KIF18A inhibitors

Abstract

Provided are compounds of formula (I) as defined herein: (I) and their synthetic intermediates, wherein the compounds of formula (I) are capable of regulating KIF18A protein, thereby affecting cell cycle and cell proliferation processes to treat cancer and cancer-related diseases. The present invention also includes pharmaceutical compositions comprising the compounds and methods for treating conditions associated with KIF18A activity.

Description

[0001] The present invention relates to the field of pharmaceutical agents, and more particularly, to compounds and compositions for regulating KIF18A, and uses and methods for managing cell proliferation and treating cancer. Background Art

[0002] Cancer is one of the most prevalent diseases afflicting humans and is a leading cause of death worldwide. In the past several decades, many groups have invested a great deal of time, energy, and financial resources in an effort to find an effective treatment or cure for one or more of the many different cancers. However, to date, only a few of the available cancer treatments and therapies have provided a significant degree of success.

[0003] Cancer is often characterized by unregulated cell proliferation. Damage to one or more genes responsible for cellular pathways (which control the progression of proliferation through the cell cycle and centrosome circulation) can cause the normal regulation of cell proliferation to be lost. These unregulated genes can encode various tumor suppressors or oncogene proteins, which participate in a series of events, leading to unchecked cell cycle progression and cell proliferation. Various kinases and kinesins have been identified to play a key role in the cell cycle and mitosis regulation and progression of normal dividing cells and cancer cells.

[0004] Kinesin is a molecular motor that plays an important role in cell division and intracellular vesicle and organelle transport. Mitotic kinesin plays a role in many aspects of spindle assembly, chromosome separation, centrosome separation and dynamics (O. Rath and F. Kozielski, Nature Review Cancer [Natural Cancer Review], 12: 527-39, 2012). Based on the sequence homology in the so-called "motor domain", human kinesin is classified into 14 subfamilies, and the ATPase activity of this domain drives unidirectional movement along microtubules (MT). The non-motor domains of these proteins are responsible for cargo attachment; "cargo" can include any of a variety of different membranous organelles, signal transduction scaffold systems and chromosomes. Kinesin uses ATP hydrolysis energy to move cargo along polarized microtubules. Therefore, kinesin is generally referred to as a "plus end" or "minus end" directional motor.

[0005] The KIF18A gene belongs to the kinesin-8 subfamily and is a plus-end directed motor. KIF18A is thought to affect the dynamics of the plus ends of the centromere microtubules to control correct chromosome positioning and spindle tension. Depletion of human KIF18A leads to longer spindles, increased chromosome oscillation in metaphase, and activation of the mitotic spindle assembly checkpoint in HeLa cervical cancer cells (MI Mayr et al., Current Biology 17, 488–98, 2007). KIF18A appears to be a viable target for cancer therapy. KIF18A is overexpressed in many types of cancer, including but not limited to colon cancer, breast cancer, lung cancer, pancreatic cancer, prostate cancer, bladder cancer, head cancer, neck cancer, cervical cancer, and ovarian cancer. In addition, in cancer cell lines, gene deletion or knockout or KIF18A inhibition affects the mitotic spindle apparatus. In particular, inhibition of KIF18A has been found to induce mitotic cell arrest, a known weakness that can promote mitotic cell death by apoptosis, mitotic catastrophe, or multiphase-driven lethality or death following mitotic slippage in interphase. Therefore, there is a strong interest in finding inhibitors of the KIF18A protein.

[0006] Therefore, inhibition of KIF18A ATPase activity is a promising approach for developing new anticancer agents. Summary of the invention

[0007] The present invention provides a novel class of compounds for use in regulating KIF18A protein alone or in a binding complex with microtubules for treating KIF18A-mediated conditions and / or diseases, including cancer, inflammation or ciliary pathology.

[0008] The compounds provided by the present invention have KIF18A regulatory activity based on MT, in particular, have KIF18A inhibitory activity. To this end, the present invention also provides the use of these compounds and pharmaceutically acceptable salts thereof in the preparation and manufacture of pharmaceutical compositions or drugs for therapeutic, preventive, acute or chronic treatment of KIF18A-mediated diseases and disorders (including but not limited to cancer). Therefore, the compounds of the present invention can be used to manufacture anticancer drugs. The present invention also provides a method for preparing a compound of formula I, and an intermediate useful in such a method.

[0009] In Example 1, the present invention provides a compound of formula (I), a compound of formula I:

[0010]

[0011] or any pharmaceutically acceptable salt thereof, wherein:

[0012] X1 N or -CR 6 ;

[0013] R 1 is -CN or a group -ZR 12 , where Z is -C 0-4 Alkyl-, -NR 11 -、-NR 11 SO2-、-SO2NR 11 -、-NR 11 -S(=O)(=NH), -S(=O)(=NH)-, -S-, -S(=O)-, -SO2-, C 0-4 Alkyl-O-, -(C=O)-, -(C=O)NR 11 -, -C=N(OH)- or -NR 11 (C=O); or

[0014] Group-ZR 12 =N = S (= O) - (R 12 )2, where two R 12 pairs may alternatively combine with their respective attached sulfur atoms to form a saturated or partially saturated 3-, 4-, 5- or 6-membered monocyclic ring containing 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S;

[0015] R 2 is halogen or group -YR 13 , where Y is -C 0-4 Alkyl-, -N(C 0-1 Alkyl)-C 0-4 Alkyl-, -C(=O)NR a R a (C 1-4 Alkyl), -OC 0-4 Alkyl-, S, S=O, S(=O)2, -SO2NR 13 or -S(=O)(=NH)-;

[0016] R 3 H, C 1-4 Alkyl or C 1-4 Haloalkyl;

[0017] R 4 H, halogen, R 4a or R 4b ;

[0018] R 5 H, halogen, C 1-8 Alkyl or C 1-4 Haloalkyl;

[0019] R6 H, halogen, C 1-8 Alkyl, C 1-4 Haloalkyl, -OC 1-8 Alkyl or -OR 6a , where R 6a is a saturated or partially saturated 3-, 4-, 5- or 6-membered monocyclic ring containing 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S;

[0020] R 7 H, halogen, C 1-8 Alkyl or C 1-4 Haloalkyl;

[0021] R 8 H, halogen, C 1-8 Alkyl, C 1-4 Haloalkyl, -OH, -OR 8a OR 8b ;

[0022] R 9 H, halogen, C 1-8 Alkyl or C 1-4 Haloalkyl;

[0023] R x Selected from the group consisting of:

[0024] R 10a , R 10b , R 10c , R 10d , R 10e , R 10f , R 10g , R 10h , R 10i and R 10j Each of H, halogen, R 10k or R 10l ;

[0025] Or alternatively, R 10a and R 10b Yes, R 10c和 R 10d Yes, R 10e和 R 10f Yes, R 10g and R 10h Yes or R 10i and R 10j Each of the pairs can independently combine with their respective attached carbon atoms to form a spiro-linked xA saturated or partially saturated 3-, 4-, 5-, 6-membered monocyclic ring; wherein the 3-, 4-, 5-, 6-membered monocyclic ring contains 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S, and further, wherein the 3-, 4-, 5-, 6-membered monocyclic ring is substituted by 0, 1, 2 or 3 groups selected from the following: F, Cl, Br, C 1-6 Alkyl, C 1-4 Haloalkyl, -OR a 、-OC 1-4 Haloalkyl, CN, -NR a R a or oxo;

[0026] R 11 H, R 11a or R 11b ;

[0027] R 12 H, R 12a or R 12b ;

[0028] R 13 For R 13a or R 13b ;

[0029] R 4a , R 8a , R 10k , R 11a , R 12a and R 13a is independently selected in each case from the group consisting of: a saturated, partially saturated or unsaturated 3-, 4-, 5-, 6- or 7-membered monocyclic ring or a 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11- or 12-membered bicyclic ring containing 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S, which is substituted by 0, 1, 2 or 3 groups selected from the group consisting of F, Cl, Br, C 1-6 Alkyl, C 1-4 Haloalkyl, -OR a 、-OC 1-4 Haloalkyl, CN, -C(=O)R b 、-C(=O)OR a 、-C(=O)NR a R a 、-C(=NR a )NR a R a 、-OC(=O)R b 、-OC(=O)NR a R a 、-OC 2-6 Alkyl NR aR a 、-OC 2-6 Alkyl OR a 、-SR a 、-S(=O)R b 、-S(=O)2R b 、-S(=O)2NR a R a 、-NR a R a 、-N(R a )C(=O)R b 、-N(R a )C(=O)OR b 、-N(R a )C(=O)NR a R a 、-N(R a )C(=NR a )NR a R a 、-N(R a )S(=O)2R b 、-N(R a )S(=O)2NR a R a 、-NR a C 2-6 Alkyl NR a R a 、-NR a C 2-6 Alkyl OR a , -C 1-6 Alkyl NR a R a , -C 1-6 Alkyl OR a , -C 1-6 Alkyl N(R a )C(=O)R b , -C 1-6 Alkyl OC(=O)R b , -C 1-6 Alkyl C(=O)NR a R a , -C 1-6 Alkyl C(=O)OR a , R 14 and oxo;

[0030] R 4b , R 8b , R 10l , R 11b , R 12b and R 13bIn each case, independently selected from the group consisting of: selected from F, Cl, Br, -OR a 、-OC 1-4 0, 1, 2, 3, 4 or 5 groups substituted with haloalkyl or CN 1-6 alkyl;

[0031] R 14 is independently selected in each case from the group consisting of: a saturated, partially saturated or unsaturated 3-, 4-, 5-, 6- or 7-membered monocyclic ring or a 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11- or 12-membered bicyclic ring containing 0, 1, 2 or 3 N atoms and 0 or 1 atom selected from O and S, which is substituted by 0, 1, 2 or 3 groups selected from the group consisting of F, Cl, Br, C 1-6 Alkyl, C 1-4 Haloalkyl, -OR a 、-OC 1-4 Haloalkyl, CN, -C(=O)R b 、-C(=O)OR a 、-C(=O)NR a R a 、-C(=NR a )NR a R a 、-OC(=O)R b 、-OC(=O)NR a R a 、-OC 2-6 Alkyl NR a R a 、-OC 2-6 Alkyl OR a 、-SR a 、-S(=O)R b 、-S(=O)2R b 、-S(=O)2NR a R a 、-NR a R a 、-N(R a )C(=O)R b 、-N(R a )C(=O)OR b ,-N(R a )C(=O)NR a R a 、-N(R a )C(=NR a )NR a R a 、-N(R a )S(=O)2R b 、-N(Ra )S(=O)2NR a R a 、-NR a C 2-6 Alkyl NR a R a 、-NR a C 2-6 Alkyl OR a , -C 1-6 Alkyl NR a R a , -C 1-6 Alkyl OR a , -C 1-6 Alkyl N(R a )C(=O)R b , -C 1-6 Alkyl OC(=O)R b , -C 1-6 Alkyl C(=O)NR a R a , -C 1-6 Alkyl C(=O)OR a and oxo;

[0032] R a is independently H or R b ;and

[0033] R b In each case independently C 1-6 Alkyl, phenyl or benzyl, where C 1-6 The alkyl group is substituted with 0, 1, 2 or 3 substituents selected from the group consisting of halogen, -OH, -OC 1-4 Alkyl, -NH2, -NHC 1-4 Alkyl, -OC(=O)C 1-4 Alkyl or -N(C 1-4 Alkyl)C 1-4 and phenyl or benzyl is substituted by 0, 1, 2 or 3 substituents selected from the group consisting of halogen, C 1-4 Alkyl, C 1-3 Haloalkyl, -OH, -OC 1-4 Alkyl, -NH2, -NHC 1-4 Alkyl, -OC(=O)C 1-4 Alkyl or -N(C 1-4 Alkyl)C 1-4 alkyl.

[0034] In embodiment 2, the present invention provides compounds wherein X 1 N; having formula (Ia):

[0035]

[0036] In embodiment 3, the present invention provides compounds wherein X 1 For-CR 6 ; having formula (Ib):

[0037]

[0038] In embodiment 4, the present invention provides compounds wherein R 3 is H or methyl; preferably, R 3 For H.

[0039] In embodiment 5, the present invention provides compounds wherein R 10c , R 10d , R 10e , R 10f , R 10g , R 10h , R 10i and R 10j Each of H, halogen, C 1-6 Alkyl or C 1-4 haloalkyl; and R 10a and R 10b Each of the pairs combines with their respective attached carbon atoms to form a spiral bond to R x a saturated 3-, 4- or 5-membered monocyclic ring; wherein the ring contains 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S.

[0040] In embodiment 6, the present invention provides compounds wherein R 10c , R 10d , R 10e , R 10f , R 10g , R 10h , R 10i and R 10j Each of is H, methyl or ethyl; and R 10a and R 10b Each of the pairs combines with their respective attached carbon atoms to form a spiral bond to R x The cyclopropyl, cyclobutyl or cyclopentyl ring.

[0041] In embodiment 7, the present invention provides a compound according to embodiments 1 to 6 or a pharmaceutically acceptable salt thereof, wherein the group Selected from

[0042] In embodiment 8, the present invention provides a compound according to embodiments 1 to 7 or a pharmaceutically acceptable salt thereof, wherein the group for

[0043] In embodiment 9, the present invention provides a compound according to embodiments 1 to 8 or a pharmaceutically acceptable salt thereof, wherein R 1 is -CN or -ZR 12 , wherein Z is a bond, -NH-, -NHSO2-, -SO2NH-, -S(=O)(=NH)-, -S-, -S(=O)-, -SO2-, -(C=O)-, -(C=O)NH-, or -NH(C=O)-; and R 12 Selected from:

[0044] (a) H;

[0045] (b) cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxiranyl, oxetanyl, tetrahydrofuranyl, azetidinyl, imidazolyl, morpholinyl, pyrrolidinyl, piperazinyl,

[0046] wherein each of the rings is substituted by 0, 1, 2 or 3 groups selected from the group consisting of OH, F, methyl, -CH2OH, -C(=O)OCH3,

[0047] -C(=O)OC(CH3)3, NH2, CN and oxo; or

[0048] (c) C substituted with 0, 1, 2 or 3 OH, F, -C(=O)OCH3, -NH2, -NH(CH3)2 1-6 alkyl.

[0049] In embodiment 10, the present invention provides a compound according to embodiments 1 to 9 or a pharmaceutically acceptable salt thereof, wherein R 1 is -CN or -ZR 12 , wherein Z is none, -NH-, -NHSO2-, -SO2NH-, -S(=O)(=NH)-, -S-, -S(=O)-, -SO2-, -(C=O)-, -(C=O)NH-, or -NH(C=O)-; and

[0050] (a)R 12 is H;

[0051] (b)R 12 is oxetane or cyclopropyl; or

[0052] (c)R 12 is C substituted with 0, 1, 2 or 3 OH groups 1-6 alkyl.

[0053] In embodiment 11, the present invention provides compounds according to embodiments 1 to 10 or pharmaceutically acceptable salts thereof, wherein the group -ZR 12 =N = S (= O) - (R 12 )2, where two R 12 The pairs may alternatively combine with their respective attached sulfur atoms to form a saturated or partially saturated 3-, 4-, 5- or 6-membered monocyclic ring containing 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S; which is selected from:

[0054]

[0055] In embodiment 12, the present invention provides a compound according to embodiments 1 to 11 or a pharmaceutically acceptable salt thereof, wherein R 1 For the group -ZR 12 , where Z is -NHSO2- or

[0056] -SO2NH-; and R 12 is oxetane, cyclopropyl, or R 12 is C substituted with 0, 1, 2 or 3 OH groups 1-6 alkyl.

[0057] In embodiment 13, the present invention provides a compound according to embodiments 1 to 12 or a pharmaceutically acceptable salt thereof, wherein R 1 For the group -ZR 12 , where Z is -NHSO2-, and R 12 It is -CH2-CH2-OH.

[0058] In embodiment 14, the present invention provides a compound according to embodiments 1 to 13 or a pharmaceutically acceptable salt thereof, wherein R 2 is halogen or group -YR 13 , where Y is a bond, -NH-, -NH-(CH2) 0-4 -or-O-(CH2) 0-4 ; and R 13 is a saturated, partially saturated or unsaturated 3-, 4-, 5-, 6- or 7-membered monocyclic ring or a 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11- or 12-membered bicyclic ring containing 0, 1, 2 or 3 N atoms and 0 or 1 atom selected from O and S, which is substituted by 0, 1, 2 or 3 groups selected from the following: F, Cl, Br, C 1-6 Alkyl, C 1-4 Haloalkyl, -OH, -OC 1-4 Haloalkyl, CN, R 14 and oxo; or

[0059] R13 is selected from F, Cl, Br, -OH, -OC 1-4 0, 1, 2, 3, 4 or 5 groups substituted with haloalkyl or CN 1-6 alkyl.

[0060] In embodiment 15, the present invention provides a compound according to embodiments 1 to 14 or a pharmaceutically acceptable salt thereof, wherein R 2 is a saturated 5- or 6-membered monocyclic ring, wherein each of said rings contains 0, 1 or 2 N atoms and 0 or 1 O atoms, and wherein each of said rings is substituted by 0, 1, 2 or 3 groups selected from the group consisting of F, Cl, Br, C 1-6 Alkyl, C 1-4 Haloalkyl, -OH, -OC 1-4 Haloalkyl, CN, R 14 and oxo.

[0061] In embodiment 16, the present invention provides a compound according to embodiments 1 to 15 or a pharmaceutically acceptable salt thereof, wherein R 2 is (a) a halogen; (b) a group -YR 13 , where Y is a bond; and R 13 is morpholinyl, piperidinyl, azetidinyl, pyrrolidinyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, piperazinyl, tetrahydrofuranyl,

[0062] wherein each of the rings is substituted by 0, 1, 2 or 3 groups selected from the group consisting of F, Cl, Br, methyl, CF3, -OH, -OCHF2, CN and oxo; or (c) a group -YR 13 , wherein Y is NH, -O-, -O-(CH2)-, -O-(CH2)-(CH2)-, or -O-(CH2)-(CH2)-(CH2)-, and wherein R 13 for or R 13 is C substituted by 0, 1, 2, 3, 4 or 5 groups selected from F, Cl, Br, methyl, CF3, -OH or CN 1-6 alkyl.

[0063] In embodiment 17, the present invention provides a compound according to embodiments 1 to 16 or a pharmaceutically acceptable salt thereof, wherein R 2 is selected from F, Cl, Br, methyl, CF 3 、-OH、-OCHF 2 , CN or oxo-substituted morpholinyl or piperidinyl.

[0064] In embodiment 18, the present invention provides a compound according to embodiments 1 to 17 or a pharmaceutically acceptable salt thereof, wherein R 2 is morpholinyl substituted by 1, 2 or 3 methyl groups.

[0065] In embodiment 19, the present invention provides a compound according to embodiments 1 to 18 or a pharmaceutically acceptable salt thereof, wherein R 2 is piperidinyl substituted by 1, 2 or 3 fluorine groups.

[0066] In embodiment 20, the present invention provides a compound according to embodiments 1 to 19 or a pharmaceutically acceptable salt thereof, wherein R 2 for

[0067]

[0068] In embodiment 21, the present invention provides a compound according to embodiments 1 to 20 or a pharmaceutically acceptable salt thereof, wherein Z is a bond, -NH-, -NHSO2-, -SO2NH-, -N=S(=O)<(R a )2(where each R 11 independently selected from the group consisting of: H, methyl or isopropyl), -S(=O)(=NH)-, -S-, -S(=O)-, -SO2-, -(C=O)-, -(C=O)NH- or -NH(C=O)-.

[0069] In embodiment 22, the present invention provides a compound according to embodiments 1 to 21 or a pharmaceutically acceptable salt thereof, wherein R 12 (a) H; (b) C substituted by 0, 1, 2 or 3 groups selected from F, Cl, Br, -OH, -OCH3 or cyclopropyl 1-6 or (c) a saturated, partially saturated or unsaturated 3-, 4-, 5-, 6- or 7-membered monocyclic ring containing 0, 1, 2 or 3 N atoms and 0 or 1 atom selected from O and S, which is substituted by 0, 1, 2 or 3 groups selected from the following: F, Cl, Br, C 1-6 Alkyl, C 1-4 Haloalkyl, -C 1-6 Alkyl OH, -OH, -OCH3, -NH2 or oxo.

[0070] In embodiment 23, the present invention provides a compound according to embodiments 1 to 22 or a pharmaceutically acceptable salt thereof, wherein R 12 is selected from cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl, azetidinyl, tetrahydrofuranyl or 1,3,4-oxathiazinyl.

[0071] In embodiment 24, the present invention provides a compound according to embodiments 1 to 23 or a pharmaceutically acceptable salt thereof, wherein R 4 is selected from (a) H; (b) C substituted with 0, 1, 2 or 3 OH groups 1-6 or (c) cyclopropyl.

[0072] In embodiment 25, the present invention provides a compound according to embodiments 1 to 24 or a pharmaceutically acceptable salt thereof, wherein R 4 It is methyl.

[0073] In embodiment 26, the present invention provides a compound according to embodiments 1 to 25 or a pharmaceutically acceptable salt thereof, wherein R 5 For H.

[0074] In embodiment 27, the present invention provides a compound according to embodiments 1 to 26 or a pharmaceutically acceptable salt thereof, wherein R 6 It is H or F.

[0075] In embodiment 28, the present invention provides a compound according to embodiments 1 to 27 or a pharmaceutically acceptable salt thereof, wherein R 7 It is H or F.

[0076] In embodiment 29, the present invention provides a compound according to embodiments 1 to 28 or a pharmaceutically acceptable salt thereof, wherein R 8 For H.

[0077] In embodiment 30, the present invention provides a compound according to embodiments 1 to 29 or a pharmaceutically acceptable salt thereof, wherein R 9 For H.

[0078] In embodiment 31, the present invention provides a compound or a pharmaceutically acceptable salt thereof selected from:

[0079]

[0080]

[0081]

[0082]

[0083]

[0084] or any pharmaceutically acceptable salt thereof.

[0085] In sub-embodiment 31a, the present invention provides N-(2-((1-hydroxy-2-methylpropan-2-yl)amino)-6-methylpyrimidin-4-yl)-4-(methylsulfonyl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide or a pharmaceutically acceptable salt thereof.

[0086] In sub-Example 31b, the present invention provides N-(2-((1-hydroxy-2-methylpropane-2-yl)amino)-6-methylpyrimidin-4-yl)-4-(N-(3-methyloxetane-3-yl)sulfamoyl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide or a pharmaceutically acceptable salt thereof.

[0087] In sub-Example 31c, the present invention provides N-(2-(2-hydroxypropan-2-yl)pyrimidin-4-yl)-4-(N-(3-methyloxetan-3-yl)sulfamoyl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide or a pharmaceutically acceptable salt thereof.

[0088] In sub-embodiment 31d, the present invention provides N-(2-(4,4-difluoropiperidin-1-yl)pyridin-4-yl)-4-(N-(3-methyloxetane-3-yl)sulfamoyl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide or a pharmaceutically acceptable salt thereof.

[0089] In sub-embodiment 31e, the present invention provides N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((1-methylcyclopropane)-1-sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzamide or a pharmaceutically acceptable salt thereof.

[0090] In sub-embodiment 31f, the present invention provides (R)-4-((2-hydroxyethyl)sulfonamido)-N-(6-methyl-2-(2-methylmorpholino)pyrimidin-4-yl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide or a pharmaceutically acceptable salt thereof.

[0091] In sub-embodiment 31g, the present invention provides N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((2-hydroxyethyl)sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzamide or a pharmaceutically acceptable salt thereof.

[0092] In sub-embodiment 31h, the present invention provides (R)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((2-hydroxy-1-methylethyl)sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzamide or a pharmaceutically acceptable salt thereof.

[0093] In sub-Example 31i, the present invention provides (S)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((2-hydroxy-1-methylethyl)sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzamide or a pharmaceutically acceptable salt thereof.

[0094] In sub-embodiment 31j, the present invention provides N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-(ethylsulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzamide or a pharmaceutically acceptable salt thereof.

[0095] In sub-embodiment 31k, the present invention provides N-(2-(3,3-difluoroazetidin-1-yl)-6-methylpyrimidin-4-yl)-4-((2-hydroxyethyl)sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzamide or a pharmaceutically acceptable salt thereof.

[0096] In sub-Example 311, the present invention provides (R)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-5-fluoro-4-((2-hydroxy-1-methylethyl)sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzamide or a pharmaceutically acceptable salt thereof.

[0097] In sub-embodiment 31m, the present invention provides (S)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-5-fluoro-4-((2-hydroxy-1-methylethyl)sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzamide or a pharmaceutically acceptable salt thereof.

[0098] In sub-embodiment 31n, the present invention provides (R)-N-(2-(3,3-difluoroazetidine-1-yl)-6-methylpyrimidin-4-yl)-4-((2-hydroxypropyl)sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzamide or a pharmaceutically acceptable salt thereof.

[0099] In sub-embodiment 31o, the present invention provides (S)-N-(2-(3,3-difluoroazetidine-1-yl)-6-methylpyrimidin-4-yl)-4-((2-hydroxypropyl)sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzamide or a pharmaceutically acceptable salt thereof.

[0100] In sub-embodiment 31p, the present invention provides N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((2-hydroxyethyl)sulfonyl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide or a pharmaceutically acceptable salt thereof.

[0101] In sub-embodiment 31q, the present invention provides (S)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((1-hydroxypropan-2-yl)sulfonyl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide or a pharmaceutically acceptable salt thereof.

[0102] In sub-embodiment 31r, the present invention provides (R)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((1-hydroxypropane-2-yl)sulfonyl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide or a pharmaceutically acceptable salt thereof.

[0103] In sub-embodiment 31s, the present invention provides N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((1-hydroxy-2-methylpropane-2-yl)sulfonyl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide or a pharmaceutically acceptable salt thereof.

[0104] In sub-embodiment 31t, the present invention provides N-(2-(4,4-difluorocyclohexyl)-6-methylpyrimidin-4-yl)-4-((2-hydroxyethyl)sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzamide or a pharmaceutically acceptable salt thereof.

[0105] In sub-example 31u, the present invention provides (R)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((2-fluoro-1-(hydroxymethyl)ethyl)sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzamide or a pharmaceutically acceptable salt thereof.

[0106] In sub-Example 31v, the present invention provides (S)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((2-fluoro-1-(hydroxymethyl)ethyl)sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzamide or a pharmaceutically acceptable salt thereof.

[0107] In sub-embodiment 31w, the invention provides N-(2-(4,4-difluoropiperidin-1-yl)pyridin-4-yl)-4-(N-(2-hydroxyethyl)sulfamoyl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide or a pharmaceutically acceptable salt thereof.

[0108] In sub-embodiment 31x, the present invention provides 2-(6-azaspiro[2.5]octan-6-yl)-4-(R-cyclopropylsulfonylimino)-N-(2-(4,4-difluoro-1-piperidinyl)-6-methyl-4-pyrimidinyl)benzamide or a pharmaceutically acceptable salt thereof.

[0109] In sub-embodiment 31y, the present invention provides 2-(6-azaspiro[2.5]octan-6-yl)-4-(S-cyclopropylsulfonylimino)-N-(2-(4,4-difluoro-1-piperidinyl)-6-methyl-4-pyrimidinyl)benzamide or a pharmaceutically acceptable salt thereof.

[0110] In sub-embodiment 31z, the present invention provides (N 1 -(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-2-(6-azaspiro[2.5]octan-6-yl)terephthalamide or a pharmaceutically acceptable salt thereof.

[0111] In sub-embodiment 31aa, the invention provides 4-(azetidin-3-ylsulfonyl)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide or a pharmaceutically acceptable salt thereof.

[0112] In sub-embodiment 31ab, the present invention provides N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((1-methylazetidin-3-yl)sulfonyl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide or a pharmaceutically acceptable salt thereof.

[0113] In embodiment 32, the present invention provides a pharmaceutical composition comprising a compound according to any one of embodiments 1 to 31 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable diluent or carrier.

[0114] In embodiment 33, the present invention provides a method for treating a disorder treatable with a KIF18a inhibitor, the method comprising administering a therapeutically effective amount of a compound according to embodiments 1 to 31 or a composition according to embodiment 31 to a patient in need thereof.

[0115] In Example 34, the present invention provides a method according to Example 33, wherein the condition is a cancer selected from the group consisting of: (a) a solid tumor or a blood-derived tumor selected from the following cancers: bladder cancer, endometrial cancer, squamous cell lung cancer, breast cancer, colon cancer, kidney cancer, liver cancer, lung cancer, small cell lung cancer, esophageal cancer, gallbladder cancer, brain cancer, head and neck cancer, ovarian cancer, pancreatic cancer, gastric cancer, cervical cancer, thyroid cancer, prostate cancer and skin cancer; (b) a hematopoietic tumor of the lymphoid system selected from the following: leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma and Burkett's lymphoma; (c) a hematopoietic neoplasm of the myeloid lineage selected from the group consisting of acute and chronic myeloid leukemias, myelodysplastic syndromes and promyelocytic leukemias; (d) a tumor of mesenchymal origin selected from the group consisting of fibrosarcoma and rhabdomyosarcoma; (e) a tumor of the central and peripheral nervous system selected from the group consisting of astrocytoma, neuroblastoma, glioma and schwannoma; or (f) a melanoma, a seminoma, a teratoma, an osteosarcoma, a xeroderma pigmentosum, a keratoacanthoma, a follicular thyroid carcinoma or a Kaposi's sarcoma.

[0116] In sub-embodiment 34a, the invention provides a method according to embodiment 33, wherein the condition is a cancer selected from the group consisting of melanoma, prostate cancer, cervical cancer, breast cancer, colon cancer, sarcoma or leukemia. See: Zhang C. et al., "Kif18A is involved in human breast carcinogenesis", Carcinogenesis, 2010.09;31(9):1676-84.doi:10.1093 / carcin / bgq134.Epub 2010Jul 1. See also: (1) https: / / www.proteinatlas.org / ENSG00000121621-KIF18A / pathology; (2) Nagahara, M. et al., “Kinesin 18A expression: clinical relevance to colorectal cancer progression,” Int. J. Cancer: 129, 2543–2552 (2011) VC 2011 UIC; and (3) Yu, Y. et al., “The Role of Kinesin Family Proteins in Tumorigenesis and Progression-Potential Biomarkers and Molecular Targets for Cancer Therapy,” Cancer 2010; 116: 5150–60. VC 2010 American Cancer Society. In sub-embodiment 34b, the invention provides a method according to embodiment 33, wherein the condition is any one of the cancers specified in embodiments (a), (b), (c), (d), (e) or (f).

[0117] In Example 35, the present invention provides a method for reducing the size of a solid tumor in a subject, the method comprising administering a therapeutically effective amount of a compound according to any one of Examples 1 to 31 or a composition according to Example 32 to a subject in need thereof.

[0118] In Example 36, the present invention provides a method for treating a cell proliferative disorder in a subject, the method comprising administering a therapeutically effective amount of a compound according to any one of Examples 1 to 31 or a composition according to Example 32 to a subject in need thereof.

[0119] In Example 37, the present invention provides a method for inhibiting KIF18A in a cell, the method comprising contacting the cell with a compound according to any one of Examples 1 to 31 or a pharmaceutically acceptable salt thereof, or a composition according to Example 32.

[0120] In embodiment 38, the present invention provides the use of a compound according to any one of embodiments 1 to 31 or a pharmaceutically acceptable salt of said compound, or a pharmaceutical composition according to embodiment 32, for the preparation of a medicament for treating a condition treatable with a KIF18a inhibitor.

[0121] In embodiment 39, the present invention provides the use according to embodiment 38, wherein the condition is a cancer selected from the group consisting of: (a) a solid tumor or a blood-derived tumor selected from the following cancers: bladder cancer, endometrial cancer, squamous cell lung cancer, breast cancer, colon cancer, kidney cancer, liver cancer, lung cancer, small cell lung cancer, esophageal cancer, gallbladder cancer, brain cancer, head and neck cancer, ovarian cancer, pancreatic cancer, gastric cancer, cervical cancer, thyroid cancer, prostate cancer and skin cancer; (b) a hematopoietic tumor of the lymphoid system selected from the following: leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma and Burkett's lymphoma; (c) a hematopoietic neoplasm of the myeloid lineage selected from the group consisting of acute and chronic myeloid leukemias, myelodysplastic syndromes and promyelocytic leukemias; (d) a tumor of mesenchymal origin selected from the group consisting of fibrosarcoma and rhabdomyosarcoma; (e) a tumor of the central and peripheral nervous system selected from the group consisting of astrocytoma, neuroblastoma, glioma and schwannoma; or (f) a melanoma, a seminoma, a teratoma, an osteosarcoma, a xeroderma pigmentosum, a keratoacanthoma, a follicular thyroid carcinoma or a Kaposi's sarcoma.

[0122] In sub-embodiment 39a, the invention provides the use according to embodiment 33, wherein the condition is any one of the cancers specified in embodiments (a), (b), (c), (d), (e) or (f).

[0123] In embodiment 40, the present invention provides the use of a compound according to any one of embodiments 1 to 31 or a pharmaceutically acceptable salt of said compound, or a pharmaceutical composition according to embodiment 32, in the preparation of a medicament for reducing the size of a solid tumor in a subject.

[0124] In embodiment 41, the present invention provides the use of a compound according to any one of embodiments 1 to 31 or a pharmaceutically acceptable salt of said compound, or a pharmaceutical composition according to embodiment 32, in the preparation of a medicament for treating a cell proliferative disorder in a subject.

[0125] In embodiment 42, the present invention provides use of a compound according to any one of embodiments 1 to 31 or a pharmaceutically acceptable salt of said compound, or a pharmaceutical composition according to embodiment 32, in the preparation of a medicament for inhibiting KIF18A in a cell.

[0126] In Example 43, the present invention provides a process for preparing a compound of formula (I) as described herein.

[0127] In Example 44, the invention provides an intermediate compound for use in the process of preparing a compound of formula (I) as described herein.

[0128] It should be understood that the above reference to any embodiment is intended to include any and all sub-embodiments thereof. For example, a reference to embodiment 31 includes a reference to sub-embodiments 31a to 31ab.

[0129] The present invention includes all pharmaceutically acceptable isotopically labeled compounds of the present invention wherein one or more atoms are replaced by an atom having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number usually found in nature.

[0130] Examples of suitable isotopes for inclusion in the compounds of the present invention include, but are not limited to, isotopes of hydrogen, such as 2 H and 3 H; isotopes of carbon, e.g. 11 C. 13 C and 14 C; isotopes of chlorine, e.g. 38 Cl; isotopes of fluorine, e.g. 18 F; isotopes of iodine, e.g. 123 I and 125 I; isotopes of nitrogen, e.g. 13 N and 15 N; isotopes of oxygen, e.g. 15 O. 17 O and 18 O; isotopes of phosphorus, e.g. 32 P and sulfur isotopes, e.g. 35 S.

[0131] Certain isotopically labeled compounds of the invention, e.g., compounds incorporating a radioactive isotope, are useful in drug and / or substrate tissue distribution studies. 3 H, and carbon-14, i.e.14 C is particularly suitable for this purpose due to its ease of incorporation and existing means of detection.

[0132] Using heavier isotopes (such as deuterium, 2 H) substitutions may offer certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements) and are therefore preferred in some circumstances.

[0133] Using positron-emitting isotopes, such as 11 C. 18 F. 15 O and 13 N substitution can be used in positron emission tomography (PET) studies to examine substrate receptor occupancy.

[0134] Isotopically labeled compounds of the present invention can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the accompanying Examples and Preparations, using an appropriate isotopically labeled reagent in place of the non-labeled reagent previously employed.

[0135] Pharmaceutically acceptable solvents according to the present invention include those in which the crystallization solvent may be isotopically substituted, such as D2O, d6-acetone, d6-DMSO.

[0136] Specific embodiments of the present invention include the compounds exemplified in the following examples, as well as their pharmaceutically acceptable salts, complexes, solvates, polymorphs, stereoisomers, metabolites, prodrugs, and other derivatives thereof.

[0137] Unless otherwise stated, the following definitions apply to terms appearing in this specification and claims:

[0138] “C α-β "Alkyl" means an alkyl group containing a minimum of α and a maximum of β carbon atoms in a branched or linear relationship or any combination of the three, where α and β represent integers. The alkyl groups described in this section may also contain one or two double or triple bonds. The designation of C0 alkyl indicates a direct bond. C 1-6 Examples of alkyl groups include, but are not limited to, the following:

[0139]

[0140] "Benzo group" alone or in combination means a divalent group C4H4=, one of which is -CH=CH-CH=CH-, which when ortho-attached to another ring forms a benzene-like ring - for example, tetralin, indole, and the like.

[0141] The terms "oxo" and "thio" refer to =0 (eg, carbonyl) and =S (eg, thiocarbonyl), respectively.

[0142] "Halo" or "halogen" means a halogen atom selected from F, Cl, Br and I.

[0143] “C α-β "Haloalkyl" means an alkyl group as described above, wherein any number (at least one) of the hydrogen atoms attached to the alkyl chain are replaced by F, Cl, Br or I.

[0144] N(R a )R a The group includes two R a The groups together form a substituent of a ring (optionally containing N, O or S atoms) and include the following groups, for example:

[0145]

[0146] Group N(C α-β Alkyl)C α-β Alkyl groups (wherein α and β are as defined above) include groups wherein two C α-β Alkyl groups are substituents that together form a ring (optionally containing N, O or S atoms) and include the following groups, for example:

[0147]

[0148] "Bicyclic" means a group having two connected rings. Bicyclic rings can be carbocyclic rings (all ring atoms are carbon atoms) or heterocyclic rings (in addition to carbon atoms, the ring atoms include, for example, 1, 2 or 3 heteroatoms, such as N, O or S). Both rings can be aliphatic (e.g., decalin and norbornane), or can be aromatic (e.g., naphthalene), or a combination of aliphatic and aromatic (e.g., tetralin). Bicyclic rings include (a) spirocyclic compounds, in which the two rings share only one single atom (the spiro atom, which is typically a quaternary carbon). Examples of spirocyclic compounds include, but are not limited to:

[0149]

[0150] (b) Fused bicyclic compounds, where the two rings share two adjacent atoms. In other words, the rings share one covalent bond, i.e., the bridgehead atoms are directly connected (e.g., α-thujene and decalin). Examples of fused bicyclic rings include, but are not limited to:

[0151]

[0152]

[0153] ; and (c) bridged bicyclic compounds, wherein the two rings share three or more atoms and the two bridgehead atoms are separated by a bridge comprising at least one atom. For example, norbornane, also known as bicyclo[2.2.1]heptane, can be considered as a pair of cyclopentane rings, each ring sharing three of their five carbon atoms. Examples of bridged bicyclic rings include, but are not limited to:

[0154]

[0155] Unless otherwise indicated, "carbocycle" or "carbocyclic" means, by itself or in combination with other terms, a ring comprising "C α-β Examples of carbocyclic rings include cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, cyclobutylene, cyclohexylene, and the like.

[0156] "Heterocycle" or "heterocyclic" means a ring containing at least one carbon atom and at least one other atom selected from N, O and S. Examples of heterocycles that may appear in the claims include, but are not limited to, the following:

[0157]

[0158]

[0159] "Pharmaceutically acceptable salts" means salts prepared by conventional means and are well known to those skilled in the art. "Pharmaceutically acceptable salts" include alkaline salts of inorganic and organic acids, including but not limited to hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, malic acid, acetic acid, oxalic acid, tartaric acid, citric acid, lactic acid, fumaric acid, succinic acid, maleic acid, salicylic acid, benzoic acid, phenylacetic acid, mandelic acid, and the like. When the compounds of the present invention contain an acidic functional group such as a carboxyl group, then suitable pharmaceutically acceptable cation pairs for the carboxyl group are well known to those skilled in the art and include alkali metals, alkaline earth metals, ammonium, quaternary ammonium cations, and the like. For additional examples of "pharmaceutically acceptable salts", see below and Berge et al., J. Pharm. Sci. [Journal of Pharmaceutical Sciences] 66: 1 (1977).

[0160] "Saturated, partially saturated or unsaturated" includes substituents saturated with hydrogen, substituents fully unsaturated with hydrogen and substituents partially saturated with hydrogen.

[0161] "Leaving group" generally refers to a group that is easily displaced by a nucleophile, such as an amine, thiol or alcohol nucleophile. Such leaving groups are well known in the art. Examples of such leaving groups include, but are not limited to, N-hydroxysuccinimide, N-hydroxybenzotriazole, halides, triflate, tosylate, etc. Preferred leaving groups are indicated herein where appropriate.

[0162] "Protective group" generally refers to a group well known in the art, which is used to prevent selected reactive groups such as carboxyl, amino, hydroxyl, sulfhydryl, etc. from undesirable reactions, such as nucleophilic, electrophilic, oxidative, reduction, etc. Preferred protective groups are indicated in this article when appropriate. Examples of amino protective groups include but are not limited to aralkyl, substituted aralkyl, cycloalkenylalkyl and substituted cycloalkenylalkyl, allyl, substituted allyl, acyl, alkoxycarbonyl, aralkyloxycarbonyl, silyl, etc. Examples of aralkyl include but are not limited to benzyl, o-methylbenzyl, trityl and diphenylmethyl, which can be optionally substituted by halogen, alkyl, alkoxy, hydroxyl, nitro, acylamino, acyl, etc. and salts (such as phosphonium salts and ammonium salts). Examples of aryl include phenyl, naphthyl, indanyl, anthracenyl, 9-(9-phenylfluorenyl), phenanthrenyl, durenyl, etc. Examples of cycloalkenylalkyl or substituted cycloalkenylalkyl groups preferably have 6 to 10 carbon atoms, including but not limited to cyclohexenylmethyl, etc. Suitable acyl, alkoxycarbonyl and aralkyloxycarbonyl include benzyloxycarbonyl, tert-butoxycarbonyl, isobutyloxycarbonyl, benzoyl, substituted benzoyl, butyryl, acetyl, trifluoroacetyl, trichloroacetyl, phthaloyl, etc. A mixture of protecting groups can be used to protect the same amino group, such as a primary amino group can be protected by both aralkyl and aralkyloxycarbonyl. Amino protecting groups can also form heterocycles with the nitrogen to which they are attached, such as 1,2-bis(methylene)benzene, phthalimide, succinimidyl, maleimide, etc., and wherein these heterocyclic groups can further include adjacent aryl and cycloalkyl rings. In addition, heterocyclic groups can be mono-, di- or tri-substituted, such as nitrophthalimide. Amino groups can also be protected from undesirable reactions, such as oxidation, by forming addition salts (e.g., hydrochloride, p-toluenesulfonic acid, trifluoroacetic acid, etc.). Many amino protecting groups are also suitable for protecting carboxyl, hydroxyl, and sulfhydryl groups. For example, aralkyl. Alkyl groups are also suitable for protecting hydroxyl and sulfhydryl groups, such as tert-butyl.

[0163] Silyl protecting groups are silicon atoms optionally substituted with one or more alkyl, aryl and aralkyl groups. Suitable silyl protecting groups include, but are not limited to, trimethylsilyl, triethylsilyl, triisopropylsilyl, tert-butyldimethylsilyl, dimethylphenylsilyl, 1,2-bis(dimethylsilyl)benzene, 1,2-bis(dimethylsilyl)ethane and diphenylmethylsilyl. The silylation of amino groups provides mono- or di-silylamino groups. The silylation of amino alcohol compounds can result in N,N,O-trimethylsilyl derivatives. Removing silyl functions from silyl ether functions is easily accomplished by treatment with, for example, metal hydroxides or ammonium fluoride reagents, which are carried out as separate reaction steps or in situ during reaction with alcohol groups. Suitable silylating agents are, for example, trimethylsilyl chloride, tert-butyl-dimethylsilyl chloride, phenyldimethylsilyl chloride, diphenylmethylsilyl chloride or a combination product thereof with imidazole or DMF. Methods for silylation of amines and removal of silyl protecting groups are well known to those skilled in the art. Methods for preparing these amine derivatives from the corresponding amino acids, amino acid amides or amino acid esters are also well known to those skilled in the art of organic chemistry, including amino acid / amino acid ester or amino alcohol chemistry.

[0164] The protecting group is removed under conditions that will not affect the remainder of the molecule. These methods are well known in the art and include acid hydrolysis, hydrogenolysis, etc. A preferred method involves removing the protecting group, for example, by utilizing palladium carbon hydrogenolysis in a suitable solvent system (e.g., alcohol, acetic acid, etc., or a mixture thereof) to remove the benzyloxycarbonyl group. The tert-butoxycarbonyl protecting group can be removed using an inorganic or organic acid (e.g., HCl or trifluoroacetic acid) in a suitable solvent system (e.g., dioxane or methylene chloride). The resulting amino salt can be easily neutralized to obtain free amine. The carboxyl protecting group (e.g., methyl, ethyl, benzyl, tert-butyl, 4-methoxyphenylmethyl, etc.) can be removed under hydrolysis and hydrogenolysis conditions well known to those skilled in the art.

[0165] It should be noted that the compounds of the present invention may contain groups that can exist in tautomeric forms, such as cyclic and acyclic amidine and guanidinium groups, heteroatom-substituted heteroaryl groups (Y'=O, S, NR), etc., which are illustrated in the following examples:

[0166]

[0167] Although one form is named, described, shown, and / or claimed herein, all tautomeric forms are intended to be inherently included in such name, description, showing, and / or claiming.

[0168] The present invention also contemplates prodrugs of the compounds of the present invention. Prodrugs are active or inactive compounds that are chemically modified to compounds of the present invention by physiological effects such as hydrolysis, metabolism, etc. in vivo after the prodrug is administered to a patient. The suitability and technology for preparing and using prodrugs are well known to those skilled in the art. For a general discussion of prodrugs involving esters, see Svensson and Tunek, Drug Metabolism Reviews [Drug Metabolism Review] 165 (1988) and Bundgaard, Design of Prodrugs [Prodrug Design], Elsevier (1985). Examples of masked carboxylate anions include a variety of esters, such as alkyl (e.g., methyl, ethyl), cycloalkyl (e.g., cyclohexyl), aralkyl (e.g., benzyl, p-methoxybenzyl) and alkylcarbonyloxyalkyl (e.g., pivaloyloxymethyl). Amines have been masked as arylcarbonyloxymethyl substituted derivatives which are cleaved in vivo by esterases, releasing the free drug and formaldehyde (Bungaard J. Med. Chem. 2503 (1989)). Similarly, drugs containing acidic NH groups (e.g., imidazoles, imides, indoles, etc.) have been masked with N-acyloxymethyl groups (Bundgaard Design of Prodrugs, Elsevier (1985)). Hydroxyl groups have been masked as esters and ethers. EP 039,051 (Sloan and Little, 4 / 11 / 81) discloses Mannich base hydroxamic acid prodrugs, their preparation and use.

[0169] This specification and claims contain generic lists (sometimes referred to as Markush groups) that use the language "selected from ... and ..." and "is ... or ...". When this language is used in this application, unless otherwise indicated, it is meant to include the group as a whole, or any individual member thereof, or any subgroup thereof. The use of this language is for shorthand purposes only and is not intended to limit in any way the removal of individual elements or subgroups as desired.

[0170] Pharmaceutical compositions, administration and routes of administration

[0171] Also provided herein are pharmaceutical compositions comprising compounds as disclosed herein and pharmaceutically acceptable excipients, such as diluents or carriers. Compounds and pharmaceutical compositions suitable for use in the present invention include those compounds and pharmaceutical compositions that can be administered in effective amounts to achieve their intended purpose. The administration of the compound will be described in more detail below.

[0172] Suitable pharmaceutical formulations can be determined by technicians based on the route of administration and the desired dosage. See, for example, Remington's Pharmaceutical Sciences, 1435-712 (18th edition, Mack Publishing Co, Easton, Pennsylvania, 1990). Formulations can affect the physical state, stability, or in vivo release rate and in vivo clearance rate of the administered medicament. Depending on the route of administration, a suitable dosage can be calculated based on body weight, body surface area, or organ size. Those of ordinary skill in the art, without excessive experimentation, will routinely perform further refinements of the calculations required to determine the appropriate therapeutic dosage, particularly based on the dosage information and assays disclosed herein and the pharmacokinetic data that can be obtained through animal or human clinical trials.

[0173] The phrase "pharmaceutically acceptable" or "pharmacologically acceptable" refers to molecular entities and compositions that do not produce adverse, allergic or other untoward reactions when administered to animals or humans. As used herein, "pharmaceutically acceptable" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. The use of such excipients for pharmaceutically active substances is well known in the art. Except in cases where any conventional medium or agent is incompatible with the therapeutic composition, its use in the therapeutic composition is contemplated. Supplementary active ingredients may also be incorporated into the composition. In an exemplary embodiment, the formulation may include corn syrup solids, high oleic safflower oil, coconut oil, soybean oil, L-leucine, tricalcium phosphate, L-tyrosine, L-proline, L-lysine acetate, DATEM (emulsifier), L-glutamine, L-valine, dipotassium phosphate, L-isoleucine, L-arginine, L-alanine, glycine, L-asparagine monohydrate, L-serine, potassium citrate, L-threonine, sodium citrate, magnesium chloride, L-histidine, L-methionine, ascorbic acid, Calcium carbonate, L-glutamic acid, L-cystine dihydrochloride, L-tryptophan, L-aspartic acid, choline chloride, taurine, m-inositol, ferrous sulfate, ascorbyl palmitate, zinc sulfate, L-carnitine, alpha-tocopheryl acetate, sodium chloride, niacinamide, mixed tocopherols, calcium pantothenate, ketone sulfate, thiamine chloride hydrochloride, vitamin A palmitate, manganese sulfate, riboflavin, pyridoxine hydrochloride, folic acid, beta-carotene, potassium iodide, phylloquinone, biotin, sodium selenate, chromium chloride, sodium molybdate, vitamin D3 and cyanocobalamin.

[0174] The compound may be present in the pharmaceutical composition as a pharmaceutically acceptable salt. As used herein, "pharmaceutically acceptable salt" includes, for example, base addition salts and acid addition salts.

[0175] Pharmaceutically acceptable base addition salts can be formed with metals or amines (e.g., alkali metals and alkaline earth metals or organic amines). Pharmaceutically acceptable salts of compounds can also be prepared with pharmaceutically acceptable cations. Suitable pharmaceutically acceptable cations are well known to those skilled in the art and include alkali metals, alkaline earth metals, ammonium and quaternary ammonium cations. Carbonates or bicarbonates are also possible. Examples of metals used as cations are sodium, potassium, magnesium, ammonium, calcium or ferric iron, etc. Examples of suitable amines include isopropylamine, trimethylamine, histidine, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, dicyclohexylamine, ethylenediamine, N-methylglucamine and procaine.

[0176] Pharmaceutically acceptable acid addition salts include inorganic or organic acid salts. Examples of suitable acid salts include hydrochlorides, formates, acetates, citrates, salicylates, nitrates, and phosphates. Other suitable pharmaceutically acceptable salts are well known to those skilled in the art and include, for example, formic acid, acetic acid, citric acid, oxalic acid, tartaric acid or mandelic acid, hydrochloric acid, hydrobromic acid, sulfuric acid or phosphoric acid; with organic carboxylic acids, sulfonic acids, sulfonic acids or phosphoric acids or N-substituted aminosulfonic acids, such as acetic acid, trifluoroacetic acid (TFA), propionic acid, glycolic acid, succinic acid, maleic acid, hydroxymaleic acid, methylmaleic acid, fumaric acid, malic acid, tartaric acid, lactic acid, oxalic acid, gluconic acid, glucuronic acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, salicylic acid, 4-aminosalicylic acid, 2-phenyl salts of 2-hydroxy-1-hydroxy-2-oxo-1-yl-2-nitro-2 ...

[0177] Pharmaceutical compositions containing the compounds disclosed herein can be manufactured in conventional manner, eg, by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or lyophilizing processes. Proper formulation depends upon the route of administration chosen.

[0178] For oral administration, suitable compositions can be easily prepared by combining the compounds disclosed herein with pharmaceutically acceptable excipients (e.g., carriers) well known in the art. Such excipients and carriers enable the compounds of the present invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, etc., for oral ingestion by patients to be treated. Pharmaceutical preparations for oral use can be obtained in the following manner: solid excipients are added to the compounds disclosed herein, the resulting mixture is optionally ground, and the granular mixture is processed (if necessary) after adding suitable adjuvants to obtain tablets or dragee cores. Suitable excipients include, for example, fillers and cellulose preparations. If necessary, disintegrants can be added. Pharmaceutically acceptable ingredients for various types of preparations are well known, and can be, for example, adhesives (e.g., natural or synthetic polymers), lubricants, surfactants, sweeteners and flavoring agents, coating materials, preservatives, dyes, thickeners, adjuvants, antimicrobial agents, antioxidants, and carriers for various types of preparations.

[0179] When a therapeutically effective amount of a compound disclosed herein is administered orally, the composition is typically in the form of a solid (eg, tablet, capsule, pill, powder or lozenge) or liquid formulation (eg, aqueous suspension, solution, elixir or syrup).

[0180] When administered in tablet form, the composition may additionally contain functional solids and / or solid carriers, such as gelatin or adjuvants.Tablets, capsules and powders may contain from about 1% to about 95% of the compound, and preferably from about 15% to about 90% of the compound.

[0181] When applied in liquid or suspension form, functional liquids and / or liquid carriers, such as water, petroleum, or oils of animal or plant origin, may be added. The liquid form of the composition may further contain physiological saline solution, sugar alcohol solution, dextrose or other sugar solution or glycol. When applied in liquid or suspension form, the composition may contain about 0.5% to about 90% of the compound disclosed herein by weight, and preferably about 1% to about 50% of the compound disclosed herein. In one embodiment considered, the liquid carrier is non-aqueous or substantially non-aqueous. For application in liquid form, the composition may be supplied as a fast-dissolving solid formulation for dissolving or suspending before being applied.

[0182] When a therapeutically effective amount of the compound disclosed herein is administered by intravenous, percutaneous or subcutaneous injection, the composition is in the form of a pyrogen-free, parenterally acceptable aqueous solution. The preparation of such parenterally acceptable solutions should take into full account pH, isotonicity, stability, etc., and is within the technical scope of the art. In addition to the compounds disclosed herein, preferred compositions for intravenous, percutaneous or subcutaneous injection typically contain isotonic vehicles. Such compositions can be prepared for use as a solution of a free alkali or a pharmacologically acceptable salt in water appropriately mixed with a surfactant (e.g., hydroxypropylcellulose). Dispersions can also be prepared in glycerol, liquid polyethylene glycol, and mixtures thereof, as well as in oils. Under common storage and use conditions, these preparations may optionally contain a preservative to prevent microbial growth.

[0183] Injectable compositions may include sterile aqueous solutions, suspensions or dispersions, and sterile powders for the extemporaneous preparation of sterile injectable solutions, suspensions or dispersions. In all embodiments, the form must be sterile, and fluidity must be such that there is easy injectability. It must be stable under manufacturing and storage conditions, and must resist the contamination of microorganisms (e.g., bacteria and fungi) by optionally including preservatives. The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, etc.), suitable mixtures thereof, and vegetable oils. In one embodiment considered, the carrier is non-aqueous or substantially non-aqueous. Appropriate fluidity may be maintained, for example, by using a coating, such as lecithin; by maintaining the desired particle size of the compound in the embodiment of the dispersion; and by using a surfactant. Prevention of microbial action may be achieved by various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, etc. In many embodiments, it will be preferred to include an isotonic agent (e.g., sugar or sodium chloride). Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.

[0184] Sterile injectable solutions are prepared by incorporating the active compound in the desired amount into an appropriate solvent optionally containing various other ingredients listed above, followed by filtration sterilization. Typically, dispersions are prepared by incorporating various sterilized active ingredients into a sterile vehicle containing a basic dispersion medium and other desired ingredients from those listed above. In the example of sterile powders for the preparation of sterile injectable solutions, preferred methods of preparation are vacuum drying and freeze drying techniques, which produce a powder of the active ingredient plus any desired other ingredients from a previously sterile filtered solution thereof.

[0185] Slow release or sustained release formulations can also be prepared to achieve controlled release of the active compound in contact with body fluids in the gastrointestinal tract, and provide substantially constant and effective active compound levels in blood plasma. For example, release can be controlled by one or more of dissolution, diffusion and ion exchange. In addition, slow release methods can promote absorption by saturable or restricted pathways in the gastrointestinal tract. For example, for this purpose, the compound can be embedded in a polymer matrix of a biodegradable polymer, a water-soluble polymer or a mixture of the two and optionally a suitable surfactant. In this case, embedding can mean incorporating microparticles in a polymer matrix. Controlled release formulations are also obtained by encapsulating dispersed microparticles or emulsified droplets via known dispersion or emulsion coating techniques.

[0186] For administration by inhalation, the compounds of the invention are conveniently delivered in the form of an aerosol spray presentation from a pressurized pack or nebulizer using a suitable propellant. In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges of, for example, gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of the compound and a suitable powder base, such as lactose or starch.

[0187] The compounds disclosed herein can be formulated for parenteral administration by injection (e.g., by bolus or continuous infusion). Injectable formulations can be presented in unit dosage form (e.g., in ampoules or in multi-dose containers) with added preservatives. The composition can take forms such as suspensions, solutions or emulsions in oily or aqueous vehicles and can contain preparatants such as suspending agents, stabilizers and / or dispersants.

[0188] Pharmaceutical formulations for parenteral administration include aqueous solutions of compounds in water-soluble form. In addition, the suspension of the compound can be prepared as a suitable oily injection suspension. Suitable lipophilic solvents or vehicles include fatty oils or synthetic fatty acid esters. Aqueous injection suspensions can contain substances that improve the viscosity of the suspension. Optionally, the suspension can also contain a suitable stabilizer or an agent that improves the solubility of the compound and allows the preparation of a highly concentrated solution. Alternatively, the composition of the present invention can be in powder form for use with a suitable vehicle (e.g., sterile pyrogen-free water) before use.

[0189] The compounds disclosed herein can also be formulated in rectal compositions, such as suppositories or retention enemas (e.g., containing conventional suppository bases). In addition to the formulations described previously, the compounds can also be formulated as long-acting preparations. Such long-acting formulations can be administered by implantation (e.g., subcutaneous or intramuscular) or by intramuscular injection. Thus, for example, the compounds can be formulated with suitable polymeric or hydrophobic materials (e.g., as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives (e.g., as sparingly soluble salts).

[0190] In particular, the compounds disclosed herein can be administered orally, buccally or sublingually in the form of tablets containing excipients (e.g., starch or lactose), or in capsules or ovules alone or mixed with excipients, or in the form of elixirs or suspensions containing flavoring agents or coloring agents. Such liquid preparations can be prepared with pharmaceutically acceptable additives (e.g., suspending agents). The compounds can also be injected parenterally, e.g., intravenously, intramuscularly, subcutaneously, or intracoronarily. For parenteral administration, the compounds are best used in the form of sterile aqueous solutions, which may contain other substances, such as salts or sugar alcohols (e.g., mannitol) or glucose, to make the solution isotonic with the blood.

[0191] For veterinary use, the compounds disclosed herein are administered as suitably acceptable formulations in accordance with normal veterinary practice. A veterinarian can readily determine the most appropriate dosing regimen and route of administration for a particular animal.

[0192] In some embodiments, all the required components for treating KIF18A-related disorders using a compound as disclosed herein (alone or in combination with another agent or intervention traditionally used to treat such a disease) can be packaged into a kit. Specifically, the present invention provides a kit for therapeutic intervention of a disease, the kit comprising a packaged set of drugs, including a compound disclosed herein and a buffer and other components for preparing a deliverable form of the drug; and / or a device for delivering such a drug; and / or any agent for combination therapy with a compound disclosed herein; and / or instructions for treating a disease packaged with the drug. The instructions can be fixed in any tangible medium, such as printed paper, or a computer-readable magnetic or optical medium, or instructions referring to a remote computer data source, such as a World Wide Web page accessible via the Internet.

[0193] "Therapeutically effective amount" means an amount that effectively treats or prevents the development of existing symptoms of the treated subject or alleviates existing symptoms. In particular, the determination of the effective amount is fully within the capabilities of those skilled in the art in light of the detailed disclosure provided herein. Generally, a "therapeutically effective dose" refers to the amount of a compound that results in the desired effect. For example, in a preferred embodiment, the therapeutically effective amount of the compound disclosed herein reduces KIF18A activity by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85% or at least 90% compared to a control group.

[0194] The amount of compound administered may depend on the subject being treated, the subject's age, health, sex, and weight, the type of concurrent treatment (if any), the severity of the condition, the nature of the desired effect, the manner and frequency of treatment, and the judgment of the prescribing physician. The frequency of administration may also depend on the pharmacodynamic effect on arterial oxygen pressure. Although individual needs vary, determination of the optimal range of effective amounts of the compound is within the skill of the art. Such doses may be administered in a single dose, or they may be divided into multiple doses.

[0195] As used herein, the terms "cancer" and "cancerous" refer to or describe a physiological condition that is typically characterized by unregulated cell growth in mammals. Examples of cancer include, but are not limited to, carcinomas, lymphomas, sarcomas, blastomas, and leukemias. More specific examples of such cancers include squamous cell carcinoma, lung cancer, pancreatic cancer, cervical cancer, bladder cancer, liver cancer, breast cancer, colon cancer, and head and neck cancer, ovarian cancer, and endometrial cancer. Although the term "cancer" as used herein is not limited to any particular form of the disease, it is believed that the methods of the present invention are particularly effective for discovering cancers that are accompanied by unregulated levels of KIF18A or that are dependent on KIF18A, which is used for proper chromosome segregation and survival in mammals.

[0196] As used herein, the terms "treat", "treating" and "treatment" refer to therapy, including but not limited to curative therapy, prophylactic therapy and preventive therapy. Preventive treatment generally involves completely preventing the onset of an individual's disease or delaying the onset of a preclinically evident stage of an individual's disorder.

[0197] As used herein, the term "patient", "subject" or "mammal" refers to any "patient", "subject" or "mammal", including humans, cows, horses, dogs and cats. In one embodiment of the present invention, the mammal is a human.

[0198] The term "comprises / comprising" is intended to be open ended, including the stated one or more components but not excluding other elements.

[0199] The term "Formula I" includes any subformulae, such as (Ia), (Ib), (Ic), (Id), etc.

[0200] Methods of using KIF18A inhibitors

[0201] The present disclosure provides compounds with general MT-based KIF18A regulatory activity, particularly inhibitory activity. In one embodiment of the present invention, a method for regulating KIF18A protein in a subject is provided, the method comprising administering an effective dose of a compound of formula I to the subject. Therefore, the compounds of the present invention can be used to treat cell proliferation disorders, including uncontrolled cell growth, abnormal cell cycle regulation, centrosome abnormalities (structural and / or quantitative, fragmentation). Other diseases or disorders associated with the accumulation of extra centrosomes (>2) include human papillomavirus (HPV) infection, including HPV-related tumors. The compound can also be used for cilia-related diseases and ablation of haploid germ cell populations that can be used as male contraceptives.

[0202] In addition, the compounds of the present invention can be used for, but not limited to, the prevention or treatment of cancer and other KIF18A-mediated diseases or conditions. For example, the compounds of the present invention can be used to treat a variety of solid tumors or blood-derived tumors, such as cancer, including but not limited to bladder cancer, breast cancer, colon cancer, kidney cancer, liver cancer, lung cancer (including squamous cell carcinoma and small cell lung cancer), esophageal cancer, gallbladder cancer, ovarian cancer, pancreatic cancer, gastric cancer, cervical cancer, thyroid cancer, prostate cancer and skin cancer (including squamous cell carcinoma); hematopoietic tumors of the lymphoid system (including leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B cell lymphoma, T cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, hair of the myeloid lineage (including acute and chronic myeloid leukemias, myelodysplastic syndromes, and promyelocytic leukemias); tumors of mesenchymal origin (including fibrosarcomas and rhabdomyosarcomas, as well as other sarcomas, such as those of soft tissue and bone); tumors of the central and peripheral nervous system (including astrocytomas, neuroblastomas, gliomas, and schwannomas); and other tumors (including melanomas, seminoma, teratocarcinoma, osteosarcoma, xeroderma pigmentosum, keratoacanthoma, follicular thyroid carcinoma, or Kaposi’s sarcoma).

[0203] The compounds of the invention may also be used to treat cancer-related indications, such as solid tumors, sarcomas (particularly Ewing's sarcoma and osteosarcoma), retinoblastoma, rhabdomyosarcoma, neuroblastoma, hematopoietic malignancies (including leukemias and lymphomas), tumor-induced pleural or pericardial effusions and malignant ascites.

[0204] Based on the ability of modulating kinesin to affect angiogenesis, the compounds of the present invention may also be used for the treatment and therapy of proliferative diseases. In particular, these compounds may be used to treat inflammatory diseases, especially the manifestations of the motor organs, such as various inflammatory rheumatoid diseases, especially chronic polyarthritis, including rheumatoid arthritis, juvenile arthritis or psoriatic arthritis; tumor-associated syndrome or tumor-induced inflammatory diseases, turbid effusions, collagen diseases, such as systemic lupus erythematosus, polymyositis, dermatomyositis, systemic scleroderma or mixed collagen diseases; post-infectious arthritis (live pathogenic organisms cannot be found in or in the affected part of the body), seronegative spondyloarthritis, such as ankylosing spondylitis; vasculitis, sarcoidosis or arthropathy; or further any combination thereof.

[0205] The compounds of the present invention can also be used as active agents for such conditions, such as arthritis, atherosclerosis, psoriasis, hemangiomas, myocardial angiogenesis, coronary artery and cerebral lateral processes, colloid ischemic angiogenesis, wound healing, peptic ulcers, Helicobacter pylori-related diseases, fractures, cat scratch fever, flushing, neovascular glaucoma, and retinopathy (e.g., retinopathy associated with diabetic retinopathy or macular degeneration). In addition, some of these compounds can be used as active agents for solid tumors, malignant ascites, hematopoietic cancers, and hyperproliferative disorders (e.g., thyroid hyperplasia (especially Graves' disease)) and cysts (e.g., ovarian stroma hypervascularization, features of polycystic ovary syndrome (Stein-Leventhal syndrome)), because such diseases require vascular cell proliferation to grow and / or metastasize.

[0206] In addition to being useful for human treatment, these compounds can also be used in veterinary treatment of companion animals, exotic animals, and farm animals, including mammals, rodents, etc. For example, animals including horses, dogs, and cats can be treated with the compounds provided herein.

[0207] combination

[0208] Although the compounds of the invention can be administered or used as the sole active pharmaceutical agent, they can also be used in combination with one or more compounds of the invention or in conjunction with other agents. When administered in combination, the therapeutic agents can be formulated into separate compositions that are administered simultaneously or sequentially at different times, or the therapeutic agents can be administered as a single composition.

[0209] In defining the use of a compound of the invention and another pharmaceutical agent, the phrase "co-therapy" (or "combination therapy") is intended to include administration of each agent in a sequential manner in a regimen that provides a beneficial pharmaceutical combination effect, and is also intended to include co-administration of the agents in a substantially simultaneous manner, such as in a single capsule having a fixed ratio of the active agents or in multiple separate capsules of each agent.

[0210] In particular, administration of the compounds of the invention can be combined with other therapies known to those skilled in the art for the prevention or treatment of cancer, such as radiation therapy, small molecule targeted agents (e.g., PARP inhibitors, kinase inhibitors), therapeutic antibodies (e.g., naked and drug-conjugated) with tumor or cytotoxic agents, immunotherapy antibodies (checkpoint inhibitors, bispecific T cell engagers).

[0211] If formulated as a fixed dose, such combination products employ the compounds of the invention within an acceptable dosage range. When a combination formulation is not appropriate, the compounds of Formula I may also be administered sequentially with known anticancer or cytotoxic agents. The invention is not limited to the order of administration; the compounds of the invention may be administered before, simultaneously with, or after the administration of a known anticancer or cytotoxic agent.

[0212] There are a large number of anticancer agents available in commercial use, clinical evaluation and preclinical development that can be selected for the treatment of tumors through combination drug chemotherapy. Such agents are divided into several major categories, such as antibiotic agents, alkylating and alkylating-like agents, antimitotic agents, small molecule targeted agents, antimetabolites, hormonal agents, immune agents, antiangiogenic agents, interferon-like agents and miscellaneous agents.

[0213] The disclosure also provides methods for combination therapy, wherein agents known to modulate other pathways or other components of the same pathway, or even overlapping sets of target enzymes, are used in combination with the compounds of the disclosure or pharmaceutically acceptable salts thereof. In one aspect, such therapy includes, but is not limited to, the combination of one or more compounds of the disclosure with chemotherapeutic agents, therapeutic antibodies, small molecule targeted agents, and radiation therapy to provide a synergistic or additive therapeutic effect.

[0214] Many chemotherapeutic agents are currently known in the art and can be used in combination with the compounds of the present disclosure. In some embodiments, the chemotherapeutic agent is selected from the group consisting of antimitotic agents, alkylating agents, antimetabolites, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, antihormones, angiogenesis inhibitors, and antiandrogens. Non-limiting examples are chemotherapeutic agents, cytotoxic agents, and non-peptide small molecules, such as (imatinib mesylate), (carfilzomib), (bortezomib), Casodex (bicalutamide), (gefitinib) and doxorubicin (Adriamycin) and various chemotherapeutic agents. Non-limiting examples of chemotherapeutic agents include alkylating agents, such as thiotepa and cyclophosphamide (CYTOXAN™); alkyl sulfonates, such as busulfan, improsulfan and piposulfan; aziridines, such as benzodopa, carboquinone, meturedopa and uredopa; ethyleneimines and methylamelamines, including hexamethylmelamine, triethylenemelamine, triethylenephosphamide, triethylenethiophosphaoramide and trimethylolomelamine; nitrogen mustards, such as chlorambucil, naphthyl nitrogen mustard, cholophosphamide, estramustine, ifosfamide, dichloromethyldiethylamine, mechlorethamine oxide hydrochloride (mechlorethamine oxide hydrochloride) hydrochloride), melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosoureas, such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine; antibiotics, such as aclacinomysin, actinomycin, authramycin, azaserine, bleomycin, cactinomycin, calicheamicin, carabicin, carminomicin, carzinophilin, CasodexTM , chromomycin, dactinomycin, daunomycin, detoxibicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin, idarubicin, marcellomycin, mitomycin, mycophenolic acid, nogamycin, olive mycin, peplomycin, potfiromycin, puromycin, quelamycin, rhodorubicin, streptomycin, streptozotocin, tuberculocidin, ubenimex, netastatin, daunorubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiopurine, thioguanine;Pyrimidine analogs, such as azacitidine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine, androgens, such as caprotestosterone, dromostanolone propionate, cyclothiosteroid, melastane, testolactone; antiadrenal agents, such as aminoglutethimide, mitotane, trilostane; folic acid supplements, such as folinic acid acid; aceglucuronide; aldehyde phosphamide glycoside; aminolevulinic acid; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; colcemid; diazocone; elfomithine; elliptonium acetate; etoglu; gallium nitrate; hydroxyurea; lentinan; lonidamine; mitoguanidine; mitoxantrone; mopidarol; nitracrine; pentostatin; phenamet; pirarubicin; podophyllinic acid acid); 2-ethylhydrazide; procarbazine; PSK; aproximine; sizoran; spirogermanamine; tricrosporin acid; triazoquinone; 2,2',2"-trichlorotriethylamine; urethan; vindesine; dacarbazine; mannomustine; dibromomannitol; dibromodulcitol; pipobroman; gacytosine; arabinoside ("Ara-C"); cyclophosphamide; thiotepa; taxanes, such as paclitaxel and docetaxel, albumin-bound paclitaxel (Nab-paclitaxel); retinoic acid; esperamicins; capecitabine; and pharmaceutically acceptable salts, acids or derivatives of any of the foregoing.

[0215] Suitable chemotherapeutic cell conditioning agents also include antihormonal agents used to modulate or inhibit the effects of hormones on tumors, such as antiestrogens, including, for example, tamoxifen (NolvadexTM), raloxifene, aromatase inhibiting 4(5)-imidazoles, 4-hydroxytamoxifen, trioxifene, keoxifene, LY 117018, onapristone and toremifene (Falutone); and antiandrogens, such as flutamide, nilutamide, bicalutamide, leuprolide and goserelin; chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs, such as cisplatin, oxaliplatin and carboplatin; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vinblastine, vincristine; vinorelbine; navelbine; novantrone; teniposide; daunomycin; aminopterin; xeloda; ibandronate; topotecan; camptothecin-11 (CPT-11); the topoisomerase inhibitor RFS 2000; and difluoromethylornithine (DMFO).

[0216] If desired, the compounds or pharmaceutical compositions disclosed herein may be used in combination with commonly prescribed anticancer drugs, e.g. Abraxane, ABVD, AVICINE, Abagovomab, Acridine carboxamide, Adecatumumab, 17-N-allylamino-17-demethoxygeldanamycin, Alpharadin, Alvocidib, 3-aminopyridine-2-carboxaldehyde thiosemicarbazide, Amonafide, Anthracenedione, Anti-CD22 immunotoxin, Antineoplastic drugs, Antitumorigenic herbs, Apaziquone, Atiprimod, Azathioprine, Belotecan, Bendamustine, BIBW 2992, Biricodar, Brostallicin, Bryostatin, Buthionine Sulfoxidesulfoximine), CBV (chemotherapy), Calyculin, cell cycle nonspecific antineoplastic agents, dichloroacetic acid, discordantolide, elsamitrucin, enocitabine, epothilone, eribulin, everolimus, exatecan, exisulind, ferruginol, forodesine, fosfestrol, ICE chemotherapy regimen, IT-101, Imexon, imiquimod, indolocarbazole, irofulven, lanicidab, Laniquidar, Larotaxel, Lenalidomide, Lucanthone, Lurtotecan, Mafosfamide, Mitozolomide, Nafoxidine, Nedaplatin, Olaparib, Talazoparib, Niraparib, Ortataxel, PAC-1, Pawpaw, Pixantrone, Proteasome Inhibitors, Rebeccamycin, Resiquimod, Rubitecan, SN-38, Salinosporamide A A), Sapacitabine, Stanford V, Swainsonine, Talaporfin, Tariquidar, Tegafur-uracil, Temodar, Tesetaxel, Triplatin tetranitratetetranitrate), tris(2-chloroethyl)amine, troxacitabine, uramustine, vadimezan, vinflunine, ZD6126 or zosuquidar, CDK4 / 6 inhibitors (palbociclib, Ibrance; ribociclib, Kisqali; abemaciclib, Verzenio).

[0217] The disclosure further relates to methods of inhibiting abnormal cell growth or treating hyperproliferative disorders in mammals using a combination of a compound or pharmaceutical composition provided herein and radiotherapy. Techniques for administering radiotherapy are known in the art, and these techniques can be used in the combination therapies described herein. The administration of the disclosed compound in such a combination therapy can be determined as described herein.

[0218] Radiotherapy can be applied by one of several methods or a combination of methods, including but not limited to external beam therapy, internal radiotherapy, implant radiation, stereotactic radiosurgery, systemic radiation therapy, radiotherapy and permanent or temporary interstitial brachytherapy. As used herein, the term "brachytherapy" refers to radiotherapy delivered by a spatially confined radioactive material that is inserted into the body at or near a tumor or other proliferative tissue diseased site. The term is intended to include exposure to radioisotopes (e.g., At-211, I-131, I-125, Y-90, Re-186, Re-188, Sm-153, Bi-212, P-32 and Lu radioisotopes) without limitation. Suitable radioactive sources used as cell conditioning agents of the present disclosure include solids and liquids. By way of non-limiting example, the radiation source can be a radionuclide, such as I-125, I-131, Yb-169, Ir-192 as a solid source, I-125 as a solid source, or other radionuclides that emit photons, beta particles, gamma radiation or other therapeutic rays. The radioactive material can also be a fluid prepared from any solution of one or more radionuclides (e.g., a solution of I-125 or I-131), or the radioactive fluid can be produced using a slurry of a suitable fluid containing small particles of solid radionuclides (e.g., Au-198, Y-90). In addition, one or more radionuclides can be embedded in a gel or radioactive microspheres.

[0219] The compounds or pharmaceutical compositions disclosed herein may be used in combination with a certain amount of one or more substances selected from the following: anti-angiogenic agents, signal transduction inhibitors, anti-proliferative agents, glycolysis inhibitors, or autophagy inhibitors.

[0220] Anti-angiogenic agents can be used in conjunction with the compounds disclosed herein and the pharmaceutical compositions described herein, and these anti-angiogenic agents are, for example, MMP-2 (matrix metalloproteinase 2) inhibitors, MMP-9 (matrix metalloproteinase 9) inhibitors, and COX-11 (cyclooxygenase 11) inhibitors. Anti-angiogenic agents include, for example, rapamycin, temsirolimus (temsirolimus, CCI-779), everolimus (RAD001), sorafenib, sunitinib, and bevacizumab. Examples of useful COX-II inhibitors include alecoxib, valdecoxib, and rofecoxib. Examples of useful matrix metalloproteinase inhibitors are described in the following patents: WO 96 / 33172, WO 96 / 27583, European Patent Publication EP 0818442, European Patent Publication EP1004578, WO 98 / 07697, WO 98 / 03516, WO 98 / 34918, WO 98 / 34915, WO 98 / 33768, WO 98 / 30566, European Patent Publication 606046, European Patent Publication 931788, WO 90 / 05719, WO 99 / 52910, WO 99 / 52889, WO 99 / 29667, WO 1999007675, European Patent Publication EP 1786785, European Patent Publication No. EP1181017, U.S. Patent Publication No. US 20090012085, US Publication US 5863949, US Publication US5861510 and European Patent Publication EP0780386, all of which are incorporated herein by reference in their entirety. Preferred MMP-2 and MMP-9 inhibitors are those with little or no activity in inhibiting MMP-1. More preferred are those that selectively inhibit MMP-2 and / or AMP-9 relative to other matrix metalloproteinases (i.e., MAP-1, MMP-3, MMP-4, MMP-5, MMP-6, MMP-7, MMP-8, MMP-10, MMP-11, MMP-12 and MMP-13). Some specific examples of MMP inhibitors that can be used in the present disclosure are AG-3340, RO 32-3555 and RS13-0830.

[0221] The compounds of the invention may also be used in co-therapy with other antineoplastic agents, such as acemannan, aclarubicin, aldesleukin, alemtuzumab, alitretinoin, hexamethylmelamine, amifostine, aminolevulinic acid, amrubicin, amsacrine, anagrelide, anastrozole, ANCER, ancestim, ARGLABIN, arsenic trioxide, BAM002 (Novelos), bexarotene, bicalutamide, bromuridine, capecitabine, cimoleukin, cetrorelix, cladribine, clotrimazole, cytarabine ocfosfate, DA 3030 (Dong-A), daclizumab, denileukin diftitox), deslorelin, dexrazoxane, dilazep, docetaxel, docosanol, doxercalciferol, doxifluridine, doxorubicin, bromocriptine, carmustine, cytarabine, fluorouracil, HIT diclofenac, interferon-α, daunomycin, doxorubicin, retinoic acid, edilfosine, edrecolomab, eflornithine, ethidium bromide, epirubicin, erythropoietin beta, etoposide phosphate, exemestane, exisulind, fadrozole, filgrastim, finasteride, fludarabine phosphate, formestane, fotemustine, gallium nitrate, gemcitabine, gemtuzumab zogamicin), gimeracil / oteracil / tegafur combination, glycopine, goserelin, heptaplatin, human chorionic gonadotropin, human fetal alpha-fetoprotein, ibandronate, idarubicin, imiquimod, interferon-α, interferon-α, natural interferon-α-2, interferon-α-2a, interferon-α-2b, interferon-α-N1, interferon-α-n3, interferon alfacon-1, interferon α, natural interferon beta, interferon beta-1a, interferon beta-1b, interferon gamma, natural interferon gamma-1a, interferon gamma-1b, interleukin-1β, iodobenzylguanidine, irinotecan, irsogladine, lanreotide, LC9018 (Yakult), leflunomide, lenograstim, lentinan sulfate, letrozole, interferon alpha, leuprorelin, levamisole + fluorouracil, liarozole, lobaplatin, lonidamine, lovastatin, masoprofen, melarsoprol, metoclopramide, mifepristone, miltefosine, milistostim, mismatched double-stranded RNA, mitoguanidine, dibromodole, mitoxantrone, moxifloxacin Molgramostim, nafarelin, naloxone + pentazocine, nartograstim, nedaplatin, nilutamide, noscapine, novel erythropoiesis stimulating protein, NSC631570 octreotide, oprelvekin, osatetron, oxaliplatin, paclitaxel, pamidronate, pegaspargase, peginterferon-α-2b, pentosan polysulfate sodium, pentostatin, picibanil, pirarubicin, rabbit anti-thymocyte polyclonal antibody, peginterferon-α-2a, porfimer sodium, raloxifene, raltitrexed, rasburiembodiment, etidronate rhenium (Re 186), RII retinamide (RII retinamide), rituximab, romotide, samarium (153Sm) lexidronam, sargramostim, sizoran, sobuzosine, sonermin, strontium-89 chloride, suramin, tasonermin, tazarotene, tegafur, temoporfin, temozolomide, teniposide, tetrachlorodecaoxide, thalidomide, thymofasin, stimulant Thyroxine alfa, topotecan, toremifene, tositumomab-iodine 131, trastuzumab, treosulfan, tretinoin, trilostane, trimetrexate, triptorelin, tumor necrosis factor alpha, natural ubenimex, bladder cancer vaccine, Maruyama vaccine, melanoma lysate vaccine, valrubicin, verteporfin, vinorelbine, virulizin, zinostatin stimalamer, or zoledronic acid; abarelix; AE 941 (Aeterna), ambamustine, antisense oligonucleotides, bcl-2 (Genta), APC8015 (Dendreon), cetuximab, decitabine, dexaminoglutethimide, diazocone, EL 532 (Elan), EM 800 (Endorecherche), eniluracil, etanidazole, fenretinide, filgrastim SD01 (Amgen), fulvestrant, galocitabine, gastrin 17 immunogen, HLA-B7 gene therapy (Vical), granulocyte-macrophage colony-stimulating factor, histamine dihydrochloride, ibritumomab tiuxetan, ilomastat, IM 862 (Cytran), interleukin-2, iproxifene, LDI 200 (Milkhaus), leridistim, lintuzumab, CA 125 MAb (Biomira), cancer MAbs (Japan Pharmaceutical Development), HER-2 and Fc MAbs (Medarex), idiotype 105AD7 MAb (CRC Technology), idiotype CEA MAb (Trilex), LYM-1-iodine 131 MAb (Techniclone), polymorphic epitheliomucin-yttrium 90 MAb (Antisoma), marimastat, menolipir, mitumomab, motexafin gadolinium, MX 6 (Galderma), nelarabine, nolatrexed, P 30 protein, pegvisomant, pemetrexed, porfiromycin, prinomastat, RL 0903 (Shire), lupinotecan, satraplatin, sodium phenylacetate, sparrosic acid, SRL 172 (SR Pharma), SU 5416 (SUGEN), TA 077 (Tanabe), tetrathiomolybdate, thaliblastine, thrombopoietin, tin ethyletiopurpurin), tirapazamine, cancer vaccine (Bamira), melanoma vaccine (New York University), melanoma vaccine (Sloan Kettering Institute), melanoma tumor lysate vaccine (New York Medical College), viral melanoma cell lysate vaccine (Royal Newcastle Hospital), or valspodar.

[0222] The compounds of the present invention may further be used together with a VEGFR inhibitor. Other compounds described in the following patents and patent applications may be used in combination therapy: US 6,258,812, US 2003 / 0105091, WO 01 / 37820, US 6,235,764, WO 01 / 32651, US 6,630,500, US 6,515,004, US 6,713,485, US 5,521,184, US 5,770,599, US 5,747,498, WO 02 / 68406, WO 02 / 66470, WO 02 / 55501, WO 04 / 05279, WO 04 / 07481, WO 04 / 07458, WO 04 / 09784, WO 02 / 59110, WO 99 / 45009, WO 00 / 59509, WO 99 / 61422, US 5,990,141, WO 00 / 12089 and WO 00 / 02871.

[0223] In some embodiments, the combination comprises a combination of the composition of the invention and at least one anti-angiogenic agent. Agents include, but are not limited to, chemical compositions, antibodies, antigen binding regions, radionuclides, and combinations and conjugates thereof prepared synthetically in vitro. Agents may be agonists, antagonists, allosteric modulators, toxins, or more generally, may be used to inhibit or stimulate their targets (e.g., receptor or enzyme activation or inhibition), and thereby promote cell death or prevent cell growth.

[0224] Exemplary anti-angiogenic agents include ERBITUX TM (IMC-C225), KDR (kinase domain receptor) inhibitors (e.g., antibodies and antigen binding regions that specifically bind to kinase domain receptors), anti-VEGF agents (e.g., antibodies or antigen binding regions that specifically bind to VEGF, or soluble VEGF receptors or their ligand binding regions) (e.g., AVASTIN TM VEGF-TRAP TM) and anti-VEGF receptor agents (e.g., antibodies or antigen-binding regions that specifically bind thereto), EGFR inhibitors (e.g., antibodies or antigen-binding regions that specifically bind thereto) (e.g., Vectibix (panitumumab), IRESSA TM (gefitinib), TARCEVA TM (erlotinib), anti-Ang1 agents and anti-Ang2 agents (e.g., antibodies or antigen binding regions that specifically bind to them or to their receptors (e.g., Tie2 / Tek)) and anti-Tie2 kinase inhibitors (e.g., antibodies or antigen binding regions that specifically bind to them). The pharmaceutical compositions of the present invention may also include one or more agents (e.g., antibodies, antigen binding regions or soluble receptors) that specifically bind to growth factors and inhibit the activity of growth factors, such as antagonists of hepatocyte growth factor (HGF, also known as scatter factor), and antibodies or antigen binding regions that specifically bind to its receptor "c-met".

[0225] Other anti-angiogenic agents include alemtuzumab (Campath), IL-8, B-FGF, Tek antagonists (Ceretti et al., U.S. Publication No. 2003 / 0162712; U.S. Pat. No. 6,413,932), anti-TWEAK agents (e.g., specific binding antibodies or antigen binding regions, or soluble TWEAK receptor antagonists; see, Wiley, U.S. Pat. No. 6,727,225), ADAM disintegrin domains antagonizing the binding of integrins to their ligands (Fanslow et al., U.S. Publication No. 2002 / 0 042368), specifically binding anti-eph receptor and / or anti-ephrin antibodies or antigen binding regions (U.S. Pat. Nos. 5,981,245, 5,728,8135,969,110, 6,596,852, 6,232,447, 6,057,124 and their family members), anti-PDGF-BB antagonists (e.g., specifically binding antibodies or antigen binding regions), and antibodies or antigen binding regions that specifically bind to PDGF-BB ligands and PDGFR kinase inhibitors (e.g., antibodies or antigen binding regions that specifically bind thereto).

[0226] Other anti-angiogenic / anti-tumor agents include: SD-7784 (Pfizer, USA); cilengitide (Merck KGaA, Germany, EPO 770622); pegaptanib octasodium (Gilead Sciences, USA); Alphastatin (BioActa, UK); M-PGA (Celgene, USA, US 5712291); ilomastat (Arriva, USA, US 5892112); emaxanib (Pfizer, USA, US 5792783; vatalanib (Novartis, Switzerland); 2-methoxyestradiol (EntreMed, USA); TLC ELL-12 (Elan, Ireland); anecortave acetate (Alcon, USA); α-D148 Mab (Amgen, USA); CEP-7055 (Cephalon, USA); anti-Vn Mab (Crucell, Netherlands) DAC: anti-angiogenic agent (ConjuChem, Canada); Angiocidin (InKine Pharmaceutical, USA); KM-2550 (Kyowa Hakko Kogyo Co., Ltd., Japan); Hakko, Japan); SU-0879 (Pfizer, USA); CGP-79787 (Novartis, Switzerland, EP 970070); ARGENT technology (Ariad, USA); YIGSR-Stealth (Johnson & Johnson, USA); Fibrinogen E fragment (BioActa, UK); Angiogenesis inhibitor (Trigen, UK); TBC-1635 (Encysive Pharmaceuticals, USA);SC-236 (Pfizer, USA); ABT-567 (Abbott, USA); metastasis (EntreMed, USA); angiogenesis inhibitor (Tripep, Sweden); serine protease inhibitor (maspin) (Sosei, Japan); 2-methoxyestradiol (Oncology Sciences Corporation, USA); ER-68203-00 (IVAX, USA); fluazifop (Lane Labs, USA); Tz-93 (Tsumura, Japan); TAN-1120 (Takeda, Japan); FR-111142 (Fujisawa, Japan, JP 02233610); platelet factor 4 (RepliGen, USA, EP 407122); vascular endothelial growth factor antagonist (Borean, Denmark); bevacizumab (pINN) (Genentech, USA); angiogenesis inhibitor (SUGEN, USA); XL 784 (Exelixis, USA); XL647 (Exelixis, USA); MAb, α5β3 integrin, second generation (Applied Molecular Evolution, USA and MedImmune, USA); gene therapy, retinopathy (Oxford Biotech, UK); BioMedica, UK); USAN (Lilly, USA); CEP 7055 (Cephalon, USA and Sanofi-Synthelabo, France); BC 1 (Genoa Institute of Cancer Research, Italy); angiogenesis inhibitors (Alchemia, Australia); VEGF antagonists (Regeneron, USA); rBPI 21 and BPI-derived antiangiogenic agents (XOMA, USA);PI 88 (Progen, Australia); cilengitide (pINN) (Merck KGaA, Germany; Munich Technical University, Germany; Scripps Clinic and Research Foundation, USA); cetuximab (INN) (Aventis, France); AVE 8062 (Ajinomoto, Japan); AS 1404 (Cancer Research Laboratory, New Zealand); SG 292 (Telios, USA); endostatin (Boston Children's Hospital, USA); ATN 161 (Attenuon, USA); angiostatin (Boston Childrens Hospital, USA); 2-methoxyestradiol (Boston Childrens Hospital, USA); ZD 6474 (AstraZeneca, UK); ZD 6126 (Angiogene Pharmaceuticals, UK); PPI 2458 (Praecis, USA); AZD 9935 (AstraZeneca, UK); AZD 2171 (AstraZeneca, UK); vatalanib (pINN) (Novartis, Switzerland and Schering Pharmaceuticals, Germany) AG, Germany); tissue factor pathway inhibitor (EntreMed, USA); pegaptanib (Pinn) (Gilead Sciences, USA); turmeric (Yonsei University, South Korea);Vaccines, gene-based, VEGF-2 (Scripps Clinic and Research Foundation, USA); SPV5.2 (Supratek, Canada); SDX 103 (University of California at San Diego, USA); PX 478 (ProlX, USA); transferstatin (EntreMed, USA); troponin I (Harvard University, USA); SU 6668 (SUGEN, USA); OXI 4503 (OXiGENE, USA); o-guanidine enhancers (Dimensional Pharmaceuticals, USA); motuporamine C (British Columbia University, Canada); CDP 791 (Celltech Group, UK); atiprimod (pINN) (GlaxoSmithKline, UK); E 7820 (Eisai, Japan); CYC 381 (Harvard University, USA); AE 941 (Aeterna, Canada); vaccines, angiogenesis inhibitors (EntreMed, USA); urokinase plasminogen activator inhibitor (Dendreon, USA); oglufanide (pINN) (Melmotte, USA); HIF-1α inhibitor (Xenova, UK); CEP 5214 (Cephalon, USA); BAY RES 2622 (Bayer, Germany); Angiocidin (InKine, USA); A6 (Angstrom, USA); KR 31372 (Korea Research Institute of Chemical Technology, South Korea); GW 2286 (GlaxoSmithKline, UK);EHT 0101 (ExonHit, France); CP 868596 (Pfizer, USA); CP 564959 (OSI, USA); CP 547632 (Pfizer, USA); 786034 (GlaxoSmithKline, UK); KRN 633 (Kirin Brewery, Japan); intraocular, 2-methoxyestradiol (EntreMed, USA); anginex (Maastricht University, Netherlands and Minnesota University, USA); ABT 510 (Abbott, USA); AAL 993 (Novartis, Switzerland); VEGI (ProteomTech, USA); TNF-α inhibitor (National Institute on Aging, USA); SU 11248 (Pfizer, USA and SUGEN, USA); ABT 518 (Abbott, USA); YH16 (Yantai Rongchang, China); S-3APG (Boston Children's Hospital, USA and EntreMed, USA); MAb, KDR (ImClone Systems, USA); MAb, α5β1 (Protein Design, USA); KDR kinase inhibitor (Celltech, UK); Group, UK) and Johnson & Johnson, USA); GFB 116 (South Florida University, USA and Yale University, USA); CS 706 (Sankyo, Japan); constatin A4 prodrug (Arizona State University, USA); chondroitinase AC (IBEX, Canada);BAY RES 2690 (Bayer, Germany); AGM 1470 (Harvard University, USA, Takeda, Japan, and TAP, USA); AG 13925 (Agouron, USA); Tetrathiomolybdate (University of Michigan, USA); GCS 100 (Wayne State University, USA); CV 247 (Ivy Medical, UK); CKD 732 (Chong Kun Dang, South Korea); MAb, vascular endothelial growth factor (Xenova, UK); irsogladine (INN) (Nippon Shinyaku Co., Ltd.); =Index (Shinyaku, Japan); RG13577 (Aventis, France); WX 360 (Wilex, Germany); squalamine (pINN) (Genaera, USA); RPI 4610 (Sirna, USA); cancer therapy (Marinova, Australia); heparanase inhibitors (InSight, Israel); KL 3106 (Kolon, South Korea); Honokiol (Emory University, USA); ZK CDK (Schering AG, Germany); ZKAngio (Schering AG, Germany); AG, Germany); ZK229561 (Novartis, Switzerland and Schering AG, Germany); XMP 300 (XOMA, USA); VGA 1102 (Taisho, Japan); VEGF receptor modulators (Pharmacopeia, USA); VE-cadherin-2 antagonists (ImClone Systems, USA);Angiostatin (National Institutes of Health, USA); Vaccine, Flk-1 (ImClone Systems, USA); TZ 93 (Tsumura, Japan); Tumstatin (Beth Israel Hospital, USA); Short soluble FLT 1 (vascular endothelial growth factor receptor 1) (Merck & Co, USA); Tie-2 ligand (Regeneron, USA); and thrombospondin 1 inhibitor (Allegheny Health, Education and Research Foundation, USA). ;

[0227] Autophagy inhibitors include, but are not limited to, chloroquine, 3-methyladenine, hydroxychloroquine (Plaquenil TM ), bafilomycin A1, 5-amino-4-imidazolecarboxamide ribonucleoside (AICAR), okadaic acid, autophagy-inhibiting algal toxins that inhibit type 2A or type 1 protein phosphatase, cAMP analogs, and drugs that increase cAMP levels such as adenosine, LY204002, N6-mercaptopurine ribonucleoside, and vinblastine. In addition, antisense or siRNA that inhibits protein expression can also be used, and these proteins include but are not limited to ATG5 (which is involved in autophagy).

[0228] Other pharmaceutically active compounds / agents that can be used to treat cancer and can be used in combination with one or more compounds of the present invention include: epoetin alfa; darbepoetin alfa; panitumumab; pegfilgrastim; palifermin; filgrastim; denosumab; anciplostim; AMG 102; AMG 386; AMG 479; AMG 655; AMG 745; AMG 951; and AMG706 or a pharmaceutically acceptable salt thereof.

[0229] In certain embodiments, the compositions provided herein are administered in combination with a chemotherapeutic agent. Suitable chemotherapeutic agents may include natural products, such as vinca alkaloids (e.g., vinblastine, vincristine, and vinorelbine), taxol, epipodophyllotoxins (e.g., etoposide and teniposide), antibiotics (e.g., dactinomycin (actinomycin D), daunomycin, doxorubicin, and idarubicin), anthracyclines, mitoxantrone, bleomycin, plicamycin (mithramycin), mitomycin, enzymes (e.g., L-asparaginase, which systemically metabolizes L-asparagine and deprive cells that do not have the ability to synthesize their own asparagine), antiplatelet agents, antiproliferative / antimitotic alkylating agents (e.g., nitrogen mustards, e.g., dichloromethyl diethylamide, cyclophosphamide and analogs, melphalan, and chlorambucil), ethyleneimines and methylmelamines (e.g., hexamethylmelaamine and thiotepa), CDK inhibitors (e.g., seliciclib, UCN-01, P1446A-05, PD-03 32991, dinaciclib, P27-00, AT-7519, RGB286638 and SCH727965), alkyl sulfonates (e.g., busulfan), nitrosoureas (e.g., carmustine (BCNU) and analogs and streptozotocin), trazenes-dacarbazinine (DTIC), antiproliferative / antimitotic antimetabolites such as folic acid analogs (e.g., methotrexate), pyrimidine analogs (e.g., The drugs include, for example, 5-fluorouracil, floxuridine, and cytarabine), purine analogs and related inhibitors (e.g., mercaptopurine, thioguanine, pentostatin, and 2-chlorodeoxyadenosine), aromatase inhibitors (e.g., anastrozole, exemestane, and letrozole), and platinum coordination complexes (e.g., cisplatin and carboplatin), procarbazine, hydroxyurea, mitotane, aminoglutethimide, histone deacetylase (HDAC) inhibitors (e.g., trichostatin, sodium butyrate, apicidan, suberoylanilide hydroxamic acid, acid, vorinostat, LBH 589, romidepsin, ACY-1215, and panobinostat), mTor inhibitors (e.g., temsirolimus, everolimus, ridaforolimus, and sirolimus), KSP (Eg5) inhibitors (e.g., Array 520), DNA binding agents (e.g., Zalypsis), PI3Kδ inhibitors (e.g., GS-1101 and TGR-1202), PI3Kδ and γ inhibitors (e.g., CAL-130), multikinase inhibitors (e.g., TG02 and sorafenib), hormones (e.g.,estrogen) and hormone agonists such as luteinizing hormone-releasing hormone (LHRH) agonists (e.g., goserelin, leuprolide and triptorelin), BAFF neutralizing antibodies (e.g., LY2127399), IKK inhibitors, p38MAPK inhibitors, anti-IL-6 (e.g., CNTO328), telomerase inhibitors (e.g., GRN 163L), Aurora kinase inhibitors (e.g., MLN8237, AMG 900, AZD-1152), cell surface monoclonal antibodies (e.g., anti-CD38 (HUMAX-CD38), anti-CS1 (e.g., elotuzumab), HSP90 inhibitors (e.g., 17AAG and KOS 953), P13K / Akt inhibitors (e.g., perifosine), Akt inhibitors (e.g., GSK-2141795), PKC inhibitors (e.g., enzastaurin), FTIs (e.g., Zarnestra, TM ), anti-CD138 (e.g., BT062), Torc1 / 2 specific kinase inhibitors (e.g., INK128), kinase inhibitors (e.g., GS-1101), ER / UPR targeting agents (e.g., MKC-3946), cFMS inhibitors (e.g., ARRY-382), JAK1 / 2 inhibitors (e.g., CYT387), PARP inhibitors (e.g., Olaparib, Talazoparib, Niraparib and veliparib (ABT-888)), BCL-2 antagonists. Other chemotherapeutic agents may include dichloromethyl diethylamine, camptothecin, ifosfamide, tamoxifen, raloxifene, gemcitabine, navelbine, sorafenib or any analog or derivative variant of the foregoing.

[0230] The compounds of the present invention may also be used in combination with radiation therapy, hormone therapy, surgery and immunotherapy, which are well known to those skilled in the art.

[0231] In certain embodiments, the pharmaceutical compositions provided herein are administered in combination with a steroid. Suitable steroids may include, but are not limited to, 21-acetoxypregnenolone, alclomethasone, algestrone, amcinonide, beclomethasone, betamethasone, budesonide, chloroprednisone, clobetasol, clocortolone, cloprednol, corticosterone, cortisone, cortivazole, deflazacort, desonide, desoximetasone, dexamethasone, diflorasone, diflucortolone, difuprednate, glycyrrhetinic acid, fluzacort, flucloronide, flumethasone, flunisolide, fluocinolone acetonide, fluocinolone acetonide, fluocortinbutyl, fluocortolone, fluorometholone, fluperolone acetate, fluperolone acetonide, fluocortin butyl, fluocortolone, fluorometholone, fluperolone acetate, fluperolone acetonide ... acetate), fluprednidene acetate, fluprednisolone, flurandrenolide, fluticasone propionate, formocortal, halcinonide, halobetasol propionate, halometasone, hydrocortisone, loteprednol etabonate, maprednione, medrysone, methylprednisone, methylprednisolone, mometasone furoate furoate, paramethasone, prednicarbate, prednisolone, prednisolone 25-diethylamino acetate, prednisolone sodium phosphate, prednisone, prednival, prednylidene, rimexolone, tixocortol, triamcinolone, triamcinolone acetonide, triamcinolone benetonide, triamcinolone hexacetonide and salts and / or derivatives thereof. In a specific embodiment, the compounds of the present invention can also be used in combination with other pharmaceutically active agents for treating nausea.Examples of agents that can be used to treat nausea include dronabinol, granisetron, metoclopramide, ondansetron, and prochlorperazine, or pharmaceutically acceptable salts thereof.

[0232] The compounds or pharmaceutical compositions disclosed herein may also be used in combination with a certain amount of one or more substances selected from the following: EGFR inhibitors, MEK inhibitors, PI3K inhibitors, AKT inhibitors, TOR inhibitors and immunotherapy, including anti-PD-1, anti-PDL-1, anti-CTLA4, anti-LAG1 and anti-OX40 agents, GITR agonists, CAR-T cells and BiTEs.

[0233] EGFR inhibitors include, but are not limited to, small molecule antagonists, antibody inhibitors, or specific antisense nucleotides or siRNA. Useful EGFR antibody inhibitors include cetuximab (Erbitux), panitumumab (Victobi), zalutumumab, nimotuzumab, and matuzumab. Small molecule antagonists of EGFR include gefitinib, erlotinib (Tarceva), and the most recent lapatinib (TykerB). See, e.g., Yan L et al., Pharmacogenetics and Pharmacogenomics In Oncology Therapeutic Antibody Development, BioTechniques 2005; 39(4): 565-8 and Paez JG et al., EGFR Mutations In Lung Cancer Correlation With Clinical Response To Gefitinib Therapy, Science 2004; 304(5676): 1497-500.

[0234] Non-limiting examples of small molecule EGFR inhibitors include any EGFR inhibitor described in the following patent publications, and all pharmaceutically acceptable salts and solvates of the EGFR inhibitors: European Patent Application EP520722, published on December 30, 1992; European Patent Application EP 566226, published on October 20, 1993; PCT International Publication WO 96 / 33980, published on October 31, 1996; U.S. Patent No. 5,747,498, issued on May 5, 1998; PCT International Publication WO 96 / 30347, published on October 3, 1996; European Patent Application EP 787772, published on August 6, 1997; PCT International Publication WO 97 / 30034, published on August 21, 1997; PCT International Publication WO 97 / 30044, published on August 21, 1997; PCT International Publication WO 97 / 38994, published on October 23, 1997; PCT International Publication WO 97 / 49688, published on December 31, 1997; European Patent Application EP 837063, published on April 22, 1998; PCT International Publication WO 98 / 02434, published on January 22, 1998; PCT International Publication WO 97 / 38983, published on October 23, 1997; PCT International Publication WO 95 / 19774, published on July 27, 1995; PCT International Publication WO 95 / 19970, published on July 27, 1995; PCT International Publication WO 97 / 13771, published on April 17, 1997; PCT International Publication WO 98 / 02437, published on January 22, 1998; PCT International Publication WO 98 / 02438, published on January 22, 1998; PCT International Publication WO 97 / 32881, published on September 12, 1997; German Application DE 19629652, published on January 29, 1998; PCT International Publication WO 98 / 33798, published on August 6, 1998; PCT International Publication WO 97 / 32880, published on September 12, 1997; PCT International Publication WO 97 / 32880, published on September 12, 1997; European Patent Application EP 682027, published on November 15, 1995; PCT International Publication WO 97 / 02266, published on January 23, 197; PCT International Publication WO 97 / 27199, published on July 31, 1997; PCT International Publication WO 98 / 07726, published on February 26, 1998; PCT International Publication WO 97 / 34895, published on September 25, 1997;PCT International Publication WO 96 / 31510', published on October 10, 1996; PCT International Publication WO 98 / 14449, published on April 9, 1998; PCT International Publication WO 98 / 14450, published on April 9, 1998; PCT International Publication WO 98 / 14451, published on April 9, 1998; PCT International Publication WO 95 / 09847, published on April 13, 1995; PCT International Publication WO 97 / 19065, published on May 29, 1997; PCT International Publication WO 98 / 17662, published on April 30, 1998; U.S. Patent No. 5,789,427, issued on August 4, 1998; U.S. Patent No. 5,650,415, issued on July 22, 1997; U.S. Patent No. 5,656,643, issued on August 12, 1997; PCT International Publication WO 99 / 35146, published on July 15, 1999; PCT International Publication WO 99 / 35132, published on July 15, 1999; PCT International Publication WO 99 / 07701, published on February 18, 1999; and PCT International Publication WO 92 / 20642, published on November 26, 1992. Other non-limiting examples of small molecule EGFR inhibitors include any of the EGFR inhibitors described in Traxler, P., 1998, Exp. Opin. Ther. Patents 8(12): 1599-1625. ;

[0235] Antibody-based EGFR inhibitors include any anti-EGFR antibody or antibody fragment that can partially or completely block activation of EGFR by its natural ligand. Non-limiting examples of antibody-based EGFR inhibitors include those described in the following literature: Modjtahedi, H. et al., 1993, Br. J. Cancer [British Journal of Cancer] 67: 247-253; Teramoto, T. et al., 1996, Cancer [Cancer] 77: 639-645; Goldstein et al., 1995, Clin. Cancer Res. [Clinical Cancer Research] 1: 1311-1318; Huang, SM et al., 1999, Cancer Res. [Cancer Research] 15: 59 (8): 1935-40; and Yang, X. et al., 1999, Cancer Res. [Cancer Research] 59: 1236-1243. Therefore, the EGFR inhibitor may be the monoclonal antibody Mab E7.6.3 (Yang, 1999, supra), or Mab C225 (ATCC Accession No. HB-8508), or an antibody or antibody fragment having the binding specificity thereof.

[0236] MEK inhibitors include, but are not limited to, CI-1040, AZD6244, PD318088, PD98059, PD334581, RDEA119, ARRY-142886, ARRY-438162, and PD-325901.

[0237] PI3K inhibitors include, but are not limited to, wortmannin, 17-hydroxywortmannin analogs described in WO 06 / 044453, 4-[2-(1H-indazol-4-yl)-6-[[4-(methylsulfonyl)piperazin-1-yl]methyl]thieno[3,2-d]pyrimidin-4-yl]morpholine (also known as GDC 0941 and described in PCT Publication Nos. WO 09 / 036,082 and WO 09 / 055,730), 2-methyl-2-[4-[3-methyl-2-oxo-8-(quinolin-3-yl)-2,3-dihydroimidazo[4,5-c]quinolin-1-yl]phenyl]propionitrile (also known as BEZ 235 or NVP-BEZ 235 and described in PCT Publication No. WO 09 / 055,730). 06 / 122806), (S)-1-(4-((2-(2-aminopyrimidin-5-yl)-7-methyl-4-morpholinothieno[3,2-d]pyrimidin-6-yl)methyl)piperazin-1-yl)-2-hydroxypropan-1-one (described in PCT Publication No. WO 2008 / 070740), LY294002 (2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one, available from Axon Medchem), PI103 hydrochloride (3-[4-(4-morpholinopyrido-[3',2':4,5]furo[3,2-d]pyrimidin-2-yl]phenol hydrochloride, available from Axon Medchem), PIK 75 (N'-[(1E)-(6-bromoimidazo[1,2-a]pyridin-3-yl)methylene]-N,2-dimethyl-5-nitrobenzenesulfonyl-hydrazide hydrochloride, available from Exon Medical Chemicals), PIK 90 (N-(7,8-dimethoxy-2,3-dihydro-imidazo[1,2-c]quinazolin-5-yl)-nicotinamide, available from Exon Medical Chemicals), GDC-0941 dimesylate (2-(1H-indazol-4-yl)-6-(4-methanesulfonyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine dimesylate, available from Exon Medical Chemicals), AS-2524 24 (5-[1-[5-(4-fluoro-2-hydroxy-phenyl)-furan-2-yl]-meth-(Z)-ylidene]-thiazolidine-2,4-dione, available from Exon Medical Chemicals) and TGX-221 (7-methyl-2-(4-morpholinyl)-9-[1-(phenylamino)ethyl]-4H-pyrido-[1,2-a]pyrimidin-4-one, available from Exon Medical Chemicals), XL-765 and XL-147.Other PI3K inhibitors include demethoxyviridin, perifosine, CAL101, PX-866, BEZ235, SF1126, INK1117, IPI-145, BKM120, XL147, XL765, Palomid 529, GSK1059615, ZSTK474, PWT33597, IC87114, TG100-115, CAL263, PI-103, GNE-477, CUDC-907, and AEZS-136.

[0238] AKT inhibitors include, but are not limited to, Akt-1-1 (inhibits Akt1) (Barnett et al. (2005) Biochem. J. [Journal of Biological Chemistry], 385 (Pt. 2), 399-408); Akt-1-1,2 (inhibits Ak1 and 2) (Barnett et al. (2005) Biochem. J. [Journal of Biological Chemistry] 385 (Pt. 2), 399-408); API-59CJ-Ome (e.g., Jin et al. (2004) Br. J. Cancer [British Cancer [Journal of Disease] 91, 1808-12); 1-H-imidazo[4,5-c]pyridinyl compounds (e.g., WO05011700); indole-3-methanol and its derivatives (e.g., U.S. Pat. No. 6,656,963; Sarkar and Li (2004) J Nutr. [Journal of Nutrition] 134(12 Supplement), 3493S-3498S); perifosine (e.g., interferes with Akt membrane localization; Dasmahapatra et al., (2004) Clin. Cancer Res. [Clinical Cancer Research] 10(15), 5242-52, 2004); phosphatidylinositol ether lipid analogs (e.g. Gills and Dennis (2004) Expert. Opin. Investig. Drugs [Expert Review of Research Drugs] 13, 787-97); and triciribine (TCN or API-2 or NCI identified agent: NSC154020; Yang et al., 2004, Cancer Res. [Cancer Research] 64, 4394-9).

[0239] TOR inhibitors include, but are not limited to, inhibitors including AP-23573, CCI-779, everolimus, RAD-001, rapamycin, temsirolimus, ATP-competitive TORC1 / TORC2 inhibitors including PI-103, PP242, PP30, and Torin 1. Other TOR inhibitors include FKBP12 enhancers; rapamycin and its derivatives, including: CCI-779 (temsirolimus), RAD001 (everolimus; WO 9409010) and AP23573; rapamycin analogs (rapalogs), such as disclosed in WO98 / 02441 and WO 01 / 14387, such as AP23573, AP23464 or AP23841; 40-(2-hydroxyethyl)rapamycin, 40-[3-hydroxy(hydroxymethyl)methylpropionate]-rapamycin (also known as CC1779), 40-epi-(tetrazolyl)-rapamycin (also known as ABT578), 32-deoxyrapamycin, 16-pentynyloxy-32(S)-dihydrorapamycin and other derivatives disclosed in WO05005434; derivatives disclosed in the following patents: U.S. Pat. Nos. 5,258,389, WO 94 / 090101, WO 92 / 05179, U.S. Pat. No. 5,118,677, U.S. Pat. No. 5,118,678, U.S. Pat. No. 5,100,883, U.S. Pat. No. 5,151,413, U.S. Pat. No. 5,120,842, WO 93 / 111130, WO 94 / 02136, WO 94 / 02485, WO 95 / 14023, WO 94 / 02136, WO 95 / 16691, WO 96 / 41807, WO 96 / 41807 and U.S. Pat. No. 5,256,790; phosphorus-containing rapamycin derivatives (e.g., WO 05016252); 4H-1-benzopyran-4-one derivatives (e.g., U.S. Provisional Application No. 60 / 528,340).

[0240] Immunotherapy includes, but is not limited to, anti-PD-1 agents, anti-PDL-1 agents, anti-CTLA-4 agents, anti-LAG1 agents, and anti-OX40 agents. Exemplary anti-PD-1 antibodies and methods of use thereof are described in Goldberg et al., Blood 110(1):186-192 (2007); Thompson et al., Clin. Cancer Res. 13(6):1757-1761 (2007); and Korman et al., International Application No. PCT / JP 2006 / 309606 (Publication No. WO 2006 / 121168A1), each of which is expressly incorporated herein by reference. Including: Yervoy TM(ipilimumab) or tremelimumab (targeting CTLA-4), galiximab (targeting B7.1), BMS-936558 (targeting PD-1), MK-3475 (targeting PD-1), AMP224 (targeting B7DC), BMS-936559 (targeting B7-H1), MPDL3280A (targeting B7-H1), MEDI-570 (targeting ICOS), AMG557 (targeting B7H2), MGA271 (targeting B7H3), IMP321 (targeting LAG-3), BMS-663513 (targeting CD137), PF-05082566 (targeting CD137), CDX-1127 (targeting CD27), anti-OX40 (Providence Health Services Services), huMAbOX40L (for OX40L), Atacicept (for TACI), CP-870893 (for CD40), Lucatumumab (for CD40), Dacilizumab (for CD40), Muromonab-CD3 (for CD3), Ipilimumab (for CTLA-4). Immunotherapy also includes genetically engineered T cells (e.g., CAR-T cells) and bispecific antibodies (e.g., BiTE).

[0241] GITR agonists include, but are not limited to, GITR fusion proteins and anti-GITR antibodies (e.g., bivalent anti-GITR antibodies), such as the GITR fusion proteins described in U.S. Pat. No. 6,111,090 box.c, European Patent No.: 090505B1, U.S. Pat. No. 8,586,023, PCT Publication Nos.: WO 2010 / 003118 and 2011 / 090754, or anti-GITR antibodies such as described in U.S. Pat. No. 7,025,962, European Patent No.: 1947183B1, U.S. Pat. No. 7,812,135, U.S. Pat. No. 8,388,967, U.S. Pat. No. 8,591,886, European Patent No.: EP 1866339, PCT Publication No.: WO 2011 / 028683, PCT Publication No.: WO 2013 / 039954, PCT Publication No.: WO 2005 / 007190, PCT Publication No.: WO 2007 / 133822, PCT Publication No.: WO 2005 / 055808, PCT Publication No.: WO 99 / 40196, PCT Publication No.: WO 2001 / 03720, PCT Publication No.: WO99 / 20758, PCT Publication No.: WO 2006 / 083289, PCT Publication No.: WO 2005 / 115451, U.S. Patent No. 7,618,632 and PCT Publication No.: WO 2011 / 051726.

[0242] The compounds described herein can be used in combination with the medicaments disclosed herein or other suitable medicaments, depending on the condition being treated. Therefore, in some embodiments, one or more compounds of the present disclosure will be co-administered with other medicaments as described above. When used in combination therapy, the compounds described herein are administered simultaneously or separately with the second medicament. This combination administration can include the simultaneous administration of two medicaments in the same dosage form, the simultaneous administration in a separate dosage form, and the separate administration. That is, the compounds described herein and any of the above-mentioned medicaments can be formulated together in the same dosage form and administered simultaneously. Alternatively, the compounds disclosed herein and any of the above-mentioned medicaments can be administered simultaneously, wherein the two medicaments are present in a separate formulation. In another alternative, any of the above-mentioned medicaments can be administered immediately after the compounds disclosed herein are administered, or vice versa. In some embodiments of the separate administration scheme, the administration of the compounds disclosed herein and any of the above-mentioned medicaments is separated by a few minutes, or a few hours, or a few days.

[0243] Since one aspect of the present invention contemplates treating diseases / disorders with combinations of pharmaceutically active compounds that can be administered separately, the present invention further relates to combining separate pharmaceutical compositions in kit form. The kit comprises two separate pharmaceutical compositions: a compound of the present invention and a second pharmaceutical compound. The kit comprises a container for containing the separate compositions, such as a separate bottle or a separate foil bag. Other examples of containers include syringes, boxes, and bags. In some embodiments, the kit comprises instructions for use of the separate components. The kit form is particularly advantageous when the separate components are preferably administered in different dosage forms (e.g., oral and parenteral), when administered at different dosage intervals, or when the prescribing medical professional requires titration of the individual components in the combination.

[0244] experiment

[0245] Abbreviations: The following abbreviations may be used in this document:

[0246]

[0247]

[0248]

[0249]

[0250] Unless otherwise noted, all materials were obtained from commercial suppliers and used without further purification. All parts are by weight and temperatures are in degrees Celsius unless otherwise noted. All microwave-assisted reactions were performed using Biotage TM Smith Synthesizer TM All compounds showed NMR spectra consistent with their assigned structures. Melting points were determined on a Buchi apparatus and are uncorrected. Mass spectral data were determined by electrospray ionization techniques. All examples were purified to a purity of >90% as determined by high performance liquid chromatography. Unless otherwise stated, reactions were performed at room temperature.

[0251] In synthesizing the compounds of the present invention, it may be desirable to use certain leaving groups. The term "leaving group" ("LG") generally refers to a group that can be displaced by a nucleophilic reagent. Such leaving groups are known in the art. Examples of leaving groups include, but are not limited to, halides (e.g., I, Br, F, Cl), sulfonates such as mesylate, toluenesulfonate), sulfides (e.g., SCH3), N-hydroxysuccinimide, N-hydroxybenzotriazole, etc. Examples of nucleophilic reagents include, but are not limited to, amines, thiols, alcohols, Grignard reagents, anionic species (e.g., alkoxides, amides, carbanions), etc.

[0252] The examples presented below illustrate specific embodiments of the present invention. These examples are representative and are not intended to limit the scope of the claims in any way.

[0253] It should be noted that when the percentage (%) is used with respect to a liquid, this is the volume percentage relative to the solution. When used with a solid, this is the percentage relative to the solid composition. Materials obtained from commercial suppliers are generally used without further purification. Reactions involving air or moisture sensitive reagents are generally carried out under a nitrogen or argon atmosphere. Purity is measured using a high performance liquid chromatography (HPLC) system with UV detection at 254nm and 215nm (System A: Agilent Zorbax Eclipse XDB-C8 4.6x150mm, 5μm, 5% to 100% CH3CN (in H2O containing 0.1% TFA), 15min, 1.5mL / min; System B: Zorbax SB-C8, 4.6x75mm, 10% to 90% CH3CN (in H2O containing 0.1% formic acid), 12min, 1.0mL / min) (Agilent Technologies, Santa Clara, CA)). Silica gel chromatography was typically performed using prepacked silica gel cartridges (Biotage, Uppsala, Sweden or Teledyne-Isco, Lincoln, NE). 1 H NMR spectra were recorded at ambient temperature on a Bruker AV-400 (400 MHz) spectrometer (Bruker Corporation, Madison, WI) or a Varian (Agilent Technologies, Santa Clara, CA) 400 MHz spectrometer. All observed protons are reported in parts per million (ppm) downfield from tetramethylsilane (TMS) or other internal reference in the appropriate solvent as specified. Data are reported as follows: chemical shift, multiplicity (s = singlet, d = doublet, t = triplet, q = quartet, br = broad, m = multiplet), coupling constant, and number of protons. Low-resolution mass spectrometry (MS) data were determined on an Agilent 1100 series (Agilent Technologies, Santa Clara, CA) LC / MS using UV detection at 254 nm and 215 nm and low-resonance electrospray mode (ESI).

[0254] For the purpose of clarity in this general synthesis section, compounds of formula (I) as defined in the Summary of the Invention may be schematically drawn to include Ar 1 Ring and Ar 2 The ring is as follows:

[0255] The group -NR 3 -(C=O)- is the linker, Ar 1 The ring is located on the left side of the joint, and Ar 2 The ring is located on the right side of the connector.

[0256] Generally, the compounds of formula (I) can be synthesized by the following three general steps:

[0257] Step 1: Preparation of CycloAr 1 Compound

[0258] Step 2: Preparation of CycloAr 2 Compound

[0259] Step 3: Ring Ar 1 Compounds with cyclic Ar 2 Compound coupling

[0260] The general schemes AE below are meant to provide guidance to the ordinary synthetic chemist, who will readily appreciate that solvents, concentrations, reagents, protecting groups, order of synthetic steps, times, temperatures, etc. can be modified as necessary and are well within the skill and judgment of the ordinary skilled artisan.

[0261] Plan A

[0262] According to Scheme A, in one embodiment, compounds of formula (I) as disclosed herein can be synthesized as follows:

[0263] Step 1a: Preparation of CycloAr 1 Compound

[0264]

[0265] Step 1a: Preparation of CycloAr 1 Compound Compound A-1 (wherein W can be reacted with water in a suitable organic solvent such as NMP, dioxane, acetonitrile, tetrahydrofuran, DMF, dichloromethane, etc.) in the presence of a suitable base. 1 is a halogen, such as fluorine or chlorine) and contains R 2The reagent of the group is reacted to form compound A-2. Compound A-1 is commercially available or can be synthesized by known methods by those skilled in the art. Examples of compound A-1 include, but are not limited to, 2-chloropyrimidine-4-amine, 2-chloro-6-methylpyrimidine-4-amine, 2-fluoro-6-methylpyridin-4-amine, 2-chloropyridin-4-amine, 2-chloro-6-methylpyridin-4-amine, 2-chloro-6-ethylpyrimidine-4-amine or 2-chloro-6-cyclopropylpyrimidine-4-amine. R 2 Examples of reagents include, but are not limited to (1) (R)-2-methylmorpholine, (2) 4,4-difluoropiperidine hydrochloride, (3) 3,3-difluoroazetidine hydrochloride or (4) 3,3,3-trifluoropropane-1-ol. Examples of bases include, but are not limited to, diisopropylethylamine, potassium carbonate or sodium hydride.

[0266] Step 1b: Preparation of CycloAr 1 Compound

[0267]

[0268] Alternatively, the reaction can be carried out by reacting with a suitable organoboron R 2 Reagent (R 2 -BY2, wherein Y is an organic functional group, such as 2-(4,4-difluorocyclohex-1-ene-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane or 2-(4-fluorocyclopent-1-ene-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane) and a suitable palladium catalyst and a base (such as PdCl2(dppf)-DCM adduct and tripotassium phosphate) are subjected to a Suzuki cross-coupling reaction to convert the compound A-1 defined in step 1a into the compound A-2 defined in step 1a. After this step, reduction is carried out with a suitable palladium catalyst and a hydrogen source (such as Pd / C) in the presence of hydrogen to form compound A-2. When R 2 The group is bonded to Ar through a carbon-carbon bond. 1 This alternative Suzuki reaction can be used when the ring is connected.

[0269] Step 2a: Preparation of CycloAr 2 Compound

[0270]

[0271] In step 2a, compound A-3 (wherein W 2 and W 3 Each of which is independently halogen, such as fluorine, chlorine, bromine or iodine) and R xA reagent (e.g., (1) 6-azaspiro[2.5]octane hydrochloride, (2) 4,4-dimethylpiperidine hydrochloride, (3) 3,4,4-trimethylpiperidine hydrochloride, (4) 4-methyl-6-azaspiro[2.5]octane hydrochloride or (5) 7-azaspiro[3.5]nonane hydrochloride) is reacted to form compound A-4.

[0272] Step 3a: Ring Ar 1 Compounds with cyclic Ar 2 Compound coupling, followed by introduction of R 1 :

[0273]

[0274] In step 3a, the compound A-4 obtained from step 2a can be reacted with an activator (e.g., acyl chloride (COCl) 2 or SOCl 2) in a suitable organic solvent (e.g., tetrahydrofuran, dichloromethane, etc.) to form an acyl chloride derivative, which can then be reacted with compound A-2 to form compound A-5. Alternatively, compound A-2 can be directly coupled with compound A-4 obtained from step 2a in the presence of a coupling reagent (e.g., N, N'-diisopropylcarbodiimide, N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide, benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate, O-(benzotriazol-1-yl)-N, N, N', N'tetramethyluronium hexafluorophosphate, carbonyldiimidazole, and polyphosphonic anhydride) in a suitable organic solvent (e.g., acetonitrile, tetrahydrofuran, DMF, dichloromethane, etc.). Those ordinary synthetic chemists will readily understand that other coupling agents can be used. The reaction can be carried out by reacting a metal catalyst and R in a suitable organic solvent (e.g., DMSO, acetonitrile, tetrahydrofuran, DMF, etc.). 1 The halogen group W is further treated by a conversion reaction in the presence of a reagent such as a metal-catalyzed sulfonamidation, sulfidation or sulfonylation. 3 To form compound (I), the R 1 Reagents such as (1) 1-methylcyclopropane-1-sulfonamide, (2) 3-methyloxetane-3-amine, (3) tert-butyl 3-mercaptoazetidine-1-carboxylate, (4) ethyl 2-sulfamoylpropionate, (5) 2-hydroxypropane-1-sulfonamide, (6) 2-hydroxyethane-1-sulfonamide, (7) ethyl iodoacetate, (8) 2-mercaptopropane-1-ol, (9) 2-mercapto-2-methylpropane-1-ol, (10) 2-aminoethan-1-ol or (11) cyclopropanethiol. Those of ordinary skill will readily appreciate that coupling reactions (e.g., as shown in step 3a) can be carried out under known conditions up to 50%.

[0275] Plan B

[0276] Step 1a or 1b: Preparation of Ring Ar 1 Compounds see Scheme A above

[0277] Step 2b: Preparation of CycloAr 2 Compound

[0278]

[0279] Scheme B provides an alternative method for forming compounds of Formula (I) disclosed herein. After step 1a or step 1b as described in Scheme A, R may alternatively be 1 The group is introduced into the ring Ar in step 2b 2 Instead of being introduced in step 3a of scheme A. According to step 2b, compound B-1 (wherein W 4 and W 5 wherein each of which is independently halogen, such as fluorine, chlorine, bromine or iodine) is reacted with an appropriate carboxylic acid protecting group (PG1 reagent, such as iodomethane) to form a methyl ester or reacted with other suitable protecting groups in a suitable organic solvent (such as NMP, acetonitrile, tetrahydrofuran, DMF, dichloromethane, etc.) to form other esters (such as benzyl esters) to form compound B-2, wherein W 4 and W 5 Each of them is as defined in compound B-1. Then, compound B-2 can be reacted with R in a suitable organic solvent (e.g., NMP, acetonitrile, tetrahydrofuran, DMF, dichloromethane, DMSO, etc.). x A reagent (eg, 6-azaspiro[2.5]octane) is reacted to form compound B-3, wherein W 5 is as defined in compound B-1. Compound B-3 can then be reacted with R by a conversion reaction (e.g., metal-catalyzed sulfonamidation, sulfidation, or sulfonylation) in the presence of a metal catalyst (e.g., copper iodide, Pd2(dba)3) in a suitable organic solvent (e.g., DMSO, acetonitrile, tetrahydrofuran, DMF, etc.). 1 The reagents are reacted to form compound B-4, which can then be further reacted with an appropriate carboxylic acid deprotecting agent to form compound B-5. Suitable carboxylic acid protecting groups and deprotecting agents are known to those skilled in the art, for example as discussed in Greene protecting groups in organic synthesis.

[0280] Step 3b: Ring Ar 1 Compounds with cyclic Ar 2 Compound coupling

[0281]

[0282] Step 3b is similar to the coupling reaction as described above in step 3a.

[0283] Plan C

[0284] Scheme C provides another alternative method for forming compounds of Formula (I) disclosed herein. According to Scheme C, step 1a can be carried out as described in Scheme A, followed by step 2b as described in Scheme B.

[0285] Step 3c: Ring Ar 1 Compounds with cyclic Ar 2 Compound coupling

[0286]

[0287] In step 3c, compound A-1a (which is compound A-1 of Scheme A, wherein X 1 N and W 1 is halogen, such as fluorine or chlorine) is reacted with compound B-5 obtained from step 2b of scheme B to form compound C-1, wherein W 1 is as defined in compound A-1a, which can then be reacted with a compound containing R in the presence of a suitable base (e.g., diisopropylethylamine, potassium carbonate or sodium hydride) in a suitable organic solvent (e.g., NMP, dioxane, acetonitrile, tetrahydrofuran, DMF, dichloromethane, etc.). 2 The reagent containing R 2 The reagents of the group are, for example, (1) (R)-2-methylmorpholine, (2) 4,4-difluoropiperidine hydrochloride, (3) 3,3-difluoroazetidine hydrochloride or (4) 3,3,3-trifluoropropan-1-ol.

[0288] Plan D

[0289] Scheme D provides another alternative method for forming compounds of Formula (I) disclosed herein. According to Scheme D, step 1a or 1b can be carried out as described in Scheme A, followed by step 2b as described in Scheme B.

[0290] Step 3d: Ring Ar 1 Compounds with cyclic Ar 2 Compound coupling

[0291]

[0292] In step 3d, compound A-1a (which is compound A-1 of Scheme A, wherein X 1 N and W 1 is a halogen, such as fluorine or chlorine) can be reacted with compound A-4 obtained from step 2a of scheme A to form compound D-1, wherein W 1 is as defined in compound A-1a and W 3is as defined in compound B-5, which can then be optionally reacted with a compound containing R in the presence of a suitable base (e.g., diisopropylethylamine, potassium carbonate or sodium hydride) in a suitable organic solvent (e.g., NMP, dioxane, acetonitrile, tetrahydrofuran, DMF, dichloromethane, etc.) 2 The reaction is carried out with a reagent (e.g., (1) (R)-2-methylmorpholine, (2) 4,4-difluoropiperidine hydrochloride, (3) 3,3-difluoroazetidine hydrochloride or (4) 3,3,3-trifluoropropane-1-ol) to form compound A-5a, i.e., compound A-5, wherein X 1 N and W 3 is as defined in compound B-5, which can then be reacted with a compound containing R by a conversion reaction (e.g., metal-catalyzed sulfonamidation, sulfidation or sulfonylation) in the presence of a metal catalyst in a suitable organic solvent (e.g., DMSO, acetonitrile, tetrahydrofuran, DMF, etc.). 1 The reagents of the group are reacted to form compounds of formula (I).

[0293] Plan E

[0294] Scheme E provides another alternative method for forming compounds of formula (I) disclosed herein. According to Scheme E, step 1a or 1b can be carried out as described in Scheme A to prepare compound A-2. Compound E-1 (wherein W 6 is a halogen, such as fluorine or chlorine, which includes but is not limited to 2-fluoro-4-nitrobenzoic acid, 2,5-difluoro-4-nitrobenzoic acid or 2,6-difluoro-4-nitrobenzoic acid) is commercially available or can be synthesized by a person skilled in the art according to a known method.

[0295] Step 3e: Reaction of the ring Ar1 compound with the ring Ar 2 Compound coupling

[0296]

[0297] In step 3e, compound A-2 can be reacted with compound E-1 in the presence of an activating agent under conditions similar to those of steps 3a and 3b above to form compound E-2, which can then be reacted with R in a manner similar to that described in step 2a. x The nitro group on compound E-3 can then be converted to an amino group by reaction with a reducing agent (which includes but is not limited to palladium carbon and hydrogen) to form compound E-4, which can then be reacted with R by a conversion reaction (e.g., metal-catalyzed sulfonamidation, sulfidation or sulfonylation) in a suitable organic solvent (such as DMSO, acetonitrile, tetrahydrofuran, DMF, etc.) in the presence of a metal catalyst. 1 The reagents react to form compound (I), wherein R 1Reagents such as (1) 1-methylcyclopropane-1-sulfonamide, (2) 3-methyloxetane-3-amine, (3) tert-butyl 3-mercaptoazetidine-1-carboxylate, (4) ethyl 2-sulfamoylpropionate, (5) 2-hydroxypropane-1-sulfonamide, (6) 2-hydroxyethane-1-sulfonamide, (7) ethyl iodoacetate, (8) 2-mercaptopropane-1-ol, (9) 2-mercapto-2-methylpropane-1-ol, (10) 2-aminoethane-1-ol or (11) cyclopropanethiol.

[0298] Examples

[0299] Preparation of synthetic intermediates

[0300] Ring AR 1 Intermediates:

[0301] Intermediate 1: (R)-2-(2-methylmorpholino)pyrimidin-4-amine

[0302]

[0303] 2-Chloropyrimidine-4-amine (60.0 g, 463 mmol, Combi-Blocks, San Diego, CA), (R)-2-methylmorpholine (65.6 g, 648 mmol, Wuxi Apptec) and DIPEA (243 mL, 1389 mmol) in NMP (600 mL) were placed in an autoclave and heated at 150 ° C for 36 h. The reaction mixture was cooled to room temperature, quenched with water (1 L) and extracted with ethyl acetate (3x500 mL). The organic layer was washed with saline solution (500 mL), dried (Na2SO4), filtered and concentrated under reduced pressure to obtain a crude material in a brownish yellow oil. The crude material was adsorbed onto a silica gel plug and purified on silica gel (60-120 mesh) by column chromatography, eluted with a gradient of 50% to 100% ethyl acetate in hexane to obtain a yellow solid. The solid was further triturated with hexanes (300 mL), filtered and dried in vacuo to give the title compound (70 g, 78% yield) as a light yellow solid. 1H NMR(400MHz,DMSO-d6)δppm 7.75(d,J=5.6Hz,1H),6.42(s,2H),5.75(d,J=5.6Hz,1H),4.26–4.49(m,2H),3.83(ddd,J=11.4,3.6,1.4 Hz,1H),3.31–3.50(m,2H),2.78(ddd,J=13.2,11.8,3.5Hz,1H),2.42–2.48(m,1H),1.11(d,J=6.2Hz,3H). m / z(ESI):195.2(M+H) + .

[0304] Intermediate 2: (R)-6-methyl-2-(2-methylmorpholino)pyrimidin-4-amine

[0305]

[0306] 2- Chloro-6-methylpyrimidine-4-amine (30.0 g, 209 mmol, Combi-Blocks, San Diego, CA), (R)-2-methylmorpholine (40.3 g, 293 mmol, Wuxi Apptec, PR China) and DIPEA (109 mL, 627 mmol) were placed in an autoclave (600 mL) and heated at 150 ° C for 12 h. The reaction mixture was quenched with water (500 mL) and extracted with ethyl acetate (2x1500 mL). The organic layer was washed with saline solution (500 mL), dried (Na2SO4), filtered and concentrated under reduced pressure. The crude residue was purified by column chromatography on silica gel (60-120 mesh) using 50% ethyl acetate in hexane as eluent to obtain the title compound (25.0 g, 57% yield) as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ6.28(s,2H),5.62(s,1H),4.45–4.31(m,2H),3.83(ddd,J=11.4,3.5,1.3Hz,1H),3.42(dd t,J=14.4,9.7,2.8Hz,2H),2.78–2.70(m,1H),2.43(dd,J=13.0,10.3Hz,1H),2.05(s,3H),1.11(d,J=6.2Hz,3H). m / z(ESI):209.2(M+H) + .

[0307] Intermediate 3: 2-(4,4-difluoropiperidin-1-yl)pyrimidin-4-amine

[0308]

[0309] 2-Chloro-4-aminopyrimidine (1.0 g, 7.7 mmol, Combi-Blocks, San Diego, CA), 4,4-difluoropiperidine hydrochloride (1.82 g, 11.58 mmol, Combi-Blocks, San Diego, CA) and DIPEA (4.04 mL, 23.2 mmol) in NMP (12 mL) were sequentially loaded into a glass microwave reaction vessel. The reaction mixture was stirred and heated in microwave at 200 ° C for 1 h. The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (30x2 mL). The organic extract was washed with brine (30xmL), dried over Na2SO4, and concentrated in vacuo to obtain a crude substance as a brown viscous liquid. The crude material was adsorbed onto a plug of silica gel and purified by flash chromatography over a Redi-Sep pre-packed silica gel column (40 g), eluting with 0% to 100% ethyl acetate in heptane to give the title compound as an off-white solid (6.02 g, 91%). 1 H NMR (300MHz, DMSO-d6) δppm7.21 (d, J = 5.6 Hz, 1H), 6.46 (br s, 2H), 5.77 (d, J = 5.6 Hz, 1H), 3.80 (t, J = 5.6 Hz, 4H), 1.85-1.90 (m, 4H). m / z(ESI):215.2(M+H) + .

[0310] Intermediate 4: 2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-amine

[0311]

[0312] 2-Chloro-6-methylpyrimidine-4-amine (46g, 320mmol, Combi-Blocks, San Diego, CA) in NMP (460mL, 10.00mL / g), 4,4-difluoropiperidine hydrochloride (76g, 481mmol, Combi-Blocks, San Diego, CA) and DIPEA (166mL, 961mmol) are placed in autoclave (1L) and heated at 180°C for 30h. The reaction mixture is cooled to room temperature and quenched with water (500mL), extracted with ethyl acetate (2x1000mL). The organic layer is washed with brine (500mL), dried (Na2SO4), filtered and concentrated under reduced pressure. The crude material is adsorbed on silica gel plug, and purified by column chromatography on silica gel (60-120 mesh), 50% to 100% ethyl acetate in hexane is used as eluent elution, to obtain product. It is redissolved in ethyl acetate (500mL), washed with water (2x500mL). The organic layer is dried (Na2SO4), filtered and concentrated under reduced pressure. The yellow solid is suspended in hexane (400mL) again, and stirred for 30min. The slurry is filtered, washed with hexane (100mL), dried in a vacuum, to provide the title compound (58g, 79% yield) as a light yellow solid. 1 HNMR (400MHz, DMSO-d6) δppm 6.33 (s, 2H), 5.63 (s, 1H), 3.80–3.78 (dd, J = 6.8, 4.7Hz, 4H), 2.06 (s, 3H), 1.95–1.85 (tt, J = 14.2, 5.7Hz, 4H). m / z(ESI):229.2(M+H) + .

[0313] Intermediate 5: 2-(3,3-difluoroazetidin-1-yl)pyrimidin-4-amine

[0314]

[0315] 2-Chloro-4-aminopyrimidine (5.0 g, 38.6 mmol, Combi-Blocks, San Diego, CA), 3,3-difluoroazetidine hydrochloride (7.50 g, 57.9 mmol, Combi-Blocks, San Diego, CA) and potassium carbonate (5.33 g, 38.6 mmol) in dioxane (25 mL) were heated at 95 ° C for 16 h. The reaction mixture was cooled to room temperature and the suspension was filtered. The crude material was adsorbed onto a silica gel plug and purified by flash chromatography over a Redi-Sep pre-packed silica gel column (40 g), eluting with 10% MeOH in DCM to provide the title compound (6.1 g, 85%) as a light brown solid. 1 H NMR (300MHz, DMSO-d6) δppm 8.29-8.66 (m, 2H), 7.81 (d, J = 7.05Hz, 1H), 6.22 (d, J = 7.26Hz, 1H), 4.61 (t, J = 12.23Hz, 4H). m / z(ESI):187.2(M+H) + .

[0316] Intermediate 6: 2-(4,4-difluorocyclohexyl)-6-methylpyrimidin-4-amine

[0317]

[0318] Step 1: To a solution of 2-chloro-6-methylpyrimidin-4-amine (70.0 g, 488 mmol, Combi-Blocks, San Diego, CA) in 1,4-dioxane (560 mL) and water (210 mL) was added 2-(4,4-difluorocyclohex-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (119 g, 488 mmol, Combi-Blocks, San Diego, CA) and tripotassium phosphate (310 g, 1463 mmol). The reaction mixture was degassed and purged with nitrogen for 5 min. PdCl2(dppf)-DCM adduct (39.8 g, 48.8 mmol) was added to the reaction mixture and stirred at 100 °C for 16 h. The product is stirred for 24 hours at 4 ℃ for 10 minutes.Then add 4- (4- difluorocyclohex-1-ene-1-yl) -6- methylpyrimidine -4- amine (70g, 64% yield) in 4- (4- difluorocyclohex-1-ene-1-yl) -6- methylpyrimidine -4- amine (70g, 64% yield) to obtain 4- (4- difluorocyclohex-1-ene-1-yl) -6- methylpyrimidine -4- amine (70g, 64% yield) in ... 1 H NMR(400MHz, DMSO-d6)δppm 6.85(br s,1H),6.59(br s,2H),6.13(s,1H),2.62–2.80(m,4H),2.19(s,3H),2.04–2.17(m,2H). m / z(ESI):226.2(M+H) + .

[0319] Step 2: To a solution of 2-(4,4-difluorocyclohex-1-en-1-yl)-6-methylpyrimidin-4-amine (70.0 g, 311 mmol) in EtOH (700 mL) was added 10% Pd on carbon (33.1 g, 155 mmol) under nitrogen. The reaction mixture was stirred at room temperature for 16 h under hydrogen pressure (1 atm). The reaction mixture was filtered through a bed and washed with a mixture of ethyl acetate and ethanol (1: 1, 500 mL). The filtrate was concentrated under reduced pressure. The crude product was purified by column chromatography on silica gel (60-120 mesh) using 50% ethyl acetate in hexane to obtain the title compound (58.5 g, 83% yield) as an off-white solid. 1 HNMR (400MHz, DMSO-d6) δppm 6.61 (s, 2H), 6.09 (s, 1H), 2.56–2.67 (m, 1H), 2.16 (s, 3H), 1.72–2.10 (m, 8H). m / z(ESI):228.1(M+H) + .

[0320] Intermediate 7: 2-methyl-6-(3,3,3-trifluoropropoxy)pyridin-4-amine

[0321]

[0322] At 0 ° C, sodium hydride (60% by weight in mineral oil, 0.79 g, 19.82 mmol) was added to a solution of 3,3,3-trifluoropropan-1-ol (1.99 g, 17.44 mmol, Combi-Blocks) in 30 mL THF. The mixture was stirred at room temperature for 30 min and then treated with 2-fluoro-6-methylpyridine-4-amine (1.00 g, 7.93 mmol, AstaTech Inc). The mixture was heated in an oil bath at 65 ° C for 5 h. It was cooled to room temperature, quenched with water (10 mL) and extracted with EtOAc (2x50 mL). The organic solution was dried over Na2SO4, filtered and concentrated in vacuo to obtain a crude substance as a yellow oil. The crude material was adsorbed onto a plug of silica gel and purified on a silica gel column (15% to 30% EtOAc in heptane) to give 2-methyl-6-(3,3,3-trifluoropropoxy)pyridin-4-amine (0.47 g, 2.13 mmol, 27% yield) as a light yellow oil. m / z (ESI): 221.1 (M+H) + .

[0323] Table 1: The following intermediates were prepared by a similar procedure to Intermediate 7:

[0324]

[0325] Intermediate 8: 2-(4,4-difluoropiperidin-1-yl)pyridin-4-amine

[0326]

[0327] A mixture of 2-chloropyridin-4-amine (2.00 g, 15.56 mmol, Combi-Blocks), DIPEA (6.03 g, 46.70 mmol, Sigma-Aldrich) and 4,4-difluoropiperidine (2.45 g, 20.22 mmol, Enamine) in NMP (6 mL) was heated in microwave at 200 ° C for 6 h. The reaction mixture was diluted with water (20 mL) and extracted with EtOAc (2x50 mL). The organic extract was concentrated. The residue was purified by silica gel chromatography (20% to 70% EtOAc in heptane) to provide 2- (4,4-difluoropiperidin-1-yl) pyridine-4-amine (2.91 g, 13.65 mmol, 88% yield) as a light yellow solid. m / z (ESI): (M+H) + 214.1.

[0328] Table 2: The following intermediates were prepared following similar procedures as described for Intermediate 8:

[0329]

[0330] Intermediate 9: 2-(4,4-difluoropiperidin-1-yl)-3-fluoro-6-methylpyridin-4-amine

[0331]

[0332] Step 1: n-Butyl lithium (2M in hexane, 20.52 mL, 41.0 mmol) was added dropwise to a solution of diisopropylamine (5.85 mL, 41.0 mmol) in tetrahydrofuran (50 mL) at -60 ° C. The reaction mixture was slowly warmed to 0 ° C and stirred at the same temperature for 45 min. In another round-bottom flask, the LDA solution prepared above was added dropwise to a solution of 2-bromo-3-fluoro-6-methylpyridine (3.9 g, 20.52 mmol) in tetrahydrofuran (50 mL) at -78 ° C. The reaction mixture obtained by the reaction was stirred at the same temperature for 45 min, and then iodine (10.42 g, 41.0 mmol) in THF (40 mL) was added dropwise. The reaction mixture was stirred at the same temperature for 1 h. After the reaction was completed, it was quenched with a saturated ammonium chloride solution and extracted with ethyl acetate. The organic layer was washed with sodium thiosulfate solution, water and brine, dried over Na2SO4, filtered and concentrated to give 2-bromo-3-fluoro-4-iodo-6-methylpyridine (6 g, 18.99 mmol, 93% yield) as a yellow solid. The product was used in the next step without further purification. 1 H NMR (400 MHz, chloroform-d) δ ppm 7.53 (d, J = 3.8 Hz, 1H), 2.52 (s, 3H). m / z (ESI): 315.8, 317.8 (M+H) + .

[0333] Step 2: A mixture of 2-bromo-3-fluoro-4-iodo-6-methylpyridine (1.5 g, 4.75 mmol), (4-methoxyphenyl)methanamine (0.782 g, 5.70 mmol), cesium carbonate (4.64 g, 14.24 mmol), Xantphos (0.549 g, 0.950 mmol) and Pd2(dba)3 (0.065 g, 0.071 mmol) in 1,4-dioxane (30 mL) was stirred at ambient temperature for 16 h. The reaction mixture was then stirred for 16 h. The filtrate was filtered through a plug, and the filtrate was diluted with EtOAc. The resulting solution was washed with water, brine, dried over Na2SO4, filtered and concentrated. The concentrate was purified by flash column chromatography using a gradient of 0% to 18% ethyl acetate in petroleum ether to obtain 2-bromo-3-fluoro-N-(4-methoxybenzyl)-6-methylpyridine-4-amine (0.7 g, 2.15 mmol, 45% yield) as a light yellow solid. 1HNMR(300MHz,DMSO-d6)δppm 7.38(s,1H),7.21–7.31(m,2H),6.84–6.95(m,2H),6.52(d,J=6.0Hz,1H),4.32(d,J=6.3Hz,2H),3.72(s,3H),2.20(s,3H). m / z(ESI):325.0,327.0(M+H) + .

[0334] Step 3: A mixture of 2-bromo-3-fluoro-N-(4-methoxybenzyl)-6-methylpyridin-4-amine (0.7 g, 2.153 mmol), 4,4-difluoropiperidine hydrochloride (0.407 g, 2.58 mmol), cesium carbonate (2.81 g, 8.61 mmol), Xantphos (0.249 g, 0.431 mmol) and Pd2(dba)3 (0.030 g, 0.032 mmol) in 1,4-dioxane (15 mL) was stirred at 100°C for 16 h in a sealed tube. The reaction mixture was then passed through The filtrate was filtered through a plug, and the filtrate was diluted with EtOAc. The resulting solution was washed with water, brine, dried over Na2SO4, filtered and concentrated. The concentrate was purified by flash column chromatography using a gradient of 0% to 6% ethyl acetate in petroleum ether to obtain 2-(4,4-difluoropiperidin-1-yl)-3-fluoro-N-(4-methoxybenzyl)-6-methylpyridine-4-amine (0.67 g, 1.83 mmol, 85% yield) as a light yellow solid. 1 HNMR(300MHz,DMSO-d6)δppm 7.19–7.29(m,2H),6.83–6.93(m,2H),6.75(m,1H),6.13(d,J=5.4Hz,1H),4. 26(d,J=6.3Hz,2H),3.72(s,3H),3.40(d,J=11.5Hz,4H),1.92–2.14(m,7H). m / z(ESI):366.1(M+H) + .

[0335] Step 4: Anisole (0.179 mL, 1.642 mmol) and TFA (1.5 mL, 19.47 mmol) were added to a solution of 2-(4,4-difluoropiperidin-1-yl)-3-fluoro-N-(4-methoxybenzyl)-6-methylpyridin-4-amine (0.3 g, 0.821 mmol) in dichloromethane (3 mL) at ambient temperature, and the reaction mixture was stirred at 50 ° C for 2.5 h. The reaction mixture was then quenched with water and the pH was adjusted to 8 with 10% sodium bicarbonate solution, which was then extracted with ethyl acetate. The organic layer was washed with water and brine, dried over Na2SO4, filtered and concentrated. The concentrate was purified by flash column chromatography using a gradient of 20% ethyl acetate in petroleum ether to give 2-(4,4-difluoropiperidin-1-yl)-3-fluoro-6-methylpyridin-4-amine (0.17 g, 0.69 mmol, 84% yield) as a yellow oil. 1 HNMR (400MHz, DMSO-d6) δppm 6.15 (d, J = 5.5 Hz, 1H), 5.79 (s, 2H), 3.40 (t, J = 5.6 Hz, 4H), 2.13 (s, 3H), 2.02 (tt, J = 14.2, 5.6 Hz, 4H). m / z(ESI):246.2(M+H) + .

[0336] Intermediate 10: 2-(6-amino-2-(4,4-difluoropiperidin-1-yl)pyrimidin-4-yl)propan-2-ol

[0337]

[0338] Step 1: (3,5-dimethoxyphenyl)methylamine (19.39 g, 116 mmol) and DIPEA (33.7 mL, 193 mmol) were added to a solution of 2,6-dichloropyrimidine-4-formic acid methyl ester (20.00 g, 97 mmol) in tetrahydrofuran (200 mL) at 0 ° C. The reaction mixture was then stirred at ambient temperature for 16 h, then quenched with water and extracted with EtOAc. The organic layer was washed with brine and dried over Na2SO4, filtered and concentrated. The concentrate was ground with DCM and hexane to obtain 2-chloro-6-((3,5-dimethoxybenzyl)amino)pyrimidine-4-formic acid methyl ester (19.5 g, 57.7 mmol, 59.8% yield) as an off-white solid. 1H NMR(400MHz,DMSO-d6)δppm 8.52(t,J=5.5Hz,1H),7.16(d,J=8.9Hz,2H),6.59(d,J=2.4Hz,1H),6.50(dd,J=8.4,2.4Hz,1H ),4.40(d,J=5.4Hz,2H),3.82(d,J=14.5Hz,6H),3.75(d,J=5.8Hz,3H).m / z(ESI):338.1(M+H) + .

[0339] Step 2: A solution of 2-chloro-6-((3,5-dimethoxybenzyl)amino)pyrimidine-4-formic acid methyl ester (3g, 8.88mmol), 4,4-difluoropiperidine hydrochloride (2.10g, 13.32mmol) and DIPEA (3.44g, 26.6mmol) in DMF (30mL) was stirred at 90 ° C for 16h in a sealed tube. The reaction mixture was then quenched with water and extracted with ethyl acetate. The organic layer was washed with brine, dried over Na2SO4, filtered and concentrated. The crude material was purified by flash column chromatography using a gradient of 50% to 60% ethyl acetate in petroleum ether to provide 2-(4,4-difluoropiperidin-1-yl)-6-((3,5-dimethoxybenzyl)amino)pyrimidine-4-formic acid methyl ester (2.6g, 6.15mmol, 69.3% yield) as a light yellow solid. 1 HNMR(300MHz,DMSO-d6)δppm 7.74(s,1H),7.13(d,J=8.3Hz,1H),6.55(d,J=2.4Hz,1H),6.47(dd,J=8.3,2.4Hz,2H) ,4.39(s,2H),3.82(s,3H),3.79(s,3H),3.73(s,3H),3.33(m,4H),1.85–2.00(m,4H).

[0340] Step 3: Sulfuric acid (0.126 mL, 2.37 mmol) was added dropwise to a solution of methyl 2-(4,4-difluoropiperidin-1-yl)-6-((3,5-dimethoxybenzyl)amino)pyrimidine-4-carboxylate (1 g, 2.367 mmol) in DCM (10 mL) at 0 ° C. The mixture was then allowed to warm to room temperature and the progress of the reaction was monitored by TLC. After completion of the starting material, the reaction mixture was quenched with ice water and the pH was adjusted to 9 by using 10% NaHCO3 solution. The reaction mixture was then extracted with ethyl acetate, the organic layer was washed with brine, dried over Na2SO4, filtered and concentrated to provide methyl 6-amino-2-(4,4-difluoropiperidin-1-yl)pyrimidine-4-carboxylate (0.45 g, 1.653 mmol, 69.8% yield) as an off-white solid. 1 HNMR(300MHz,DMSO-d6)δppm 6.90(br s,2H),6.39(s,1H),3.85–3.83(m,4H),3.79(s,3H),1.99-1.35(m,4H).m / z(ESI):273.1(M+H) + .

[0341] Step 4: Methylmagnesium bromide (2.0M diethyl ether) (2.07 mL, 4.13 mmol) was added to a solution of 6-amino-2-(4,4-difluoropiperidin-1-yl)-pyrimidine-4-carboxylic acid methyl ester (0.45 g, 1.653 mmol) in tetrahydrofuran (5 mL) at 0 ° C and stirred at room temperature for 2 h. After the starting material was consumed, the reaction mixture was quenched with ice water and extracted with ethyl acetate. The organic layer was washed with brine, dried over Na2SO4, filtered and concentrated. The concentrate was purified by flash column chromatography using a gradient of 0% to 90% ethyl acetate in petroleum ether to provide 2-(6-amino-2-(4,4-difluoropiperidin-1-yl)pyrimidin-4-yl)propan-2-ol (0.35 g, 1.28 mmol, 78% yield) as a light yellow solid. m / z(ESI):273.1(M+H) + .

[0342] Table 3: The following intermediates were prepared following a similar procedure as described for Intermediate 10

[0343]

[0344] Intermediate 11: 2-(4-amino-6-methylpyrimidin-2-yl)propan-2-ol

[0345]

[0346] Step 1: A mixture of 2,4-dichloro-6-methylpyrimidine (3.0 g, 18.40 mmol, Aldrich, St. Louis, MO, USA), bis(4-methoxybenzyl)-amine (7.10 g, 27.6 mmol, Combi-Blocks Inc., San Diego, CA, USA), potassium carbonate (7.63 g, 55.2 mmol, Aldrich, St. Louis, MO, USA) in tetrahydrofuran (100 mL) was stirred at room temperature for 72 h. The reaction mixture was diluted with water (50 mL) and then extracted with EtOAc (2 x 100 mL). The combined organic extracts were then dried over MgSO4 and concentrated in vacuo. The residue was purified by chromatography (silica gel, 0% to 100% EtOAc / heptane) to provide 2-chloro-N,N-bis(4-methoxybenzyl)-6-methylpyrimidin-4-amine (3.11 g, 8.10 mmol, 44.0%) as an off-white solid. 1 H NMR(DMSO-d6)δppm 7.16(br d,J=6.2Hz,4H),6.89(d,J=8.7Hz,4H),6.60(s,1H),4.39-4.84(m,4H),3.73(s,6H),2.20(s,3H).m / z(ESI):384.2(M+H)+.

[0347] Step 2: 2-Chloro-N,N-bis(4-methoxybenzyl)-6-methylpyrimidin-4-amine (1.0 g, 2.61 mmol), 1,3-bis(diphenylphosphino)propane (64.5 mg, 0.156 mmol, Aldrich, St. Louis, MO, USA), diethyl oxalate (0.529 mL, 3.91 mmol, Aldrich, St. Louis, MO, USA), trans-dichlorobis(triphenylphosphine)palladium(ii) (54.9 mg, 0.078 mmol, Strem Chemicals, Newburyport, MA, USA) in ethanol (0.5 mL) were added. Inc., Newburyport, MA, USA)) and 4-(dimethylamino)pyridine (477 mg, 3.91 mmol, Aldrich, St. Louis, MO, USA)) were subjected to microwave irradiation at 140 ° C for 20 min. The reaction mixture was then diluted with water (50 mL) and then extracted with EtOAc (2x50 mL). The combined organic extracts were then dried over MgSO4 and concentrated in vacuo. The residue was purified by chromatography (silica gel, 0% to 100% EtOAc / heptane) to provide 4-(bis(4-methoxybenzyl)amino)-6-methylpyrimidine-2-carboxylic acid ethyl ester (274 g, 0.650 mmol, 24.95% yield) as a light yellow solid. 1 HNMR(methanol-d4)δppm 7.29(br s, 4H), 6.97 (d, J = 8.5Hz, 4H), 6.68 (s, 1H), 4.78-4.93 (m, 4H), 4.54 (q, J = 7.2Hz, 2H), 3.88 (s, 6H), 2.43 (s, 3H), 1.53 (t, J = 7.0Hz, 3H). m / z(ESI):422.1(M+H)+.

[0348] Step 3: Methylmagnesium bromide (3.4M in 2-methyltetrahydrofuran) (0.829 mL, 2.82 mmol, Aldrich, St. Louis, MO, USA) was added dropwise to a solution of ethyl 4-(bis(4-methoxybenzyl)amino)-6-methylpyrimidine-2-carboxylate (396 mg, 0.940 mmol) in 2-methyltetrahydrofuran (7 mL) at 0°C under N2. After addition, the mixture was then stirred at 0°C for 3.5 h. The mixture was then quenched with saturated NH4Cl (10 mL) and then extracted with EtOAc (2x50 mL). The combined organic extracts were then dried over MgSO4 and concentrated in vacuo. The residue was purified by chromatography (silica gel, 0% to 100% EtOAc / heptane) to provide 2-(4-(bis(4-methoxybenzyl)amino)-6-methylpyrimidin-2-yl)propan-2-ol (307 mg, 0.753 mmol, 80% yield) as a yellow solid. m / z (ESI): 408.2 (M+H)+.

[0349] Step 4: A solution of 2-(4-(bis(4-methoxybenzyl)amino)-6-methylpyrimidin-2-yl)propan-2-ol (300 mg, 0.736 mmol) in trifluoroacetic acid (10 mL, Aldrich, St. Louis, MO, USA) was subjected to microwave irradiation at 110 °C for 30 min. The mixture was then concentrated under reduced pressure. The crude product was then dissolved in DCM (10 mL) and then quenched with saturated Na2CO3 (15 mL). The mixture was then extracted with EtOAc (2x50 mL). The combined organic extracts were then dried over MgSO4 and concentrated in vacuo. Chromatographic purification of the residue (silica gel, 0% to 100% EtOAc:EtOH (3:1) / heptane) provided 2-(4-amino-6-methylpyrimidin-2-yl)propan-2-ol (123 mg) as a light yellow solid. m / z(ESI):168.2(M+H)+.

[0350] Cyclic Ar 2 Preparation of intermediates

[0351] Intermediate 12: 4-iodo-2-(6-azaspiro[2.5]oct-6-yl)benzoic acid

[0352]

[0353] 6-azaspiro [2.5] octane hydrochloride (216 g, 1.47 mol, Wuxi AppTec) was added to a solution of 2-fluoro-4-iodobenzoic acid (300 g, 1.13 mol, Combi-Blocks, San Diego, CA) in DMSO (2.10 L) at 20 ° C. K2CO3 (468 g, 3.38 mol) was then added, and the reaction solution was stirred at 140 ° C for 48 h under N2. The reaction solution was slowly poured into ice water (4.20 L), and then extracted with hexane (2.00 L x 3). The aqueous phase was separated and pH = 6 was adjusted with HCl (2.00 mol / L, aqueous solution). Solids were precipitated and collected. The solids were washed with water (700 mL x 3) and filtered. The wet solids were spread on a large observation glass and dried in air at 25 ° C for 72 h. 4-iodo-2-(6-azaspiro[2.5]octan-6-yl)benzoic acid (280 g, 777 mmol, 68.9% yield) was obtained as a light yellow solid. 400 MHz DMSO-d6 δ ppm 8.07 (s, 1H), 7.76-7.66 (m, 2H), 3.10 (t, J=5.2 Hz, 4H), 1.55 (br s, 4H), 0.41 (s, 4H).

[0354] Table 4: The following intermediates were prepared by a similar procedure to Intermediate 12:

[0355]

[0356] Intermediate 13: 4-(Methylsulfonyl)-2-(6-azaspiro[2.5]oct-6-yl)benzoic acid

[0357]

[0358] Step 1: benzyl bromide (78.1g, 454.0mmol) and sodium carbonate (52.5g, 495mmol) were added to a solution of 2-fluoro-4-(methylsulfonyl)benzoic acid (90.0g, 412.1mmol) in N,N-dimethylformamide (1.0L) at 0°C. The reaction mixture was stirred at room temperature for 12h. The reaction mixture was quenched with water (1L) and extracted with MTBE (3x1L). The combined organic layers were washed with brine (1L), dried over Na2SO4, filtered and concentrated under reduced pressure. The crude residue was purified by column chromatography on silica gel using 0% to 30% ethyl acetate in hexane as eluent to obtain 2-fluoro-4-(methylsulfonyl)benzoic acid benzyl ester (100g, 79% yield) as a white solid. 1HNMR (300MHz, DMSO-d6) δppm 8.16 (dd, J=8.2, 6.9Hz, 1H), 7.98–7.86 (m, 2H), 7.48–7.31 (m, 5H), 5.40 (s, 2H), 3.33 (s, 3H).

[0359] Step 2: DIPEA (57.6g, 446mmol) is added to the solution of 2-fluoro-4-(methylsulfonyl)benzyl benzoate (55g, 178mmol) in dimethyl sulfoxide (550mL), 6-azaspiro [2.5] octane (29.8g, 268mmol) is subsequently added, and the reaction mixture is stirred at 100 ℃ for 24h. The reaction mixture is quenched with water (1L) and extracted with MTBE (3x1L). The organic layer merged is washed with saline solution (1L), through Na2SO4 drying, filtered and concentrated under reduced pressure. The crude product is purified by column chromatography on silica gel (230-400 mesh) using 0% to 10% ethyl acetate in hexane as eluent, to obtain 4-(methylsulfonyl)-2-(6-azaspiro [2.5] octane-6-yl)benzyl benzoate (55g, 77% yield) as a white solid. 1 HNMR(400MHz,DMSO-d6)δppm 7.76(d,J=8.0Hz,1H),7.52–7.45(m,4H),7.43–7.35(m,3H),5.35(s,2H) ,3.25(s,3H),3.05(t,J=5.3Hz,4H),1.36(t,J=5.3Hz,4H),0.30(s,4H). m / z(ESI):400.1(M+H) + .

[0360] Step 3: 1N aqueous sodium hydroxide solution (407mL, 407mmol) was added to a solution of 4-(methylsulfonyl)-2-(6-azaspiro[2.5]oct-6-yl)benzyl benzoate (65g, 163mmol) in tetrahydrofuran (108mL) and methanol (36mL) at 60°C, and the reaction mixture was stirred for 12h. The reaction mixture was concentrated under reduced pressure to remove THF and methanol. The remaining aqueous solution was acidified to pH~2 with 1.5N HCl solution. The precipitated solid was filtered, washed with water (200mL), then washed with hexane (200mL), and dried in vacuo for 12h to obtain 4-(methylsulfonyl)-2-(6-azaspiro[2.5]oct-6-yl)benzoic acid (42g, 83% yield) as an off-white solid. 1H NMR(400MHz,DMSO-d6)δppm 16.13(s,1H),8.04(d,J=8.0Hz,1H),7.99(s,1H),7.75(d,J=8.0Hz,1H),3.29(s,3H),3.17(bs,4H),1.55(bs,4H),0.41(s,4H). m / z(ESI):310.1(M+H) + .

[0361] Intermediate 14: 4-((1-methylcyclopropane)-1-sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzoic acid

[0362]

[0363] Step 1: Sodium carbonate (31.5 g, 297 mmol) was added to a solution of 4-bromo-2-fluorobenzoic acid (50.0 g, 228 mmol, F Chemicals, China) in DMF (500 mL) at 0 ° C, followed by benzyl bromide (43.0 g, 251 mmol), and the reaction mixture was stirred at room temperature for 24 h. The reaction mixture was quenched with water (1000 mL) and extracted with ethyl acetate (3x2000 mL). The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude residue was purified by column chromatography on silica gel using 10% ethyl acetate in hexane as eluent to obtain 4-bromo-2-fluorobenzoic acid benzyl ester (65 g, 92% yield) as a colorless viscous oil. 1 H NMR (400MHz, chloroform-d) δppm7.91(t,J=8.4Hz,1H),7.47(dt,J=6.0,1.5Hz,2H),7.43(dd,J=6.8,1.8Hz,1 H),7.41(q,J=1.5Hz,1H),7.39–7.35(m,1H),7.02(dd,J=8.7,2.1Hz,1H),6.79(s,1H),5.38(s,2H). m / z(ESI):310.2(MH) + .

[0364] Step 2: 6-azaspiro [2.5] octane (32.4, 291mmol, Wuxi Apptec) in DMSO (200mL) and DIPEA (30g, 291mmol) was added to a solution of 4-bromo-2-fluorobenzoic acid benzyl ester (60.0g, 194mmol) in DMSO (200mL), and stirred at 100°C for 12h. The reaction mixture was quenched with water (2000mL) and extracted with ethyl acetate (3x2000mL). The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude residue was purified by column chromatography on silica gel using 10% ethyl acetate in hexane to obtain 4-bromo-2- (6-azaspiro [2.5] octane-6-yl) benzyl benzoate (70g, 90% yield) as a yellow oil. 1 H NMR (400MHz, chloroform-d) δppm 7.60(dd,J=8.4,1.9Hz,1H),7.50–7.45(m,2H),7.49–7.28(m,3H),7.20(d,J=1.9Hz,1H),7.07(dd ,J=8.3,1.9Hz,1H),5.36(s,2H),3.12–3.02(m,4H),1.47(t,J=5.3Hz,4H),0.33(d,J=1.8Hz,4H). m / z(ESI):398.1,400.1(M+H) + .

[0365] Step 3: 4-bromo-2-(6-azaspiro[2.5]n-oct-6-yl)benzyl benzoate (9 g, 22.48 mmol), 1-methylcyclopropane-1-sulfonamide (3.95 g, 29.2 mmol, Combi-Blocks, San Diego, CA) and K2CO3 (6.21 g, 45.0 mmol) in 1,4-dioxane (90 mL) were added to a 250 mL sealed tube, the reaction was degassed and purged with nitrogen for 5 min. Xantphos (1.301 g, 2.248 mmol) was added to the reaction mixture, followed by Pd2(dba)3 (1.03 g, 1.12 mmol), the sealed tube was closed and stirred at 110 ° C for 18 h. The reaction mixture was quenched with water (250 mL) and extracted with ethyl acetate (2x150 mL). The organic layer was washed with water (100 mL), dried over Na2SO4, filtered and concentrated under reduced pressure. The crude residue was purified by column chromatography on silica gel eluting with a gradient of 0% to 15% EtOAc in hexanes to afford benzyl 4-((1-methylcyclopropane)-1-sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzoate (6.1 g, 59% yield) as an orange oil.1 HNMR(400MHz,DMSO-d6)δppm10.06(s,1H),7.62(d,J=8.5Hz,1H),7.48–7.31(m,5H),6.98(s,1H),6.81(d,J=8.5Hz,1H),5. 27(s,2H),2.92(t,J=4.96Hz,4H),1.40–1.30(m,7H),1.16(dd,J=6.4,4.7Hz,2H),0.81(dd,J=6.4,4.7Hz,2H),0.28(s,4H). m / z(ESI):455.2(M+H) + .

[0366] Step 4: To a solution of benzyl 4-((1-methylcyclopropane)-1-sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzoate (2.1 g, 4.62 mmol) in methanol (20 mL) and ethyl acetate (10 mL) was added 10% Pd-C (1.05 g, 50% wt / wt) under nitrogen atmosphere. The reaction mixture was degassed and stirred under hydrogen pressure (1 atm, balloon pressure) for 4 h. The reaction mixture was filtered through a bed of 4-nitropropane and washed with methanol (20 mL). The filtrate was concentrated under reduced pressure. The residue was ground with EtO (50 mL) to give 4-((1-methylcyclopropane)-1-sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzoic acid (1.2 g, 71% yield) as an off-white solid. 1 H NMR(400MHz,DMSO-d6)δppm 13.20(s,1H),10.33(s,1H),7.95(d,J=8.6Hz,1H),7.41(s,1H),7.20(d,J=8.6Hz,1H),2.99 (s, 4H), 1.56 (s, 4H), 1.39 (s, 3H), 1.18 (t, J = 4.8Hz, 2H), 0.83 (t, J = 4.7Hz, 2H), 0.42 (s, 4H). m / z(ESI):363.2(M+H) + .

[0367] Intermediate 15: 4-(N-(3-methyloxetan-3-yl)sulfamoyl)-2-(6-azaspiro[2.5]octane-6-yl)sulfamoyl benzoic acid

[0368]

[0369] Step 1: 4-(chlorosulfonyl)-2-fluorobenzoic acid methyl ester (12.37 g, 49.0 mmol) was added to a solution of 3-methyl-3-oxetanes hydrochloride (5.50 g, 44.5 mmol) and N,N-diisopropylethylamine (23.26 mL, 134 mmol) in DCM (200 mL) at 0 ° C, and the mixture was stirred from 0 ° C to room temperature for 1 h. The mixture was diluted with 1.0 N HCl (200 mL) and extracted with dichloromethane (150 mL x 2). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a crude product. The crude product was purified using a Biotage SNAP 100 g column eluting with 0% to 30% 3:1 EtOAc-EtOH in heptane to afford methyl 2-fluoro-4-(N-(3-methyloxetan-3-yl)sulfamoyl)benzoate (13.59 g, 44.8 mmol, 100% yield) as a white solid. 1 HNMR(500MHz,DMSO-d6)δppm 8.67(s,1H),8.11(t,J=7.37Hz,1H),7.76-7.82(m,1H),7.69-7.76(m,1H) ,4.56(d,J=6.23Hz,2H),4.18(d,J=6.75Hz,2H),3.90(s,3H),1.42(s,3H).

[0370] Step 2: A mixture of N,N-diisopropylethylamine (16.23 mL, 93 mmol), 6-azaspiro[2.5]octane (6.22 g, 55.9 mmol) and methyl 2-fluoro-4-(N-(3-methyloxetan-3-yl)sulfamoyl)benzoate (14.13 g, 46.6 mmol) in anhydrous 1,4-dioxane was stirred at 100 °C for 20 h. The mixture was cooled to room temperature, quenched with water and extracted with ethyl acetate. The combined organic phases were washed with brine, dried and evaporated to dryness under reduced pressure. The crude product was purified using a Biotage SNAP 340g column eluting with 0% to 40% 3:1 EtOAc-EtOH in heptane to afford methyl 4-(N-(3-methyloxetan-3-yl)sulfamoyl)-2-(6-azaspiro[2.5]oct-6-yl)benzoate (14.15 g, 35.9 mmol, 77% yield) as an off-white solid. 1HNMR(500MHz,DMSO-d6)δppm 8.42(s,1H),7.72(d,J=8.04Hz,1H),7.47(d,J=1.56Hz,1H),7.36(dd,J=1.82,8.04Hz,1H),4.55(d,J=5.97H z, 2H), 4.14 (d, J = 6.49Hz, 2H), 3.85 (s, 3H), 3.02-3.09 (m, 4H), 1.44-1.50 (m, 4H), 1.42 (s, 3H), 0.35 (s, 4H).

[0371] Step 3: A mixture of 4-(N-(3-methyloxetane-3-yl)sulfamoyl)-2-(6-azaspiro[2.5]octyl-6-yl)benzoic acid methyl ester (14.15 g, 35.9 mmol) and lithium hydroxide monohydrate (22.58 g, 538 mmol) in THF-water-MeOH (1:1:1, 300 mL) was stirred at room temperature overnight. The mixture was concentrated under reduced pressure to partially remove the organic solvent. The solution was acidified to pH <3 with 2N HCl. The precipitated solid was filtered and dried in air to obtain 4-(N-(3-methyloxetane-3-yl)sulfamoyl)-2-(6-azaspiro[2.5]octyl-6-yl)benzoic acid (9.94 g, 26.1 mmol, 72.8% yield) as a white solid. 1 H NMR(500MHz,DMSO-d6)δppm 8.51(s,1H),8.04(d,J=8.04Hz,1H),7.89(d,J=1.30Hz,1H),7.66(dd,J=1.69,8.17Hz,1H),4.55(d,J=6. 23Hz, 2H), 4.09-4.17 (m, 2H), 3.06-3.19 (m, 4H), 1.56 (t, J = 5.19Hz, 4H), 1.40 (s, 3H), 0.36-0.46 (s, 4H).

[0372] Intermediate 16: 4-((1-(tert-butoxycarbonyl)azetidin-3-yl)sulfonyl)-2-(6-azaspiro[2.5] 6-Octyl)benzoic acid

[0373]

[0374] Step 1: A mixture of methyl 4-bromo-2-(6-azaspiro[2.5]octan-6-yl)benzoate (10.50 g, 32.4 mmol, intermediate 12-2), DIPEA (8.37 mL, 64.8 mmol), Xantphos (1.874 g, 3.24 mmol) and Pd2(dba)3 (2.97 g, 3.24 mmol) in 1,4-dioxane was bubbled with argon, and then tert-butyl 3-mercaptoazetidine-1-carboxylate (7.66 mL, 40.5 mmol) was added. The mixture was stirred at 100° C. for 18 h. The mixture was cooled to room temperature, concentrated and purified by Biotage SNAP eluting with a gradient of 0% to 25% 3:1 EtOAc-EtOH in heptane to afford tert-butyl 3-((4-(methoxycarbonyl)-3-(6-azaspiro[2.5]octan-6-yl)phenyl)thio)azetidine-1-carboxylate (13.85 g, 32.0 mmol, 99% yield) as a light yellow sticky solid. 1 H NMR(500MHz,DMSO-d6)δppm 7.55(d,J=8.04Hz,1H),6.79(s,1H),6.76(d,J=8.14Hz,1H),4.34-4.43(m,2H),4.27-4.34(m,1H),3.79(s ,3H),3.70(dd,J=4.67,8.56Hz,2H),2.97-3.04(m,4H),1.41-1.51(m,4H),1.38(s,9H),0.29-0.37(m,4H).

[0375] Step 2: To a solution of tert-butyl 3-((4-(methoxycarbonyl)-3-(6-azaspiro[2.5]octan-6-yl)phenyl)thio)azetidine-1-carboxylate (13.85 g, 32.0 mmol) in 1,4-dioxane (300 mL) was added Oxone monopersulfate (39.4 g, 64.0 mmol) in 150 mL of water. The mixture was stirred at room temperature for 5 h, and 150 mL of ethyl acetate and 150 mL of water were added, and the mixture was stirred for 10 min. The organic layer was separated, and the aqueous layer was extracted with ethyl acetate. The combined organics were washed with brine, dried, filtered and concentrated. The crude product was purified by Biotage SNAP 340 g column eluting with a gradient of 0% to 25% EtOAc-EtOH (3:1) in heptane to afford tert-butyl 3-((4-(methoxycarbonyl)-3-(6-azaspiro[2.5]oct-6-yl)phenyl)sulfonyl)azetidine-1-carboxylate (12.77 g, 27.5 mmol, 86% yield) as an off-white solid. 1H NMR(500MHz,DMSO-d6)δ7.75(d,J=8.04Hz,1H),7.44-7.50(m,2H),4.48-4.55(m,1H),4.09(br s,2H),3.97-4.02(m,2H),3.86(s,3H),3.04-3.17(m,4H),1.42-1.51(m,4H),1.38(s,9H),0.35(s,4H).

[0376] Step 3: A mixture of tert-butyl 3-((4-(methoxycarbonyl)-3-(6-azaspiro[2.5]oct-6-yl)phenyl)sulfonyl)azetidine-1-carboxylate (12.77 g, 27.5 mmol) and lithium hydroxide monohydrate (11.53 g, 275 mmol) in THF-water-MeOH (1:1:1, 230 mL) was stirred at room temperature for 15 h. The mixture was concentrated under reduced pressure to remove some organic solvents. The solution was acidified to pH <3 with 2N HCl. The precipitated solid was filtered and dried to give 4-((1-(tert-butoxycarbonyl)azetidine-3-yl)sulfonyl)-2-(6-azaspiro[2.5]oct-6-yl)benzoic acid (10.6 g, 23.53 mmol, 86% yield) as an off-white solid. 1 H NMR(500MHz,DMSO-d6)δppm 8.03(d,J=8.30Hz,1H),7.93(d,J=1.82Hz,1H),7.72(dd,J=1.69,8.17Hz,1H),4.48-4.60(m,1H),4 .10(br.s.,2H),3.99-4.06(m,3H),3.14-3.22(m,4H),1.49-1.59(m,4H),1.38(s,9H),0.41(s,4H).

[0377] With AR 1 and AR 2 Cyclic intermediate compounds

[0378] Intermediate 17: N-(2-chloro-6-methylpyrimidin-4-yl)-4-iodo-2-(6-azaspiro[2.5]octan-6-yl)benzoyl amine

[0379]

[0380] 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphine 2,4,6-trioxide (50wt% solution in EtOAc, 12.50mL, 21.00mmol) and triethylamine (2.93mL, 21.00mmol) were added to a suspension of 4-iodo-2-(6-azaspiro[2.5]octan-6-yl)benzoic acid (3.0g, 8.40mmol, Intermediate 12) and 2-chloro-6-methylpyrimidin-4-amine (1.45g, 10.08mmol, Aurum Pharmtech Inc.) in DCE (20mL). The mixture was heated at 85°C for 24h and then cooled to room temperature. Water (10mL) was added, the layers were separated, and the aqueous layer was extracted with DCM (1x10mL). The combined organic extracts were dried over anhydrous MgSO4, filtered and concentrated in vacuo to give a solid. The solid was suspended in 1:1 EtOAc / heptane and filtered to provide N-(2-chloro-6-methylpyrimidin-4-yl)-4-iodo-2-(6-azaspiro[2.5]octan-6-yl)benzamide (3.61 g, 7.48 mmol, 89% yield) as an off-white solid. 1 H NMR (400MHz, chloroform-d) δppm 13.53 (br s,1H)8.11(s,1H)7.92(d,J=8.29Hz,1H)7.70(s,1H)7.65-7.69(m,1H)3.05(t,J=5.39Hz,4H)2.53(s,3H)1.61-1.88(m,4H)0.44(s,4H). m / z(ESI):483.0(M+H) + .

[0381] Intermediate 18: (R)-4-Bromo-N-(6-methyl-2-(2-methylmorpholino)pyrimidin-4-yl)-2-(6-azaspiro [2.5]oct-6-yl)benzamide

[0382]

[0383] Step 1: To a 100 mL round bottom flask was added 4-bromo-2-(6-azaspiro[2.5]octyl-6-yl)benzoic acid (0.9 g, 2.90 mmol, Intermediate 12-1), pyridine (0.657 mL, 8.12 mmol) and perfluorophenyl 2,2,2-trifluoroacetate (0.717 g, 3.77 mmol) in DCM (8 mL). The resulting mixture was stirred at room temperature for 16 h, and the solvent was removed in vacuo to give the crude product, which was used in the next step without purification. m / z (ESI): 476 and 478 (M+1).

[0384] Step 2: (R)-6-methyl-2-(2-methylmorpholino)pyrimidine-4-amine (0.223 g, 1.15 mmol, intermediate 2) dissolved in N,N-dimethylformamide (6 mL) was added to a 50 mL round-bottom flask, followed by sodium hydride (0.084 g, 2.1 mmol) at room temperature under a nitrogen atmosphere. The reaction mixture was stirred for 10 min, then treated with 4-bromo-2-(6-azaspiro[2.5]octane-6-yl)benzoic acid perfluorophenyl ester (0.5 g, 1.050 mmol) at room temperature under a nitrogen atmosphere. The resulting reaction mixture was stirred for another 2 h at room temperature. The reaction mixture was extracted with DCM (2x30 mL), separated, dried over anhydrous sodium sulfate and evaporated to dryness to obtain a crude material. It was then adsorbed onto a plug of silica gel and purified by flash column chromatography on silica gel eluting with 30% to 50% EtOAc / hexanes to provide the title compound (0.20 g, 0.40 mmol, 38% yield) as a white solid. 1 H NMR(300MHz,DMSO-d6)δppm 13.19(bs,1H),7.99(d,J=8.4Hz,1H),7.69(s,1H),7.55(d,J=8.4Hz,1H),7.36(s,1H),4.45(t,J=12Hz,2H),3.8 8(m,1H),3.52-3.48(m,2H)3.10-2.90(m,6H),2.30(s,3H),1.71-1.62(m,4H),1.14(d,J=6Hz,3H),0.36(s,4H). m / z(ESI):500 and 502(M+1).

[0385] Intermediate 19: N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-iodo-2-(6-azaspiro [2.5]oct-6-yl)benzamide

[0386]

[0387] Under argon, 4-iodo-2-(6-azaspiro [2.5] octane-6-yl) benzoic acid (150.0g, 420mmol, intermediate 12) is suspended in dichloromethane (1000mL). Catalytic DMF (1.0mL) is added, followed by dropwise addition of a solution of thionyl chloride (54.6g, 28mL, 459mmol, Sigma-Aldrich Corporation) in dichloromethane (500mL) in 10min. After stirring at ambient temperature for 30min, the mixture is evaporated to dryness under reduced pressure. The crude product is azeotroped with toluene (2x300mL), and is suspended in dichloromethane (300mL) under argon. Potassium phosphate (267g, 1.26mol, Sigma-Aldrich Corporation) was added, followed by a solution of 2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidine-4-amine (100g, 438mmol, intermediate 4) and N,N-diisopropylethylamine (200mL, 1.14mol, Sigma-Aldrich Corporation) in DCM (300mL, added within 5min). The yellow mixture was stirred for 3h at ambient temperature and then evaporated to dryness under reduced pressure. The crude solid was suspended in dichloromethane (1L) and stirred for 10min. The mixture was filtered through a glass frit, and the solid was washed with additional dichloromethane (2x100mL). The solid was discarded, and the filtrate was evaporated to dryness under reduced pressure. The crude residue was suspended in acetonitrile (750mL) and stirred for 15min at ambient temperature. The suspension was filtered through a glass frit and the solid was washed with additional acetonitrile (75 mL). The solid was dried under a stream of nitrogen to give N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-iodo-2-(6-azaspiro[2.5]octan-6-yl)benzamide (186 g, 328 mmol, 78% yield). 1 HNMR(400MHz,DMSO-d6)δppm 13.38(br s,1H)7.72-7.87(m,3H)7.39(s,1H)3.91(br s,4H)2.99-3.06(m,4H)2.32(s,3H)1.92-2.07(m,4H)1.62-1.85(m,4H)0.38(s,4H). m / z(ESI):568.0(M+H) + .

[0388] Intermediate 20: 4-Bromo-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-2-fluoro-6-(6-nitrogen Heterospiro[2.5]octan-6-yl)benzamide

[0389]

[0390] Step 1: To a solution of 4-bromo-2,6-difluorobenzoic acid (3.0 g, 12.7 mmol, Apollo Scientific Ltd.) in THF (50 mL) was added oxalyl chloride (1.7 mL, 19.0 mmol) followed by 1 drop of DMF. The mixture was stirred for 1 h and then the solvent was removed in vacuo to give a solid which was used directly in the next stage without further characterization. The solid was dissolved in DCM (50 mL) and anhydrous pyridine (4.31 mL, 50.6 mmol) followed by the addition of 2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-amine (2.89 g, 12.7 mmol, Intermediate 4) and the mixture was stirred at room temperature for 16 h. EtOAc (200 mL) was added and the mixture was washed with saturated NH4Cl (1x), water (1x), brine (1x), dried over anhydrous MgSO4, filtered and concentrated in vacuo to give an oil. The oil was purified by silica gel chromatography eluting with 0% to 40% EtOAc / heptane to provide 4-bromo-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-2,6-difluorobenzamide (1.36 g, 3.04 mmol, 24% yield) as a white solid. 1 HNMR (400MHz, DMSO-d6) δppm 11.24 (s, 1H) 7.64 (d, J = 7.05Hz, 2H) 7.19-7.37 (m, 1H) 3.86 (br s, 4H) 2.32 (s, 3H) 1.98 (br d, J = 11.40Hz, 4H). m / z(ESI):447.0,449.0(M+H) + .

[0391] Step 2: 4-Bromo-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-2,6-difluorobenzamide (0.65 g, 1.45 mmol), 6-azaspiro[2.5]octane (0.18 g, 1.60 mmol, Wuxi App Tech) and DIPEA (0.31 mL, 1.74 mmol) in DMSO (2.5 mL) were heated to 100° C. for 8 h and then cooled to room temperature. Water was added, the resulting suspension was filtered, and the obtained solid was dried. The solid was purified by silica gel chromatography eluting with 0% to 15% EtOAc / heptane to provide 4-bromo-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-2-fluoro-6-(6-azaspiro[2.5]octan-6-yl)benzamide (0.23 g, 0.43 mmol, 28.2% yield) as a white solid.1 H NMR (400MHz, DMSO-d6) δppm 10.86 (br s, 1H) 7.30 (br s, 1H) 7.21 (br d, J = 9.12Hz, 1H) 7.10 (br s, 1H) 3.88 (br s, 4H) 3.05 (br s, 4H) 2.32 (br s,3H)1.77-2.04(m,4H)1.36(brs,4H)0.27(s,4H). 19 F NMR (376MHz, DMSO-d6) δppm-94.88(s,1F)-113.60(s,1F). m / z(ESI):538.2,540.2(M+H) + .

[0392] Intermediate 21: 4-amino-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-5-fluoro-2-(6- Azaspiro[2.5]octan-6-yl)benzamide

[0393]

[0394] Step 1: Triethylamine (3.11 mL, 22.2 mmol) and 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphine 2,4,6-trioxide (50 wt% in EtOAc, 13.2 mL, 22.2 mmol) were added to a solution of 2,5-difluoro-4-nitrobenzenecarboxylic acid (1.5 g, 7.39 mmol, Combi-Blocks Inc.) and 2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-amine (1.69 g, 7.39 mmol, Intermediate 4) in DCE (15 mL), the mixture was heated to 85 °C for 2 h, then cooled to room temperature. Water (15 mL) was added, the resulting biphasic mixture was separated, the organic layer was dried over anhydrous MgSO4, filtered and concentrated in vacuo to give a solid. The solid was suspended in DCM (15 mL), filtered and dried to afford N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-2,5-difluoro-4-nitrobenzamide (3.05 g, 4.55 mmol, 62% yield) as a yellow solid. 1 HNMR(400MHz, DMSO-d6)δppm 11.15(s,1H)8.27(dd,J=8.81,5.91Hz,1H)7.99(dd,J=10.57,5.39Hz,1H)7.23(br s,1H)3.85(br s,3H)2.33(s,4H)1.89-2.05(m,4H). 19F NMR (376MHz, DMSO-d6) δppm-95.11(s,1F)-123.35(s,1F)-123.40(s,1F). m / z(ESI):414.2(M+H) + .

[0395] Step 2: Place N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-2,5-difluoro-4-nitrobenzamide (1.0 g, 2.42 mmol) and palladium (10 wt %, 0.45 g, 0.42 mmol on activated carbon) under argon atmosphere, then add EtOH (15 mL) followed by ammonium formate (0.76 g, 12.1 mmol). Stir the mixture at 75 °C for 10 min and then cool to room temperature. Use The palladium was filtered off and the filtrate was concentrated in vacuo. The resulting residue was dissolved in EtOAc (10 mL) and the solution was washed with water (2 x 10 mL), brine (1 x 10 mL), dried over anhydrous MgSO4, filtered, and concentrated in vacuo to provide 4-amino-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-2,5-difluorobenzamide (0.93 g, 2.19 mmol, 91% yield) as a white solid. 1 HNMR(400MHz,DMSO-d6)δppm 9.68-9.80(m,1H)7.40(dd,J=11.61,6.84Hz,1H)7.28(s,1H)6.54(dd,J=13. 48,7.26Hz,1H)6.26(s,2H)3.83-3.90(m,4H)2.30(s,3H)1.92-2.05(m,4H). 19 F NMR (376MHz, DMSO-d6) δppm-95.07(s,1F)-116.10(s,1F)-140.24(s,1F). m / z(ESI):384.2(M+H) + .

[0396] Step 3: 4-amino-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-2,5-difluorobenzamide (0.80 g, 2.09 mmol), 6-azaspiro[2.5]octane (0.70 g, 6.26 mmol, Wuxi App Tech) and DIPEA (1.1 mL, 6.26 mmol) in NMP (4 mL) were heated to 200 °C for 4 h in a microwave reactor. Water (4 mL) was added and the resulting suspension was stirred for 30 min, filtered and the collected solid was dried to give a white solid. The solid was dissolved in DCM, melted on silica gel and purified by silica gel chromatography eluting with 0% to 50% EtOAc / heptane to provide 4-amino-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-5-fluoro-2-(6-azaspiro[2.5]octan-6-yl)benzamide (702 g, 1.48 mmol, 70.9% yield) as a white solid. 1 HNMR (400MHz, DMSO-d6) δppm 13.77 (s, 1H) 7.64 (d, J = 12.85Hz, 1H) 7.38 (s, 1H) 6.83 (d, J = 8.09Hz, 1H) 6.04 (s, 2H) 3.90 (br t, J = 5.39Hz, 4H) 2.91 (br s, 4H) 2.29 (s, 3H) 1.88-2.04 (m, 4H) 0.37 (s, 4H). (Note: 4 protons were not observed) 19 F NMR (376MHz, DMSO-d6) δppm-94.73(s,1F)-138.31(s,1F). m / z(ESI):475.2(M+H) + .

[0397] Table 5: Intermediates 21-1 to 21-20 were prepared according to similar procedures to Intermediates 17 to 21:

[0398]

[0399]

[0400]

[0401] Intermediate 22: Ethyl 2-sulfamoylpropionate

[0402] Step 1: nBuLi (1.6M in hexane, 608.0mL, 973.0mmol) was slowly added to a solution of N,N-bis(4-methoxybenzyl)ethanesulfonamide (200.0g, 572.0mmol) in tetrahydrofuran (4000mL) at -78°C, and stirred for 30min. Ethyl chloroformate (92.0mL, 973.0mmol) in THF (50mL) was added to the reaction mixture, and stirred for 1h at -78°C. The reaction mixture was quenched with HCl (1.5N, 3000mL) and extracted with EtOAc (2x3000mL). The organic extract was dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude substance of ethyl 2-(N,N-bis(4-methoxybenzyl)sulfamoyl)propanoate (250.0g, 60% purity) as a yellow oil. 1 H-NMR showed the expected peak and was carried to the next step without any purification.

[0403] Step 2: To a solution of 2-(N,N-bis(4-methoxybenzyl)sulfamoyl)ethyl propionate (600.0g, 1.4mol) in trifluoroacetic acid (2.50L, 32.45mol), anisole (500.0mL, 4.57mol) was added and stirred at room temperature for 3h. The reaction mixture was concentrated under reduced pressure, quenched with 10% cold NaHCO3 aqueous solution (3L) and extracted with EtOAc (2x3L). The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude residue was purified by column chromatography on silica gel using 25% ethyl acetate in hexane to obtain a light yellow solid (168g), which was dissolved in DCM (1L) and precipitated by adding hexane (3000mL). The solid was filtered and dried in vacuo to obtain the title compound (109.0g, 42% yield) as a white solid. 1 HNMR (400MHz, DMSO-d6) δppm7.14 (s, 2H), 4.15 (q, J = 7.1Hz, 2H), 3.98 (q, J = 7.0Hz, 1H), 1.45 (d, J = 7.0Hz, 3H), 1.21 (t, J = 7.1Hz, 3H). m / z(ESI):180.1(M+H) + .

[0404] Intermediate 23: 2-Hydroxypropane-1-sulfonamide

[0405]

[0406] Step 1: Methanesulfonyl chloride (1.73mL, 22.3mmol) was added dropwise to a 0°C solution of bis(4-methoxybenzyl)amine (5.0g, 19.4mmol, Combi-Blocks Inc.) and triethylamine (8.12mL, 58.3mmol) in DCM (40mL) over 5min. The mixture was then stirred at room temperature for 2h before 1N HCl (50mL) was added. The layers were separated and the organic layer was washed with brine (1x50mL), dried over anhydrous MgSO4, filtered and then concentrated in vacuo to give a brown oil. The oil was dissolved in MeOH (50mL) and partially concentrated in vacuo until a thick suspension was formed. The suspension was stirred for 30min, filtered and the collected solid was dried in vacuo to provide N,N-bis(4-methoxybenzyl)methanesulfonamide (5.11g, 15.2mmol, 78% yield) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ ppm 7.19 (d, J = 8.50 Hz, 4H) 6.90 (d, J = 8.50 Hz, 4H) 4.19 (s, 4H) 3.75 (s, 6H) 2.89 (s, 3H).

[0407] Step 2: n-Butyl lithium (4.10 mL, 6.56 mmol) was added dropwise to a solution of N,N-bis(4-methoxybenzyl)methanesulfonamide (2.0 g, 5.96 mmol) in THF (15 mL) at -78 °C. The mixture was stirred for 10 min before acetaldehyde (0.37 mL, 6.56 mmol) was added dropwise. The mixture was stirred at -78 °C for 5 min before replacing the -78 °C bath with a 0 °C bath. The mixture was stirred for 15 min and then the reaction was quenched with saturated NH4Cl. EtOAc was added, the resulting two-phase mixture was separated, and the organic layer was dried over anhydrous MgSO4, filtered and concentrated in vacuo to give an oil. The oil was purified by silica gel chromatography eluting with a 0% to 70% EtOAc / heptane gradient to provide 2-hydroxy-N,N-bis(4-methoxybenzyl)propane-1-sulfonamide (1.84 g, 4.85 mmol, 81% yield) as an oil. 1HNMR(400MHz,DMSO-d6)δppm 7.16(d,J=8.50Hz,4H)6.80-6.92(m,4H)4.99(d,J=5.18Hz,1H)4.15-4.28(m,4H)4.01-4.13(m, 1H)3.74(s,6H)3.16(dd,J=13.99,6.53Hz,1H)3.04(dd,J=13.89,5.39Hz,1H)1.12-1.27(m,3H). m / z(ESI):402.2(M+Na) + .

[0408] Step 3: A mixture of 2-hydroxy-N,N-bis(4-methoxybenzyl)propane-1-sulfonamide (1.84 g, 4.85 mmol) and anisole (1.06 mL, 9.70 mmol) in TFA (10 mL) was stirred at room temperature for 2 h, and then the volatiles were removed in vacuo. The resulting oil was purified by silica gel chromatography eluting with a 0% to 100% EtOAc / heptane gradient to provide 2-hydroxypropane-1-sulfonamide (592 g, 4.25 mmol, 88% yield) as a colorless oil. 1 H NMR (400MHz, DMSO-d6) δppm 6.70 (s, 2H) 4.02-4.13 (m, 1H) 3.50-3.25 (br s, 1H) 3.07-3.15 (m, 1H) 2.98-3.06 (m, 1H) 1.20 (d, J = 6.22Hz, 3H).

[0409] Example 1: N-(2-((1-hydroxy-2-methylpropane-2-yl)amino)-6-methylpyrimidin-4-yl)-4-(methylsulfonyl)- acyl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide

[0410]

[0411] The mixture of N-(2-chloro-6-methylpyrimidine-4-yl)-4-(methylsulfonyl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide (200mg, 0.46mmol, intermediate 21-19), 2-amino-2-methyl-1-propanol (180uL, 1.80mmol, St. Louis, Missouri Sigma Aldrich (Aldrich, St.Louis, MO)) and DIPEA (200uL, 1.14mmol) in NMP (1mL) is heated at 130 DEG C for 60h.The reaction mixture is cooled to room temperature and quenched with water (10mL), extracted with ethyl acetate (2x10mL).The organic layer is washed with brine (500mL), dried (NaSO), filtered and concentrated under reduced pressure. The crude material was adsorbed onto a plug of silica gel and purified by column chromatography on silica gel (60-120 mesh) eluting with 0% to 70% ethyl acetate in hexanes to give the title compound (111 mg, 49%) as an off-white solid. 1 H NMR(400MHz,DMSO-d6)δppm11.65-11.96(m,1H),8.07-8.25(m,1H),7.82- 7.87(m,1H),7.75-7.81(m,1H),7.32-7.41(m,1H),6.13-6.24(m,1H),4.7 3-4.91(m,1H),3.42-3.54(m,2H),3.30-3.33(m,3H),3.01-3.16(m,4H),2 .18-2.28(m,3H),1.55-1.75(m,4H),1.25-1.42(m,6H),0.27-0.40(m,4H). m / z(ESI):487.4(M+H) + .

[0412] Table 6: Examples 1-1 to 1-7 were prepared following a similar procedure as described for Example 1:

[0413]

[0414]

[0415] Example 2: N-(2-(2-hydroxypropane-2-yl)pyrimidin-4-yl)-4-(N-(3-methyloxetane-3-yl)amino sulfonyl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide

[0416]

[0417] To a solution of 4-(N-(3-methyloxetan-3-yl)sulfamoyl)-2-(6-azaspiro[2.5]octan-6-yl)benzoic acid (150 mg, 0.394 mmol, Intermediate 15) and 2-(4-aminopyrimidin-2-yl)propan-2-ol (91 mg, 0.591 mmol, AstaTech, Bristol, PA, USA) in dichloromethane (2.6 mL) at 0 °C was added 1-propanephosphonic anhydride (50% in ethyl acetate, 0.469 mL, 0.789 mmol, Aldrich) followed by DIPEA (0.207 mL, 1.18 mmol 1, Aldrich (Aldrich)). The resulting mixture is then stirred at room temperature overnight. The mixture is then diluted with saturated NaHCO3 aqueous solution (2mL), followed by saturated NH4Cl (7mL). The mixture is then extracted with EtOAc (2x15mL). The combined organic extracts are then dried over MgSO4 and concentrated. The residue is purified by chromatography (silica gel, 0% to 100% EtOAc / heptane) to provide N-(2-(2-hydroxypropane-2-yl)pyrimidine-4-yl)-4-(N-(3-methyloxetane-3-yl)sulfamoyl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide (45mg, 0.087mmol) as a light yellow solid. 1 H NMR(DMSO-d6)δppm 13.52(br s,1H),8.76(d,J=5.6Hz,1H),8.54(br d,J=3.5Hz,1H),8.24(d,J=8.1Hz,1H),8.08(br d,J=5.4Hz,1H),7.86(d,J=1.0Hz,1H),7.74(dd,J=8.1,1.5Hz,1H),4.96(s,1H),4.55(d,J=6.0Hz,2H),4.13(d,J=6.4Hz,2H),3.08(br t,J=5.2Hz,4H),1.70(br s,4H),1.51(s,6H),1.41(s,3H),0.38(s,4H). m / z(ESI):516.2(M+H)+.

[0418] Table 7: Examples 2-1 to 2-8 were prepared following a similar procedure as described for Example 2:

[0419]

[0420]

[0421] Example 3: (R)-4-((2-hydroxyethyl)sulfonamido)-N-(6-methyl-2-(2-methylmorpholino)pyrimidine-4- 2-(6-azaspiro[2.5]octan-6-yl)benzamide

[0422]

[0423] Potassium phosphate (9.89 g, 46.6 mmol, Sigma-Aldrich), copper (I) iodide (0.710 g, 3.73 mmol, Sigma-Aldrich), 2-hydroxyethane-1-sulfonamide (1.166 g, 9.32 mmol, Wuxi Apptec, China), sarcosine (0.830 g, 9.32 mmol) and (R)-4-iodo-N-(6-methyl-2-(2-methylmorpholino)pyrimidin-4-yl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide (5.1 g, 9.32 mmol, Intermediate 21-2) were combined in a three-necked flask under argon. Dry, degassed DMF (20 mL) was added and the mixture was heated to 110 °C for 45 min with overhead stirring. The reaction mixture is cooled to ambient temperature, and saturated ammonium chloride (75mL), water (200mL) and ethyl acetate (200mL) are added. Each phase is mixed and separated, and the organic layer is dried with saline (75mL), then evaporated to dryness under reduced pressure. The crude solid is stirred for 10m in boiling ethanol (15mL), then cooled to ambient temperature, and filtered by sintered glass frit. The solid is dried on glass frit, then suspended in water (75mL), and heated to 80 DEG C. After 10min, the mixture is cooled to ambient temperature, and filtered by sintered glass frit. The solid is dried under nitrogen stream, to obtain the title compound (3.3g, 6.06mmol, 65.0% yield) in off-white solid. 1 H NMR(400MHz,DMSO-d6)δppm 13.23(bs,1H),10.26(bs,1H),8.05(m,1H),7.37(s,1H),7.26(s,1H),7.1 3(s,1H),4.95(bs,1H),4.50-4.42(m,2H),3.84(m,1H),3.76-3.74(m,2H), 3.51-3.45(m,2H),3.00-2.82(m,6H),2.61-2.58(m,2H),2.31(s,3H),1.91 -1.65(m,4H),1.17(d,J=6.0Hz,3H),0.39(s,4H).m / z(ESI):545.2(M+H)+.

[0424] Example 4: N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((2-hydroxyethyl)sulfonamide 2-(6-azaspiro[2.5]octan-6-yl)benzamide

[0425]

[0426] A mixture of 2-hydroxyethane-1-sulfonamide (1.28 g, 10.3 mmol, Wuxi AppTec), copper (I) iodide (0.49 g, 2.56 mmol), tripotassium phosphate (5.44 g, 25.6 mmol) and sarcosine (0.48 g, 5.13 mmol) in a 100 mL round-bottom flask was placed under an argon atmosphere. Anhydrous DMF (20 mL) was added, and the mixture was warmed to 50 ° C for 5 min. N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-iodo-2-(6-azaspiro[2.5]octan-6-yl)benzamide (2.91 g, 5.13 mmol, Intermediate 19) was added as a solid, and the mixture was heated to 100 ° C and stirred for 2 h, then cooled to room temperature. EtOAc (20mL) and water (20mL) are added, the obtained two-phase mixture is separated, and the water layer is extracted with EtOAc (3x).Then the combined organic extracts are washed with water (2x), 9:1 NH4Cl / NH4OH (aqueous solution), brine, dried over anhydrous MgSO4, filtered and concentrated in a vacuum to obtain oil. The oil is purified by silica gel chromatography, eluted with a 0% to 50% EtOAc / heptane gradient, and then eluted with 50% EtOAc / heptane isocratic, to provide an off-white solid. The solid is suspended in methanol, filtered, and dried to obtain a white solid. The solid was then suspended in water, stirred for 24 h, filtered and dried in vacuo to afford N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((2-hydroxyethyl)sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzamide (1.55 g, 2.75 mmol, 54% yield) as a white solid. 1 HNMR(400MHz,DMSO-d6)δppm 13.37(s,1H)10.03-10.52(m,1H)8.06(d,J=8.71Hz,1H)7.41(s,1H)7.28 (d,J=1.87Hz,1H)7.15(dd,J=8.71,1.87Hz,1H)4.73-5.14(m,1H)3.92(br t,J=5.39Hz,4H)3.77(t,J=6.43Hz,2H)3.34-3.40(m,2H)2.98(br t,J=4.56Hz,4H)2.32(s,3H)1.93-2.07(m,4H)1.58-1.85(m,4H)0.40(s,4H). 19F NMR (376MHz, DMSO-d6) δppm-94.74 (s, 1F). m / z(ESI):565.2(M+H) + .

[0427] Examples 5-1 and 5-2: (R)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((2-hydroxy (S)-N-(2-(4,4-difluoro-1-yl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide Piperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((2-hydroxy-1-methylethyl)sulfonamido)-2-(6-azaspiro[2.5] 6-octyl)benzamide

[0428]

[0429] Step 1: A mixture of ethyl 2-sulfamoylpropionate (1.44 g, 7.93 mmol, Intermediate 22), copper (I) iodide (0.503 g, 2.64 mmol, Strem), sarcosine (0.47 g, 5.29 mmol, Sigma-Aldrich Corporation) and potassium phosphate (4.49 g, 21.2 mmol) in DMF (15 mL) was placed under an argon atmosphere and warmed to 50°C for 5 min. N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-iodo-2-(6-azaspiro[2.5]octan-6-yl)benzamide (3.0 g, 5.29 mmol, Intermediate 19) was added and the mixture was heated to 100°C for 3 h and then cooled to room temperature. EtOAc (50mL), IPA (5mL) and water (50mL) are added, and the mixture is stirred vigorously for 5min. The gained two-phase mixture is transferred to a separating funnel and each layer is separated. The aqueous layer is extracted with EtOAc (2x20mL), then the combined extracts are washed with water (2x50mL), 9:1NH4Cl / NH4OH (1x50mL), dried over anhydrous MgSO4, filtered and concentrated in a vacuum to obtain oil. The crude oil was purified by silica gel chromatography using a Redi-Sep prepacked silica gel column (80 g) eluting with a 0% to 50% EtOAc / heptane gradient to afford ethyl 2-(N-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)carbamoyl)-3-(6-azaspiro[2.5]oct-6-yl)phenyl)sulfamoyl)propanoate (2.76 g, 4.45 mmol, 84% yield) as a white solid. 1HNMR(400MHz,DMSO-d6)δppm 13.35(s,1H)10.69(br s,1H)8.07(d,J=8.71Hz,1H)7.40(s,1H)7.31(d,J=1.87Hz,1H)7.17(dd,J=8.60,1.97Hz,1H)4.06(qd,J=7.08,4.87Hz,2H)3.92(br t,J=5.49Hz,4H)2.98(br t,J=4.77Hz,4H)2.32(s,3H)1.85-2.06(m,5H)1.73(br s,4H)1.48(d,J=6.84Hz,3H)1.14(t,J=7.05Hz,3H)0.39(s,4H). 19 F NMR (376MHz, DMSO-d6) δppm-94.75 (s, 1F). m / z(ESI):621.2(M+H) + .

[0430] Step 2: To a 250 mL round bottom flask was added ethyl 2-(N-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)carbamoyl)-3-(6-azaspiro[2.5]octan-6-yl)phenyl)sulfamoyl)propanoate (10.39 g, 16.74 mmol) and lithium borohydride solution (2.0 M in THF, 16.7 mL, 33.5 mmol, Sigma-Aldrich Corporation) in THF (100 mL). Methanol (4.29 mL, 134 mmol) was added slowly over 5 min and the resulting solution was stirred at room temperature for 30 min. 1 N HCl (20 mL) was added slowly followed by EtOAc (20 mL) and the resulting biphasic mixture was transferred to a separatory funnel and the phases were separated. The aqueous layer was extracted with EtOAc (1x25 mL), and the combined extracts were washed with saturated NaHCO (1x50 mL), brine (1x50 mL), dried over anhydrous MgSO, filtered and concentrated to give 8.9 g of a racemic mixture. The material was separated by preparative SFC using a Chiral Tech AD column (250X30 mm, 5 mm) with a flow rate of 150 mL / min of 85% liquid CO and a mobile phase of 15% MeOH with 0.2% TEA to give:

[0431] Example 5-1: (R)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((2-hydroxy-1-methyl

[0063] 1-(6-azaspiro[2.5]octan-6-yl)benzamide. First eluting peak (3.50 g, 6.05 mmol, 36.1% yield, >99% ee). 1HNMR(400MHz,DMSO-d6)δppm 13.36(s,1H)8.05(d,J=8.50Hz,1H)7.40(s,1H)7.31(d,J=1.87Hz,1H)7.17(dd,J=8.71,2.07Hz,1H)3.88-3.97(m,4H)3.84(dd ,J=10.99,4.35Hz,1H)3.37-3.54(m,1H)3.25-3.30(m,1H)2.97(brt,J=4.77Hz,4H)2.32(s,3H)1.84-2.06(m,4H)1.57-1.84(br s, 4H) 1.30 (d, J = 6.84 Hz, 3H) 0.39 (s, 4H). Two exchangeable protons were not observed. 19 F NMR (376MHz, DMSO-d6) δppm-94.74 (s, 1F). m / z(ESI):579.2(M+H) + .

[0432] Example 5-2: (S)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((2-hydroxy-1-methyl

[0063] 1-(6-azaspiro[2.5]octan-6-yl)benzamide. Second eluting peak (2.66 g, 4.60 mmol, 27.5% yield, 98.9% ee). 1 HNMR(400MHz,DMSO-d6)δppm 13.35(s,1H)8.05(d,J=8.50Hz,1H)7.40(s,1H)7.31(d,J=2.07Hz,1H)7.17(dd,J=8.60,1.97Hz,1H)3.88-3.97(m,4H)3.84(dd,J= 10.99,4.35Hz,1H)3.50(dd,J=10.99,7.46Hz,1H)3.25-3.32(m,1H)2.97(brt,J=4.77Hz,4H)2.31(s,3H)1.83-2.06(m,4H)1.73(br s, 4H) 1.30 (d, J = 6.84 Hz, 3H) 0.39 (s, 4H). Two exchangeable protons were not observed. 19 F NMR (376MHz, DMSO-d6) δppm-94.75 (s, 1F). m / z(ESI):579.2(M+H) + The stereochemistry is arbitrarily determined.

[0433] Table 8: Examples 6-1 to 6-40 were prepared according to similar procedures as Examples 3 to 5-2:

[0434]

[0435]

[0436]

[0437]

[0438]

[0439]

[0440]

[0441]

[0442] Example 7: N-(2-(3,3-difluoroazetidin-1-yl)-6-methylpyrimidin-4-yl)-4-((2-hydroxyethyl) Sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzamide

[0443]

[0444] Step 1: A solution of N-(2-chloro-6-methylpyrimidin-4-yl)-4-nitro-2-(6-azaspiro[2.5]octan-6-yl)benzamide (0.3 mg, 0.747 mmol, intermediate 21-18), 3,3-difluoroazetidine hydrochloride (0.145 g, 1.120 mmol, Combi-Blocks) and DIPEA (0.261 mL, 1.49 mmol) in DMF (0.5 mL) and ethanol (1 mL) was heated at 80 ° C for 4 h. The reaction mixture was then quenched with water (50 mL) and extracted with ethyl acetate (3x50 mL). The combined organic layer was washed with brine (50 mL), dried over Na2SO4, filtered and concentrated. The concentrate was purified by flash column chromatography eluting with 30% to 50% ethyl acetate in petroleum ether to afford N-(2-(3,3-difluoroazetidin-1-yl)-6-methylpyrimidin-4-yl)-4-nitro-2-(6-azaspiro[2.5]octan-6-yl)benzamide (180 g, 0.393 mmol, 52.6% yield) as a yellow solid. 1 HNMR(400MHz,chloroform-d)δppm 8.47(d,J=8.7Hz,1H),8.25(d,J=2.2Hz,1H),8.17(dd,J=8.7,2.2Hz,1H),7.68(s,1 H), 3.79–3.66 (m, 4H), 3.17 (t, J = 5.4Hz, 4H), 2.60 (s, 3H), 1.28 (s, 4H), 0.50 (s, 4H). m / z(ESI):459.2(M+H) + .

[0445] Step 2: To a solution of N-(2-(3,3-difluoroazetidin-1-yl)-6-methylpyrimidin-4-yl)-4-nitro-2-(6-azaspiro[2.5]octan-6-yl)benzamide (0.18 g, 0.39 mmol) in ethanol (8 mL) and water (8 mL) were added iron powder (0.066 g, 1.18 mmol) and ammonium chloride (0.063 g, 1.18 mmol). The mixture was then heated at 90 °C for 3 h and then passed through The bed was filtered and washed with ethyl acetate (3×100 mL). The filtrate was washed with brine, dried over Na 2 SO 4 , filtered and concentrated to provide 4-amino-N-(2-(3,3-difluoroazetidine-1-yl)-6-methylpyrimidin-4-yl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide (0.12 g, 0.280 mmol, 71.3% yield) as a light yellow solid. It was used directly in the next step without further purification. m / z(ESI):429.2(M+H) + .

[0446] Step 3: To a solution of 4-amino-N-(2-(3,3-difluoroazetidin-1-yl)-6-methylpyrimidin-4-yl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide (0.120 g, 0.280 mmol) in DCM (5 mL) was added Et3N (0.078 mL, 0.560 mmol) and methyl 2-(chlorosulfonyl)acetate (0.058 g, 0.336 mmol, Combi-Blocks) at 0°C. The mixture was stirred at room temperature for 4 h, then it was quenched with water (50 mL) and extracted with DCM (3×50 mL). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, filtered and concentrated to give methyl 2-(N-(4-((2-(3,3-difluoroazetidin-1-yl)-6-methylpyrimidin-4-yl)carbamoyl)-3-(6-azaspiro[2.5]octan-6-yl)phenyl)sulfamoyl)acetate (140 g, 0.25 mmol, 89% yield) as a light yellow solid. It was used in the next step without further purification. m / z(ESI):565.2(M+H) + .

[0447] Step 4: A solution of methyl 2-(N-(4-((2-(3,3-difluoroazetidine-1-yl)-6-methylpyrimidin-4-yl)carbamoyl)-3-(6-azaspiro[2.5]octan-6-yl)phenyl)sulfamoyl)acetate (130 mg, 0.230 mmol) in THF (5 mL) was treated with LiBH4 (230 μl, 0.460 mmol) at -30 °C. The reaction mixture was stirred at 0 °C for 30 min, then quenched with saturated NH4Cl aqueous solution (50 mL) at 0 °C and extracted with EtOAc (2x10 mL). The combined organic layers were washed with brine (5 mL), dried over Na2SO4, filtered and concentrated. The concentrate was purified by flash column chromatography eluting with a gradient of 20% to 100% EtOAc in hexanes to afford N-(2-(3,3-difluoroazetidin-1-yl)-6-methylpyrimidin-4-yl)-4-((2-hydroxyethyl)sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzamide (60 mg, 0.112 mmol, 48.6% yield) as a white solid. 1 HNMR(400MHz,DMSO-d6)δppm 13.55(s,1H),10.28(s,1H),8.05(d,J=8.7Hz,1H),7.51(s,1H),7.28(d,J=2.2Hz,1H),7.14(dd,J=8.6,2.2Hz,1H),4.95(s ,1H),4.45(d,J=12.3Hz,4H),3.76(t,J=6.4Hz,2H),3.64(s,1H),3.57(s,1H),2.97(t,J=5.4Hz,4H),2.34(s,3H),1.74(br s,4H),0.40(s,4H). m / z(ESI):537.2(M+H) + .

[0448] Examples 8-1 and 8-2: (R)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-5-fluoro-4- ((2-Hydroxy-1-methylethyl)sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzamide and (S)-N-(2-(4, 4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-5-fluoro-4-((2-hydroxy-1-methylethyl)sulfonamido)-2-(6- Azaspiro[2.5]octan-6-yl)benzamide

[0449]

[0450] Step 1: To a solution of 4-amino-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-5-fluoro-2-(6-azaspiro[2.5]octan-6-yl)benzamide (0.41 g, 0.86 mmol, intermediate 21) and triethylamine (0.18 mL, 1.30 mmol) in DCM (3 mL) at 0 °C was added methyl 2-(chlorosulfonyl)propanoate (177 mg, 0.95 mmol, Enamine). The mixture was stirred for 1 h and then concentrated in vacuo and purified by silica gel chromatography using a Redi-Sep prepacked silica gel column (12 g) eluting with a 25% EtOAc / heptane gradient to afford methyl 2-(N-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)carbamoyl)-2-fluoro-5-(6-azaspiro[2.5]octan-6-yl)phenyl)sulfamoyl)propanoate (0.54 g, 0.48 mmol, 54.8% yield) as a white solid. 1 HNMR(400MHz,DMSO-d6)δppm13.71(s,1H)10.66(br s,1H)7.88(d,J=11.61Hz,1H)7.60(d,J=7.26Hz,1H)7.39(s,1H)4.38(d,J=7.05Hz,1H)3.88-3.96(m,4H)3.57(s,3H)2.98(br t,J=4.56Hz,4H)2.33(s,3H)1.92-2.07(m,4H)1.57-1.91(m,4H)1.52(d,J=6.84Hz,3H)0.40(s,4H). 19 F NMR (376MHz, DMSO-d6) δppm-94.77(s,1F)-126.39(s,1F). m / z(ESI):625.2(M+H) + .

[0451] Step 2: To a solution of methyl 2-(N-(4-((2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)carbamoyl)-2-fluoro-5-(6-azaspiro[2.5]octan-6-yl)phenyl)sulfamoyl)propanoate (192 mg, 0.31 mmol) and lithium borohydride (2.0 M in THF, 0.35 mL, 0.69 mmol) in THF (2.5 mL) was added methanol (0.10 mL, 2.52 mmol) dropwise. The mixture was stirred for 30 min, then aqueous NH4Cl was added. The product was extracted into EtOAc (2x), and the combined extracts were dried over anhydrous MgSO4, filtered and concentrated in vacuo to give the racemic product as a solid. The material was separated by preparative SFC using an OD column (250×21 mm, 5 mm) and an OD column (150×21 mm, 5 mm) with a mobile phase of 90% liquid CO and 10% EtOH / 0.2% triethylamine at a flow rate of 80 mL / min to give:

[0452] Example 8-1: (R)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-5-fluoro-4-((2-hydroxy

[0063] The invention relates to 2-(6-azaspiro[2.5]octan-6-yl)benzamide. First eluting peak (94 mg, 0.16 mmol, 33.2% yield, >99% ee). 1 H NMR(400MHz,DMSO-d6)δppm 13.74(s,1H)7.82(d,J=11.82Hz,1H)7.58(d,J=7.26Hz,1H)7.39(s,1H)3.77-3.96(m,6H)3 .37-3.54(m,1H)3.22-3.30(m,1H)2.88-3.05(m,4H)2.32(s,3H)1.92-2.05(m,4H)1.73(br s,4H)1.31(d,J=6.84Hz,3H)0.40(s,4H). 19 F NMR (376MHz, DMSO-d6) δppm-94.77(s,1F)-127.36(s,1F). m / z(ESI):597.2(M+H) + .

[0453] Example 8-2: (S)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-5-fluoro-4-((2-hydroxy

[0063] The invention relates to 2-(6-azaspiro[2.5]octan-6-yl)benzamide. Second eluting peak (102 mg, 0.17 mmol, 36.1% yield, >98.4% ee). 1HNMR(400MHz,DMSO-d6)δppm 13.74(s,1H)7.82(d,J=11.82Hz,1H)7.58(d,J=7.26Hz,1H)7.39(s,1H)3.77-3.96(m,6H)3 .37-3.54(m,1H)3.22-3.30(m,1H)2.88-3.05(m,4H)2.32(s,3H)1.92-2.05(m,4H)1.73(br s,4H)1.31(d,J=6.84Hz,3H)0.40(s,4H). 19 F NMR (376MHz, DMSO-d6) δppm-94.76(s,1F)-127.73(s,1F). m / z(ESI):597.2(M+H) + The stereochemistry is arbitrarily determined.

[0454] Table 9: Examples 9-1 to 9-2 were prepared according to similar procedures as Examples 8-1 and 8-2:

[0455]

[0456] Examples 10-1 and 10-2: (R)-N-(2-(3,3-difluoroazetidin-1-yl)-6-methylpyrimidin-4-yl)-4- ((2-Hydroxypropyl)sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzamide and (S)-N-(2-(3,3-difluoronitrogen) Heterocyclobutane-1-yl)-6-methylpyrimidin-4-yl)-4-((2-hydroxypropyl)sulfonamido)-2-(6-azaspiro[2.5]octane- 6-amino)benzamide

[0457]

[0458] Step 1: A solution of N-(2-chloro-6-methylpyrimidin-4-yl)-4-iodo-2-(6-azaspiro[2.5]octan-6-yl)benzamide (2.0 mg, 4.14 mmol, intermediate 17), 3,3-difluoroazetidine hydrochloride (1.07 g, 8.29 mmol, Combi-Blocks Inc.) and potassium carbonate (1.72 g, 12.4 mmol, Combi-Blocks Inc.) in NMP (10 mL) was heated to 90 ° C for 24 h. The mixture was cooled to room temperature, EtOAc (10 mL) was added, and the mixture was then washed with water (1x10 mL), 1N HCl (1x10 mL) and brine (1x10 mL). The mixture was then dried over anhydrous MgSO4, filtered and concentrated in vacuo to give a solid. The solid was then suspended in MeOH, filtered and dried in vacuo to afford N-(2-(3,3-difluoroazetidin-1-yl)-6-methylpyrimidin-4-yl)-4-iodo-2-(6-azaspiro[2.5]octan-6-yl)benzamide (1.16 g, 2.15 mmol, 51.9% yield) as a light brown solid. 1HNMR(400MHz,DMSO-d6)δppm 13.53(br s,1H)7.79-7.86(m,2H)7.73-7.77(m,1H)7.49(s,1H)4.43(t,J=12.44Hz,4H)3.02(br t,J=5.08Hz,4H)2.31-2.38(m,3H)1.65-1.82(m,4H)0.39(s,4H). 19 F NMR (376MHz, DMSO-d6) δppm-99.09 (s, 1F). m / z(ESI):540.0(M+H) + .

[0459] Step 2: 2-Hydroxypropane-1-sulfonamide (206 mg, 1.48 mmol, intermediate 23), copper (I) iodide (71 mg, 0.37 mmol), sarcosine (66 mg, 0.74 mmol) and potassium phosphate (787 mg, 3.71 mmol) were placed under argon atmosphere, dissolved in anhydrous DMF (3 mL) and warmed to 50 ° C for 5 min. N-(2-(3,3-difluoroazetidin-1-yl)-6-methylpyrimidin-4-yl)-4-iodo-2-(6-azaspiro[2.5]octan-6-yl)benzamide (0.40 g, 0.74 mmol) was added in one portion and the mixture was heated to 100 ° C for 2.5 h and then cooled to room temperature. Water was added and the product was extracted into EtOAc (2x). The combined extracts were then washed with water (2x), 9:1 saturated NH4Cl / NH4OH (1x), dried over anhydrous MgSO4, filtered and concentrated in vacuo to give the racemic product as an oil. The material was separated by preparative SFC using an IF column (250×301 mm, 5 mm) with a mobile phase of 75% liquid CO2 and 25% MeOH at a flow rate of 130 mL / min to give:

[0460] Example 10-1: (R)-N-(2-(3,3-difluoroazetidine-1-yl)-6-methylpyrimidin-4-yl)-4-((2-hydroxy

[0063] The invention relates to 2-(6-azaspiro[2.5]octan-6-yl)benzamide. First eluting peak (88 mg, 0.16 mmol, 21.6% yield, >99% ee). 1HNMR (400MHz, DMSO-d6) δppm 13.54 (s, 1H) 8.05 (d, J = 8.71 Hz, 1H) 7.51 (s, 1H) 7.27 (d, J = 1.87 Hz, 1H) 7.14 (dd, J = 8.71, 1.87 Hz, 1H) 4.44 (t, J = 12.44 Hz, 4H) 4.07-4.15 (m, 1H) 3.22-3.29 (m, 2H) 2.97 (br t, J = 4.87 Hz, 4H) 2.34 (s, 3H) 1.74 (br s, 4H) 1.19 (d, J = 6.22 Hz, 3H) 0.40 (s, 4H). Two exchangeable protons were not observed. 19 F NMR (376MHz, DMSO-d6) δppm-99.08 (s, 1F). m / z(ESI):551(M+H) + .

[0461] Example 10-2: (S)-N-(2-(3,3-difluoroazetidine-1-yl)-6-methylpyrimidin-4-yl)-4-((2-hydroxy

[0063] The invention relates to 2-(6-azaspiro[2.5]octan-6-yl)benzamide. Second eluting peak (89 mg, 0.162 mmol, 21.8% yield, >99% ee). 1 HNMR (400MHz, DMSO-d6) δppm 13.55 (s, 1H) 8.05 (d, J = 8.71 Hz, 1H) 7.51 (s, 1H) 7.26 (d, J = 1.87 Hz, 1H) 7.14 (dd, J = 8.71, 1.87 Hz, 1H) 4.43 (t, J = 12.44 Hz, 4H) 4.11 (d, J = 6.01 Hz, 1H) 3.21-3.31 (m, 2H) 2.97 (br t, J = 4.87 Hz, 4H) 2.34 (s, 3H) 1.53-2.01 (m, 4H) 1.19 (d, J = 6.43 Hz, 3H) 0.40 (s, 4H). Two exchangeable protons were not observed. 19 FNMR (376MHz, DMSO-d6) δppm-99.09 (s, 1F). m / z(ESI):551(M+H) + The stereochemistry is arbitrarily determined.

[0462] Table 10: Examples 11-1 to 11-83 were prepared according to a similar procedure to Example 10:

[0463]

[0464]

[0465]

[0466]

[0467]

[0468]

[0469]

[0470]

[0471]

[0472]

[0473]

[0474]

[0475]

[0476] Example 12: N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((2-hydroxyethyl)sulfonyl 2-(6-azaspiro[2.5]octan-6-yl)benzamide

[0477]

[0478] Step 1: In dimethyl sulfoxide (3 mL), N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-iodo-2-(6-azaspiro[2.5]octan-6-yl)benzamide (500 mg, 0.881 mmol, intermediate 19), triphenylphosphine (34.7 mg, 0.132 mmol, Aldrich, St. Louis, MO USA), 1,10-phenanthroline (23.82 mg, 0.132 mmol, Aldrich, St. Louis, MO USA), palladium(II) acetate (9.89 mg, 0.044 mmol, Strem Chemicals Inc., Newburyport, MA USA) were added. USA)), sodium formate (132mg, 1.939mmol, Thermo Fisher Scientific, Grand Island, NY USA) and tetrabutylammonium bromide (426mg, 1.322mmol, Aldrich, St. Louis, MO USA)) were stirred at 70 ° C for 45min under N2. Then, the mixture was cooled to room temperature and ethyl iodoacetate (0.157mL, 1.322mmol, Aldrich, St. Louis, MO USA) was added. The mixture was then stirred at room temperature for 10min. The reaction mixture was then diluted with water (20mL) and then extracted with EtOAc (2x40mL). The combined organic extracts were then dried over MgSO4 and concentrated. The residue was purified by chromatography (silica gel, 0% to 100% EtOAc / heptane) to provide ethyl 2-((4-((2-(4,4-difluoropiperidin-l-yl)-6-methylpyrimidin-4-yl)carbamoyl)-3-(6-azaspiro[2.5]oct-6-yl)phenyl)sulfonyl)acetate (330 mg, 0.558 mmol) as a light yellow solid. 1H NMR(DMSO-d6)δ13.14(br s,1H),8.27(br d,J=8.1Hz,1H),7.97(s,1H),7.84(br d,J=7.7Hz,1H),7.40(s,1H),4.80(s,2H),4.05(q,J=7.2Hz,2H),3.92(br s,4H),3.03-3.13(m,4H),2.34(s,3H),1.90-2.07(m,4H),1.71(br s, 4H), 1.07 (t, J = 7.0Hz, 3H), 0.39 (s, 4H). m / z(ESI):592.3(M+H)+.

[0479] Step 2: To a solution of ethyl 2-((4-((2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)carbamoyl)-3-(6-azaspiro[2.5]octan-6-yl)phenyl)sulfonyl)acetate (320 mg, 0.541 mmol) in 2-methyltetrahydrofuran (3.5 mL) was added dropwise a solution of lithium borohydride (2.0 M in tetrahydrofuran, 0.541 mL, 1.082 mmol, Aldrich, St. Louis, MO USA) at 0 °C under N2. After addition, the mixture was stirred at room temperature overnight. Then, the mixture was quenched with saturated NH4Cl (18 mL) and stirred at room temperature for 15 min. The mixture was then extracted with EtOAc (2x30 mL). The combined organic extracts were then dried over MgSO4 and concentrated. The residue was purified by chromatography (silica gel, 0% to 100% EtOAc / heptane) to provide N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((2-hydroxyethyl)sulfonyl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide (60 mg, 0.109 mmol, 20% yield) as a white solid. 1H NMR(DMSO-d6)δppm 13.17(br s,1H),8.25(br d,J=8.3Hz,1H),7.93(br s,1H),7.82(br d,J=8.3Hz,1H),7.40(br s,1H),4.91(br t,J=5.0Hz,1H),3.92(br s,4H),3.68-3.77(m,2H),3.52-3.60(m,2H),3.09(br s, 4H), 2.34 (s, 3H), 1.87-2.08 (m, 4H), 1.70 (brd, J = 1.0Hz, 4H), 0.39 (s, 4H). 19 F NMR(DMSO-d6)δppm-94.76(s,2F). m / z(ESI):550.1(M+H)+.

[0480] Examples 13-1 and 13-2: (S)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((1-hydroxy (R)-N-(2-(4,4-difluoropiperidin-1-yl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide 6-((1-hydroxypropane-2-yl)sulfonyl)-2-(6-azaspiro[2.5]octan-6-yl)benzoyl amine

[0481]

[0482] Step 1: A solution of N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-iodo-2-(6-azaspiro[2.5]octan-6-yl)benzamide (1.08 g, 1.90 mmol, Intermediate 19), 2-mercaptopropan-1-ol (0.48 g, 5.21 mmol, Enamine) and potassium carbonate (0.237 mL, 3.91 mmol) in 4 mL DMSO was heated at 90 °C for 4 h in a sealed bottle. The mixture was cooled to room temperature, 50 mL ethyl acetate and 10 mL brine were added. The organic layer was separated, washed with brine, dried and evaporated. The resulting product was adsorbed onto a plug of silica gel and purified by silica gel chromatography (0% to 30% EtOAc in heptane) to afford N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((1-hydroxypropan-2-yl)thio)-2-(6-azaspiro[2.5]octan-6-yl)benzamide as a yellow solid. 1HNMR(400MHz, chloroform-d)δppm0.39-0.44(m,4H)1.37-1.43(m,3H)1.55-1.60(m,4H)1.96-2.04(m,4H)2.35-2.41(m,3H)3.01-3 .11(m,4H)3.46-3.78(m,3H)3.96-4.05(m,4H)7.28-7.36(m,2H)7.48-7.53(m,1H)8.12-8.32(m,1H)13.01-13.37(m,1H). m / z(ESI):531.4(M+H) + .

[0483] Step 2: To N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((1-hydroxypropan-2-yl)thio)-2-(6-azaspiro[2.5]octan-6-yl)benzamide (0.62 g, 1.17 mmol) in 15 mL of THF was added oxone(r) monopersulfate compound (0.72 g, 1.17 mmol) in 5 mL of water. After stirring for 1.5 h, LCMS showed that a mixture of sulfone and sulfoxide was formed. An additional 0.4 g of oxone in 3 mL of water was added. After stirring for another 2 h, EtOAc (50 mL) and brine (20 mL) were added to the reaction mixture and the organic layer was collected, washed with brine, dried and evaporated. The crude product was purified by preparative SFC using a (S,S) Whelk-01 (250×21 mm, 5 mm) column with a mobile phase of 60% liquid CO and 40% MeOH at a flow rate of 80 mL / min to give N-2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((1-hydroxypropane-2-yl)sulfonyl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide (337 mg, 0.58 mmol, 50% yield). 1 H NMR (400MHz, chloroform-d) δppm 0.30-0.57(m,4H)1.27-1.37(m,3H)1.59-1.72(m,4H)1.94-2.10(m,4H)2.40-2.54(m,3H)2.55-2.97(m,1H)3 .05-3.23(m,4H)3.28-3.41(m,1H)3.84-4.06(m,6H)7.63-7.86(m,2H)8.13-8.37(m,1H)11.08-11.59(m,1H). m / z(ESI):598.3(M+H) +The racemic mixture was separated by preparative SFC using an OD (250×21 mm, 5 mm) with a mobile phase of 85% liquid CO 2 and 15% iPrOH at a flow rate of 90 mL / min to yield:

[0484] Example 13-1: (S)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((1-hydroxypropyl)- alkyl-2-yl)sulfonyl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide. First eluting peak (85 mg, >99% ee). 1 HNMR(400MHz,chloroform-d)δppm 12.74-13.01(m,1H),8.36-8.54(m,1H),7.69-8.03(m,2H),7.41-7.58(m,1H),3.83-4.06(m,6H),3.25-3.43(m,1H),3.03-3. 17(m,4H),2.50-2.81(m,1H),2.31-2.42(m,3H),1.93-2.10(m,4H),1.61-1.90(m,4H),1.28-1.36(m,3H),0.35-0.50(m,4H). m / z(ESI):563.2(M+H) + .

[0485] Example 13-2: (R)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((1-hydroxypropyl)- alkyl-2-yl)sulfonyl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide. Second eluting peak (84 mg, 97% ee). 1 HNMR(400MHz,chloroform-d)δppm 12.71-13.05(m,1H),8.39-8.56(m,1H),7.74-8.03(m,2H),7.38-7.54(m,1H),3.81-4.04(m,6H),3.26-3.39(m,1H),3.03-3. 19(m,4H),2.48-2.83(m,1H),2.33-2.41(m,3H),1.92-2.10(m,4H),1.60-1.90(m,4H),1.28-1.33(m,3H),0.36-0.46(m,4H). m / z(ESI):563.2(M+H) + The stereochemistry is arbitrarily determined.

[0486] Example 14: N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((1-hydroxy-2-methylpropane alkyl-2-yl)sulfonyl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide

[0487]

[0488] Step 1: N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-iodo-2-(6-azaspiro[2.5]octan-6-yl)benzamide (0.55 g, 0.967 mmol, Intermediate 19), 4,5-bis(diphenylphosphino)-9,9-dimethyl-xanthene (0.034 g, 0.058 mmol), tris(dibenzylideneacetone)dipalladium(0)chloroform adduct (0.035 g, 0.034 mmol), DIPEA (0.4 mL, 2.29 mmol) and 2-mercapto-2-methylpropan-1-ol (0.134 g, 1.29 mmol) in 3 mL of dioxane were bubbled with N in a sealed tube for 25 min. The mixture was heated at 90° C. for 3 h and cooled to room temperature. The crude product was adsorbed onto a plug of silica gel and purified by silica gel chromatography (0% to 7% EtOAc in DCM) to afford N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((1-hydroxy-2-methylpropan-2-yl)thio)-2-(6-azaspiro[2.5]octan-6-yl)benzamide. 1 HNMR (400MHz, chloroform-d) δppm 0.38-0.47 (m, 4H) 1.25-1.31 (m, 6H) 1.56-1.56 (m, 4H) 1.94-2.04 (m, 4H) 2.35-2.41 (m, 3H) 3.03-3.12 (m, 4H) 3.31-3.38 (m, 2H) 3.95-4.04 (m, 4H) 7.41-7.47 (m, 2H) 7.48-7.52 (m, 1H) 8.14-8.32 (m, 1H) 12.96-13.41 (m, 1H). 4H overlaps with water peak. m / z (ESI): 546.2 (M+H) + .

[0489] Step 2: To N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((1-hydroxy-2-methylpropan-2-yl)thio)-2-(6-azaspiro[2.5]octan-6-yl)benzamide (0.36 g, 0.66 mmol) in THF (15 mL) cooled to 0°C was added oxone(r) monopersulfate compound (0.52 g, 0.85 mmol) in water (5 mL). The mixture was stirred from 0°C to room temperature for 2.5 h. An additional 0.35 g of oxone was added. After 1 h, Icms showed that the sulfoxide was almost consumed. Ethyl acetate (40 mL) and brine (20 mL) were added, the organic layer was separated, dried and evaporated. The crude mixture was purified by preparative SFC using (S,S) Whelk-01 (250x21 mm, 5 mm) with a mobile phase of 60% liquid CO2 and 40% MeOH at a flow rate of 80 mL / min to give N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((1-hydroxy-2-methylpropan-2-yl)sulfonyl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide. 1 H NMR (400MHz, chloroform-d) δppm 8.15-8.73(m,1H),7.64-7.90(m,2H),7.41-7.52(m,1H),3.94-4.07(m,4H),3.70-3.83(m,2H),3.04-3.2 0(m,4H),2.34-2.47(m,3H),1.96-2.12(m,4H),1.47-1.95(m,4H),1.30-1.41(m,6H),0.37-0.52(m,4H). 19 F NMR (376MHz, chloroform-d) δppm-96.68 (br s, 1F). m / z (ESI): 578.2 (M+H) + .

[0490] Table 11: Examples 14-1 to 14-9 were prepared according to similar procedures as Examples 12 to 14:

[0491]

[0492]

[0493] Example 15: N-(2-(4,4-difluorocyclohexyl)-6-methylpyrimidin-4-yl)-4-((2-hydroxyethyl)sulfonamide 2-(6-azaspiro[2.5]octan-6-yl)benzamide

[0494]

[0495] A mixture of 4-bromo-N-(2-(4,4-difluorocyclohexyl)-6-methylpyrimidin-4-yl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide (0.055 g, 0.106 mmol, Intermediate 21-13), 2-hydroxyethane-1-sulfonamide (0.020 g, 0.159 mmol, Wuxi), tripotassium phosphate (0.045 g, 0.212 mmol), copper (I) iodide (0.020 g, 0.106 mmol) and (1R,2R)-N,N'-dimethyl-1,2-cyclohexanediamine (7.53 mg, 0.053 mmol, Combi-Blocks) in DMF (1.5 mL) was heated at 90 °C for 16 h. The reaction mixture was then added by The mixture was filtered through a pad of a pad, and the filtrate was diluted with EtOAc. The resulting solution was washed with water and brine, dried over Na2SO4, filtered and concentrated. The residue was purified by reverse phase HPLC using a 60% ACN gradient in water (0.1% TFA) to obtain N-(2-(4,4-difluorocyclohexyl)-6-methylpyrimidine-4-yl)-4-((2-hydroxyethyl)sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzamide (0.025 g, 0.044 mmol, 42% yield) as a white solid. 1 HNMR(400MHz,DMSO-d6)δppm13.70(s,1H),8.04(d,J=8.8Hz,1H),7.92(s,1H),7.25(d,J=2.2Hz,1H),7.16–7.08(m,1H),3.7 5(t,J=6.3Hz,2H),3.03–2.85(m,6H),2.44(d,J=4.5Hz,5H),2.05(s,5H),1.92(d,J=11.7Hz,4H),1.72(s,4H),0.38(s,4H). m / z(ESI):564.1(M+H)+.

[0496] Examples 16-1 and 16-2: (S)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((2- Fluoro-1-(hydroxymethyl)ethyl)sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzamide and (R)-N-(2-(4, 4-((2-fluoro-1-(hydroxymethyl)ethyl)sulfonamido)-2-(6-nitrogen Heterospiro[2.5]octan-6-yl)benzamide

[0497]

[0498]

[0499] Step 1: To a solution of 1-(benzyloxy)-3-((tert-butyldimethylsilyl)oxy)propane-2-sulfonamide (0.803 g, 2.23 mmol) in THF was added tetrabutylammonium fluoride solution (2.75 mL, 2.75 mmol, 1 M in THF) at room temperature. The reaction was stirred for 1 h and then concentrated under reduced pressure. The resulting material was used immediately in the next step.

[0500] Step 2: To a pressure release bottle containing the sulfonamide from the previous step was added copper (I) iodide (0.196 g, 1.03 mmol), methylglycine (0.128 g, 1.440 mmol), tripotassium phosphate (0.934 g, 4.40 mmol) and N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-iodo-2-(6-azaspiro[2.5]octan-6-yl)benzamide (0.975 g, 1.72 mmol, Intermediate 19). The bottle was sealed and evacuated / backfilled with nitrogen, then DMF (7 mL) was added. The lid was closed and the reaction was stirred in a preheated 100 °C oil bath for 16 h. The reaction mixture was partitioned between saturated NH4Cl:NH4OH (9:1) and EtOAc. The organic phase was separated, washed with brine and concentrated in vacuo. The material was purified by silica gel chromatography (20% to 100% EtOAc in heptane) to give 4-((2-(benzyloxy)-1-(hydroxymethyl)ethyl)sulfonamido)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-2-(6-azaspiro[2.5]octyl-6-yl)benzamide (0.635 g, 0.927 mmol, 54.0% yield). 1 HNMR (400MHz, chloroform-d) δppm 13.11 (br dd, J=4.66, 2.38Hz, 1H) 8.14 (d, J=8.50Hz, 1H) 7.49 (br s,1H)7.31-7.44(m,5H)7.13(d,J=1.87Hz,1H)6.83(dd,J=8.60,2.18Hz,2H)4.49-4. 62(m,2H)4.08(dt,J=11.77,5.83Hz,1H)3.94-4.03(m,5H)3.88-3.93(m,1H)3.95(br s,1H)3.45-3.55(m,1H)2.96(brt,J=4.98Hz,4H)2.29-2.49(m,4H)1.95-2.07(m,4H)1.57(br s,4H)1.18-1.35(m,2H)0.85-0.91(m,1H)0.40(s,4H). m / z(ESI,+ve ion):683.8(M+H) + .

[0501] Step 3: To a solution of xtalfluor-m (0.351 g, 1.446 mmol) in DCM (6 mL) was added triethylamine trihydrofluoride (0.262 mL, 1.61 mmol) dropwise via an addition funnel in a brine / ice bath, followed by 4-((2-(benzyloxy)-1-(hydroxymethyl)ethyl)sulfonamido)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-2-(6-azaspiro[2.5]octyl-6-yl)benzamide (0.55 g, 0.803 mmol) in DCM (10 mL). The reaction was gradually warmed to room temperature and stirred for 16 h. The reaction was quenched with saturated sodium bicarbonate (aq.) and diluted with water and DCM. The organic phase was separated, washed with brine, dried over magnesium sulfate and concentrated in vacuo. The crude material was purified by silica gel chromatography (20% to 100% EtOAc in heptane) to give a 1:0.8 mixture of rac-4-((2-(benzyloxy)-1-(fluoromethyl)ethyl)sulfonamido)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-2-(6-azaspiro[2.5]oct-6-yl)benzamide and 4-((1-((benzyloxy)methyl)vinyl)sulfonamido)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-2-(6-azaspiro[2.5]oct-6-yl)benzamide (265 mg). This mixture was carried forward without further purification.

[0502] Step 4: To a suspension of 265 mg of the product mixture from the previous step and palladium hydroxide on carbon (0.135 g, 0.193 mmol) in EtOH (20 mL) was added AcOH (0.033 mL, 0.579 mmol). The reaction was hydrogenated at room temperature under 55 psi hydrogen for 24 h. After flushing the reaction with nitrogen, an additional portion of palladium hydroxide on carbon (0.135 mL, 0.193 mmol) was added, followed by additional AcOH (0.033 mL, 0.579 mmol). The reaction vessel was flushed with N2, and then the atmosphere was replaced with 55 psi hydrogen. The reaction was continued at room temperature for an additional 48 h. The reaction was flushed with nitrogen, and then heated to 40 °C. Filter and concentrate the filtrate in vacuo. The racemic mixture was separated by preparative SFC using IE (250x21 mm, 5 mm) with a mobile phase of 80% liquid CO2 and 20% MeOH (flow rate of 80 mL / min) to give:

[0503] Example 16-1: (S)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((2-fluoro-1-(hydroxy (6-azaspiro[2.5]octan-6-yl)benzamide. First eluting peak 1H NMR(400MHz,DMSO-d6)δppm 13.35(s,1H),10.21-10.84(m,1H),8.06(d,J=8.50Hz,1H)7.40(s,1H),7.29(d,J=2.07Hz,1H),7.16(dd,J=8.50,2.07Hz,1H),5.1 2-5.38(m,1H),4.88-4.97(m,1H)4.71-4.84(m,1H),3.85-4.01(m,5H),3.74(dd,J=11.30,7.98Hz,1H),3.51-3.63(m,1H)2.98(br t, J = 4.77 Hz, 4H), 2.32 (s, 3H), 1.93-2.07 (m, 4H), 1.51-1.91 (m, 4H), 0.40 (s, 4H). m / z (ESI, +ve ion): 597.2 (M+H) + .

[0504] Example 16-2: (R)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((2-fluoro-1-(hydroxy (6-azaspiro[2.5]octan-6-yl)benzamide. Second eluting peak. 1 H NMR(400MHz,DMSO-d6)δppm 13.35(s,1H)10.32-10.70(m,1H)8.05(d,J=8.71Hz,1H)7.40(s,1H)7.29(d,J=2.07Hz,1H)7.16(dd,J=8.71,2.07H z,1H)5.14-5.40(m,1H)4.85-4.97(m,1H)4.73-4.84(m,1H)3.87-3.98(m,5H),3.74(dd,J=11.09,7.98Hz,1H)3.48. m / z(ESI,+ve ion):597.2(M+H) + The stereochemistry is arbitrarily determined.

[0505] Example 17: N-(2-(4,4-difluoropiperidin-1-yl)pyridin-4-yl)-4-(N-(2-hydroxyethyl)sulfamoyl)- 2-(6-Azaspiro[2.5]oct-6-yl)benzamide

[0506]

[0507] In a glass tube, N-(2-(4,4-difluoropiperidin-1-yl)pyridin-4-yl)-4-iodo-2-(6-azaspiro[2.5]octan-6-yl)benzamide (212 mg, 0.384 mmol, Intermediate 21-14), 1,4-diazabicyclo[2.2.2]octane bis(sulfur dioxide) adduct (DABSO) (55 mg, 0.23 mmol, Sigma-Aldrich Corporation), diacetoxypalladium (13 mg, 0.06 mmol, Strem), ((3R,5R,7R)-adamantan-1-yl)((3S,5S,7S)adamantan-1-yl)(butyl)phosphine were added in IPA (3 mL). A mixture of 4-nitropropene (28 mg, 0.08 mmol, Strem) and triethylamine (107 uL, 0.77 mmol) was degassed for 3 min. The tube was sealed and then heated in an oil bath at 85 ° C for 3 h. The heterogeneous mixture was cooled to room temperature, treated with 2-aminoethane-1-ol (47 mg, 0.77 mmol, Sigma-Aldrich), followed by sodium hypochlorite solution (10 wt%, 571 mg, 0.77 mmol, Sigma-Aldrich), and stirred at room temperature for 18 h. The mixture was treated with 2-aminoethane-1-ol (23 mg), followed by sodium hypochlorite solution (10 wt%, 275 mg), and then stirred at room temperature for 5 h. EtOAc (20 mL) and water (5 mL) were added to the heterogeneous mixture, and the insoluble solids were filtered out. The filter cake is washed with water (2x2mL), then washed with EtOAc (2x4mL). Take organic solution and concentrate in vacuo. The residue is purified by silica gel chromatography (10% to 60% EtOAc in heptane) to obtain N- (2- (4,4- difluoropiperidin-1-yl) pyridin-4-yl) -4- (N- (2- hydroxyethyl) sulfamoyl) -2- (6- azaspiro [2.5] octane -6- yl) benzamide (100mg, 0.18mmol, 47% yield) as off-white solid. 1H NMR (400MHz, methanol-d4) δppm 7.99-8.14(m,2H),7.79(s,1H),7.68(d,J=7.88Hz,1H),7.47(s,1H),7.01(d,J=4.77Hz,1H),3.76(t,J=5.29Hz,4H), 3.58(t,J=5.91Hz,2H),3.16(t,J=5.08Hz,4H),3.02(t,J=5.80Hz,2H),1.98-2.11(m,4H),1.62(s,4H),0.42(s,4H). m / z(ESI):(M+H) + 550.1.

[0508] Table 12: Examples 17-1 to 17-8 were prepared according to the similar procedure of Example 17:

[0509]

[0510]

[0511] Examples 18-1 and 18-2: 2-(6-azaspiro[2.5]octan-6-yl)-4-(R-cyclopropylsulfonylimino)-N-(2- (4,4-difluoro-1-piperidinyl)-6-methyl-4-pyrimidinyl)benzamide and 2-(6-azaspiro[2.5]octan-6-yl)-4-(S- Cyclopropylsulfonylimino)-N-(2-(4,4-difluoro-1-piperidinyl)-6-methyl-4-pyrimidinyl)benzamide

[0512]

[0513] Step 1: N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-iodo-2-(6-azaspiro[2.5]octan-6-yl)benzamide (1.00 g, 1.762 mmol, Intermediate 19), tris(dibenzylideneacetone)dipalladium(0) (0.161 g, 0.176 mmol) and 4,5-bis(diphenylphosphino)-9,9-dimethyl-xanthene (0.102 g, 0.176 mmol) followed by 1,4-dioxane (10 mL) were placed in a 20 mL microwave vessel. The resulting mixture was stirred and purged with nitrogen for 5 min, then 1,1'-dimethyltriethylamine (0.616 mL, 3.52 mmol) was added under nitrogen followed by cyclopropanethiol (0.142 mL, 1.939 mmol). The container was sealed and subjected to microwave conditions (10 h, 90 ° C) again. The crude mixture was directly loaded onto a silica gel pre-column and combi-flash column chromatography was performed on a 40-g ISCO gold column, eluted twice with MeOH / DCM (5 min at 0%, 25 min from 0% to 6%) to obtain 4-(cyclopropylsulfur)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide (0.92 g, 1.791 mmol, 102% yield) as an off-white solid.1 HNMR (400MHz, dichloromethane-d2) δppm 13.33 (s, 1H), 8.15 (d, J = 8.29 Hz, 1H), 7.48 (s, 1H), 7.22-7.35 (m, 2H), 3.91-4.09 (m, 4H), 3.06 (br t, J = 5.18 Hz, 4H), 2.35 (s, 3H), 2.17-2.28 (m, 1H), 1.62-2.10 (m, 6H), 1.52 (s, 2H), 1.13-1.21 (m, 2H), 0.68-0.76 (m, 2H), 0.40 (s, 4H). m / z (ESI): 514.1 (M+H) + .

[0514] Step 2: To a stirred solution of 4-(cyclopropylthio)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide (0.89 g, 1.733 mmol) and ammonium carbonate (0.250 g, 2.60 mmol) in MeOH (4.5 mL) and dichloromethane (9.0 mL) was added (acetoxy)(phenyl)-iodoacetate (1.284 g, 3.99 mmol) as a solid in one portion. The resulting mixture was stirred at room temperature under open air for 18 h. The resulting mixture was loaded directly onto a silica gel pre-column (25 g) and subjected to combi-flash column chromatography on a 40-g ISCO gold column, eluting with MeOH / DCM (3 min at 0%, 25 min from 0% to 14%) to afford a racemic mixture of 4-(cyclopropanesulfonylimino)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide as an off-white solid (0.95 g, 1.744 mmol, 101% yield). Enantiomers were separated by preparative SFC using Regis (S,S) Whelk-01 (250X21 mm, 5 mm) with a mobile phase of 50% liquid CO2 and 50% MeOH at a flow rate of 60 mL / min to produce:

[0515] Example 18-1: 2-(6-azaspiro[2.5]octan-6-yl)-4-(R-cyclopropylsulfonylimino)-N-(2-(4,4-di fluoro-1-piperidinyl)-6-methyl-4-pyrimidinyl)benzamide. The first eluting peak, 1H NMR (400MHz, chloroform-d) δppm13.20 (br d,J=3.73Hz,1H),8.44(d,J=8.29Hz,1H),7.96(d,J=1.45Hz,1H),7.87(dd,J=1.66,8.29Hz,1H),7.52(s,1H),4.03(br s,4H),3.14(t,J=5.29Hz,4H),2.53-2.63(m,1H),2.44(br s,3H),1.95-2.10(m,4H),1.53-1.89(m,5H),1.45(tdd,J=5.08,6.92,10.29H z,1H),1.20-1.30(m,1H),1.07-1.17(m,1H),0.93-1.03(m,1H),0.44(s,4H). m / z(ESI):545.2(M+H) + .

[0516] Example 18-2: 2-(6-azaspiro[2.5]octan-6-yl)-4-(S-cyclopropylsulfonylimino)-N-(2-(4,4-di fluoro-1-piperidinyl)-6-methyl-4-pyrimidinyl)benzamide. Second eluting peak. 1 H NMR (400MHz, chloroform-d) δppm13.20 (br d,J=3.73Hz,1H),8.44(d,J=8.29Hz,1H),7.96(d,J=1.45Hz,1H),7.87(dd,J=1.66,8.29Hz,1H),7.52(s,1H),4.03(br s,4H),3.14(t,J=5.29Hz,4H),2.53-2.63(m,1H),2.44(br s,3H),1.95-2.10(m,4H),1.53-1.89(m,5H),1.45(tdd,J=5.08,6.92,10.29H z,1H),1.20-1.30(m,1H),1.07-1.17(m,1H),0.93-1.03(m,1H),0.44(s,4H). m / z(ESI):545.2(M+H) + The stereochemical determination is arbitrary.

[0517] Table 13: Examples 19-1 to 19-9 were prepared according to the procedures described for Examples 18-1 and 18-2:

[0518]

[0519]

[0520]

[0521] Example 20: (N 1 -(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-2-(6-azaspiro[2.5] 6-octan-1,2-dicarboxylic acid terephthalamide

[0522]

[0523] Step 1: To a solution of 4-bromo-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide (250 mg, 0.48 mmol, intermediate 21-3) in DMF (2.5 mL) was added Pd(PPh3)4 (6 mg, 4.8 μmol) and Zn(CN)2 (113 mg, 0.961 mmol), and the reaction mixture was stirred at 100°C for 16 h. The reaction mixture was then diluted with EtOAc and filtered. The filtrate was washed with water and brine, dried over Na2SO4, filtered and concentrated, and purified by flash column chromatography using a 30% EtOAc gradient in petroleum ether to provide 4-cyano-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide (160 mg, 0.343 mmol, 71.4% yield) as an off-white solid. 1 HNMR(400MHz,DMSO-d6)δppm 13.20(s,1H),8.18(d,J=8.1Hz,1H),8.03(s,1H),7.79(d,J=8.1Hz,1H),7.39(s,1H),4.00–3.80( m, 4H), 3.07 (t, J = 5.2Hz, 4H), 2.34 (s, 3H), 1.99 (tt, J = 13.3, 5.7Hz, 4H), 1.69 (s, 4H), 0.38 (s, 4H). m / z(ESI):467.2(M+H) + .

[0524] Step 2: To a solution of 4-cyano-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide (110 mg, 0.236 mmol) in dimethyl sulfoxide (2 mL) was added KCO (6.52 mg, 0.047 mmol) and H0 (103 μL, 1.179 mmol) at 0 °C and the reaction mixture was stirred for 1 h before being quenched with water and extracted with EtOAc (2 x 20 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, concentrated and purified by flash column chromatography using a 10% methanol gradient in dichloromethane to provide N 1-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-2-(6-azaspiro[2.5]octan-6-yl)terephthalamide (108 mg, 0.223 mmol, 95% yield). 1 H NMR(400MHz,DMSO-d6)δppm 13.75(s,1H),8.26(m,1H),8.17(m,1H),8.00(s,1H),7.89–7.78(m,1H),7.61(s,1H),7.42(s,1H),3.94(d,J=5.8H z, 4H), 3.06 (d, J = 6.0Hz, 4H), 2.34 (d, J = 2.3Hz, 3H), 2.01 (q, J = 8.5, 8.1Hz, 4H), 1.74 (s, 4H), 0.41 (d, J = 2.3Hz, 4H). m / z(ESI):485.2(M+H) + .

[0525] Example 21: 4-(azetidin-3-ylsulfonyl)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidine- 4-yl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide

[0526]

[0527] To a solution of 4-((1-(tert-butoxycarbonyl)azetidin-3-yl)sulfonyl)-2-(6-azaspiro[2.5]octan-6-yl)benzoic acid (2.54 g, 5.64 mmol, Intermediate 16) and HATU (3.22 g, 8.46 mmol, ChemPep) in DMF (35 mL) was added 2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-amine (1.93 g, 8.46 mmol, Intermediate 4) and DIPEA (2.46 mL, 14.09 mmol). The mixture was stirred at room temperature for 18 h. The mixture was diluted with saturated NaCO and EtOAc. The organics were separated and washed with NaCO, water and brine, dried over NaSO and concentrated in vacuo. The crude product was purified by silica gel chromatography: 0% to 30% to 60% EtOAc in heptane. m / z (ESI): 661.3 (M+H) + . The residue was treated with DCM (8 mL) and TFA (4 mL) for 30 min at room temperature and concentrated in vacuo. The resulting solid was suspended in EtOAc and washed with 1N NaOH solution, and the mixture was extracted with EtOAc. The organic extract was washed with water and brine, dried over Na2SO4 and concentrated in vacuo. The crude product was purified by silica gel chromatography: 0% to 20% MeOH in DCM with 2% NH4OH to obtain the title compound as a white solid. 1H NMR (400MHz, methanol-d4) δppm 8.35(d,J=8.09Hz,1H),7.90(d,J=1.24Hz,1H),7.81(dd,J=1.66,8.09Hz,1H),7.46(s,1H),4.46-4.60(m,1H),3.97 -4.06(m,6H),3.76-3.86(m,2H),3.10-3.21(m,4H),2.37(s,3H),1.93-2.01(m,4H),1.73-1.88(m,4H),0.45(s,4H). m / z(ESI):561.2(M+H) + .

[0528] Example 22: 4-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((1-methylazetidine alkyl-3-yl)sulfonyl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide

[0529]

[0530] To a mixture of 4-(azetidine-3-ylsulfonyl)-N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-2-(6-azaspiro[2.5]octan-6-yl)benzamide (0.045 g, 0.080 mmol, Example 21), MeOH (1 mL) and formaldehyde (0.016 g, 0.482 mmol, Fisher) with 10% to 15% MeOH was added AcOH (0.037 mL, 0.642 mmol, Aldrich), followed by sodium triacetoxyborohydride (0.204 g, 0.963 mmol, Aldrich). The mixture was stirred at room temperature for 18 h and concentrated in vacuo. The acid was neutralized with 1 N NaOH solution and the mixture was extracted with EtOAc. The organic phase was washed with water and brine, dried over Na2SO4 and concentrated in vacuo. Purification by silica gel chromatography: 0% to 100% EtOAc / EtOH (3 / 1) in heptane gave the title compound as a white solid. 1 HNMR (400MHz, chloroform-d) δppm 12.82 (br s,1H),8.44(d,J=8.29Hz,1H),7.82(d,J=1.66Hz,1H),7.73(dd,J=1.76,8.19Hz,1H),7.46(s,1H),3.97-4.11(m ,5H),3.60-3.90(m,4H),3.12(t,J=5.29Hz,4H),2.44-2.57(m,3H),2.39(s,3H),1.68-2.06(m,8H),0.43(s,4H). m / z(ESI):575.3(M+H)+ .

[0531] Biological Examples

[0532] The following assays were used to test exemplary compounds of the invention. The data for those examples tested according to the following procedure are shown in Table A below.

[0533] KIF18A enzyme assay: The KIF18A enzyme activity after treatment with the compounds was measured using a microtubule-stimulated ATPase activity assay. The compounds were serially diluted 2-fold in DMSO (Sigma Inc) over a range of 22 concentration points. Recombinant human KIF18A (1-467His-tagged) protein was expressed using a baculovirus system and purified by affinity chromatography at Amgen Inc. ADP-Glo ​​was used. TM The concentrations of KIF18A protein, microtubules (MTs), and ATP in the reaction were optimized by the Kinase / ATPase Assay Kit (Promega Inc) for standardized homozygous enzyme assays. The assay measures the ADP formed by the ATPase reaction. Prepare reaction buffer [(15 mM Tris, pH 7.5 (Teknova Inc), 10 mM MgCl2 (JT Baker Inc), 0.01% Pluronic F-68 (Life Technologies Inc), 1 μM Paclitaxel (Cytoskeleton Inc), and 30 μg / mL porcine microtubules (Cytoskeleton Inc)]. Add compounds and KIF18A protein (30 nM) to the prepared reaction buffer and incubate at room temperature for 15 min, then add ATP (K m , 75 μM) and incubate for another 15 min at room temperature. TM The reagent was mixed with 2.5 μl of reaction mixture and incubated at room temperature for 40 min. 10 μl of ADP-Glo ​​was added TM The reagent was added and incubated at room temperature for 40 min. Luminescence was read using an EnVision microplate reader with a superluminescent module (Perkin Elmer Inc). Concentration-response curve fitting and IC were performed using Genedata Screener software (version 15.0.1, Genedata Inc) with a four-parameter logistic regression fitting model. 50 Sure.

[0534] Table A provides data on compounds exemplified in the present application and its priority documents as representative compounds of the present invention, as follows: Compound name and biological data (IC 50 In μM. Example # refers to the example number)

[0535] Table A: Biological Data

[0536]

[0537]

[0538]

[0539]

[0540]

[0541]

[0542]

[0543]

[0544]

[0545]

[0546]

[0547]

[0548] For the purpose of clarity and understanding, the foregoing invention has been described in detail by way of illustration and example. It is understood by those skilled in the art that changes and modifications may be made within the scope of the appended claims. Therefore, it should be understood that the above description is intended to be illustrative rather than restrictive. Therefore, the scope of the present invention should not be determined with reference to the description above, but should be determined with reference to the full scope of the equivalent schemes given by the following appended claims and the claims.

[0549] All patents, patent applications, and publications cited herein are hereby incorporated by reference in their entirety for all purposes to the same extent as if each individual patent, patent application or publication was so individually denoted.

Claims

1. A compound of formula I: or any pharmaceutically acceptable salt thereof, wherein: X 1 N or -CR 6 ; R 1 For the group -ZR 12 , where Z is -NR 11 SO2-、-SO2NR 11 -, -S(=O)(=NH)-, -SO2- or -(C=O)NR 11 -; R 2 For the group -YR 13 , where Y is a bond, -NH-, -NH-(CH2) 1-4 -, -O-, or -OC 1-4 alkyl-; R 3 is H; R 4 is H, C substituted by 0, 1, 2 or 3 OH groups 1-6 Alkyl or R 4a ; R 5 is H; R 6 is H or halogen; R 7 is H or halogen; R 8 H, halogen, C 1-8 Alkyl or -OR 8b ; R 9 is H; R x for R 10a , R 10b , R 10c , R 10d , R 10e , R 10f , R 10g , R 10h , R 10i and R 10j Each of H, halogen, R 10k or R 10l ; Or alternatively, R 10a and R 10b Yes, R 10c and R 10d Yes, R 10e and R 10f Yes, R 10g and R 10h Yes or R 10i and R 10j Each of the pairs can independently combine with their respective attached carbon atoms to form a spiro-linked x a saturated or partially saturated 3-, 4-, 5-, 6-membered monocyclic ring; wherein the 3-, 4-, 5-, 6-membered monocyclic ring contains 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from the group consisting of O and S, and further, wherein the 3-, 4-, 5-, 6-membered monocyclic ring is substituted by 0, 1, 2 or 3 groups selected from the group consisting of F, Cl, Br, C 1-6 Alkyl, C 1-4 Haloalkyl, -OR a 、-OC 1-4 Haloalkyl, CN, -NR a R a and oxo; R 11 is H; R 12 H, R 12a or R 12b ; R 13 For R 13a or R 13b ; R 4a , R 10k , R 12a and R 13a is independently in each case: a saturated, partially saturated or unsaturated 3-, 4-, 5-, 6- or 7-membered monocyclic ring or a 5-, 6-, 7-, 8-, 9-, 10-, 11- or 12-membered bicyclic ring containing 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from the group consisting of O and S, which is substituted by 0, 1, 2 or 3 groups selected from the group consisting of F, Cl, Br, C 1-6 Alkyl, C 1-4 Haloalkyl, -OR a 、-OC 1-4 Haloalkyl, CN, -C(=O)R b 、-C(=O)OR a 、-C(=O)NR a R a 、-C(=NR a )NR a R a 、-OC(=O)R b 、-OC(=O)NR a R a 、-OC 2-6 Alkyl NR a R a 、-OC 2-6 Alkyl OR a 、-SR a 、-S(=O)R b 、-S(=O)2R b 、-S(=O)2NR a R a 、-NR a R a 、-N(R a )C(=O)R b 、-N(R a )C(=O)OR b 、-N(R a )C(=O)NR a R a 、-N(R a )C(=NR a )NR a R a 、-N(R a )S(=O)2R b 、-N(R a )S(=O)2NR a R a 、-NR a C 2-6 Alkyl NR a R a 、-NR a C 2-6 Alkyl OR a , -C 1-6 Alkyl NR a R a , -C 1-6 Alkyl OR a , -C 1-6 Alkyl N(R a )C(=O)R b , -C 1-6 Alkyl OC(=O)R b , -C 1-6 Alkyl C(=O)NR a R a , -C 1-6 Alkyl C(=O)OR a , R 14 and oxo; R 8b , R 10l , R 12b and R 13b In each case, independently selected from the group consisting of: selected from F, Cl, Br, -OR a 、-OC 1-4 0, 1, 2, 3, 4 or 5 groups substituted with the group consisting of haloalkyl and CN 1-6 alkyl; R 14 is independently in each case: a saturated, partially saturated or unsaturated 3-, 4-, 5-, 6- or 7-membered monocyclic ring or a 5-, 6-, 7-, 8-, 9-, 10-, 11- or 12-membered bicyclic ring containing 0, 1, 2 or 3 N atoms and 0 or 1 atom selected from the group consisting of O and S, which is substituted by 0, 1, 2 or 3 groups selected from the group consisting of F, Cl, Br, C 1-6 Alkyl, C 1-4 Haloalkyl, -OR a 、-OC 1-4 Haloalkyl, CN, -C(=O)R b 、-C(=O)OR a 、-C(=O)NR a R a 、-C(=NR a )NR a R a 、-OC(=O)R b 、-OC(=O)NR a R a 、-OC 2-6 Alkyl NR a R a 、-OC 2-6 Alkyl OR a 、-SR a 、-S(=O)R b 、-S(=O)2R b 、-S(=O)2NR a R a 、-NR a R a 、-N(R a )C(=O)R b 、-N(R a )C(=O)OR b 、-N(R a )C(=O)NR a R a 、-N(R a )C(=NR a )NR a R a 、-N(R a )S(=O)2R b 、-N(R a )S(=O)2NR a R a 、-NR a C 2-6 Alkyl NR a R a 、-NR a C 2-6 Alkyl OR a , -C 1-6 Alkyl NR a R a , -C 1-6 Alkyl OR a , -C 1-6 Alkyl N(R a )C(=O)R b , -C 1-6 Alkyl OC(=O)R b , -C 1-6 Alkyl C(=O)NR a R a , -C 1-6 Alkyl C(=O)OR a and oxo; R a is independently H or R b ;and R b In each case independently C 1-6 alkyl, phenyl or benzyl, wherein the C 1-6 The alkyl group is substituted with 0, 1, 2 or 3 substituents selected from the group consisting of halogen, -OH, -OC 1-4 Alkyl, -NH2, -NHC 1-4 Alkyl, -OC(=O)C 1-4 Alkyl and -N(C 1-4 Alkyl)C 1-4 alkyl; and the phenyl or benzyl is substituted with 0, 1, 2 or 3 substituents selected from the group consisting of halogen, C 1-4 Alkyl, C 1-3 Haloalkyl, -OH, -OC 1-4 Alkyl, -NH2, -NHC 1-4 Alkyl, -OC(=O)C 1-4 Alkyl and -N(C 1-4 Alkyl)C 1-4 alkyl.

2. The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein the compound of formula (I) is a compound of formula (Ia):

3. The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein the compound of formula (I) is a compound of formula (Ib):

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 10c , R 10d , R 10e , R 10f , R 10g , R 10h , R 10i and R 10j Each of H, halogen, C 1-6 Alkyl or C 1-4 haloalkyl; and R 10a and R 10b Each of the pairs combines with their respective attached carbon atoms to form a spiral bond to R x a saturated 3-, 4- or 5-membered monocyclic ring; wherein the ring contains 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from the group consisting of O and S.

5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 10c , R 10d , R 10e , R 10f , R 10g , R 10h , R 10i and R 10j Each of is H, methyl or ethyl; and R 10a and R 10b Each of the pairs combines with their respective attached carbon atoms to form a spiral bond to R x The cyclopropyl, cyclobutyl or cyclopentyl ring.

6. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the group for:

7. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the group for 8. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, wherein Z is -NHSO2-, -SO2NH-, -S(=O)(=NH)-, or -(C=O)NH-; and R 12 Selected from the group consisting of: (a) H; (b) cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxiranyl, oxetanyl, tetrahydrofuranyl, azetidinyl, imidazolyl, morpholinyl, pyrrolidinyl, piperazinyl, wherein each of said rings is substituted with 0, 1, 2 or 3 groups selected from the group consisting of OH, F, methyl, -CH2OH, -C(=O)OCH3, -C(=O)OC(CH3)3, NH2, CN and oxo; and (c) 0, 1, 2 or 3 OH or F groups.

9. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, wherein Z is -NHSO2-, -SO2NH-, -S(=O)(=NH)-, -SO2- or -(C=O)NH-; and (a)R 12 is H; (b)R 12 is oxetanyl or cyclopropyl; or (c)R 12 is C substituted with 0, 1, 2 or 3 OH groups 1-6 alkyl.

10. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, wherein Z is -NHSO2- or -SO2NH-; and R 12 is oxetane, cyclopropyl or C substituted with 0, 1, 2 or 3 OH groups 1-6 alkyl.

11. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, wherein Z is -NHSO2-, and R 12 It is -CH2-CH2-OH.

12. The compound or pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 7, wherein Y is a bond, -NH-(CH2) 0-4 -or-O-(CH2) 0-4 -;and R 13 is a saturated, partially saturated or unsaturated 3-, 4-, 5-, 6- or 7-membered monocyclic ring or a 5-, 6-, 7-, 8-, 9-, 10-, 11- or 12-membered bicyclic ring containing 0, 1, 2 or 3 N atoms and 0 or 1 atom selected from the group consisting of O and S, which is substituted by 0, 1, 2 or 3 groups selected from the group consisting of F, Cl, Br, C 1-6 Alkyl, C 1-4 Haloalkyl, -OH, -OC 1-4 Haloalkyl, CN, R 14 and oxo; or R 13 is selected from F, Cl, Br, -OH, -OC 1-4 0, 1, 2, 3, 4 or 5 groups substituted with the group consisting of haloalkyl and CN 1-6 alkyl.

13. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, wherein R 2 is a saturated 5- or 6-membered monocyclic ring, wherein each of said rings contains 0, 1 or 2 N atoms and 0 or 1 O atoms, and wherein each of said rings is substituted by 0, 1, 2 or 3 groups selected from the group consisting of: F, Cl, Br, C 1-6 Alkyl, C 1-4 Haloalkyl, -OH, -OC 1-4 Haloalkyl, CN, R 14 and oxo.

14. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, wherein Y is a bond; and R 13 is morpholinyl, piperidinyl, azetidinyl, pyrrolidinyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, piperazinyl, tetrahydrofuranyl, wherein each of said rings is substituted by 0, 1, 2 or 3 groups selected from the group consisting of: F, Cl, Br, methyl, CF3, -OH, -OCHF2, CN and oxo; or Y is NH, -O-, -O-(CH2)-, -O-(CH2)-(CH2)-, or -O-(CH2)-(CH2)-(CH2)-, and wherein R 13 for or C substituted by 0, 1, 2, 3, 4 or 5 groups selected from the group consisting of F, Cl, Br, methyl, CF3, -OH and CN 1-6 alkyl.

15. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, wherein R 2 is morpholinyl or piperidinyl substituted by 0, 1, 2 or 3 groups selected from the group consisting of F, Cl, Br, methyl, CF3, -OH, -OCHF2, CN and oxo.

16. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, wherein R 2 is morpholinyl substituted by 1, 2 or 3 methyl groups.

17. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, wherein R 2 is piperidinyl substituted by 1, 2 or 3 fluorine groups.

18. A compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof, wherein R 2 for 19. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, wherein Z is -NHSO2-, -SO2NH-, -S(=O)(=NH)-, -SO2- or -(C=O)NH-.

20. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, wherein R 12 (a) H; (b) C substituted by 0, 1, 2 or 3 groups selected from the group consisting of F, Cl, Br, -OH and -OCH3 1-6 or (c) a saturated, partially saturated or unsaturated 3-, 4-, 5-, 6- or 7-membered monocyclic ring containing 0, 1, 2 or 3 N atoms and 0 or 1 atom selected from the group consisting of O and S, which is substituted by 0, 1, 2 or 3 groups selected from the group consisting of F, Cl, Br, C 1-6 Alkyl, C 1-4 Haloalkyl, -C 1-6 Alkyl OH, -OH, -OCH3, -NH2 and oxo.

21. A compound or a pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 7, wherein R 12 Selected from the group consisting of cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl, azetidinyl, tetrahydrofuranyl and 1,3,4-oxathiazinyl.

22. A compound or a pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 7, wherein R 4 Selected from the group consisting of: (a) H; (b) C 1-6 alkyl; and (c) cyclopropyl.

23. A compound or a pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 7, wherein R 4 It is H or methyl.

24. The compound of claim 1 or 3 or a pharmaceutically acceptable salt thereof, wherein R 6 It is H or F.

25. A compound as described in any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof, wherein R 7 It is H or F.

26. A compound or a pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 7, wherein R 8 For H.

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

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

29. A pharmaceutical composition comprising a compound according to any one of claims 1 to 28 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable diluent or carrier.

30. Use of the compound of any one of claims 1 to 28 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 29 in the preparation of a medicament for inhibiting KIF18A in a cell.

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