate salt). 1 H NMR (300 MHz, DMSO-d6) δ 8.22(s, 1H), 8.10 (s,3H), 7.29 - 7.25(m, 1H), 7.17 -7.11 (m, 1H), 4.44- 4.30 (m, 4H),3.87 - 3.82 (m,3H), 3.78 - 3.72(m, 1H), 3.69 -3.54 (m, 5H), 2.94- 2.82 (m, 2H),2.00 - 1.93 (m,1H), 1.93 - 1.81(m, 1H), 1.72 -1.52 (m, 4H). MS(ESI, m / e) [M+H] + 656.10.

[0326] Example 38: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((3-methyl-1-(methylsulfonyl)azetidin-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chemical Structure

[0327] Step 1: (3-Methyl-1-(methylsulfonyl)azetidin-3-yl)methanol

Chemical Structure

[0328] At 0 °C, methanesulfonyl chloride (0.23 mL, 2.91 mmol), anhydrous pyridine (0.47 mL, 5.82 mmol), and DMAP (356 mg, 2.91 mmol) were added dropwise to a solution of (3-methylazetidin-3-yl)methanol hydrochloride (400 mg, 2.91 mmol) in DCM (10 mL). Then, the reaction mixture was warmed to room temperature for 16 h. After completion, the reaction mixture was concentrated to obtain a crude product. The residue was purified by silica gel column chromatography, eluting with 50% ethyl acetate in petroleum ether to 2% methanol in DCM, to obtain the crude title product (360 mg, 1.0 mmol, purity = 50%).

[0329] Example 38 was prepared by replacing 1-(3-(hydroxymethyl)-3-methylazetidin-1-yl)ethan-1-one with (3-methyl-1-(methylsulfonyl)azetidin-3-yl)methanol by the same procedure as described in Example 13 to obtain the title product (34 mg). 11H NMR (500 MHz, CD3OD) δ 8.12 (s, 1H), 7.23 - 7.20 (m, 1H), 7.03 - 7.00 (m, 1H), 4.66 - 4.46 (m, 4H), 4.00 - 3.98 (m, 4H), 3.83 - 3.75 (m, 2H), 3.68 - 3.67 (m, 2H), 3.01 (s, 3H), 2.00 (s, 4H), 1.46 (s, 3H). MS (ESI, m / e) [M + 1] + 694.5.

[0330] Example 39: 4-(2-((1-Acetyl-3-methoxyazetidin-3-yl)methoxy)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chemical Structure

[0331] Example 39 was prepared by replacing (3-methoxytetrahydrofuran-3-yl)methanol with 1-(3-(hydroxymethyl)-3-methoxyazetidin-1-yl)ethan-1-one according to the same procedure as described in Example 3 to obtain the title product (28.74 mg). 1 1H NMR (500 MHz, CD3OD) δ 8.13 (s, 1H), 7.22 - 7.20 (m, 1H), 7.03 - 6.99 (m, 1H), 4.78 - 4.71 (m, 2H), 4.59 - 4.56 (m, 1H), 4.49 - 4.47 (m, 1H), 4.31 - 4.22 (m, 2H), 4.04 - 3.97 (m, 2H), 3.73 - 3.65 (m, 4H), 3.40 (s, 3H), 1.90 (s, 3H), 1.90 - 1.78 (m, 4H). MS (ESI, m / e) [M + H] + 674.4.

[0332] Example 40: 4-(2-((2-Acetyl-2-azabicyclo[2.1.1]hexan-4-yl)methoxy)-4-(3,8-diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chem.

[0333] Example 40 was prepared by replacing (3-methoxytetrahydrofuran-3-yl)methanol with 1-(4-(hydroxymethyl)-2-azabicyclo[2.1.1]hexan-2-yl)ethan-1-one according to the same procedure as described in Example 3 to obtain the title product (19.66 mg). 1 H NMR (500 MHz, CD3OD) δ 8.12 (s, 1H), 7.22 - 7.20 (m, 1H), 7.03 - 6.99 (m, 1H), 4.76 - 4.68 (m, 3H), 4.59 - 4.48 (m, 2H), 3.75 - 3.66 (m, 4H), 3.59 (s, 1H), 3.43 (s, 1H), 2.13 - 2.04 (m, 5H), 1.92 - 1.87 (m, 4H), 1.64 - 1.57 (m, 2H). MS (ESI, m / e) [M+H] + 671.0.

[0334] Example 41: 4-(2-((2-Oxabicyclo[2.1.1]hexan-4-yl)methoxy)-8-fluoro-4-((1S,4R)-2-((2-methoxyethyl)amino)-7-azabicyclo[2.2.1]heptan-7-yl)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chem.

[0335] Step 1: (1S,4R)-2-((2-Methoxyethyl)amino)-7-azabicyclo[2.2.1]heptane-7-carboxylic acid tert-butyl

Chem.

[0336] A solution of (1S,4R)-2-oxo-7-azabicyclo[2.2.1]heptane-7-carboxylic acid tert-butyl (2.11 g, 10 mmol) and 2-methoxyethan-1-amine (750 mg, 10 mmol) in methanol (40 mL) was stirred at 50 °C for 15 h. Then, NaBH4 (1520 mg, 40 mmol) was added at 0 °C and the reaction mixture was stirred at 25 °C for 3 h. After completion, the reaction mixture was diluted with EtOAc (50 mL) and washed with saturated NaCl aqueous solution (15 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to obtain a residue. The residue was purified by silica gel (petroleum ether:EtOAc = 1:1) to obtain the title product (2.3 g, 85.2%). MS (ESI, m / e) [M+1] + 271.5.

[0337] Step 2: (1S,4R)-2-(((Benzyloxy)carbonyl)(2-methoxyethyl)amino)-7-azabicyclo[2.2.1]heptane-7-carboxylic acid tert-butyl

Chem.

[0338] A mixture of tert-butyl (1S,4R)-2-((2-methoxyethyl)amino)-7-azabicyclo[2.2.1]heptane-7-carboxylate (1.68 g, 6.0 mmol), CbzCl (1.02 g, 6.0 mmol) and DIPEA (1.55 g, 12.0 mmol) in DCM (20 mL) was stirred at 25 °C for 3 h. After completion, the reaction mixture was diluted with EtOAc (40 mL) and washed with saturated aqueous NaCl solution (15 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to give a residue. The residue was purified by silica gel (petroleum ether:EtOAc = 5:1) to give the title product (1.6 g, 66.1%). MS (ESI, m / e) [M+1] + 405.5.

[0339] Step 3: Benzyl ((1S,4R)-7-azabicyclo[2.2.1]heptan-2-yl)(2-methoxyethyl)carbamate hydrochloride

Chemical formula

[0340] A solution of tert-butyl (1S,4R)-2-(((benzyloxy)carbonyl)(2-methoxyethyl)amino)-7-azabicyclo[2.2.1]heptane-7-carboxylate (606 mg, 1.5 mmol) in HCl / dioxane solution (4N, 10 mL) was stirred at room temperature for 4 h. After completion, the reaction mixture was concentrated. The residue was washed with ethyl ether and used directly in the next step without further purification. MS (ESI, m / e) [M+1] + 305.4.

[0341] Step 4: Benzyl ((1S,4R)-7-(7-bromo-2-chloro-8-fluoro-6-(trifluoromethyl)quinazolin-4-yl)-7-azabicyclo[2.2.1]heptan-2-yl)(2-methoxyethyl)carbamate

Chemical formula

[0342] Under a nitrogen atmosphere, a mixture of 7-bromo-2,4-dichloro-8-fluoro-6-(trifluoromethyl)quinazoline (364 mg, 1.0 mmol), benzyl ((1S,4R)-7-azabicyclo[2.2.1]heptan-2-yl)(2-methoxyethyl)carbamate hydrochloride (340 mg, 1.0 mmol), and DIPEA (516 mg, 4.0 mmol) in DCM (10 mL) was stirred at 25 °C for 1 hour. After completion, the reaction mixture was diluted with DCM (50 mL) and washed with water (15 mL × 3). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to obtain a residue. The residue was purified by silica gel (petroleum ether:EtOAc = 3:1) to obtain the title product (444 mg, 70.5%). MS (ESI, m / e) [M+1] + 631.4.

[0343] Step 5: Benzyl ((1S,4R)-7-(2-((2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-7-bromo-8-fluoro-6-(trifluoromethyl)quinazolin-4-yl)-7-azabicyclo[2.2.1]heptan-2-yl)(2-methoxyethyl)carbamate

Chem.

[0344] A solution of benzyl ((1S,4R)-7-(7-bromo-2-chloro-8-fluoro-6-(trifluoromethyl)quinazolin-4-yl)-7-azabicyclo[2.2.1]heptan-2-yl)(2-methoxyethyl)carbamate (444 mg, 0.704 mmol), (2-oxabicyclo[2.1.1]hexan-4-yl)methanol (160 mg, 1.4 mmol) and KF (204 mg, 3.5 mmol) in DMSO (15 mL) was stirred at 100 °C for 4 h. After completion, the reaction mixture was diluted with EtOAc (50 mL) and washed with water (15 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to give a residue. The residue was purified by silica gel (petroleum ether:EtOAc = 3:1) to give the title product (290 mg, 58.0%). MS (ESI, m / e) [M+1] + 709.5,711.5.

[0345] Step 6: ((1S,4R)-7-(2-((2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-6-(trifluoromethyl)quinazolin-4-yl)-7-azabicyclo[2.2.1]heptan-2-yl)(2-methoxyethyl)carbamate

Chem.

[0346] Benzyl ((1S,4R)-7-(2-((2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-7-bromo-8-fluoro-6-(trifluoromethyl)quinazolin-4-yl)-7-azabicyclo[2.2.1]heptan-2-yl)(2-methoxyethyl)carbamate (213 mg, 0.3 mmol), tert-butyl (3-cyano-7-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[b]thiophen-2-yl)carbamate (502 mg, 1.2 mmol), DPEphosPdCl2 (107 mg, 0.15 mmol), and Cs2CO3 (293 mg, 0.9 mmol) in toluene (50 mL) were stirred at 80 °C for 4 h. After completion, the reaction mixture was concentrated and purified by silica gel (petroleum ether:EtOAc = 3:1) to give the title product (110 mg, 40%). MS (ESI, m / e) [M+1] + 921.5.

[0347] Step 7: tert-Butyl (4-(2-((2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-8-fluoro-4-((1S,4R)-2-((2-methoxyethyl)amino)-7-azabicyclo[2.2.1]heptan-7-yl)-6-(trifluoromethyl)quinazolin-7-yl)-3-cyano-7-fluorobenzo[b]thiophen-2-yl)carbamate

Chem.

[0348] Under an H2 atmosphere, a mixture of ((1S,4R)-7-(2-((2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-6-(trifluoromethyl)quinazolin-4-yl)-7-azabicyclo[2.2.1]heptan-2-yl)(2-methoxyethyl)carbamic acid benzyl (100 mg, 0.109 mmol) and 10% Pd / C (100 mg) in methanol (10 mL) was stirred at 20 °C for 4 hours. After completion, the reaction mixture was filtered and used directly in the next step without further purification. MS (ESI, m / e) [M+1] + 787.5.

[0349] Step 8: 4-(2-((2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-8-fluoro-4-((1S,4R)-2-((2-methoxyethyl)amino)-7-azabicyclo[2.2.1]heptan-7-yl)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

[0350] At room temperature, TFA (4 mL) was added to a solution of tert-butyl (4-(2-((2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-8-fluoro-4-((1S,4R)-2-((2-methoxyethyl)amino)-7-azabicyclo[2.2.1]heptan-7-yl)-6-(trifluoromethyl)quinazolin-7-yl)-3-cyano-7-fluorobenzo[b]thiophen-2-yl)carbamate (70 mg, 0.089 mmol) in DCM (4 mL). The resulting mixture was stirred at room temperature for 3 hours. After completion, the reaction mixture was concentrated to obtain a residue. The residue was purified by prep-HPLC to obtain the title product (2.0 mg, 3.3%). 11H NMR (500 MHz, CD3OD) δ 8.28 (s, 1H), 7.24 - 7.20 (m, 1H), 7.04 - 7.00 (m, 1H), 5.10 - 5.06 (m, 1H), 5.03 - 4.96 (m, 1H), 4.83 - 4.79 (m, 2H), 4.57 (s, 2H), 3.73 (s, 3H), 3.65 - 3.62 (m, 2H), 3.44 - 3.40 (m, 4H), 2.25 - 1.86 (m, 6H), 1.70 - 1.53 (m, 4H). MS (ESI, m / e) [M+1] + 687.4.

[0351] Example 42: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)quinazolin-7-yl)naphthalen-2-ol

Chemical Structure

[0352] Step 1: tert-Butyl 3-(7-bromo-8-fluoro-2-((1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

Chemical Structure

[0353] A solution of (1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methanol (85 mg, 0.66 mmol) in THF (5 mL) at 0 °C was added sodium hydride (27 mg, 0.66 mmol). The reaction mixture was stirred at 0 °C for 0.5 h. Then tert-butyl 3-(7-bromo-2,8-difluoroquinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (200 mg, 0.44 mmol) was added to the reaction mixture and stirred at room temperature for 2 h. After completion, the reaction mixture was quenched with water (10 mL), extracted with EtOAc (30 mL x 3), and washed with saturated aqueous NaCl solution. The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to give the crude title product (240 mg, 0.43 mmol). MS (ESI, m / e) [M+1] + 563.1.

[0354] Step 2: tert-Butyl 3-(8-fluoro-7-(3-(methoxymethoxy)naphthalen-1-yl)-2-((1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate [Chemical formula]

[0355] At room temperature, to a solution of tert-butyl 3-(7-bromo-8-fluoro-2-((1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (100 mg, 0.18 mmol), 2-(3-(methoxymethoxy)naphthalen-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (110 mg, 0.36 mmol) and PdCl2(dtbpf) (46 mg, 0.07 mmol) in 1,4-dioxane (3 mL) and H2O (0.6 mL) was added K3PO4 (115 mg, 0.54 mmol). The reaction mixture was then stirred at 90 °C for 2 h. After completion, the reaction mixture was quenched with water (30 mL), extracted with EtOAc (30 mL x 3), and washed with saturated aqueous NaCl solution. The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to give a residue. The residue was purified by silica gel column chromatography, eluting with 0% - 60% ethyl acetate in petroleum ether to give the title product (90 mg, 0.13 mmol). MS (ESI, m / e) [M+1] + 671.3.

[0356] Step 3: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)quinazolin-7-yl)naphthalen-2-ol

[0357] At 0 °C, TFA (4.5 mL) was added to a solution of tert-butyl 3-(8-fluoro-7-(3-(methoxymethoxy)naphthalen-1-yl)-2-((1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (70 mg, 0.10 mmol) in DCM (24 mL). The resulting mixture was stirred at room temperature for 2 h. After completion, the reaction mixture was concentrated to give a residue. It was dissolved in 3 mL of DCM and adjusted to pH ~10 using 7N NH3 in MeOH. After concentration, the residue was purified by pre-HPLC to give the title product (3 mg, formate). 1 H NMR (500 MHz, CD3OD) δ 8.50 (s, 0.5H), 7.87 - 7.73 (m, 2H), 7.46 - 7.38 (m, 2H), 7.36 - 7.28 (m, 1H), 7.26 - 7.17 (m, 2H), 7.09 (s, 1H), 4.74 (s, 2H), 4.63 - 4.55 (m, 2H), 3.96 (s, 2H), 3.80 (s, 2H), 3.70 - 3.67 (m, 2H), 2.06 - 2.04 (m, 4H), 1.82 - 1.80 (m, 2H), 1.65 - 1.62 (m, 2H), 1.43 (s, 3H). MS (ESI, m / e) [M+1]+ 527.4.

[0358] Example 43: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-2-((4-(difluoromethyl)tetrahydro-2H-pyran-4-yl)methoxy)-8-fluoro-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chemical Structure

[0359] Example 43 was prepared by replacing 3-(hydroxymethyl)tetrahydrofuran-3-ol with (4-(difluoromethyl)tetrahydro-2H-pyran-4-yl)methanol according to the same procedure as described in Example 6 to obtain the title product (12 mg, 26.7% yield). 1 H NMR (500 MHz, CD3OD) δ 8.12 (s, 1H), 7.25 - 7.21 (m, 1H), 7.04 - 7.01 (m, 1H), 6.08 - 5.86 (m, 1H), 4.85 - 4.62 (m, 4H), 4.20 - 4.12 (m, 2H), 3.90 - 3.69 (m, 6H), 2.14 - 2.03 (m, 4H), 1.90 - 1.85 (m, 2H), 1.72 - 1.69 (m, 2H). MS (ESI, m / e) [M+1] + 681.6.

[0360] Example 44: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((3-fluorooxetan-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chem.

[0361] Step 1: tert-Butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-2-((3-fluorooxetan-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

Chem.

[0362] To a sealed tube (20 mL) was added (3-fluorooxetan-3-yl)methanol (70 mg, 0.660 mmol), tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-2,8-difluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (50 mg, 0.068 mmol), THF (6 mL) and NaH (8 mg, 0.2 mmol, 60%). The mixture was stirred at room temperature for 16 h. The reaction was quenched with saturated aqueous NH4Cl (20 mL) and extracted with EtOAc (40 mL x 3). The combined organic phases were dried over anhydrous Na2SO4 and concentrated in vacuo to give the crude title product (40 mg, 89.6% yield). MS (ESI, m / e) [M+H] + 821.5.

[0363] Step 2: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((3-fluorooxetan-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

[0364] To a 25 mL round-bottom flask were added tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-2-((3-fluorooxetan-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (40 mg, 0.049 mmol), TFA (4 mL), and DCM (2 mL). The reaction mixture was stirred at room temperature for 16 h. The reaction mixture was concentrated under vacuum. The residue was dissolved in EtOAc (40 mL), and saturated aqueous Na2CO3 solution (20 mL) was added. The solution was stirred at room temperature for 10 min and extracted with EtOAc (40 mL x 2). The combined organic phases were washed with brine (20 mL), dried over anhydrous Na2SO4, and then concentrated under vacuum. The residue was purified by Prep-HPLC (column: XBridge Shield RP18 OBD column, mobile phase A: water (10 mmol / L NH4HCO3 + 0.1% NH3 . H2O), mobile phase B: CH3CN, gradient: 20% B - 75% B for 9 min) to give the title product (7 mg, 23.0% yield). 1 1H NMR (500 MHz, CD3OD) δ 8.13 (s, 1H), 7.25 - 7.19 (m, 1H), 7.03 - 7.00 (m, 1H), 4.78 - 4.58 (m, 7H), 3.99 (s, 2H), 3.86 - 3.77 (m, 3H), 2.00 (s, 4H). MS (ESI, m / e) [M+H] + 621.3.

[0365] Example 45: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((1-methoxycyclopropyl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chemical Structure

[0366] Example 45 was prepared by replacing (3-fluorooxetan-3-yl)methanol with (1-methoxycyclopropyl)methanol according to the same procedure as described in Example 44 to give the title product (11.8 mg, 39.0% yield). 1 H NMR (500 MHz, CD3OD) δ 8.11 (s, 1H), 7.23 - 7.22 (m, 1H), 7.04 - 7.00 (m, 1H), 4.68 - 4.59 (m, 4H), 4.23 (s, 2H), 3.92 - 3.83 (m, 2H), 3.41 (s, 3H), 2.14 - 2.13 (m, 4H), 0.92 (s, 2H), 0.80 (s, 2H). MS (ESI, m / e) [M+H] + 617.4.

[0367] Example 46: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((3-methyl-1-(oxazol-2-yl)azetidin-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chemical formula

[0368] Example 46 was prepared by replacing azetidin-3-ylmethanol hydrochloride and 2-iodo-1-methyl-1H-imidazole with (3-methylazetidin-3-yl)methanol hydrochloride and 2-iodooxazole according to the same procedure as described in Example 24 to give the title product (12 mg, 26.7% yield). 11H NMR (500 MHz, CD3OD) δ 8.13 (s, 1H), 7.53 (s, 1H), 7.22 - 7.19 (m, 1H), 7.11 - 7.08 (m, 1H), 7.04 - 7.01 (m, 1H), 4.77 - 4.74 (m, 1H), 4.67 - 4.64 (m, 1H), 4.59 - 4.54 (m, 2H), 4.30 - 4.24 (m, 4H), 4.01 - 3.99 (m, 2H), 3.91 - 3.80 (m, 2H), 2.20 - 2.07 (m, 4H), 1.51 (s, 3H). MS (ESI, m / e) [M+1] + 683.6.

[0369] Example 47: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-2-((1-(cyclopropoxymethyl)-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-8-fluoro-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chemical formula

[0370] Example 47 was prepared by replacing sodium methoxide with sodium cyclopropoxide according to the same procedure as described in Example 11 to obtain the title product (1 mg, 16.1% yield). 11H NMR (500 MHz, CD3OD) δ 8.12 (s, 1H), 7.28 - 7.13 (m, 1H), 7.02 - 6.99 (m, 1H), 4.81 - 4.69 (m, 2H), 4.63 - 4.54 (m, 2H), 3.94 - 3.88 (m, 2H), 3.82 - 3.80 (m, 2H), 3.79 - 3.76 (m, 1H), 3.75 - 3.73 (m, 2H), 3.44 - 3.39 (m, 2H), 2.03 - 1.92 (m, 4H), 1.91 - 1.84 (m, 2H), 1.72 - 1.60 (m, 2H), 0.60 - 0.50 (m, 2H), 0.50 - 0.41 (m, 2H). MS (ESI, m / e) [M+1] + 699.6.

[0371] Example 48: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-2-((3-(difluoromethyl)oxetan-3-yl)methoxy)-8-fluoro-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chemical Structure

[0372] Example 48 was prepared by replacing (3-ethyloxetan-3-yl)methanol with (3-(difluoromethyl)oxetan-3-yl)methanol according to the same procedure as described in Example 32 to obtain the title product (101 mg), which was further separated by chiral HPLC (Prep-HPLC instrument: Prep-HPLC-Gilson, column: IE, 20 mm × 250 mm, 5 μm, mobile phase A: n-hexane, mobile phase B: EtOH (0.2% 2M NH3 in MeOH), flow rate: 18 mL / min, temperature: 25 °C, gradient: 40% B for 14 minutes, retention time - isomer 1: 6.5 minutes, retention time - isomer 2: 11.5 minutes) to obtain several atropisomers. Isomer 1 (42.0 mg, 30.4%): 11H NMR (500 MHz, CD3OD) δ 8.13 (s, 1H), 7.23 - 7.20 (m, 1H), 7.03 - 6.99 (m, 1H), 6.37 - 6.15 (m, 1H), 4.81 (s, 2H), 4.78 - 4.77 (m, 2H), 4.70 - 4.65 (m, 2H), 4.59 - 4.57 (m, 1H), 4.50 - 4.47 (m, 1H), 3.74 - 3.66 (m, 4H), 1.90 - 1.81 (m, 4H); MS (ESI, m / e) [M+1] + 653.7. Isomer 2 (40.0 mg, 29.0%): 1 1H NMR (500 MHz, CD3OD) δ 8.13 (s, 1H), 7.23 - 7.20 (m, 1H), 7.03 - 6.99 (m, 1H), 6.38 - 6.15 (m, 1H), 4.80 (s, 2H), 4.78 - 4.77 (m, 2H), 4.70 - 4.65 (m, 2H), 4.58 - 4.56 (m, 1H), 4.49 - 4.47 (m, 1H), 3.73 - 3.65 (m, 4H), 1.90 - 1.81 (m, 4H); MS (ESI, m / e) [M+1] + 653.6.

[0373] Example 49: 3 - ((((7 - (2 - Amino - 3 - cyano - 7 - fluorobenzo[b]thiophen - 4 - yl) - 4 - (3,8 - diazabicyclo[3.2.1]octan - 3 - yl) - 8 - fluoro - 6 - (trifluoromethyl)quinazolin - 2 - yl)oxy)methyl)oxetan - 3 - carbonitrile [Chemical Structure Diagram]

[0374] Example 49 was prepared by replacing (3 - methoxytetrahydrofuran - 3 - yl)methanol with 3 - (hydroxymethyl)oxetan - 3 - carbonitrile in the same procedure as described in Example 3 to obtain the title product (24.91 mg). 11H NMR (500 MHz, CD3OD) δ 8.14 (s, 1H), 7.23 - 7.20 (m, 1H), 7.03 - 6.99 (m, 1H), 4.97 - 4.95 (m, 4H), 4.78 - 4.77 (m, 2H), 4.61 - 4.58 (m, 1H), 4.51 - 4.48 (m, 1H), 3.74 - 3.66 (m, 4H), 1.89 - 1.78 (m, 4H). MS (ESI, m / e) [M+H] + 628.4.

[0375] Example 50: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((4-(hydroxymethyl)tetrahydro-2H-pyran-4-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chem.

[0376] Step 1: tert-Butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-2-((4-(hydroxymethyl)tetrahydro-2H-pyran-4-yl)methoxy)-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

Chem.

[0377] At room temperature, sodium hydride (15 mg, 0.60 mmol) was added to a solution of (tetrahydro-2H-pyran-4,4-diyl)dimethanol (44 mg, 0.3 mmol) in THF (10 mL). The resulting mixture was stirred at room temperature for 1 hour. Then, tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-2,8-difluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (73.4 mg, 0.10 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature for 1 hour. After completion, the reaction mixture was diluted with EtOAc (50 mL) and washed with saturated aqueous NaCl solution (15 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to obtain a residue. The residue was purified by Prep-TLC (petroleum ether:EtOAc = 1:1) to obtain the title product (40 mg, 46.5%). MS (ESI, m / e) [M+1] + 860.5.

[0378] Step 2: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((4-(hydroxymethyl)tetrahydro-2H-pyran-4-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

[0379] At room temperature, TFA (2 mL) was added to a solution of tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-2-((4-(hydroxymethyl)tetrahydro-2H-pyran-4-yl)methoxy)-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (40 mg, 0.0465 mmol) in DCM (2 mL). The resulting mixture was stirred at room temperature for 3 hours. After completion, the reaction mixture was concentrated to obtain a residue. The residue was purified by prep-HPLC to obtain the title product (15.0 mg, 48.9%).1 1H NMR (500 MHz, CD3OD) δ 8.11 (s, 1H), 7.24 - 7.19 (m, 1H), 7.04 - 7.00 (m, 1H), 4.71 - 4.68 (m, 1H), 4.60 - 4.58 (m, 1H), 4.51 - 4.45 (m, 2H), 4.06 - 4.02 (m, 2H), 3.90 - 3.70 (m, 6H), 3.64 (s, 2H), 2.09 - 2.01 (m, 4H), 1.69 - 1.58 (m, 4H). MS (ESI, m / e) [M+1] + 661.5.

[0380] Example 51: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-2-((3-(cyanomethyl)oxetan-3-yl)methoxy)-8-fluoro-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chemical Structure

[0381] Example 51 was prepared by replacing 3-(hydroxymethyl)tetrahydrofuran-3-ol with 2-(3-(hydroxymethyl)oxetan-3-yl)acetonitrile according to the same procedure as described in Example 6 to obtain the title product (25 mg, 40.2% yield, formate). 1 1H NMR (500 MHz, CD3OD) δ 8.47 (s, 1H), 8.13 (s, 1H), 7.23 - 7.20 (m, 1H), 7.04 - 7.00 (m, 1H), 4.79 - 4.75 (m, 2H), 4.70 - 4.68 (m, 3H), 4.63 - 4.61 (m, 3H), 4.01 - 3.95 (m, 2H), 3.85 - 3.77 (m, 2H), 3.06 (s, 2H), 2.04 - 1.95 (m, 4H). MS (ESI, m / e) [M+1] + 642.5.

[0382] Example 52: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-6-ethyl-8-fluoro-2-((1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile [Chemical formula]

[0383] Step 1: tert-Butyl 3-(7-bromo-2-chloro-8-fluoro-6-iodoquinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate [Chemical formula]

[0384] To a mixture of 7-bromo-2,4-dichloro-8-fluoro-6-iodoquinazoline (0.2 g, 0.47 mmol) and DIPEA (0.12 g, 0.94 mmol) in DCM (10 mL) cooled to 0 °C, tert-butyl 3,8-diazabicyclo[3.2.1]octane-8-carboxylate was added. The mixture was stirred at 0 °C for 2 hours, diluted with DCM (10 mL), the operation was repeated 3 times, washed with brine (10 mL), filtered, the filtrate was concentrated, and purified by a silica chromatography column to obtain the title product (0.23 g, 82.1%). MS (ESI, m / e) [M+1] + 598.4.

[0385] Step 2: tert-Butyl 3-(7-bromo-2,8-difluoro-6-iodoquinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate [Chemical formula]

[0386] A solution of tert-butyl 3-(7-bromo-2-chloro-8-fluoro-6-iodoquinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (0.1 g, 0.16 mmol) in DMSO (5 mL) was added with KF (0.039 g, 0.64 mmol) and stirred at 90 °C for 5 - 6 h. The resulting mixture was cooled to room temperature, diluted with EtOAc (10 mL), the operation was repeated 3 times, washed with brine (10 mL), and filtered. The filtrate was concentrated and purified by a silica chromatography column to obtain the title product (0.08 g, 82.4%). MS (ESI, m / e) [M+1] + 581.3.

[0387] Step 3: tert-Butyl 3-(7-bromo-2,8-difluoro-6-vinylquinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

Chemical Structure

[0388] To a mixture of tert-butyl 3-(7-bromo-2,8-difluoro-6-iodoquinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (620 mg, 1.06 mmol), trifluoro(vinyl)-l4-borane, potassium salt (428 mg, 3.2 mmol), NaHCO3 (268 mg, 3.2 mmol), dioxane (20 mL) and H2O (2 mL) was added Pd(dppf)Cl2DCM (173 mg, 0.21 mmol), degassed 3 times with N2, stirred at 70 °C for 2 h, the resulting mixture was filtered, the filtrate was concentrated and purified by column chromatography to obtain the title product (450 mg, 87.8%). MS (ESI, m / e) [M+H] + 481.1.

[0389] Step 4: tert-Butyl 3-(7-bromo-6-ethyl-2,8-difluoroquinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate [Chemistry]

[0390] To a solution of tert-butyl 3-(7-bromo-2,8-difluoro-6-vinylquinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (250 mg, 0.51 mmol) in EtOAc (15 mL) was added Pt / C (10%) (25 mg). The mixture was stirred at room temperature for 48 h, and the resulting mixture was filtered. The filtrate was concentrated and purified by column chromatography to give the title product (160 mg, 64.2%). MS (ESI, m / e) [M+H] + 483.1.

[0391] Step 5: tert-Butyl 3-(7-bromo-6-ethyl-8-fluoro-2-((1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate [Chemistry]

[0392] A solution of (1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methanol (100 mg, 0.78 mmol) in THF (30 mL) was cooled to 0 °C, and NaH (29 mg, 0.74 mmol) was added in one portion. The mixture was stirred at 0 °C for 30 min and then added to tert-butyl 3-(7-bromo-6-ethyl-2,8-difluoroquinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (160 mg, 0.33 mmol). The resulting mixture was stirred at room temperature for 4 - 5 h, then quenched with H2O (20 mL), extracted three times with EtOAc (30 mL), washed with brine (20 mL), dried over Na2SO4, filtered, and the filtrate was concentrated and purified by column chromatography to give the title product (150 mg, 76.9%). MS (ESI, m / e) [M+H] + 591.3.

[0393] Step 6: tert-Butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-6-ethyl-8-fluoro-2-((1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate [Chemical formula]

[0394] To a mixture of tert-butyl 3-(7-bromo-6-ethyl-8-fluoro-2-((1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (50 mg, 0.084 mmol), tert-butyl (3-cyano-7-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[b]thiophen-2-yl)carbamate (140 mg, 0.034 mmol), Cs2CO3 (82 mg, 0.25 mmol), and DPEphosPdCl2 (12 mg, 0.017), toluene (10 mL) was added. The mixture was degassed three times with N2 and stirred at 100 °C for 5 hours. The resulting mixture was filtered, the filtrate was concentrated, and purified by column chromatography to obtain the title product (25 mg, 36.7%). MS (ESI, m / e) [M+H] + 803.6.

[0395] Step 7: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-6-ethyl-8-fluoro-2-((1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

[0396] At room temperature, TFA (1 mL) was added to a solution of tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-6-ethyl-8-fluoro-2-((1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (80 mg, 0.1 mmol) in DCM (5 mL). The resulting mixture was stirred at room temperature for 2 hours. After completion, the reaction mixture was concentrated to obtain a residue. The residue was purified by prep-HPLC to obtain the title product (3.0 mg, 5.0%, formate). 1 H NMR (500 MHz, CD3OD) δ 8.49 (s, 0.5H), 7.61 (s, 1H), 7.18 - 7.15 (m, 1H), 7.05 - 7.02 (m, 1H), 4.74 - 4.68 (m, 2H), 4.58 - 4.56 (m, 1H), 4.52 - 4.49 (m, 1H), 4.00 (s, 2H), 3.79 (s, 2H), 3.73 - 3.66 (m, 2H), 2.62 - 2.42 (m, 2H), 2.08 - 2.04 (m, 4H), 1.81 - 1.80 (m, 2H), 1.62 - 1.61 (m, 2H), 1.42 (s, 3H), 1.10 - 1.07 (m, 3H). MS (ESI, m / e) [M+1] + 603.4.

[0397] Example 53: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((3-(fluoromethyl)oxetan-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chem.

[0398] Example 53 was prepared by replacing 3-(hydroxymethyl)tetrahydrofuran-3-ol with (3-(fluoromethyl)oxetan-3-yl)methanol according to the same procedure as described in Example 6 to obtain the title product (17 mg, 36.2% yield). 1 H NMR (500 MHz, CD3OD) δ 8.13 (s, 1H), 7.23 - 7.20 (m, 1H), 7.04 - 6.99 (m, 1H), 4.83 - 4.81 (m, 1H), 4.75 - 4.71 (m, 3H), 4.69 - 4.66 (m, 2H), 4.63 - 4.60 (m, 3H), 4.54 - 4.51 (m, 1H), 3.81 - 3.70 (m, 4H), 1.97 - 1.83 (m, 4H). MS (ESI, m / e) [M+1] + 635.5.

[0399] Example 54: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((3-methyl-1-(1-methyl-1H-imidazol-2-yl)azetidin-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chemical Structure

[0400] Example 54 was prepared by replacing azetidin-3-ylmethanol hydrochloride with (3-methylazetidin-3-yl)methanol hydrochloride according to the same procedure as described in Example 46 to obtain the title product (4 mg, 16.7% yield). 11H NMR (500 MHz, CD3OD) δ 8.12 (s, 1H), 7.22 - 7.20 (m, 1H), 7.03 - 7.00 (m, 1H), 6.77 - 6.73 (m, 2H), 4.67 - 4.55 (m, 4H), 4.22 - 4.19 (m, 2H), 3.97 - 3.89 (m, 4H), 3.83 - 3.75 (m, 2H), 3.50 (s, 3H), 2.00 - 1.92 (m, 4H), 1.51 (s, 3H). MS (ESI, m / e) [M+1] + 696.5.

[0401] Example 55: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((6-methyl-2-oxaspiro[3.3]heptan-6-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chemical formula

[0402] Example 55 was prepared by replacing (1-(difluoromethyl)-2-oxabicyclo[2.1.1]hexan-4-yl)methanol with (6-methyl-2-oxaspiro[3.3]heptan-6-yl)methanol according to the same procedure as described in Example 1 (Steps 6 and 7), to give the title product (15 mg, 48.3%). 1 1H NMR (500 MHz, CD3OD) δ 8.10 (s, 1H), 7.21 - 7.20 (m, 1H), 7.03 - 6.99 (m, 1H), 4.74 - 4.71 (m, 4H), 4.59 - 4.57 (m, 1H), 4.49 - 4.46 (m, 1H), 4.22 (s, 2H), 3.79 - 3.67 (m, 4H), 2.40 - 2.37 (m, 2H), 2.07 - 2.05 (m, 2H), 1.89 (s, 4H), 1.19 (s, 3H). MS (ESI, m / e) [M+H] + 657.5

[0403] Example 56: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((1-(methoxymethyl)-3,3-dimethyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chem.

[0404] Step 1: Methyl 1-(2-hydroxypropan-2-yl)-3-methylenecyclobutane-1-carboxylate

Chem.

[0405] At -78 °C, n-BuLi (4.4 mL, 2.5 N, 0.011 mol) was added dropwise to a mixture of diisopropylamine (1.11 g, 0.011 mol) in THF (50 mL). Then it was warmed to 0 °C for 30 minutes. After re-cooling to -78 °C, methyl 3-methylenecyclobutane-1-carboxylate (1.27 g, 0.01 mol) was added to the reaction mixture and stirred at -78 °C for 1 hour. Then, acetone (1.17 g, 0.02 mol) was added dropwise to the reaction mixture and warmed to room temperature for 2 hours. After completion, the reaction mixture was quenched with 50 mL of H2O and extracted with EtOAc (50 mL x 3). The combined organic layers were washed with saturated aqueous NaCl solution (100 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to obtain the crude title product (1.15 g, 6.25 mmol). 1 H NMR (500 MHz, CDCl3) δ 4.84 (s, 2H), 3.77 (s, 3H), 3.11 (s, 1H), 3.07 - 3.04 (m, 2H), 2.93 - 2.90 (m, 2H), 1.24 (s, 6H).

[0406] Step 2: Methyl 1-(iodomethyl)-3,3-dimethyl-2-oxabicyclo[2.1.1]hexane-4-carboxylate [Chemical formula]

[0407] At room temperature, iodine (1.79 g, 7.06 mmol) and sodium bicarbonate (594 mg, 7.06 mmol) were added to a mixture of methyl 1-(2-hydroxypropan-2-yl)-3-methylenecyclobutane-1-carboxylate (650 mg, 3.53 mmol) in tert-butyl methyl ether (20 mL) and H2O (10 mL). Then it was stirred at room temperature for 4 hours. After completion, the reaction mixture was diluted with tert-butyl methyl ether (100 mL) and quenched with a saturated aqueous solution of Na2S2O3 (100 mL). The combined organic layers were washed with a saturated aqueous solution of NaCl (100 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to obtain the crude title product (720 mg, 2.32 mmol). 1 1H NMR (500 MHz, CDCl3) δ 3.72 (s, 3H), 3.38 (s, 2H), 2.23 - 2.22 (m, 2H), 2.05 - 2.04 (m, 2H), 1.43 (s, 6H).

[0408] Step 3: 1-(Methoxymethyl)-3,3-dimethyl-2-oxabicyclo[2.1.1]hexane-4-carboxylic acid [Chemical formula]

[0409] A mixture of methyl 1-(iodomethyl)-3,3-dimethyl-2-oxabicyclo[2.1.1]hexane-4-carboxylate (600 mg, 1.94 mmol) in HPLC grade MeOH (30 mL) at 0 °C was added sodium methoxide (3.14 g, 58.2 mmol). Then it was heated to reflux for 24 h. After completion, the reaction mixture was concentrated. The residue was diluted with H2O (100 mL). The aqueous layer was extracted with tert-butyl methyl ether (50 mL x 2). Then the aqueous layer was adjusted to pH = 1 - 2 with 1N aqueous HCl and extracted with DCM (70 mL x 3). The combined organic layers were washed with saturated aqueous NaCl (100 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to give the crude title product (243 mg, 1.22 mmol, purity % = 75%) as a brown solid. 1 H NMR (500 MHz, CDCl3) δ 3.58 (s, 2H), 3.40 (s, 3H), 2.25 - 2.24 (m, 2H), 2.11 - 2.05 (m, 2H), 1.46 (s, 6H).

[0410] Step 4: (1-(Methoxymethyl)-3,3-dimethyl-2-oxabicyclo[2.1.1]hexan-4-yl)methanol

Chemical formula

[0411] To a mixture of 1-(methoxymethyl)-3,3-dimethyl-2-oxabicyclo[2.1.1]hexane-4-carboxylic acid (190 mg, 0.95 mmol) in THF (5 mL) at 0 °C was added dropwise BH3·THF (1.9 mL, 1N, 1.90 mmol). Then it was stirred at room temperature for 3 h. After completion, the reaction mixture was quenched with MeOH (10 mL) at 0 °C and concentrated to give the crude product. It was purified by silica gel column chromatography, eluting with 0 - 1.2% MeOH in DCM to give the title product (60 mg, 0.32 mmol) as a colorless oil. 11H NMR (500 MHz, CDCl3) δ 3.79 (s, 2H), 3.58 (s, 2H), 3.40 (s, 3H), 1.89 - 1.88 (m, 2H), 1.71 - 1.70 (m, 2H), 1.32 (s, 6H).

[0412] Step 5: tert-Butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-2-((1-(methoxymethyl)-3,3-dimethyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

Chemical formula

[0413] At 0 °C, to a mixture of tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-2,8-difluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (70.0 mg, 0.095 mmol) in THF (5 mL) were added (1-(methoxymethyl)-3,3-dimethyl-2-oxabicyclo[2.1.1]hexan-4-yl)methanol (35 mg, 0.19 mmol) and sodium hydride (11.4 mg, 0.29 mmol). The reaction mixture was stirred at room temperature for 2 hours. After completion, the reaction mixture was quenched with H2O (30 mL) and extracted with EtOAc (30 mL x 3). The combined organic layers were washed with saturated NaCl aqueous solution (100 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated to obtain the crude title product. It was purified by silica gel column chromatography, eluting with 0 - 20% EtOAc in petroleum ether, to give the title product (45 mg, 0.05 mmol) as a yellow solid. MS (ESI, m / e) [M+1] + 901.7.

[0414] Step 6: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((1-(methoxymethyl)-3,3-dimethyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

[0415] At room temperature, TFA (3 mL) was added to a solution of tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-2-((1-(methoxymethyl)-3,3-dimethyl-2-oxabicyclo[2.1.1]hexan-4-yl)methoxy)-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (45 mg, 0.05 mmol) in DCM (3 mL). The resulting mixture was stirred at room temperature for 2 h. After completion, the reaction mixture was concentrated to give a residue. The residue was purified by prep-HPLC to give the title product (25 mg, 71% yield) as a yellow solid. 1 H NMR (500 MHz, CD3OD) δ 8.11 (s, 1H), 7.23 -7.20 (m, 1H), 7.03 - 7.00 (m, 1H), 4.72 - 4.54 (m, 4H), 4.02 (s, 2H), 3.84 - 3.81 (m, 1H), 3.78 - 3.76 (m, 1H), 3.55 (s, 2H), 3.36 (s, 3H), 2.02 (s, 4H), 1.97 - 1.96 (m, 2H), 1.84 (s, 2H), 1.36 (s, 6H). MS (ESI, m / e) [M+1] + 701.4.

[0416] Example 57: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((3-(hydroxymethyl)oxetan-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

Chemical formula

[0417] Step 1: tert-Butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-2-((3-(hydroxymethyl)oxetan-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

Chemical formula

[0418] At room temperature, NaH (10 mg, 0.25 mmol) was added to a solution of tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-2,8-difluoro-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (150 mg, 0.20 mmol) and 3,3-dihydroxymethyloxetane (110 mg, 0.93 mmol) in THF (3 mL). The resulting mixture was stirred at room temperature for 4 hours. After completion, the reaction mixture was diluted with EtOAc (50 mL) and washed with saturated aqueous NaCl solution (15 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to obtain a residue. It was purified by silica gel column chromatography, eluting with 0 - 100% EtOAc in petroleum ether to obtain the title product (100 mg, 0.12 mmol). MS (ESI, m / e) [M+1] + 833.5.

[0419] Step 2: 4-(4-(3,8-Diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-((3-(hydroxymethyl)oxetan-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-7-yl)-2-amino-7-fluorobenzo[b]thiophene-3-carbonitrile

[0420] At room temperature, TFA (2 mL) was added to a solution of tert-butyl 3-(7-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorobenzo[b]thiophen-4-yl)-8-fluoro-2-((3-(hydroxymethyl)oxetan-3-yl)methoxy)-6-(trifluoromethyl)quinazolin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (50 mg, 0.06 mmol) in DCM (4 mL). The reaction mixture was stirred at room temperature for 4 hours. The solvent was concentrated. The residue was dissolved in EtOAc (20 mL), and saturated aqueous Na2CO3 solution (20 mL) was added. The solution was stirred at room temperature for 10 minutes and extracted with EtOAc (40 mL x 2). The combined organic phases were washed with brine (20 mL), dried over Na2SO4, and then concentrated under vacuum. It was purified by prep-HPLC to give the title product (20 mg, 53% yield). 1 H NMR (500 MHz, CD3OD) δ 8.49 (s, 0.5H), 8.12 (s, 1H), 7.23 - 7.20 (m, 1H), 7.03 - 7.00 (m, 1H), 4.73 - 4.68 (m, 3H), 4.64 - 4.63 (m, 2H), 4.56 - 4.55 (m, 3H), 3.91 (s, 4H), 3.83 - 3.74 (m, 2H), 1.97 (s, 4H). MS (ESI, m / e) [M+1] + 633.4.

[0421] Assay KRAS Probe Displacement Assay

[0422] This assay was used to identify compounds that bind to GDP-loaded KRAS protein and can displace the biotinylated probe occupying the KRAS binding site. GST-tagged GDP-loaded WT KRAS (amino acids 1-188) and GST-tagged GDP-loaded KRAS G12D (amino acids 1-188) were expressed in E. coli and purified in-house. All proteins and reaction solutions were prepared in assay buffer containing 50 mM HEPES pH 7.5, 50 mM NaCl, 1 mM MgCl2, 1 mM TCEP, 0.01% BSA, and 0.008% Brij-35. Purified WT KRAS (final concentration 3 nM) or KRAS G12D protein (final concentration 0.5 nM) was incubated with serially diluted compounds in assay plates (384-well microplates, black, Corning). The plates were incubated at 24 °C for 1 hour. After incubation, biotinylated probe 1 (final assay concentration 60 nM) for WT KRAS and biotinylated probe 2 (final assay concentration 4 nM) for KRAS G12D were added to the assay plates respectively. After incubation at 24 °C for 1 hour, Mab Anti-GST-Tb cryptate (Cisbio) and Streptavidin-XL665 (Cisbio) were added and incubated at 24 °C for an additional 1 hour. The TR-FRET signal (excitation 337 nm, emission 665 nm / 620 nm) was read on a BMG PHERAstar FSX instrument. The percentage inhibition of KRAS protein binding to the biotinylated probe was calculated based on the ratio of fluorescence at 665 nm to fluorescence at 620 nm while increasing the concentration of the compound. The IC 50 value for each compound was calculated by fitting the data to a four-parameter logistic model by Dotmatics.

[0423] pERK assay In this study, the AsPC-1 cell line was used. The cells were maintained in RPMI-1640 supplemented with 10% fetal bovine serum (Thermo Fisher), 50 units / mL of penicillin and streptomycin (Thermo Fisher), and kept at 37 °C in a humidified atmosphere of 5% CO2 in air. The cells were restored to their original state from a frozen stock established within 30 passages from the originally purchased cells. 30,000 cells per well were seeded into a 96-well plate and incubated overnight. The cells were treated with a 10-step dilution series. The final compound concentration was 0 - 10 μM. After 2 hours of compound treatment, the cells were lysed, and the levels of pERK1 / 2 (THR202 / TYR204) in the cell lysates were detected using an HTRF kit (Cisbio). Briefly, a total of 16 μL of cell lysate was transferred from each well of the 96-well plate to a 384-well white assay plate. The lysates in each well were incubated overnight at room temperature in the dark with 2 μL of Eu3+-cryptate (donor)-labeled anti-phospho-ERK1 / 2 antibody and 2 μL of D2 (acceptor)-labeled anti-phospho-ERK1 / 2 antibody (Cisbio). When the donor and acceptor are in proximity, fluorescence resonance energy transfer (FRET) from the donor excited by a laser occurs towards the acceptor, and then the acceptor emits fluorescence at a wavelength of 655 nm. The FRET signal was measured using a PHERAstar FSX reader (BMG Labtech). IC 50 50 was determined using Dotmatics by fitting a curve of the inhibition rate against the logarithm of the inhibitor concentration.

[0424] Activity table Each compound in Table 2 was tested in one or more of the biochemical assays shown herein and was found to be active in those assays.

[0425]

Table 3-1

Table 3-2

Table 3-3

[0426] As demonstrated by the data in Table 2, the inventors have surprisingly and unexpectedly discovered that the exemplary compounds of Table 2 modulate or inhibit the activity of KRAS G12D.

[0427] Although numerous references are cited, the entire disclosures thereof are hereby incorporated by reference.

Claims

1. A compound having the formula (I): 【Chemical 1】 or a pharmaceutically acceptable salt, tautomer, isotopic molecular species, stereoisomer, or prodrug thereof (wherein R ５ is H, -Cl, -CF ３ , or unsubstituted C １－４ alkyl, and R ８ is unsubstituted or substituted alkyl, unsubstituted or substituted cycloalkyl, unsubstituted or substituted cycloalkylalkyl, unsubstituted or substituted aryl, unsubstituted or substituted aralkyl, unsubstituted or substituted heterocyclyl, unsubstituted or substituted heterocyclylalkyl, unsubstituted or substituted heteroaryl, unsubstituted or substituted heteroaralkyl, unsubstituted or substituted spirocyclic ring, or unsubstituted or substituted -alkyl-spirocyclic ring, and R １１ is 3,8-diazabicyclo[3.2.1]oct-3-yl, ​ 3,8-diazabicyclo[3.2.1]oct-8-yl, 【Chemical Formula 3】 3-oxa-7,9-diazabicyclo[3.3.1]non-9-yl, [Chemical Formula 4] 3-oxa-7,9-diazabicyclo[3.3.1]non-7-yl, or 【Chemical Formula 5】 and L １ is -O- or a bond).

2. L １ The compound according to claim 1, wherein L is -O-.

3. R ５ is -CF ３ and R １１ is 【Chemical Formula 6】 and R ８ The compound according to claim 2, wherein R is unsubstituted or substituted heterocyclic alkyl.

4. The compound according to claim 3, wherein the heterocyclylalkyl contains at least one oxygen.

5. R ８ is -L ８ａ -R ８ａ and L ８ａ is -(CH ２ ) ｎ - and n is an integer of 1, 2, or 3, R ８ａ is oxetanyl, tetrahydrofuryl, tetrahydro-2H-pyranyl, dihydro-2H-pyranyl, oxabicyclo[2.1.1]hexyl, oxabicyclo[2.2.1]heptyl, oxaspiro[3.3]heptyl, oxabicyclo[3.2.1]octyl, oxabicyclo[2.2.2]octyl, oxaspiro[3.5]nonyl, or oxaspiro[3.4]octyl, R ８ａ is the compound according to claim 4, which is optionally substituted.

6. R ８ is [Chemical Formula 7] The compound according to claim 5, wherein

7. The compound according to claim 3, wherein the heterocyclylalkyl contains at least one nitrogen.

8. R ８ is -L ８ａ -R ８ａ and L ８ａ is -(CH ２ ) ｎ - and n is an integer of 1, 2, or 3, R ８ａ is azetidyl, pyridyl, isoxazolyl, oxazolyl, dihydro-2H-pyranyl, tetrahydro-2H-pyranyl, pyrrolidinonyl, azaspiro[3.3]heptyl, azabicyclo[2.1.1]hexyl, pyrrolidyl, 1H-pyrazolyl, R ８ａ is the compound according to claim 7, which is optionally substituted.

9. R ８ is 【Chemical 8】 The compound according to claim 8, wherein

10. L １ is a bond, R ５ is -CF ３ and R １１ is 【Chemical Formula 9】 and R ８ The compound according to claim 1, wherein R is unsubstituted or substituted heterocyclyl, unsubstituted or substituted heterocyclylalkyl, unsubstituted or substituted cycloalkyl, or unsubstituted or substituted cycloalkylalkyl.

11. R ８ is -L ８ａ -R ８ａ and L ８ａ is -(CH ２ ) ｎ - and n is an integer of 0, 1, 2, or 3, R ８ａ is cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or azetidyl, and R ８ａ is the compound according to claim 10, which is optionally substituted.

12. R ８ is 【Chemical Formula 10】 The compound according to claim 11, wherein

13. R ５ is -CF ３ and L １ is -O- and R １１ is 3,8-diazabicyclo[3.2.1]oct-8-yl, 【Chemical 11】 3-oxa-7,9-diazabicyclo[3.3.1]non-9-yl, 【Chemical 12】 3-oxa-7,9-diazabicyclo[3.3.1]non-7-yl, or 【Chemical 13】 and R ８ is 【Chemical Formula 14】 and R ８ is the compound according to claim 1, which is optionally substituted.

14. The compound is 【Chemical Formula 15】 The compound according to claim 13, wherein

15. R ５ is -Cl, L １ is -O- and R ８ is 【Chemical 16】 and R ８ is the compound according to claim 1, which is optionally substituted.

16. The compound is 【Chemical 17】 The compound according to claim 15, wherein

17. R ５ is unsubstituted C １－４ alkyl, and R １１ is 【Chemical 18】 and L １ is -O- and R ８ is 【Chemical Formula 19】 and R ８ is the compound according to claim 1, which is optionally substituted.

18. The compound is 【Chemical 20】 The compound according to claim 17, wherein

19. R ５ is -CF ３ and R １１ is 3-oxa-7,9-diazabicyclo[3.3.1]non-9-yl, 【Chemical 21】 3-oxa-7,9-diazabicyclo[3.3.1]non-7-yl, L １ is -O- and R ８ is 【Chemical 22】 and R ８ is the compound according to claim 1, which is optionally substituted.

20. The compound is 【Chemical 23】 The compound according to claim 19, wherein

21. The compound according to any one of claims 1 to 20, wherein the compound is selected from Table 2.

22. A pharmaceutical composition comprising an effective amount of the compound according to any one of claims 1 to 21, or a pharmaceutically acceptable salt, tautomer, isotopic molecular species, stereoisomer, or prodrug thereof, and a pharmaceutically acceptable carrier, excipient, or vehicle.

23. A method for inhibiting the activity of a KRAS mutant protein in a cell, comprising contacting the cell with an effective amount of a compound according to any one of claims 1 to 21, or a pharmaceutically acceptable salt, tautomer, isotopic molecular species, stereoisomer, or prodrug thereof, wherein optionally, the KRAS mutant protein is a KRAS G12D mutant protein, said method.

24. A method for treating or preventing cancer, comprising administering to a subject in need of treatment or prevention of cancer an effective amount of a compound according to any one of claims 1 to 21, or a pharmaceutically acceptable salt, tautomer, isotopic molecular species, stereoisomer, or prodrug thereof, wherein optionally, the cancer is mediated by a KRAS mutation, preferably a KRAS G12D mutation, said method.

Images