KIF18A inhibitor

Abstract

Compounds of formula (I) as defined herein which are capable of regulating KIF18A protein and thereby affecting the processes of cell cycle and cell proliferation, and treating cancer and cancer-related diseases: TIFF2022542319000083.tif45170 and synthetic intermediates thereof. The present invention also includes pharmaceutical compositions containing the compounds and methods for treating conditions associated with KIF18A activity.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceuticals, and more specifically, to compounds and compositions useful for modulating KIF18A, and to uses and methods for controlling cell proliferation and treating cancer.

Background Art

[0002] Cancer is one of the most prevalent diseases afflicting humanity and a major cause of death worldwide. In attempts over the past 20 to 30 years to discover effective treatments or cures for one or more of the many types of cancer, many groups have expended a great deal of time, effort, and financial resources. However, to date, only a few of the available cancer treatments and therapies have achieved great success.

[0003] Cancer is often characterized by disordered cell proliferation. Damage to one or more genes that govern cell pathways, which control the progression of proliferation throughout the cell cycle and the centrosome cycle, can cause the loss of normal regulation of cell proliferation. These disordered genes may encode various tumor suppressor or oncogene proteins involved in a cascade of events, resulting in unregulated cell cycle progression and cell proliferation. Various kinases and kinesin proteins that play important roles in cell cycle and mitosis regulation, as well as the progression of normal and cancerous cells, have been identified.

[0004] Kinesin is a molecular motor that plays an important role in cell division and the transport of intracellular vesicles and organelles. Mitotic kinesins play roles in spindle assembly, chromosome segregation, centrosome separation, and several aspects of motility (discussed in O. Rath and F. Kozielski, “Nature Review Cancer” 12:527-39, 2012). Human kinesins are classified into 14 subfamilies based on sequence homology within the so-called “motor domain,” and the ATPase activity of this domain drives unidirectional movement along microtubules (MTs). The non-motor domains of these proteins mediate cargo binding, and the “cargo” can include any one of a variety of membranous organelles, signal transduction scaffolding systems, and chromosomes. Kinesins use the energy of ATP hydrolysis to move cargo along polar microtubules. Therefore, kinesins are often called “plus-end” or “minus-end” directed motors.

[0005] The KIF18A gene belongs to the kinesin-8 subfamily and is a plus-end directed motor. KIF18A is thought to affect the dynamics at the plus ends of kinetochore microtubules, which control correct chromosome positioning and spindle tension. Depletion of human KIF18A results in longer spindles, increased chromosomal oscillations during metaphase of mitosis, 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 various types of cancer, including but not limited to colon cancer, breast cancer, pancreatic cancer, prostate cancer, bladder cancer, head cancer, neck cancer, cervical cancer, and ovarian cancer. Furthermore, gene deletion or knockdown, or inhibition of KIF18A, results in the formation of the mitotic spindle apparatus in cancer cell lines. Specifically, inhibition of KIF18A has been found to induce lethality or death driven by mitotic cell arrest, apoptosis-mediated cell death during mitosis, a known vulnerability, mitotic catastrophe, or multipolarity after mitotic slippage during interphase. Therefore, there is strong interest in discovering inhibitors of the KIF18A protein.

Prior Art Documents

Non-Patent Documents

[0006]

Non-Patent Document 1

Non-Patent Document 2

Summary of the Invention

Problems to be Solved by the Invention

[0007] Therefore, inhibition of the ATPase activity of KIF18A is a promising approach for developing novel anti-cancer agents.

Means for Solving the Problems

[0008] One aspect of the present invention is a novel class of compounds useful for regulating KIF18A protein, either alone or in a binding complex with microtubules, for treating KIF18A-mediated symptoms and / or diseases including cancer, inflammation, or ciliopathologies.

[0009] The compounds provided by the present invention have MT-based KIF18A regulatory activity, and in particular KIF18A inhibitory activity. For this purpose, the present invention also provides the use of these compounds, and their pharmaceutically acceptable salts, in the preparation and manufacture of pharmaceutical compositions or agents for the therapeutic, prophylactic, acute, or chronic treatment of KIF18A-mediated diseases and disorders including, but not limited to, cancer. Thus, the compounds of the present invention are useful in the manufacture of anti-cancer agents. The present invention also provides a process for producing a compound of formula I, and intermediates useful in such a process.

[0010] In Embodiment 1, the present invention provides a compound of formula (I):

Chemical formula

Chemical formula

[0011] In Embodiment 2, the present invention provides that L is -NR 8 -(C=O), and has the formula (Ia):

Chemical formula

[0012] In Embodiment 3, the present invention provides that L is -(C=O)-NR 8 -, and has the formula (Ib): [Chem.] having, wherein said R Xa and R Xb pairs, in combination with the carbon atom to which each is attached, can form a saturated or partially saturated 3-membered, 4-membered, 5-membered, 6-membered monocyclic ring that is spiro to the piperidinyl ring, where the 3-membered, 4-membered, 5-membered, 6-membered monocyclic ring contains 0 N, O and S atoms, and further, the 3-membered, 4-membered, 5-membered, 6-membered monocyclic ring is substituted with 0, 1, 2 or 3 groups selected from F, Cl, Br, C 1~6 alk, C 1~4 haloalk, -OR a , -OC 1~4 haloalk, CN, -NR a R a , or oxo, and provides a compound.

[0013] In Embodiment 4, the present invention provides a compound wherein R 8 is H or methyl.

[0014] In Embodiment 5, the present invention provides a compound wherein each of R Xc , R Xd , R Xe , R Xf , R Xg , R Xh , R Xi , R Xj , R Xk , and R Xl is each H, halo, C 1~6 alk, or C 1~4 haloalk, and each pair of R Xa and R Xb , in combination with the carbon atom to which each is attached, forms a saturated 3-membered, 4-membered, or 5-membered monocyclic ring that is spiro to the piperidinyl ring, where the ring contains 0, 1, 2 or 3 N atoms and 0 or 1 atom selected from O and S, and provides the compound according to any of the above embodiments, or a pharmaceutically acceptable salt thereof.

[0015] In Embodiment 6, the present invention provides a compound wherein RXc , R Xd , R Xe , R Xf , R Xg , R Xh , R Xi , R Xj , R Xk , and R Xl each of which is H, methyl, or ethyl, and R Xa and R Xb each pair of which, in combination with the carbon atom to which each is attached, forms a cyclopropyl, cyclobutyl, or cyclopentyl ring that is spiro to the piperidinyl ring, a compound according to any of the above embodiments, or a pharmaceutically acceptable salt thereof.

[0016] In Embodiment 7, the present invention provides a compound according to any of the above embodiments, or a pharmaceutically acceptable salt thereof, wherein R X is (a) ethynyl substituted with isobutyl or cyclopentyl, (b) phenyl substituted with 0, 1, 2, or 3 F, Cl, CF3, CH2OCH3, methyl, ethyl, propyl, isopropyl, tert-butyl, or cyclopropyl groups, (c) or pyrazolyl substituted with 0, 1, 2, or 3 F, Cl, CF3, CH2OCH3, methyl, ethyl, propyl, isopropyl, tert-butyl, or cyclopropyl groups, (d)

Chemical formula

[0017] In Embodiment 8, the present invention provides a compound according to any of the above embodiments, or a pharmaceutically acceptable salt thereof, wherein R X is

Chemical formula

[0018] In embodiment 9, the present invention provides a compound comprising R X but, [ka]

[0023] The present invention provides a compound according to any of the above embodiments, wherein:

[0019] In embodiment 10, the present invention provides a compound according to any of the above embodiments, wherein Z is absent, -NH-, -NHSO2-, -SO2NH-, or -CH2SO2-, or a pharma- ceutically acceptable salt thereof.

[0020] In embodiment 11, the present invention provides a compound comprising R 9 but, a) H, (b) C substituted with 0, 1, 2 or 3 groups selected from F, Cl, Br, -CF3, or -OH. 1~6 alk, or (c) containing 0, 1, 2 or 3 N atoms and 0 or 1 atom selected from O and S, and further including F, Cl, Br, methyl, ethyl, -CF3, -CH2OH, -OH, -OCH3, -NH 2、 or oxo; or a pharma- ceutically acceptable salt thereof.

[0021] In embodiment 12, the present invention provides a compound comprising R 9 is selected from cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl, azetidinyl, diazirinyl, oxazolidinyl, isothiazolidinyl, pyrazolyl, or pyrrolidinyl, or a pharma- ceutically acceptable salt thereof.

[0022] In embodiment 13, the present invention provides a compound comprising R 1 Base-ZR 12 where R 1 Base-ZR9 wherein Z is absent, -NH-, -NHSO2-, -SO2NH-, or -CH2SO2-, and R 9 is cyclopropyl or oxazolinyl substituted with 0, 1, 2 or 3 OH, -CH2OH, methyl, or oxo groups, or R 9 is C 1~6 alk substituted with 0, 1, 2 or 3 OH or CF3 groups, and provides a compound according to any of the above embodiments, or a pharmaceutically acceptable salt thereof.

[0023] In embodiment 14, the present invention provides a compound according to any of the above embodiments, or a pharmaceutically acceptable salt thereof, wherein R 1 is the group -Z-R 9 wherein Z is -NHSO2- and R 9 is -CH2-CH2-OH.

[0024] In embodiment 15, the present invention provides a compound according to any of the above embodiments, or a pharmaceutically acceptable salt thereof, wherein R 2 is the group -Y-R 10 wherein Y is absent, -NH-, -NHSO2-, SO2NH-, or SO2-, R 10 contains 0, 1, 2 or 3 N atoms and 0 or 1 atom selected from O and S, and is a saturated, partially saturated or unsaturated 3-membered, 4-membered, 5-membered, 6-membered, or 7-membered monocyclic ring substituted with 0, 1, 2 or 3 groups selected from F, Cl, Br, methyl, ethyl, CF3, CH2OH, -OH, -OCH3, -NH2, -NH(CH3), and oxo, or R 10 is C 1~6 alk substituted with 0, 1, 2, 3, 4, or 5 groups selected from F, Cl, Br, -OH, or -CF3.

[0025] In embodiment 16, the present invention provides a compound according to any of the above embodiments, or a pharmaceutically acceptable salt thereof, wherein R 2is a morpholinyl ring, a thiomorpholinyl ring, a cyclobutyl ring, a cyclopentyl ring, or a piperidinyl ring, wherein each of said rings is substituted with 0, 1, 2 or 3 groups selected from F, Cl, Br, methyl, CF3, -OH, -OCHF2, or oxo, a compound according to any of the above embodiments, or a pharmaceutically acceptable salt thereof.

[0026] In Embodiment 17, the present invention provides a compound according to any of the above embodiments, or a pharmaceutically acceptable salt thereof, wherein R 2 is morpholinyl or piperidinyl substituted with 0, 1, 2 or 3 selected from F or methyl.

[0027] In Embodiment 18, the present invention provides a compound according to any of Embodiments 1 to 17, or a pharmaceutically acceptable salt thereof, wherein R 3 is H or F.

[0028] In Embodiment 19, the present invention provides a compound according to any of the above embodiments, or a pharmaceutically acceptable salt thereof, wherein R 4 is H.

[0029] In Embodiment 20, the present invention provides a compound according to any of the above embodiments, or a pharmaceutically acceptable salt thereof, wherein R 5 is H.

[0030] In Embodiment 21, the present invention provides a compound according to any of the above embodiments, or a pharmaceutically acceptable salt thereof, wherein R 6 is H, F, or methyl.

[0031] In Embodiment 22, the present invention provides a compound according to any of the above embodiments, or a pharmaceutically acceptable salt thereof, wherein R 7 is H.

[0032] In Embodiment 23, the present invention provides N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(4-methylpiperidin-1-yl)pyrazine-2-carboxamide N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-3-(4,4-difluoropiperidin-1-yl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)pyrazine-2-carboxamide N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(4-(trifluoromethyl)piperidin-1-yl)pyrazine-2-carboxamide N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-3-(4-ethylpiperidin-1-yl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)pyrazine-2-carboxamide N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-3-(4,4-dimethylpiperidin-1-yl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)pyrazine-2-carboxamide N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(4-isopropylpiperidin-1-yl)pyrazine-2-carboxamide 3-(4-(tert-Butyl)piperidin-1-yl)-N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)pyrazine-2-carboxamide 3-(4-(sec-Butyl)piperidin-1-yl)-N-(3-(N-(tert-butyl)sulfamoyl)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)pyrazine-2-carboxamide, N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(4-propylpiperidin-1-yl)pyrazine-2-carboxamide, N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-3-(4-cyclopropylpiperidin-1-yl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)pyrazine-2-carboxamide, N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(4-(methoxymethyl)piperidin-1-yl)pyrazine-2-carboxamide, N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(2-azaspiro[3.3]heptan-2-yl)pyrazine-2-carboxamide, N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(2-azaspiro[3.5]nonan-2-yl)pyrazine-2-carboxamide, N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(3-oxa-9-azaspiro[5.5]undecan-9-yl)pyrazine-2-carboxamide, N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(7-azaspiro[3.5]nonan-7-yl)pyrazine-2-carboxamide, N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-3-(2,2-difluoro-7-azaspiro[3.5]nonan-7-yl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)pyrazine-2-carboxamide, N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(8-azaspiro[4.5]dec-8-yl)pyrazine-2-carboxamide, N-(3-(N-(tert-Butyl)sulfamoyl)-5-methylphenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide, 5-((1-Hydroxy-2-methylpropan-2-yl)amino)-3-(6-azaspiro[2.5]octan-6-yl)-N-(3-((1,1,1-trifluoro-2-methylpropan-2-yl)sulfonyl)phenyl)pyrazine-2-carboxamide, N-(3-(tert-Butylsulfonyl)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide, 5-((1-Hydroxy-2-methylpropan-2-yl)amino)-N-(3-((1-methylcyclobutyl)sulfonyl)phenyl)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide, N-(3-(N-(tert-Butyl)sulfamoyl)-5-fluorophenyl)- 5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide, N-(3-(N-(tert-Butyl)sulfamoyl)-2-fluorophenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide, N-(3-(1,1-Dioxidothiomorpholino)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide 5-((1-hydroxy-2-methylpropan-2-yl)amino)-N-(3-morpholinophenyl)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide 5-((1-hydroxy-2-methylpropan-2-yl)amino)-N-(3-((1-hydroxy-2-methylpropan-2-yl)amino)phenyl)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide (S)-N-(3-((3,3-difluorocyclopentyl)sulfonyl)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide (R)-N-(3-((3,3-difluorocyclopentyl)sulfonyl)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide N-(3-(cyclopentylsulfonyl)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide 5-((1-hydroxy-2-methylpropan-2-yl)amino)-N-(3-(piperidin-1-ylsulfonyl)phenyl)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide N-(3-(cyclopentylsulfonyl)phenyl)-5-((1-(hydroxymethyl)cyclopropyl)amino)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-5-((1-(hydroxymethyl)cyclopropyl)amino)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide, 5-((2-Hydroxy-2-methylpropyl)amino)-N-(3-(piperidin-1-ylsulfonyl)phenyl)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide, N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-3-(4-chlorophenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)pyrazine-2-carboxamide, N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-3-(4-fluorophenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)pyrazine-2-carboxamide, N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(4-(trifluoromethyl)phenyl)pyrazine-2-carboxamide, N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-3-(4-ethylphenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)pyrazine-2-carboxamide, N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(4-isopropylphenyl)pyrazine-2-carboxamide, (R)-N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-3-(6-azaspiro[2.5]octan-6-yl)-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)pyrazine-2-carboxamide, (S)-N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-3-(6-azaspiro[2.5]octan-6-yl)-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)pyrazine-2-carboxamide, or N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-5-((1,3-dihydroxy-2-methylpropan-2-yl)amino)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide Provided is a compound according to any of the above embodiments, or any pharmaceutically acceptable salt thereof, selected from the group consisting of:

[0033] In Embodiment 24, the present invention provides a compound selected from the following, or a pharmaceutically acceptable salt thereof:

[0034] [Table 1]

[0035] [Table 2]

[0036] [Table 3] or any pharmaceutically acceptable salt thereof.

[0037] Another aspect of the present invention is a composition comprising a novel class of compounds or pharmaceutically acceptable salts thereof useful for modulating KIF18A protein, either alone or in a binding complex with microtubules.

[0038] Embodiment 25 In this case, the present invention provides a pharmaceutical composition comprising a compound according to any one of Embodiments 1 to 24 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable diluent or carrier.

[0039] Yet another aspect of the present invention is a method of treating a condition treatable with a KIF18A inhibitor, the method comprising administering to a patient in need thereof a therapeutically effective amount of a novel class of compounds useful for modulating KIF18A protein, alone or in a binding complex with microtubules, or a pharmaceutically acceptable salt thereof.

[0040] In Embodiment 26, the present invention provides a method of treating a condition treatable with a KIF18A inhibitor, the method comprising administering to a patient in need thereof a therapeutically effective amount of a compound according to Embodiments 1-24 or a composition according to Embodiment 25.

[0041] In Embodiment 27, the present invention provides the method of Embodiment 26, wherein the above-mentioned condition is (a) a solid tumor selected from 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, or a hematologically derived tumor, (b) a lymphoid hematopoietic tumor selected from leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B cell lymphoma, T cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, hairy cell lymphoma, and Burkitt lymphoma, (c) a myeloid hematopoietic tumor selected from acute and chronic myelogenous leukemia, myelodysplastic syndrome, and promyelocytic leukemia, (d) a mesenchymal-derived tumor selected from fibrosarcoma and rhabdomyosarcoma, (e) a central and peripheral nervous system tumor selected from astrocytoma, neuroblastoma, glioma, and schwannoma, or (f) a cancer selected from the group consisting of melanoma, seminoma, teratoma, osteosarcoma, xeroderma pigmentosum, keratoacanthoma, follicular thyroid cancer, or Kaposi sarcoma.

[0042] In Sub - embodiment 27a, the present invention provides the method of Embodiment 32, wherein the above - mentioned symptoms are cancers 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 Sep; 31(9):1676 - 84. doi:10.1093 / carcin / bgq134. Epub 2010 Jul 1. (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.

[0043] In Embodiment 28, the present invention provides a method for reducing the size of a solid tumor in a subject, the method comprising administering to the subject in need thereof a therapeutically effective amount of the compound according to Embodiments 1 - 24, or the composition according to Embodiment 25.

[0044] In Embodiment 29, the present invention provides a method for treating a cell growth disorder in a subject, the method comprising administering to the subject in need thereof a therapeutically effective amount of the compound according to Embodiments 1 - 24, or the composition according to Embodiment 25.

[0045] In Embodiment 30, the present invention provides a method for inhibiting KIF18A in cells, the method comprising contacting the cells with a compound described in Embodiments 1 to 24, or a pharmaceutically acceptable salt thereof, or a composition described in Embodiment 25.

[0046] Yet another aspect of the present invention is a method for preparing a novel class of compounds or pharmaceutically acceptable salts thereof useful for regulating KIF18A protein, either alone or in a binding complex with microtubules.

[0047] In Embodiment 31, the present invention provides a method for preparing a compound of formula (I) described herein.

[0048] In Embodiment 32, the present invention provides an intermediate compound used in the method for preparing a compound of formula (I) described herein.

Modes for Carrying Out the Invention

[0049] Detailed Description of the Invention The present invention includes all pharmaceutically acceptable isotope-labeled compounds of the present invention in which one or more atoms are replaced by atoms having the same atomic number but a different atomic mass or mass number than the atom that predominates in nature.

[0050] Examples of isotopes suitable for inclusion in the compounds of the present invention include 2 H and 3 H and other hydrogens, 11 C 、13 C and 14 C and other carbons, 38 Cl and other chlorines, 18 F and other fluorines, 123 I and 125 I and other iodines, 13 N and 15 N and other nitrogens, 15 O, 17 O and 18 O and other oxygens, 32 P and other phosphors, as well as 35 S and other sulfur isotopes, but are not limited thereto.

[0051] Certain isotope-labeled compounds of the present invention, for example, those incorporating radioisotopes, are useful in the study of drug and / or substrate tissue distribution. Tritium, a radioisotope, i.e., 3 H, and carbon-14, i.e., 14 C, are particularly useful for this purpose from the viewpoints of ease of incorporation and ease of detection means.

[0052] Substitution with heavier isotopes such as deuterium, i.e., 2 H, etc., may be preferred in some situations because specific therapeutic advantages can be obtained, such as an extended in vivo half-life or a reduced required dose, resulting from increased metabolic stability.

[0053] 11 C, 18 F, 15 O and 13 substitution with positron-emitting isotopes such as N can be useful in positron emission tomography (PET) studies for examining the receptor occupancy of substrates.

[0054] The isotope-labeled compounds of the present invention can generally be prepared by methods similar to those described in the accompanying examples and preparations, by the prior art known to those skilled in the art, or using appropriate isotope-labeled reagents in place of the unlabeled reagents previously used.

[0055] Examples of pharmaceutically acceptable solvates according to the present invention include those in which the crystallization solvent can be substituted with isotopes, such as D2O, d6-acetone, d6-DMSO.

[0056] Specific embodiments of the present invention include the compounds exemplified in the following examples, and their pharmaceutically acceptable salts, complexes, solvates, polymorphs, stereoisomers, metabolites, prodrugs, and other derivatives.

[0057] Unless otherwise indicated, the following definitions apply to the terms found in this specification and claims.

[0058] 「C α~β alk」 means an alkyl group that includes a branched or straight-chain relationship, or any three combinations thereof, with a minimum of α and a maximum of β carbon atoms, where α and β represent integers. The alkyl groups described in this section may also include one or two double or triple bonds. The notation C0alk refers to a direct bond. C 1~6 Examples of alkyl include, but are not limited to, the following:

Chemical formula

[0059] The term "benzo group", alone or in combination, means the divalent radical C4H4=, one of whose representations, when bonded adjacent to another ring, forms a benzene-like ring such as tetrahydronaphthylene, indole, etc., which is -CH=CH-CH=CH-.

[0060] The terms "oxo" and "thioxo" represent the groups =O (such as in a carbonyl) and =S (such as in a thiocarbonyl), respectively.

[0061] "Halo" or "halogen" means a halogen atom selected from F, Cl, Br, and I.

[0062] 「C α~β "C haloalk" means the above alk group in which any number (at least one) of the hydrogen atoms bonded to the alk chain is replaced by F, Cl, Br, or I.

[0063] Groups such as N(R a )R a include substituents in which the two R a groups together optionally form a ring containing an N, O, or S atom, and examples of such groups include

Chemical formula

[0064] Base N(C α~β alk)C α~β alk (where α and β are as defined above), examples of which include substituents where two C α~β alk groups together optionally form a ring containing an N, O, or S atom. Such examples include

Chemical formula

[0065] The term "bicyclic ring" means a group characterized by two joined rings. The bicyclic ring may be carbocyclic (where all ring atoms are carbon) or heterocyclic (where the ring atoms include, in addition to carbon atoms, for example, 1, 2, or 3 heteroatoms, such as N, O, or S). Both of the two rings may be aliphatic (such as decalin and norbornane), or aromatic (such as naphthalene), or a combination of aliphatic and aromatic (such as tetralin). Examples of bicyclic rings include (a) spiro ring compounds, where the two rings share only a single atom, which is usually a quaternary carbon atom, i.e., the spiro atom. Examples of spiro ring compounds include

Chemical formula

Chemical formula

[0066] "Carbocyclic" or "carbocyclic ring" means, by itself or in combination with other terms, unless otherwise specified, a ring including the cyclic version of "C α~β alk". Examples of carbocyclic rings include cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, cyclobutylene, cyclohexylene, etc.

[0067] "Heterocyclic" or "heterocyclic ring" means a ring containing at least one carbon atom and at least one other atom selected from N, O, and S. Examples of heterocyclic rings that can be found in the claims of this patent include [Chem.] include, but are not limited to, these.

[0068] "Pharmaceutically acceptable salt" means a salt prepared by conventional means, which are well known to those skilled in the art. Examples of "pharmacologically acceptable salts" include, but are not limited to, basic salts of inorganic and organic acids such as 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, etc. When the compound of the present invention contains an acidic functional group such as a carboxy group, suitable pharmaceutically acceptable cationic pairs for the carboxy group are well known to those skilled in the art, and examples thereof include alkali, alkaline earth, ammonium, quaternary ammonium cations, etc. Additional examples of "pharmacologically acceptable salts" are referred to below and to Berge et al., J. Pharm. Sci. 66:1 (1977).

[0069] "Saturated, partially saturated, or unsaturated" includes substituents saturated with hydrogen, substituents not saturated with hydrogen at all, and substituents partially saturated with hydrogen.

[0070] The "leaving group" generally refers to a group that can be easily substituted 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, triflates, tosylates, etc. Preferred leaving groups are indicated herein as necessary.

[0071] "Protecting group" generally refers to a group well-known in the art that is used to prevent selected reactive groups such as carboxy, amino, hydroxy, mercapto, etc. from undergoing unwanted reactions such as nucleophilic reactions, electrophilic reactions, oxidation reactions, reduction reactions, etc. Preferred protecting groups are indicated herein as necessary. Examples of amino protecting groups include, but are not limited to, aralkyl, substituted aralkyl, cycloalkenylalkyl and substituted cycloalkenylalkyl, allyl, substituted allyl, acyl, alkoxycarbonyl, aralkoxycarbonyl, silyl, etc. Examples of aralkyl include, but are not limited to, benzyl, orthomethylbenzyl, trityl and benzhydryl, which may be optionally substituted with halogen, alkyl, alkoxy, hydroxy, nitro, acylamino, acyl, etc., and salts such as phosphonium and ammonium salts. Examples of aryl groups include phenyl, naphthyl, indanyl, anthracenyl, 9-(9-phenylfluorenyl), phenanthrenyl, durenyl, etc. Examples of preferably cycloalkenylalkyl or substituted cycloalkenylalkyl radicals having 6 to 10 carbon atoms include, but are not limited to, cyclohexenylmethyl. Suitable acyl, alkoxycarbonyl and aralkoxycarbonyl groups include benzyloxycarbonyl, t-butoxycarbonyl, iso-butoxycarbonyl, benzoyl, substituted benzoyl, butyryl, acetyl, trifluoroacetyl, trichloroacetyl, phthaloyl, etc. Mixtures of protecting groups can be used to protect the same amino group, such that a primary amino group can be protected by both an aralkyl group and an aralkoxycarbonyl group. Amino protecting groups can also form heterocycles with the nitrogen to which they are attached, such as 1,2-bis(methylene)benzene, phthalimidyl, succinimidyl, maleimidyl, etc., and these heterocyclic groups can further include adjacent aryl and cycloalkyl rings. In addition, the heterocyclic groups can be mono-, di-, or trisubstituted, such as nitrophthalimidyl. An amino group can also be protected from unwanted reactions such as oxidation by forming addition salts such as hydrochloric acid, toluenesulfonic acid, trifluoroacetic acid, etc.Many amino acid protecting groups are also suitable for the protection of carboxy, hydroxy, and mercapto groups. For example, aralkyl groups. Alkyl groups such as tert-butyl are also suitable for the protection of hydroxy and mercapto groups.

[0072] A silyl protecting group is a silicon atom optionally substituted by 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. Silylation of an amino group gives a mono- or disilylamino group. Silylation of an amino alcohol compound can result in an N,N,O-trisilyl derivative. Removal of the silyl functional group from a silyl ether functional group is readily accomplished, as a separate reaction step or in situ during reaction with an alcohol group, for example, by treatment with a metal hydroxide or ammonium fluoride reagent. Suitable silylating agents are, for example, trimethylsilyl chloride, tert-butyl-dimethylsilyl chloride, phenyldimethylsilyl chloride, diphenylmethylsilyl chloride, or their combination products with imidazole or DMF. Methods for the silylation of amines and removal of silyl protecting groups are well known to those skilled in the art. Also, methods for preparing these amine derivatives from the corresponding amino acids, amino acid amides, or amino acid esters are well known to those skilled in organic chemistry including amino acid / amino acid ester, or amino alcohol chemistry.

[0073] Protecting groups are removed under conditions that do not affect the remainder of the molecule. These methods are well known in the art and include, for example, acid hydrolysis, hydrogenolysis, etc. Preferred methods involve removal of protecting groups such as removal of the benzyloxycarbonyl group by hydrogenolysis using palladium on carbon in a suitable solvent system such as alcohol, acetic acid, or mixtures thereof. The t-butoxycarbonyl protecting group can be removed using an inorganic or organic acid such as HCl or trifluoroacetic acid in a suitable solvent system such as dioxane or methylene chloride. The resulting amino salt can be easily neutralized to obtain the free amine. Carboxy protecting groups such as methyl, ethyl, benzyl, tert-butyl, 4-methoxyphenylmethyl, etc. can be removed under hydrolysis and hydrogenolysis conditions well known to those skilled in the art.

[0074] The compounds of the present invention may contain groups that can exist in tautomeric forms such as cyclic and acyclic amidine and guanidine groups, heteroatom-substituted heteroaryl groups (Y' = O, S, NR), etc., exemplified by the following examples:

Chemical formula

[0075] Prodrugs of the compounds of the present invention are also contemplated by the present invention. A prodrug is an active or inactive compound that, after administration of the prodrug to a patient, is chemically modified by in vivo physiological actions such as hydrolysis and metabolism to become the compound of the present invention. Compatibility and techniques associated with the manufacture and use of prodrugs are well known to those of ordinary skill in the art. For an overview of prodrugs involving esters, see Svensson and Tunek Drug Metabolism Reviews 165(1988) and Bundgaard Design of Prodrugs, Elsevier(1985). Examples of masked carboxylate anions include various esters such as alkyl (e.g., methyl, ethyl), cycloalkyl (e.g., cyclohexyl), aralkyl (e.g., benzyl, p-methoxybenzyl), and alkylcarbonyloxyalkyl (e.g., pivaloyloxymethyl). Amines are masked as arylcarbonyloxymethyl-substituted derivatives that are cleaved by esterases in vivo to release the free drug and formaldehyde (Bungaard J.Med.Chem.2503(1989)). Also, drugs containing acidic NH groups such as imidazole, imide, and indole are masked with N-acetyloxymethyl groups (Bundgaard Design of Prodrugs, Elsevier(1985)). Hydroxy groups are masked as esters and ethers. European Patent No. 039,051 (Sloan and Little, 4 / 11 / 81) discloses Mannich base hydroxamic acid prodrugs, their preparation, and use.

[0076] This specification and the claims include listings of chemical species (sometimes referred to as Markush groups) using the expressions "selected from ~ and ~" and "is ~ or ~". When this term is used in this application, unless otherwise specified, it is intended to include the entire group, or any single component thereof, or any subgroup thereof. The use of this expression is for the purpose of mere abbreviation and in no way limits the exclusion of individual elements or subgroups if necessary.

[0077] Pharmaceutical Compositions, Administration, and Routes of Administration Furthermore, the present specification also provides pharmaceutical compositions comprising the compounds disclosed herein, together with pharmaceutically acceptable excipients such as, for example, diluents or carriers. Suitable compounds and pharmaceutical compositions for use in the present invention include those in which the compound can be administered in an effective amount to achieve its intended purpose. The administration of the compound will be described in more detail below.

[0078] Suitable pharmaceutical formulations can be determined by those skilled in the art according to the route of administration and the desired dosage. See, for example, Remington’s Pharmaceutical Sciences, 1435 - 712 (18th ed., Mack Publishing Co, Easton, Pennsylvania, 1990). The formulation can affect the physical state, stability, in vivo release rate, and in vivo elimination rate of the administered drug. Depending on the route of administration, the suitable dosage can be calculated according to body weight, body surface area, or organ size. Further fine - tuning of the calculations necessary to determine the appropriate therapeutic dosage is mechanically performed by those skilled in the art without undue experimentation, particularly in light of the dosage information and assays disclosed herein and the pharmacokinetic data obtained from animal or human clinical trials.

[0079] The terms "pharmaceutically acceptable" or "pharmacologically acceptable" refer to molecular entities and compositions that do not produce adverse allergic or other untoward reactions when administered to an animal or a human. As used herein, "pharmaceutically acceptable excipients" include, for example, any solvent, dispersion medium, coating, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. The use of such excipients for pharmaceutically active substances is known in the art. Its use in therapeutic compositions is contemplated, except where any conventional medium or agent is incompatible with the therapeutic composition. Auxiliary active ingredients can also be incorporated into the compositions. 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, copper sulfate, thiamine chloride hydrochloride, vitamin A palmitate, manganese sulfate, riboflavin, pyridoxine hydrochloride, folic acid, beta-carotene, potassium iodide, phylloquinone, biotin, sodium selenate, chromium chloride, sodium molybdate, vitamin D3, and cyanocobalamin.

[0080] The compounds can be present in pharmaceutical compositions as pharmaceutically acceptable salts. As used herein, "pharmaceutically acceptable salts" include, for example, base addition salts and acid addition salts.

[0081] Pharmaceutically acceptable base addition salts can be formed with metals or amines such as alkali metals, alkaline earth metals or organic amines. Pharmaceutically acceptable salts of the 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, alkaline earth, ammonium and quaternary ammonium cations. Carbonates or bicarbonates are also possible. Examples of metals used as cations are sodium, potassium, magnesium, ammonium, calcium or iron, etc. Examples of suitable amines include isopropylamine, trimethylamine, histidine, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, dicyclohexylamine, ethylenediamine, N-methylglucamine and procaine.

[0082] Pharmaceutically acceptable acid addition salts include salts of inorganic acids or organic acids. Examples of suitable acid salts include hydrochloride, formate, acetate, citrate, salicylate, nitrate, and phosphate. Other suitable pharmaceutically acceptable salts are well known to those skilled in the art and include, for example, formic acid, acetic acid, citric acid, oxalic acid, tartaric acid or mandelic acid, hydrochloric acid, hydrobromic acid, sulfuric acid or phosphoric acid; organic carboxylic acids, sulfonic acids, sulfinic acids or phosphonic acids or N-substituted sulfamic acids, such as acetic acid, trifluoroacetic acid (TFA), propionic acid, glycolic acid, succinic acid, maleic acid, hydroxymaleic acid, methylmaleic acid, fumaric acid, malic acid, tartaric acid, lactic acid, oxalic acid, gluconic acid, glucaric acid, glucuronic acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, salicylic acid, 4-aminosalicylic acid, 2-phenoxybenzoic acid, 2-acetoxybenzoic acid, embonic acid, nicotinic acid or isonicotinic acid; and with the 20 alpha amino acids involved in the synthesis of natural proteins, such as glutamic acid or aspartic acid, and also phenylacetic acid, methanesulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, ethane 1,2-disulfonic acid, benzenesulfonic acid, 4-methylbenzenesulfonic acid, naphthalene 2-sulfonic acid, naphthalene 1,5-disulfonic acid, 2- or 3-phosphoglyceric acid, glucose 6-phosphate, N-cyclohexylsulfamic acid (involving the formation of cyclamate) or salts with other acid organic compounds such as ascorbic acid.

[0083] Pharmaceutical compositions containing the compounds disclosed herein can be prepared by conventional methods, for example, by conventional mixing, dissolving, granulating, sugar coating, powdering, emulsifying, encapsulating, entrapping or lyophilization processes. Suitable formulations depend on the chosen route of administration.

[0084] For oral administration, suitable compositions can be readily formulated by combining the compounds disclosed herein with pharmaceutically acceptable excipients such as carriers well known in the art. Such excipients and carriers enable the compounds of the invention to be formulated as tablets, pills, dragees, capsules, solutions, gels, syrups, slurries, suspensions, etc. for oral ingestion by the patient being treated. Pharmaceutical preparations for oral use can be obtained by a process which comprises adding the compounds disclosed herein together with a solid excipient, optionally grinding the resulting mixture, and treating the granule mixture, after adding suitable auxiliaries if necessary, to obtain tablets or dragee cores. Suitable excipients include, for example, fillers and cellulose preparations. Disintegrants can be added if necessary. Pharmaceutically acceptable components are well known for various types of formulations and include, for example, binders (e.g., natural or synthetic polymers), lubricants, surfactants, sweeteners and flavorants, coating agents, preservatives, dyes, thickeners, adjuvants, antibacterial agents, antioxidants, and carriers for various formulation types.

[0085] When a therapeutically effective amount of the compounds disclosed herein is administered orally, the composition is typically in the form of a solid (e.g., tablet, capsule, pill, powder or troche) or a liquid formulation (e.g., aqueous suspension, solution, elixir or syrup).

[0086] When administered in tablet form, the composition can further comprise a functional solid and / or a functional solid carrier such as gelatin or an adjuvant. Tablets, capsules and powders can contain from about 1% to about 95% of the compound, preferably from about 15% to about 90% of the compound.

[0087] When administered in the form of a solution or suspension, functional liquids and / or functional liquid carriers such as water, petroleum, or oils of animal or plant origin can be added. The solution form of the composition can further comprise an aqueous physiological saline solution, a sugar alcohol solution, a dextrose or other saccharide solution, or a glycol. When administered in the form of a solution or suspension, the composition can comprise from about 0.5 to about 90% by weight of the compounds disclosed herein, preferably from about 1 to about 50% of the compounds disclosed herein. In one possible embodiment, the liquid carrier is non-aqueous or substantially non-aqueous. For administration in liquid form, the composition can be provided as a rapidly soluble solid formulation that dissolves or suspends immediately prior to administration.

[0088] When a therapeutically effective amount of the compounds disclosed herein is administered by intravenous, dermal, or subcutaneous injection, the composition is in the form of a pyrogen-free parenterally acceptable aqueous solution. The preparation of such parenterally acceptable solutions, with due consideration of pH, isotonicity, stability, etc., is within the scope of the art. Preferred compositions for intravenous, dermal, or subcutaneous injection typically comprise an isotonic vehicle in addition to the compounds disclosed herein. Such compositions can be prepared for administration as a solution of the free base or a pharmaceutically acceptable salt in water, appropriately mixed with a surfactant such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof, and in oils. Under normal conditions of storage and use, these preparations can optionally contain preservatives to prevent the growth of microorganisms.

[0089] Injectable compositions can include sterile aqueous solutions, suspensions or dispersions and sterile powders for immediate preparation of sterile injectable solutions, suspensions or dispersions. In all embodiments, this form must be sterile and fluid to the extent of having easy syringeability. It must be stable under manufacturing and storage conditions and, optionally by including preservatives, must counter the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium including, for example, water, ethanol, polyols (such as glycerol, propylene glycol and liquid polyethylene glycols, etc.), suitable mixtures thereof and vegetable oils. In one contemplated embodiment, the carrier is non-aqueous or substantially non-aqueous. Appropriate fluidity can be maintained, for example, by the use of coating agents such as lecithin, by maintaining the particle size of the compounds required in the dispersion embodiments and by the use of surfactants. Prevention of microbial action can be brought about by various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, etc. In many embodiments, it is preferred to include an isotonic agent such as sugar or sodium chloride. Sustained absorption of the injectable composition can be brought about by the use in the composition of agents that delay absorption such as aluminum monostearate and gelatin.

[0090] Sterile injectables are prepared by incorporating the required amount of the active compound into a suitable solvent, along with the various other necessary ingredients listed above, followed by filtration sterilization. Generally, dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle containing a basic dispersion medium and the desired other ingredients from those listed above. In the embodiment of sterile powders for preparing sterile injectable solutions, the preferred methods of preparation are vacuum drying techniques and freeze-drying techniques where the powder with any additional desired ingredients added to the active ingredient is obtained from its pre-sterilized solution.

[0091] Sustained or controlled release formulations can also be prepared to effect a controlled release of an active compound in contact with body fluids in the gastrointestinal tract and to provide a substantially constant and effective level of the active compound in the plasma. For example, release can be controlled by one or more of dissolution, diffusion, and ion exchange. Further, the controlled release approach can enhance absorption via a saturable or restricted pathway in the gastrointestinal tract. For example, the compound can be encapsulated in a polymeric matrix of a biodegradable polymer, a water-soluble polymer, or a mixture of both, optionally with a suitable surfactant. Encapsulation can, in this context, mean incorporating microparticles into the matrix of the polymer. Controlled release formulations can also be obtained by encapsulation of dispersed microparticles or emulsified microdroplets via known dispersion or emulsion coating techniques.

[0092] For administration by inhalation, the compounds of the invention are conveniently delivered in the form of an aerosol spray from a pressurized pack or a nebulizer using a suitable propellant. In the case of a pressurized aerosol, the dosage unit can 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 can be formulated to contain a powder mixture of the compound and a suitable powder base such as lactose or starch.

[0093] The compounds disclosed herein can be formulated for parenteral administration (e.g., by bolus injection or continuous infusion). Formulations for injection can be presented in unit dosage form (e.g., in ampoules or multi-dose containers) with added preservatives. The composition can take such forms as suspensions, solutions, or emulsions in oily or aqueous vehicles, and can contain formulating agents such as suspending, stabilizing, and / or dispersing agents.

[0094] Pharmaceutical preparations for parenteral administration include an aqueous solution of the compound in water-soluble form. Further, a 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. The aqueous injection suspension can contain substances that increase the viscosity of the suspension. Optionally, the suspension can also contain suitable stabilizers or agents that increase the solubility of the compound and enable the preparation of highly concentrated solutions. Alternatively, the compositions of the present invention can be in powder form for constitution with a suitable vehicle (e.g., sterile pyrogen-free water) before use.

[0095] 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 above, the compounds can also be formulated as depot preparations. Such long-acting preparations can be administered by injection (e.g., subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the compounds can be formulated with suitable polymers or hydrophobic materials (e.g., as an emulsion in an acceptable oil) or ion exchange resins, or can be formulated as poorly soluble derivatives, e.g., as poorly soluble salts.

[0096] In particular, the compounds disclosed herein can be administered orally, buccally or sublingually in the form of tablets containing excipients such as starch or lactose, or in capsules or ovules alone or in admixture with excipients, or in the form of elixirs or suspensions containing flavoring or coloring agents. Such liquid preparations can be prepared using pharmaceutically acceptable additives such as suspending agents. The compounds can also be administered parenterally, for example, by intravenous, intramuscular, subcutaneous or intracoronary injection. For parenteral administration, the compounds are most often used in the form of a sterile aqueous solution that can contain other substances, such as salts or sugar alcohols such as mannitol or glucose, to render the solution isotonic with blood.

[0097] For veterinary use, the compounds disclosed herein are administered as a suitably acceptable formulation in accordance with normal veterinary practice. A veterinarian can readily determine the most appropriate dosing regimen and route of administration for a particular animal.

[0098] In some embodiments, for the treatment of KIF18A-related disorders, the compounds disclosed herein are used alone or in combination with another agent or intervention conventionally used for the treatment of such a disorder. All the components necessary for such treatment can be packaged in a kit. Specifically, the present invention provides a kit for use in a therapeutic intervention for a disease, comprising a compound disclosed herein, an agent comprising a buffer and other components for preparing a deliverable form of the agent, and / or an instrument for delivering such an agent, and / or any agent used in combination therapy with a compound disclosed herein, and / or a packaged set of instructions for the treatment of a disease packaged with the agent. The instructions can be fixed on any tangible medium such as printed paper or a computer-readable magnetic or optical medium, or can be instructions that refer to a data source of a remote computer such as a web page of the World Wide Web accessible via the Internet.

[0099] "Therapeutically effective amount" means an amount effective to treat, prevent the progression of, or alleviate existing symptoms in a subject being treated. Determination of an effective amount is well within the ability of one of ordinary skill in the art, especially in light of the detailed disclosure provided herein. Generally, "therapeutically effective dose" refers to the amount of a compound that produces the desired effect. For example, in a preferred embodiment, a therapeutically effective amount of a 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.

[0100] The amount of the compound administered may depend on the subject being treated, the subject's age, health, sex and weight, the type of co-treatment (if any), the severity of the disease, the nature of the desired effect, the mode 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. Individual requirements vary, but determination of the optimal range of the effective amount of the compound is within the skill of the art. Such dosages may be administered as a single dose or divided into multiple doses.

[0101] As used herein, the terms "cancer" and "cancerous" typically refer to or describe a mammalian physiological condition characterized by unregulated cell growth. Examples of cancers 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, hepatocellular carcinoma, breast cancer, colon cancer, and head and neck cancer, ovarian cancer, and endometrial cancer. The term "cancer" as used herein is not limited to any one specific form of disease, but the methods of the present invention are particularly effective for cancers that have been found to be associated with an unregulated level of KIF18A or that are dependent on KIF18A for proper chromosome segregation and survival in mammals.

[0102] As used herein, the terms "treat", "treating", and "treatment" refer to therapies including, but not limited to, curative therapies, prophylactic therapies, and palliative therapies. Prophylactic treatment generally comprises either preventing the onset of the overall disorder in an individual or delaying the onset of the disorder at a stage prior to its clinical manifestation.

[0103] As used herein, the terms "patient", "subject", or "mammal" refer to any "patient", "subject", or "mammal" including humans, cows, horses, dogs, and cats. In one embodiment of the invention, the mammal is a human.

[0104] The term "comprising" means that the indicated components are included, but it is an open-ended one that does not exclude other elements.

[0105] The term "Formula I" includes any sub-formula.

[0106] Methods of using KIF18A inhibitors The present disclosure generally provides compounds having MT-based KIF18A regulatory activity, specifically inhibitory activity. In one embodiment of the invention, a method of modulating KIF18A protein in a subject is provided, the method comprising administering to the subject an effective dosage of a compound of Formula I. Thus, the compounds of the present invention can be used in the treatment of cell proliferation disorders including uncontrolled cell growth, abnormal cell cycle regulation, and centrosome abnormalities (structural and / or numerical, fragmentation). Other diseases or disorders associated with the accumulation of extra centrosomes (>2) include human papillomavirus (HPV) infections including HPV-related neoplasia. The compounds can also be useful for cilia-related diseases and for excising the haploid germ cell population that can be used as male contraceptives.

[0107] In addition, the compounds of the present invention are useful for preventing or treating, but not limited to, cancer and other KIF18A-mediated diseases or disorders. For example, the compounds of the present invention are useful for treating various solid and blood-derived tumors such as carcinomas including 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 lymphatic system (including leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B cell lymphoma, T cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, hairy cell lymphoma, and Burkitt lymphoma); hematopoietic tumors of the myeloid cell lineage (including acute and chronic myeloid leukemia, myelodysplastic syndrome, and promyelocytic leukemia); mesenchymal-derived tumors (including fibrosarcoma and rhabdomyosarcoma, and other sarcomas including soft tissue and bone); tumors of the central and peripheral nervous systems (including astrocytoma, neuroblastoma, glioma, and schwannoma); and other tumors (including melanoma, seminoma, teratoma, osteosarcoma, xeroderma pigmentosum, keratoacanthoma, follicular thyroid cancer, and Kaposi sarcoma).

[0108] The compounds of the present invention are also useful for treating cancer-related indications such as solid tumors, sarcomas (particularly Ewing sarcoma and osteosarcoma), retinoblastoma, rhabdomyosarcoma, neuroblastoma, hematopoietic malignancies including leukemia and lymphoma, tumor-induced pleural or pericardial effusion, and malignant ascites.

[0109] Based on the ability to modulate kinesins that affect angiogenesis, the compounds of the present invention are also useful for the treatment and therapy of proliferative diseases. Specifically, these compounds are useful for the treatment of various inflammatory rheumatic-like diseases, particularly symptoms in the locomotor organs, in particular, rheumatoid arthritis, juvenile arthritis, or psoriatic arthritis including chronic polyarthritis; paraneoplastic syndromes or tumor-induced inflammatory diseases, turbid exudates, collagen diseases such as systemic lupus erythematosus, polymyositis, dermatomyositis, systemic sclerosis, or mixed collagen disease; post-infectious arthritis (where no pathogen is found in or within the affected part of the body), seronegative spondyloarthritis such as ankylosing spondylitis; vasculitis, sarcoidosis or arthrosis; or any further arbitrary combination thereof, etc., and can be used for the treatment of inflammatory diseases.

[0110] The compounds of the present invention can also be used as active agents for conditions such as arthritis, atherosclerosis, psoriasis, hemangioma, myocardial angiogenesis, coronary and cerebral collateral arteries, ischemic peripheral angiogenesis, wound healing, Helicobacter pylori-related diseases of peptic ulcers, fractures, cat scratch fever, rubellosis, neovascular glaucoma and retinopathy such as those 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 such as thyroid hyperplasia (particularly Graves' disease), and cysts (e.g., vascular hyperplasia of ovarian stroma, a characteristic of polycystic ovary syndrome (Stein-Leventhal syndrome)), because these diseases require the proliferation of vascular cells for growth and / or metastasis.

[0111] In addition to being useful for the treatment of humans, these compounds are useful for veterinary treatment of companion animals, exotic animals and livestock including mammals, rodents, etc. For example, animals including horses, dogs, and cats can be treated with the compounds provided by the present invention.

[0112] Combination The compounds of the present invention can be taken or administered as a single active pharmaceutical, but these may be used in combination with one or more compounds of the present invention, or in combination with other agents. When administered as a combination agent, the therapeutic agent may be formulated as separate compositions to be administered sequentially at the same time or at different times, or the therapeutic agent may be given as a single composition.

[0113] In defining the use of the compounds of the present invention and another pharmaceutical, the term "co-therapy" (or "combination-therapy") encompasses the administration of each agent sequentially in a dosing regimen that provides a beneficial effect of the combination of agents, and also encompasses the co-administration of these agents substantially simultaneously, such as in a single capsule having a fixed ratio of these active agents or in individual capsules of each of the plurality of agents.

[0114] Specifically, the administration of the compounds of the present invention may be combined with additional therapies known to those skilled in the art in the field of cancer prevention or treatment, using radiotherapy, small molecule targeting agents (e.g., PARP inhibitors, kinase inhibitors), therapeutic antibodies (e.g., naked and drug conjugates), immunotherapy antibodies (checkpoint inhibitors, bispecific T cell engagers), and anti-neoplastic or cytotoxic agents.

[0115] When formulated as a fixed dose, such combination products use the compounds of the present invention within the acceptable dosage range. If the combination formulation is inappropriate, the compound of formula I may also be administered sequentially with a known anti-cancer or cytotoxic agent. The present invention is not limited in the order of administration, and the compounds of the present invention may be administered before, simultaneously with, or after the administration of a known anti-cancer or cytotoxic agent.

[0116] There are numerous anti-cancer agents available for commercial use, clinical evaluation, and pre-clinical development that are selected for the treatment of neoplasia by combination drug chemotherapy. Such agents are divided into several major classifications such as antibiotic-type agents, alkylating and alkylating-like agents, anti-mitotic agents, targeted small molecule agents, antimetabolites, hormonal agents, immunological agents, anti-angiogenic agents, interferon-type agents, and other agent classifications.

[0117] The present disclosure also provides methods of combination therapy in which agents that are known to modulate other pathways or other components of the same pathway or further overlapping sets of target enzymes are used in combination with the compounds of the present disclosure or pharmaceutically acceptable salts thereof. In one aspect, such treatments include, but are not limited to, combinations of one or more compounds of the present disclosure with chemotherapeutic agents, therapeutic antibodies, targeted small molecule agents, and radiation therapy to provide synergistic or additive therapeutic effects.

[0118] Currently, many chemotherapeutic agents are 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 an anti-mitotic agent, an alkylating agent, an antimetabolite, an intercalating antibiotic, a growth factor inhibitor, a cell cycle inhibitor, an enzyme, a topoisomerase inhibitor, a biological response modifier, an antihormonal agent, an angiogenesis inhibitor, and an antiandrogen agent. Non-limiting examples include chemotherapeutic agents, cytotoxic agents, and non-peptide small molecules (e.g., Gleevec® (imatinib mesylate), Kyprolis® (carfilzomib), Velcade® (bortezomib), Casodex (bicalutamide), Iressa® (gefitinib), and adriamycin), and are hosts for 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, carboquone, meturedopa, and uredopa; ethyleneimine and methylamelamine including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolomelamine; nitrogen mustards such as chlorambucil, chloronaphazine, chlorophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembicin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine;Antibiotics such as aclacinomycin, actinomycin, aurostatin, azaserine, bleomycin, cactinomycin, calicheamicin, carabicin, calminomycin, cardinophilin, Casodex (trademark), chromomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin, idarubicin, marcellomycin, mitomycin, mycophenolic acid, nogalamycin, olivomycin, peplomycin, potfiromycin, puromycin, keramycin, rhodomycin, streptozocin, streptozocin, tubercidin, ubenimex, dinostatin, zorubicin; Antimetabolites such as methotrexate and 5-fluorouracil (5-FU); Folic acid analogs such as denopterin, methotrexate, pteropterin, trimethoprim; Purine analogs such as fludarabine, 6-mercaptopurine, thiampurine, thioguanine; Pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, didoxuridine, doxifluridine, enocitabine, floxuridine; Androgens such as calusterone, drostanolone propionate, epithiostanol, mepithiostane, testolactone; Anti-adrenals such as aminoglutethimide, mitotane, trilostane; Folic acid supplements such as folinic acid; Aceglatone; Aldophosphamide glycoside; Aminolevulinic acid; Amsacrine; Bestrabucil; Bisantrene; Edatraxate; Defofamine; Dexamethasone; Diazquone; Elfornithine; Elliptinium acetate; Etoglucid; Gallium nitrate; Hydroxyurea; Lentinan; Lonidamine; Mitoguazone; Mitoxantrone; Mopidamol; Nitracrine; Pentostatin; Phenamet; Pirarubicin; Podophyllinic acid; 2-ethylhydrazide; Procarbazine; PSK; Razoxane; Schizophyllan; Spirogermanium; Tenuazonic acid; Triaziquone; 2,2’,2’’-trichloroethylamine; Urethane; Vinblastine; Dacarbazine; Mannomustine; Mitobronitol; Mitolactol; Pipobroman; Gacitabine; Arabinoside (“Ara-C”); Cyclophosphamide; Thiotepa;Taxanes, such as paclitaxel and docetaxel, Nab-paclitaxel; retinoic acid; esperamicin; capecitabine; and any pharmaceutically acceptable salts, acids or derivatives thereof are included.

[0119] Also included as suitable chemotherapy cell conditioners are antihormonal agents, such as antiestrogens that act to modulate or inhibit the hormonal action on tumors, for example, tamoxifen, (Nolvadex (trademark)), raloxifene, aromatase inhibitor 4(5)-imidazole, 4-hydroxytamoxifen, trioxifene, keoxifene, LY 117018, onapristone and toremifene (Fareston); and antiandrogens such as flutamide, nilutamide, bicalutamide, leuprolide and goserelin; chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin, oxaliplatin, carboplatin; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vinblastine; vincristine; vinorelbine; navelbine; novantrone; teniposide; daunomycin; aminopterin; xeloda; ibandronate; topotecan; camptothecin-11 (CPT-11); topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO).

[0120] Optionally, the compounds or pharmaceutical compositions of the present disclosure can be used in combination with commonly prescribed anti-cancer drugs such as Herceptin®, Avastin®, Erbitux®, Rituxan®, Taxol®, Abraxane, Arimidex®, Taxotere®, ABVD, AVICINE, abagovomab, acridine carboxamide, adecatumumab, 17-N-allylamino-17-demethoxygeldanamycin, alpharadin, arbociclib, 3-aminopyridine-2-carboxaldehyde thiosemicarbazone, amonafide, anthracenedione, anti-CD22 immunotoxin, anti-neoplastic agent, anti-tumor herb, apaziquone, atiprimod, azathioprine, belotecan, bendamustine, BIBW2992, bilicodar, brostallicin, bryostatin, buthionine sulfoximine, CBV (chemotherapy), caliculin, cell cycle non-specific anti-neoplastic agent, dichloroacetic acid, discodermolide, elsamitrucin, enocitabine, epothilone, eribulin, everolimus, exatecan, exisulind, feruginol, foldecsin, phosphoestrol, ICE chemotherapy regimen, IT-101, imexon, imiquimod, indolocarbazole, irofulven, lanquidar, larotaxel, lenalidomide, lucanthone, lutotecan, mafosfamide, mitozolomide, naphthoxidine, nedaplatin, olaparib, talazoparib, niraparib, ortataxel, PAC-1, porfiromycin, pixantrone, proteasome inhibitor, rebeccamycin, resiquimod, rubitecan, SN-38, salinosporamide A, sapacitabine, Stanford V, swainsonine, talaporfin, talquidar, tegafur-uracil, temodal, tesetaxel, triplatin tetranitrate, tris(2-chloroethyl)amine, troxacitabine, uramustine, vadimezan, vinflunine, ZD6126 or zosuquidar, CDK4 / 6 inhibitors (palbociclib, Ibrance; ribociclib, Kisqali; abemaciclib, Verzenio).

[0121] The present disclosure further relates to a method of combining a compound or pharmaceutical composition provided herein with radiation therapy to inhibit abnormal cell growth in a mammal or to treat a proliferative disorder. Techniques for administering radiation therapy are known in the art and those techniques can be used in the combination therapies described herein. Administration of the compounds of the present disclosure in this combination therapy can be determined as described herein.

[0122] Radiation therapy can be administered by one or a combination of several methods including, but not limited to, external beam therapy, internal radiation therapy, interstitial irradiation, stereotactic radiosurgery, total body radiation therapy, radiotherapy, and permanent or temporary brachytherapy. As used herein, the term "brachytherapy" refers to radiation therapy delivered by a spatially restricted radioactive material inserted into the body at or near a tumor or other proliferative tissue disease site. This term is intended to include, but not be limited to, exposure to radioisotopes (e.g., radioisotopes of At-211, I-131, I-125, Y-90, Re-186, Re-188, Sm153, Bi-212, P-32, and Lu). Suitable radiation sources for use as cell conditioners in the present disclosure include both solids and liquids. By way of non-limiting example, the radiation source can be a radioisotope such as I-125, I-131, Yb-169, Ir-192 as a solid source, or other radioisotopes that emit radioactive nuclides or photons, beta particles, gamma rays, or other therapeutic rays such as I-125 as a solid source. The radioactive material can be a fluid made from any solution of a radioisotope, such as a solution of I-125 or I-131, or the radioactive fluid can be produced using a suitable fluid slurry containing microparticles of a solid radioisotope such as Au-198, Y-90. Additionally, the radioisotope can be embodied as a gel or radioactive microspheres.

[0123] The compounds or pharmaceutical compositions of the present disclosure can be used in combination with a certain amount of one or more substances selected from anti-angiogenic agents, signal transduction inhibitors, anti-proliferative agents, glycolysis inhibitors, or autophagy inhibitors.

[0124] Anti-angiogenic agents such as MMP-2 (matrix metalloproteinase 2) inhibitors, MMP-9 (matrix metalloproteinase 9) inhibitors, and COX-11 (cyclooxygenase 11) inhibitors can be used in combination with the disclosed compounds and pharmaceutical compositions described herein. Anti-angiogenic agents include, for example, rapamycin, temsirolimus (CCI-779), everolimus (RAD001), sorafenib, sunitinib, and bevacizumab. Examples of useful COX-II inhibitors include celecoxib, valdecoxib, and rofecoxib. Examples of useful matrix metalloproteinase inhibitors are described in WO 96 / 33172, WO 96 / 27583, EP 0818442, EP 1004578, WO 98 / 07697, WO 98 / 03516, WO 98 / 34918, WO 98 / 34915, WO 98 / 33768, WO 98 / 30566, EP 606046, EP 931788, WO 90 / 05719, WO 99 / 52910, WO 99 / 52889, WO 99 / 29667, WO 99007675, EP 1786785, EP 1181017, US 20090012085, US 5863949, US 5861510, and EP 0780386, all of which are incorporated herein by reference in their entirety. Preferred MMP-2 and MMP-9 inhibitors have little or no activity to inhibit MMP-1. More preferably, they selectively inhibit MMP-2 and / or MMP-9 with respect to other matrix metalloproteinases (i.e., MMP-1, MMP-3, MMP-4, MMP-5, MMP-6, MMP-7, MMP-8, MMP-10, MMP-11, MMP-12, and MMP-13).Some specific examples of MMP inhibitors useful in the present disclosure are AG-3340, RO32-3555, and RS13-0830.

[0125] The compounds of the present invention are acemannan, aclarubicin, aldeferon, alemtuzumab, alitretinoin, altretamine, amifostine, aminolevulinic acid, amrubicin, amsacrine, anagrelide, anastrozole, ANCER, ansetim, ARGLABIN, arsenic acid, BAM002 (Novelos), bexarotene, bicalutamide, broxuridine, capecitabine, sermolukin, cetrorelix, cladribine, clotrimazole, cytarabine ocfosfate, DA3030 (Dong-A), daclizumab, denileukin diftitox, deslorelin, dexrazoxane, dirazep, docetaxel, docosanol, doxercalciferol, doxifluridine, doxorubicin, bromocriptine, carmustine, cytarabine, fluorouracil, HIT diclofenac, interferon alpha, daunorubicin, doxorubicin, tretinoin, edelfosine, edrecolomab, eflornithine, emitefur, epirubicin, epoetin beta, etoposide phosphate, exemestane, exisulind, fadrozole, filgrastim, finasteride, fludarabine phosphate, formestane, fotemustine, gallium nitrate, gemcitabine, gemtuzumab ozogamicin, the combination of gimeracil / oteracil / tegafur, glycopyrronium, goserelin, heptaplatin, human chorionic gonadotropin, human fetal alpha-fetoprotein, ibandronic acid, idarubicin, (imiquimod, interferon alpha, interferon alpha, natural, interferon alpha-2, interferon alpha-2a, interferon alpha-2b, interferon alpha-N1, interferon alpha-n3, interferon alphacon-1, interferon alpha, natural, interferon beta, interferon beta-1a, interferon beta-1b, interferon gamma, natural interferon gamma-1a, natural interferon gamma-1b, interleukin-1 beta, iobenguane, irinotecan, ilsogladine, lanreotide, LC9018 (Yakult), leflunomide, lenograstim, lentinan sulfate, letrozole, leukocyte alpha interferon, leuprolide,Levamisole + Fluorouracil, Riarozole, Lobaplatin, Lonidamine, Lovastatin, Masoprocol, Melarsoprol, Metoclopramide, Mifepristone, Miltefosine, Milimostim, Mismatch Double-stranded RNA, Mitoguazone, Mitolactol, Mitoxantrone, Molgramostim, Nafarelin, Naloxone + Pentazocine, Nartograstim, Nedaplatin, Nilutamide, Noscapine, Novel Erythropoiesis-stimulating Protein, NSC631570 Octreotide, Oprelvekin, Osaterone, Oxaliplatin, Paclitaxel, Pamidronic Acid, Pegaspargase, Peg Interferon Alpha-2b, Pentosan Polysulfate Sodium, Pentostatin, Picibanil, Pirarubicin, Rabbit Anti-thymocyte Polyclonal Antibody, Polyethylene Glycol Interferon Alpha-2a, Porfimer Sodium, Raloxifene, Raltitrexed, Rasburicase, Rhenium Re186 Etidronate, RII Retinamid, Rituximab, Romurtide, Samarium (153Sm) Lexidronam, Sargramostim, Schizophyllan, Sobuzoxane, Sonermin, Strontium-89 Chloride, Suramin, Tasonermin, Tazarotene, Tegafur, Temoporfin, Temozolomide, Teniposide, Tetrachlorodecaoxide, Thalidomide, Timalphasin, Thyrotropin Alpha, Topotecan, Toremifene, Tositumomab-Iodine 131, Trastuzumab, Treosulfan, Tretinoin, Trilostane, Trimethoprim, Triptorelin, Tumor Necrosis Factor Alpha, Natural, Ubenimex, Bladder Cancer Vaccine, Maruyama Vaccine, Melanoma Lysate Vaccine, Barbisine, Verteporfin, Vinorelbine, VIRULIZIN, Dinostatin Stimalamer or Zoledronic Acid; Abarelix; AE941 (Aeterna), Ambamustine, Antisense Oligonucleotide, bcl-2 (Genta), APC8015 (Dendreon), Cetuximab, Decitabine, Dexaminoglutethimide, Diadicon, EL532 (Elan), EM800 (Endorecherche), Eniluracil, Ethanidazole, Fenretinide, Filgrastim SD01 (Amgen), Fulvestrant, Galocitabine, Gastrin 17 Immunogen,HLA-B7 gene therapy (Vical), granulocyte macrophage colony-stimulating factor, histamine dihydrochloride, ibritumomab tiuxetan, iromostat, IM862 (Cytran), interleukin 2, ibuprofen, LDI200 (Milkhaus), leridistim, lym-1-iodine 131 MAb (Techniclone), polymorphic epithelial mucin yttrium 90 MAb (Antisoma), marimastat, menogaril, mitumomab, motexafin gadolinium, MX6 (Galderma), nelarabine, nolatrexed, P30 protein, pegvisomant, pemetrexed, porfiromycin, prinomastat, RL0903 (Shire), rubitecan, satraplatin, sodium phenylacetate, strontium ranelate, SRL172 (SR Pharma), SU5416 (SUGEN), TA077 (Tanabe), tetrathiomolybdate, taliblastine, thrombopoietin, tin ethyl etiopurpurin, tirapazamine, cancer vaccine (Biomira), 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 it may also be used in combination therapy with other antineoplastic agents such as balsopodar.

[0126] The compounds of the present invention can be further used in combination with a VEGFR inhibitor. Other compounds described in the following patents and patent applications can be used in combination therapies: U.S. Patent No. 6,258,812, U.S. Patent Application Publication No. 2003 / 0105091, International Publication No. 01 / 37820 Pamphlet, U.S. Patent No. 6,235,764, International Publication No. 01 / 32651 Pamphlet, U.S. Patent No. 6,630,500, U.S. Patent No. 6,515,004, U.S. Patent No. 6,713,485, U.S. Patent No. 5,521,184, U.S. Patent No. 5,770,599, U.S. Patent No. 5,747,498, International Publication No. 02 / 68406 Pamphlet, International Publication No. 02 / 66470 Pamphlet, International Publication No. 02 / 55501 Pamphlet, International Publication No. 04 / 05279 Pamphlet, International Publication No. 04 / 07481 Pamphlet, International Publication No. 04 / 07458 Pamphlet, International Publication No. 04 / 09784 Pamphlet, International Publication No. 02 / 59110 Pamphlet, International Publication No. 99 / 45009 Pamphlet, International Publication No. 00 / 59509 Pamphlet, International Publication No. 99 / 61422 Pamphlet, U.S. Patent No. 5,990,141, International Publication No. 00 / 12089 Pamphlet, and International Publication No. 00 / 02871 Pamphlet.

[0127] In some embodiments, the combination drug comprises a composition of the present invention combined with at least one anti-angiogenic agent. The agents include, but are not limited to, chemically synthesized compositions, antibodies, antigen-binding regions, radionuclides, and combinations and conjugates thereof prepared in vitro. The agents can be agonists, antagonists, allosteric regulators, toxins, or more generally, can act to inhibit or stimulate their targets (e.g., activation or inhibition of a receptor or enzyme), thereby promoting cell death or arresting cell growth.

[0128] Exemplary anti-angiogenic agents include ERBITUX (trademark) (IMC-C225), KDR (kinase domain receptor) inhibitors (e.g., antibodies and antigen-binding regions that specifically bind to the kinase domain receptor), anti-VEGF agents such as AVASTIN (trademark) or VEGF-TRAP (trademark) (e.g., antibodies or antigen-binding regions that specifically bind to VEGF or soluble VEGF receptor or its ligand-binding region), and anti-VEGF receptor drugs (e.g., antibodies or antigen-binding regions that specifically bind thereto), Vectibix (panitumumab), IRESSA (trademark) (gefitinib), TARCEVA (trademark) (erlotinib) and other EGFR inhibitors (e.g., antibodies or antigen-binding regions that specifically bind thereto), anti-Ang1 and anti-Ang2 agents (e.g., to them, or their receptors, e.g., Tie2 / Tek), and anti-Tie2 kinase inhibitors (e.g., antibodies or antigen-binding regions that specifically bind thereto). The pharmaceutical composition of the present invention may also include one or more agents that specifically bind and inhibit the activity of growth factors (e.g., antibodies, antigen-binding regions or soluble receptors), for example, antagonists and antibodies or antigen-binding regions of hepatocyte growth factor (HGF, also known as scatter factor) that specifically binds to the receptor "c-met".

[0129] Other anti-angiogenic agents include Campath, IL-8, B-FGF, Tek antagonists (Ceretti et al., US Patent Application Publication No. 2003 / 0162712; US Patent No. 6,413,932), anti-TWEAK agents (e.g., specifically binding antibodies or antigen-binding regions, or soluble TWEAK receptor antagonists; see Wiley, US Patent No. 6,727,225), ADAM disintegrin domains that antagonize the binding of integrin to its ligand (Fanslow et al., US Patent Application Publication No. 2002 / 0042368), anti-eph receptor and / or anti-ephrin antibodies or antigen-binding regions that specifically bind (US Patent No. 5,981,245; US Patent No. 5,728,813; US Patent No. 5,969,110; US Patent No. 6,596,852; US Patent No. 6,232,447; US Patent No. 6,057,124, and members of their patent families), and anti-PDGF-BB antagonists (e.g., specifically binding antibodies or antigen-binding regions), as well as antibodies or antigen-binding regions that specifically bind to the PDGF-BB ligand, and PDGFR kinase inhibitors (e.g., specifically binding antibodies or antigen-binding regions thereto).

[0130] Additional anti-angiogenic / anti-tumor agents include the following: SD-7784 (Pfizer, USA); Sirengetide (Merck KGaA, Germany, EPO 770622); Pegaptanib octasodium (Gilead Sciences, USA); Alphastatin (BioActa, UK); M-PGA (Celgene, USA, US Patent No. 5,712,291); Irormastat (Arriva, USA, US Patent No. 5,892,112); Emaxanib (Pfizer, USA, US Patent No. 5,792,783); Batranib (Novartis, Switzerland); 2-Methoxyestradiol (EntreMed, USA); TLC ELL-12 (Elan, Ireland); Anecortave acetate (Alcon, USA); α-D148Mab (Amgen, USA); CEP-7055 (Cephalon, USA); Anti-Vn Mab (Crucell, Netherlands) DAC: Anti-angiogenic agent (ConjuChem, Canada); Angiostatin (InKine Pharmaceutical, USA); KM-2550 (Kyowa Hakko, Japan); SU-0879 (Pfizer, USA); CGP-79787 (Novartis, Switzerland, European Patent No. 970,070); ARGENT technology (Ariad, USA); YIGSR-Stealth (Johnson & Johnson, USA); Fibrinogen E fragment (BioActa, UK); Angiogenesis inhibitor (Trigen, UK); TBC-1635 (Encysive Pharmaceuticals, USA); SC-236 (Pfizer, USA); ABT-567 (Abbott, USA); Metastatin (EntreMed, USA); Angiogenesis inhibitor (Tripep, Sweden); Maspin (Sosei, Japan); 2-Methoxyestradiol (Oncology Sciences Corporation, USA); ER-68203-00 (IVAX, USA); Benefin (Lane Labs, USA); Tz-93 (Tsumura, Japan); TAN-1120 (Takeda, Japan);FR-111142 (Fujisawa, Japan; Japanese Patent Publication No. 02233610); Platelet Factor 4 (RepliGen, USA; European Patent No. 407122); Vascular Endothelial Growth Factor Antagonist (Borean, Denmark); Bevacizumab (pINN) (Genentech, USA); Angiogenesis Inhibitor (SUGEN, USA); XL784 (Exelixis, USA); XL647 (Exelixis, USA); MAb, α5β3 Integrin, Second Generation (Applied Molecular Evolution, USA and MedImmune, USA); Gene Therapy, Retinopathy (Oxford BioMedica, UK); Enzastaurin Hydrochloride (USAN), (Lilly, USA); CEP7055 (Cephalon, USA and Sanofi-Synthelabo, France); BC1 (Genoa Institute of Cancer Research, Italy); Angiogenesis Inhibitor (Alchemia, Australia); VEGF Antagonist (Regeneron, USA); rBPI21 and BPI-derived Anti-angiogenic Agent (XOMA, USA); PI88 (Progen, Australia); Sirengeptide (pINN), (Merck KGaA,; Munich Technical University, Scripps Clinic and Research Foundation, USA); Cetuximab (INN), (Aventis, France); AVE8062 (Ajinomoto, Japan); AS1404 (Cancer Research Laboratory, New Zealand); SG292 (Telios, USA); Endostatin (Boston Children's Hospital, USA); ATN161 (Attenuon, USA); Angiostatin (Boston Children's Hospital, USA); 2-Methoxyestradiol (Boston Children's Hospital, USA); ZD6474 (AstraZeneca, UK); ZD6126 (Angiogene Pharmaceuticals, UK); PPI2458 (Praecis, USA); AZD9935 (AstraZeneca, UK);AZD2171 (AstraZeneca, UK); Batranib (pINN) (Novartis, Switzerland and Schering AG, Germany); Tissue Factor Pathway Inhibitor (EntreMed, USA); Pegaptanib (Pinn) (Gilead Sciences, USA); Xantholol (Yonsei University, South Korea); Gene-based Vaccine, VEGF-2 (Scripps Clinic and Research Foundation, USA); SPV5.2 (Supratek, Canada); SDX103 (University of California, San Diego, USA); PX478 (ProlX, USA); Metastatin (EntreMed, USA); Troponin I (Harvard University, USA); SU6668 (SUGEN, USA); OXI4503 (OXiGENE, USA); o-Guanidine (Dimensional Pharmaceuticals, USA); Motuporamine C (British Columbia University, Canada); CDP791 (Celltech Group, UK); Actiprimod (pINN) (GlaxoSmithKline, UK); E7820 (Eisai, Japan); CYC381 (Harvard University, USA); AE941 (Aeterna, Canada); Vaccine, Angiogenesis (EntreMed, USA); Urokinase Plasminogen Activator Inhibitor (Dendreon, USA); Oglufanide (pINN) (Melmotte, USA); HIF-1 Alpha Inhibitor (Xenova, UK); CEP5214 (Cephalon, USA); BAY RES2622 (Bayer, Germany); Angiostatin (InKine, USA); A6 (Angstrom, USA); KR31372 (Korea Research Institute of Chemical Technology, South Korea); GW2286 (GlaxoSmithKline, UK); EHT0101 (ExonHit, France); CP868596 (Pfizer, USA);CP564959 (OSI, USA); CP547632 (Pfizer, USA); 786034 (GlaxoSmithKline, UK); KRN633 (Kirin Brewery, Japan); Drug delivery system, intraocular, 2-methoxyestradiol (EntreMed, USA); Anginex (Maastricht University, Netherlands and Minnesota University, USA); ABT510 (Abbott, USA); AAL993 (Novartis, Switzerland); VEGI (ProteomTech, USA); Tumor necrosis factor-α inhibitor (National Institute on Aging, USA); SU11248 (Pfizer, USA and SUGEN, USA); ABT518 (Abbott, USA); YH16 (Yantai Rongchang, China); S-3APG (Boston Childrens Hospital, USA and EntreMed, USA); MAb, KDR (ImClone Systems, USA); MAb, α5β1 (Protein Design, USA); KDR kinase inhibitor (Celltech Group, UK and Johnson&Johnson, USA); GFB116 (South Florida University, USA and Yale University, USA); CS706 (Sankyo, Japan); Combretastatin A4 prodrug (Arizona State University, USA); Chondroitinase AC (IBEX, Canada); BAY RES2690 (Bayer, Germany); AGM1470 (Harvard University, USA, Takeda, Japan and TAP, USA); AG13925 (Agouron, USA); Tetrathiomolybdate (University of Michigan, USA); GCS100 (Wayne State University, USA) CV247 (Ivy Medical, UK); CKD732 (Chong Kun Dang, South Korea); MAb, vascular endothelial growth factor (Xenova, UK); Iloprost (INN) (Nippon Shinyaku, Japan);RG13577 (Aventis, France); WX360 (Wilex, Germany); squalamine (pINN) (Genaera, USA); RPI4610 (Sirna, USA); cancer therapy (Marinova, Australia); heparinase inhibitor (InSight, Israel); KL3106 (Kolon, South Korea); honokiol (Emory University, USA); ZK CDK (Schering AG, Germany); ZK angio (ZK Angio) (Schering AG, Germany); ZK229561 (Novartis, Switzerland and Schering AG, Germany); XMP300 (XOMA, USA); VGA1102 (Taisho, Japan); VEGF receptor regulator (Pharmacopeia, USA); VE-cadherin-2 antagonist (ImClone Systems, USA); batostatin (National Institutes of Health, USA); vaccine, Flk-1 (ImClone Systems, USA); TZ93 (Tsumura, Japan); tamastatin (Beth Israel Hospital, USA); truncated soluble FLT1 (vascular endothelial growth factor receptor 1) (Merck & Co, USA); Tie-2 ligands (Regeneron, USA); and thrombospondin 1 inhibitor (Allegheny Health, Education and Research Foundation, USA).;

[0131] Examples of autophagy inhibitors include, but are not limited to, chloroquine, 3-methyladenine, hydroxychloroquine (Plaquenil™), bafilomycin A1, 5-amino-4-imidazolecarboxamide riboside (AICAR), okadaic acid, autophagy-inhibiting algal toxins that inhibit type 2A or type 1 protein phosphatases, cAMP analogs, and drugs that increase cAMP levels such as adenosine, LY204002, N6-mercaptopurine riboside, and vinblastine. In addition, antisense or siRNA that inhibits the expression of proteins, including but not limited to ATG5 (involved in autophagy), can also be used.

[0132] Additional pharmaceutically active compounds / drugs that can be used in the treatment of cancer and can be used in combination with one or more compounds of the present invention include epoetin α; darbepoetin α; panitumumab; pegfilgrastim; palifermin; filgrastim; denosumab; ancetastim; AMG102; AMG386; AMG479; AMG655; AMG745; AMG951; and AMG706, or pharmaceutically acceptable salts thereof.

[0133] In certain embodiments, the compositions provided herein are administered in combination with a chemotherapeutic agent. Suitable chemotherapeutic agents include natural products such as vinca alkaloids (e.g., vinblastine, vincristine, and vinorelbine), paclitaxel, epipodophyllotoxins (e.g., etoposide and teniposide), antibiotics (e.g., dactinomycin (actinomycin D), daunorubicin, doxorubicin, and idarubicin), anthracyclines, mitoxantrone, bleomycin, plicamycin (mithramycin), mitomycin, enzymes (e.g., L-asparaginase which metabolizes L-asparagine systemically and removes cells that do not have the ability to synthesize their own asparagine), antiplatelet agents, nitrogen mustards (e.g., mechlorethamine, cyclophosphamide and analogs, melphalan, and chlorambucil), ethyleneimine and methylmelamine (e.g., hexamethylmelamine and thiotepa), CDK inhibitors (e.g., seliciclib, UCN-01, P1446A-05, PD-0332991, dinaciclib, P27-00, AT-7519, RGB286638, and SCH727965), alkyl sulfonates (e.g., busulfan), nitrosoureas (e.g., carmustine (BCNU) and analogs, and streptozocin), temozolomide, folic acid analogs (e.g., methotrexate), pyrimidine analogs (e.g., fluorouracil, floxuridine, and cytarabine), purine analogs and related inhibitors (e.g., mercaptopurine, thioguanine, pentostatin, and 2-chlorodeoxyadenosine), aromatase inhibitors (e.g., anastrozole, exemestane, and letrozole), and platinum coordination complexes (e.g., cisplatin and carboplatin), procarbazine, hydroxyurea, mitotane, aminoglutethimide, histone deacetylase (HDAC) inhibitors (e.g., trichostatin, sodium butyrate, apicidin, suberoylanilide hydroxamic acid, vorinostat, LBH589, romidepsin, ACY-1215 and panobinostat), mTor inhibitors (e.g., temsirolimus, everolimus, ridaforolimus and sirolimus), KSP (Eg5) inhibitors (e.g., Array 520), DNA binders (e.g., zalypsis), PI3K delta inhibitors (e.g., GS-1101 and TGR-1202), PI3K delta and gamma inhibitors (e.g., CAL-130), multi-kinase inhibitors (e.g., TG02 and sorafenib), hormones (e.g., estrogen) and luteinizing hormone-releasing hormone (LHRH) agonists (e.g., goserelin, leuprorelin and triptorelin) such as hormone agonists, BAFF neutralizing antibodies (e.g., LY2127399), IKK inhibitors, p38MAPK inhibitors, anti-IL-6 (e.g., CNTO328), telomerase inhibitors (e.g., GRN163L), aurora kinase inhibitors (e.g., MLN8237, 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), FTI (e.g., Zarnestra (trademark)), anti-CD138 (e.g., BT062), Torc1 / 2 specific kinase inhibitors (e.g., INK128), kinase inhibitors (e.g., GS-1101), ER / UPR targeting agents (e.g., MKC-3946), cFMS inhibitors (e.g., ARRY-382), JAK1 / 2 inhibitors (e.g., CYT387), PARP inhibitors (e.g., olaparib, talazoparib, niraparib, veliparib (ABT-888)), BCL-2 antagonists may be mentioned. Other chemotherapeutic agents may include mechlorethamine, camptothecin, ifosfamide, tamoxifen, raloxifene, gemcitabine, navelbine, sorafenib, or any of the above analogs or induced mutants.

[0134] The compounds of the present invention can also be used in combination with radiation therapy, hormone therapy, surgery, and immunotherapy, which are well known to those skilled in the art.

[0135] In certain embodiments, the pharmaceutical compositions provided herein are administered in combination with a steroid. Suitable steroids include, but are not limited to, 21-acetoxypregnenolone, alclometasone, algestone, amcinonide, beclomethasone, betamethasone, budesonide, chloroprednisone, clobetasol, clocortolone, cloprednol, corticosterone, cortisone, cortivazol, deflazacort, desonide, desoxymethasone, dexamethasone, diflorasone, diflucortolone, difluprednate, enoxolone, fluazacort, flucloronide, flumethasone, flunisolide, fluocinonide acetonide, fluocinonide, fluocortin butyl, fludrocortisone, fluorometholone, fluprednolone acetate, fluprednidene acetate, fluprednisolone, flurandrenolide, fluticasone propionate, formocortal, halcinonide, halobetasol propionate, halomethasone, hydrocortisone, loteprednol etabonate, madipredone, medrysone, meprednisolone, methylprednisolone, mometasone furoate, paramethasone, prednicarbate, prednisolone, prednisolone 25-diethylaminoacetate, prednisolone sodium phosphate, prednisone, prednival, prednylidene, remexolone, tixocortol, triamcinolone, triamcinolone acetonide, triamcinolone benetonide, triamcinolone hexacetonide, and salts and / or derivatives thereof. In certain embodiments, the compounds of the present invention can also be used in combination with additional pharmaceutically active agents for treating nausea. Examples of agents that can be used to treat nausea include, but are not limited to, dronabinol; granisetron; metoclopramide; ondansetron; and prochlorperazine; or pharmaceutically acceptable salts thereof.

[0136] The compounds or pharmaceutical compositions of the present disclosure can also be used in combination with a certain amount of one or more substances selected from EGFR inhibitors, MEK inhibitors, PI3K inhibitors, AKT inhibitors, TOR inhibitors, and anti-immunotherapies (including those containing PD-1, anti-PDL-1, anti-CTLA4, anti-LAG1, and anti-OX40 agents), GITR agonists, CAR-T cells, and BiTEs.

[0137] EGFR inhibitors include, but are not limited to, small molecule antagonists, antibody inhibitors, or specific antisense nucleotides or siRNAs. Useful antibody inhibitors of EGFR include cetuximab (Erbitux), panitumumab (Vectibix), zalutumumab, nimotuzumab, and matuzumab. Small molecule antagonists of EGFR include gefitinib, erlotinib (Tarceva), and more recently lapatinib (TykerB). See, for example, Yan L, et.al., Pharmacogenetics and Pharmacogenomics In Oncology Therapeutic Antibody Development, BioTechniques 2005;39(4):565-8 and Paez J G, et.al., EGFR Mutations In Lung Cancer Correlation With Clinical Response To Gefitinib Therapy, Science 2004;304(5676):1497-500.

[0138] Non-limiting examples of small molecule EGFR inhibitors include any of the EGFR inhibitors described in the following patent publications and all pharmaceutically acceptable salts and solvates of said EGFR inhibitors: European Patent Application Publication No. 520722, published December 30, 1992; European Patent Application Publication No. 566226, published October 20, 1993; International Publication No. 96 / 33980, published October 31, 1996; U.S. Patent No. 5,747,498, issued May 5, 1998; International Publication No. 96 / 30347, published October 3, 1996; European Patent Application Publication No. 787772, published August 6, 1997; International Publication No. 97 / 30034, published August 21, 1997; International Publication No. 97 / 30044, published August 21, 1997; International Publication No. 97 / 38994, published October 23, 1997; International Publication No. 97 / 49688, published December 31, 1997; European Patent Application Publication No. 837063, published April 22, 1998; International Publication No. 98 / 02434, published January 22, 1998; International Publication No. 97 / 38983, published October 23, 1997; International Publication No. 95 / 19774, published July 27, 1995; International Publication No. 95 / 19970, published July 27, 1995; International Publication No. 97 / 13771, published April 17, 1997; International Publication No. 98 / 02437, published January 22, 1998; International Publication No. 98 / 02438, published January 22, 1998; International Publication No. 97 / 32881, published September 12, 1997; German Patent Application Publication No. 19629652, published January 29, 1998; International Publication No. 98 / 33798, published August 6, 1998; International Publication No. 97 / 32880, published September 12, 1997; International Publication No. 97 / 32880, published September 12, 1997; European Patent Application Publication No. 682027, published November 15, 1995; International Publication No. 97 / 02266, published January 23, 1997; International Publication No. 97 / 27199, published July 31, 1997;International Publication Pamphlet No. WO 98 / 07726 published on February 26, 1998; International Publication Pamphlet No. WO 97 / 34895 published on September 25, 1997; International Publication Pamphlet No. WO 96 / 31510 published on October 10, 1996; International Publication Pamphlet No. WO 98 / 14449 published on April 9, 1998; International Publication Pamphlet No. WO 98 / 14450 published on April 9, 1998; International Publication Pamphlet No. WO 98 / 14451 published on April 9, 1998; International Publication Pamphlet No. WO 95 / 09847 published on April 13, 1995; International Publication Pamphlet No. WO 97 / 19065 published on May 29, 1997; International Publication Pamphlet No. 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; International Publication Pamphlet No. WO 99 / 35146 published on July 15, 1999; International Publication Pamphlet No. WO 99 / 35132 published on July 15, 1999; International Publication Pamphlet No. WO 99 / 07701 published on February 18, 1999 and International Publication Pamphlet No. WO 92 / 20642 published on November 26, 1992. Additional 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.;

[0139] Examples of antibody-based EGFR inhibitors include any anti-EGFR antibody or antibody fragment that can partially or completely block EGFR activation by its natural ligand. Non-limiting examples of antibody-based EGFR inhibitors include those described in Modjtahedi, H., et al., 1993, Br. J. Cancer 67:247-253; Teramoto, T., et al., 1996, Cancer 77:639-645; Goldstein et al., 1995, Clin. Cancer Res. 1:1311-1318; Huang, S.M., et al., 1999, Cancer Res. 15:59(8):1935-40; and Yang, X., et al., 1999, Cancer Res. 59:1236-1243. Thus, the EGFR inhibitor can be the monoclonal antibody Mab E7.6.3 (Yang, 1999, supra), or Mab C225 (ATCC accession number HB-8508), or an antibody or antibody fragment having its binding specificity.

[0140] Examples of MEK inhibitors include, but are not limited to, CI-1040, AZD6244, PD318088, PD98059, PD334581, RDEA119, ARRY-142886, ARRY-438162, and PD-325901.

[0141] Examples of PI3K inhibitors include 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 GDC0941 and described in 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 BEZ235 or NVP-BEZ235 and described in WO 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 WO 2008 / 070740), LY294002 (2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one available from Axon Medchem), PI103 hydrochloride (3-[4-(4-morpholinylpyrido-[3’,2’:4,5]furo[3,2-d]pyrimidin-2-yl]phenol hydrochloride available from Axon Medchem), PIK75 (N’-[(1E)-(6-bromoimidazo[1,2-a]pyridin-3-yl)methylene]-N,2-dimethyl-5-nitrobenzenesulfonohydrazide hydrochloride available from Axon Medchem), PIK90 (N-(7,8-dimethoxy-2,3-dihydro-imidazo[1,2-c]quinazolin-5-yl)-nicotinamide available from Axon Medchem), GDC-0941 bismesylate (2-(1H-indazol-4-yl)-6-(4-methanesulfonyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d] pyrimidine bis mesylate), AS-252424 (5-[1-[5-(4-fluoro-2-hydroxy-phenyl)-furan-2-yl]-meth-(Z)-ylidene]-thiazolidine-2,4-dione available from Axon Medchem), and TGX-221 (7-methyl-2-(4-morpholinyl)-9-[1-(phenylamino)ethyl]-4H-pyrido-[1,2-a]pyrimidin-4-one available from Axon Medchem), XL-765, and XL-147, among others, but not limited to these. Other PI3K inhibitors include demethoxybirudin, 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.

[0142] Examples of AKT inhibitors include Akt-1-1 (which inhibits Akt1) (Barnett et al. (2005) Biochem. J., 385 (Pt. 2), 399-408); Akt-1-1,2 (which inhibits Ak1 and 2) (Barnett et al. (2005) Biochem. J. 385 (Pt. 2), 399-408); API-59CJ-Ome (e.g., Jin et al. (2004) Br. J. Cancer 91, 1808-12); 1-H-imidazo[4,5-c]pyridinyl compounds (e.g., WO 05011700 pamphlet); indole-3-carbinol and its derivatives (e.g., U.S. Pat. No. 6,656,963; Sarkar and Li (2004) J Nutr. 134 (12 Suppl), 3493S-3498S); perifosine (e.g., which prevents the membrane localization of Akt; Dasmahapatra et al. (2004) Clin. Cancer Res. 10 (15), 5242-52, 2004); phosphatidylinositol ether lipid analogs (e.g., Gills and Dennis (2004) Expert. Opin. Investig. Drugs 13, 787-97); and triciribine (TCN or API-2 or NCI identifier: NSC 154020; Yang et al. (2004) Cancer Res. 64:4394-9), but are not limited thereto.

[0143] Examples of TOR inhibitors include, but are not limited to, AP-23573, CCI-779, everolimus, RAD-001, rapamycin, temsirolimus, and ATP-competitive TORC1 / TORC2 inhibitors (including PI-103, PP242, PP30, and torin1). Other TOR inhibitors in FKBP12 enhancers; rapamycin and its derivatives including the following: CCI-779 (temsirolimus), RAD001 (everolimus; WO 94 / 09010 pamphlet), and AP23573; rapalogs such as those disclosed in WO 98 / 02441 pamphlet and WO 01 / 14387 pamphlet, such as AP23573, AP23464, or AP23841; 40-(2-hydroxyethyl)rapamycin, 40-[3-hydroxy(hydroxymethyl)methylpropanoate]-rapamycin (also called CC1779), 40-epi-(tetrazolyl)-rapamycin (also called ABT578), 32-deoxorapamycin, 16-pentynyloxy-32(S)-dihydrorapamycin, and other derivatives disclosed in WO 05 / 005434 pamphlet; derivatives disclosed in US Patent No. 5,258,389, WO 94 / 090101 pamphlet, WO 92 / 05179 pamphlet, US Patent No. 5,118,677, US Patent No. 5,118,678, US Patent No. 5,100,883, US Patent No. 5,151,413, US Patent No. 5,120,842, WO 93 / 111130 pamphlet, WO 94 / 02136 pamphlet, WO 94 / 02485 pamphlet, WO 95 / 14023 pamphlet, WO 94 / 02136 pamphlet, WO 95 / 16691 pamphlet, WO 96 / 41807 pamphlet, WO 96 / 41807 pamphlet, and US Patent No. 5,256,790; phosphorus-containing rapamycin derivatives (e.g., WO 05 / 016252 pamphlet); 4H-1-benzopyran-4-one derivatives (e.g., US Provisional Patent Application No. 60 / 528,340).

[0144] Immunotherapies include, but are not limited to, anti-PD-1 agents, anti-PDL-1 agents, anti-CTLA-4 agents, anti-LAG1 agents, and anti-OX40 agents. Exemplary anti-PD-1 antibodies and methods of using them are described in Goldberg et al., Blood 110(1):186-192(2007), Thompson et al., Clin. Cancer Res. 13(6):1757-1761(2007) and Korman et al., International Application No. PCT / JP2006 / 309606 (International Publication No. 2006 / 121168A1 Pamphlet), which are each hereby expressly incorporated by reference herein. These include the following: Yervoy (trademark) (ipilimumab) or tremelimumab (to CTLA-4), galiximab (to B7.1), BMS-936558 (to PD-1), MK-3475 (to PD-1), AMP224 (to B7DC), BMS-936559 (to B7-H1), MPDL3280A (to B7-H1), MEDI-570 (to ICOS), AMG557 (to B7H2), MGA271 (to B7H3), IMP321 (to LAG-3), BMS-663513 (to CD137), PF-05082566 (to CD137), CDX-1127 (to CD27), anti-OX40 (Providence Health Services), huMAbOX40L (to OX40L), atacicept (to TACI), CP-870893 (to CD40), lucatumumab (to CD40), dacetuzumab (to CD40), muromonab-CD3 (to CD3), ipilimumab (to CTLA-4). Immunotherapies also include genetically engineered T cells (e.g., CAR-T cells) and bispecific antibodies (e.g., BiTE).

[0145] Examples of GITR agonists include GITR fusion proteins described in US Patent No. 6,111,090 box.c, European Patent No. 090505 B1, US Patent No. 8,586,023, International Publication No. 2010 / 003118 pamphlet and No. 2011 / 090754 pamphlet, or, for example, US Patent No. 7,025,962, European Patent No. 1947183 B1, US Patent No. 7,812,135, No. 8,388,967, No. 8,591,886, European Patent No. 1866339, International Publication No. 2011 / 028683 pamphlet, International Publication No. 2013 / 039954 pamphlet, International Publication No. 2005 / 007190 pamphlet, International Publication No. 2007 / 133822 pamphlet, International Publication No. 2005 / 055808 pamphlet, International Publication No. 99 / 40196 pamphlet, International Publication No. 2001 / 03720 pamphlet, International Publication No. 99 / 20758 pamphlet, International Publication No. 2006 / 083289 pamphlet, International Publication No. 2005 / 115451 pamphlet, US Patent No. 7,618,632, and International Publication No. 2011 / 051726 pamphlet, such as GITR fusion proteins and anti-GITR antibodies (e.g., bivalent anti-GITR antibodies), but are not limited thereto.

[0146] The compounds described herein can be used in combination with the agents disclosed herein or other suitable agents, depending on the condition being treated. Thus, in some embodiments, one or more compounds of the disclosure will be co-administered with the other agents described above. When used in combination therapy, the compounds described herein are administered simultaneously with or separately from the second agent. Such co-administration can include simultaneous administration of the two agents in the same dosage form, simultaneous administration of separate dosage forms, and separate administrations. That is, the compounds described herein and any of the above agents can be formulated together in the same dosage form and administered simultaneously. Alternatively, the compounds of the disclosure and any of the above agents can be administered simultaneously, where both agents are present in separate formulations. In another alternative, any of the above agents can be administered subsequent to or in the reverse order of administration of the compounds of the disclosure. In some embodiments of the separate administration protocol, the compounds of the disclosure and any of the above optional agents are administered at intervals of minutes, hours, or days.

[0147] As one aspect of the invention, since the treatment of a disease / condition by a combination of pharmaceutically active compounds that can be administered separately is being considered, the invention further relates to combining individual pharmaceutical compositions in the form of a kit. The kit comprises two separate pharmaceutical compositions: a compound of the invention and a second pharmaceutical compound. The kit includes a container for containing the individual compositions, such as a divided bottle or a divided foil pouch. Other examples of containers include syringes, boxes, and bags. In some embodiments, the kit includes instructions for use of the individual components. The kit form is particularly advantageous when the individual components are preferably administered in different dosage forms (e.g., oral and parenteral), when administered at different dosing intervals, or when titration of the individual components of the combination is desired by the medical professional prescribing the treatment.

Examples

[0148] Experiment Abbreviations: The following abbreviations can be used herein.

[0149]

Table 4

[0150]

Table 5

[0151] Unless otherwise noted, all substances were obtained from commercial suppliers and used without further purification. Unless otherwise indicated, all parts are by weight and temperatures are in degrees Celsius. All microwave-assisted reactions were carried out using a Smith Synthesizer from Biotage (trademark). All compounds showed NMR spectra consistent with their assigned structures. Melting points were determined on a Buchi instrument and not corrected. Mass spectral data were measured by electrospray ionization technique. All examples were determined by high performance liquid chromatography and purified to >90%. Unless otherwise specified, reactions were carried out at room temperature.

[0152] In the synthesis of the compounds of the present invention, the use of certain leaving groups may be desirable. The term "leaving group" ("LG") generally refers to a group that can be replaced by a nucleophile. Such leaving groups are well known in the art. Examples of leaving groups include, but are not limited to, halides (e.g., I, Br, F, Cl), sulfonates (e.g., mesylate, tosylate), sulfides (e.g., SCH3), N-hydroxysuccinimide, N-hydroxybenzotriazole, etc. Examples of nucleophiles include, but are not limited to, amines, thiols, alcohols, Grignard reagents, anionic species (e.g., alkoxides, amides, carbanions), etc.

[0153] In the examples shown below, specific embodiments of the present invention are described. These examples are meant to be representative and are not intended to limit the scope of the claims in any way.

[0154] When percentages (%) are used with respect to liquids, it should be noted that these are volume percentages with respect to the solution. When used with respect to solids, these are solid composition percentages. Materials obtained from commercial vendors were typically used without further purification. Reactions involving air- or moisture-sensitive reagents were typically carried out under a nitrogen or argon atmosphere. Purity was measured using a high-performance liquid chromatography (HPLC) system equipped with UV detection at 254 nm and 215 nm (System A: Agilent Zorbax Eclipse XDB-C8 4.6×150 mm, 5 μm, 5 - 100% CH3CN in H2O containing 0.1% TFA, 1.5 mL / min for 15 min; System B: Zorbax SB-C8, 4.6×75 mm, 10 - 90% CH3CN in H2O containing 0.1% formic acid, 1.0 mL / min for 12 min) (Agilent Technologies, Santa Clara, CA). Silica gel chromatography was generally carried out using prepacked silica gel cartridges (Biotage, Uppsala, Sweden or Teledyne-Isco, Lincoln, NE). 1 1H NMR was 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 as parts per million (ppm) downfield from tetramethylsilane (TMS) or other internal standard in the appropriate solvent 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 equipped with UV detection at 254 nm and 215 nm and low-resolution electrospray mode (ESI).

[0155] General synthetic scheme Unless otherwise indicated, the starting materials and reagents used in preparing these compounds are available from commercial suppliers such as Aldrich Chemical Co. (Milwaukee, Wis.), Bachem (Torrance, Calif.), Sigma (St. Louis, Mo.), or are prepared by methods known to those skilled in the art according to procedures described in references such as Fieser and Fieser’s Reagents for Organic Synthesis, Volumes 1-17 (John Wiley and Sons, 1991); Rodd’s Chemistry of Carbon Compounds, Volumes 1-5 and Supplementals (Elsevier Science Publishers, 1989); Organic Reactions, Volumes 1-40 (John Wiley and Sons, 1991), March’s Advanced Organic Chemistry, (John Wiley and Sons, 4th Edition) and Larock’s Comprehensive Organic Transformations (VCH Publishers Inc., 1989). These schemes are merely illustrative of some of the ways in which the compounds of the invention can be synthesized, and various modifications can be made to these schemes, which would be suggested to those skilled in the art upon reference to the present disclosure. The starting materials and intermediates, as well as the final products of the reactions, can be isolated and purified, if necessary, using conventional techniques including, but not limited to, filtration, distillation, crystallization, chromatography, etc. Such materials can be characterized using conventional means including physical constants and spectral data.

[0156] Unless otherwise indicated, the reactions described herein are carried out at atmospheric pressure in a temperature range of about -78 °C to about 150 °C, more preferably in a temperature range of about 0 °C to about 125 °C, and most preferably at approximately room (or ambient) temperature, e.g., about 20 °C.

[0157] For the sake of clarity in this section on general synthesis, the compounds of formula (I) as defined in the summary of the invention can be schematically depicted as follows with respect to ring Ar 1 and ring Ar 2 as including:

Chemical formula

[0158] Generally, the compounds of formula (I) can be synthesized via the following three general steps: Step 1: Preparation of the ring Ar 1 compound. Step 2: Preparation of the ring Ar 2 compound. Step 3: Coupling of the ring Ar 1 compound to the ring Ar 2 compound.

[0159] The following general schemes A - D are intended to provide guidance to a synthetic chemist of ordinary skill, who can readily understand that the solvent, concentration, reagents, protecting groups, order of synthetic steps, time, temperature, etc. can be modified as necessary within the scope of the skill and judgment of one of ordinary skill in the art.

[0160] In one embodiment, the following schemes A - C provide a general preparation of compounds of formula (I) represented by the following formula (I - A):

Chemical formula

[0161] Examples of the compounds of formula (Ia) include, but are not limited to, (I - A - 1) (wherein, preferably, R 5(wherein W is H) can be mentioned.

Chemical formula

[0162] Scheme A: Preparation of Compound (I-A): According to Scheme A, in one embodiment, the compound of formula (I) disclosed herein can be synthesized as follows.

[0163] Step A-1: Ring Ar 1 Preparation of the compound:

Chemical formula

[0164] Alternatively, compound A-1 (wherein W 1 is F) can be converted into the corresponding sulfonyl chloride compound intermediate by a two-step procedure, which is treatment with benzyl mercaptan followed by oxidative chlorination with 1,3-dichloro-5,5-dimethylhydantoin.

[0165] Compound A-1 is commercially available or can be synthesized by methods known to those skilled in the art. Examples of compound A-1 include, but are not limited to, 1-methyl-3-nitrobenzene, 1-chloro-4-nitrobenzene, 1-methoxy-4-nitrobenzene, 1-methoxy-3-nitrobenzene, or 3,5-difluoronitrobenzene.

[0166] In a suitable organic solvent such as NMP, dioxane, acetonitrile, tetrahydrofuran, DMF, methylene chloride, etc., in the presence of a suitable base, the sulfonyl chloride intermediate compound obtained above is further reacted with an R containing an amine 10 group to give sulfonamide A-2 (wherein R 2 =SO2R 10is obtained. Examples of the base include, but are not limited to, diisopropylethylamine, potassium carbonate, or sodium hydride. R 10 Examples of the amine include, but are not limited to, tert-butylamine, cyclopropylamine, cyclohexylamine, piperidine, or 4,4-difluoropiperidine.

[0167] Next, compound A-2 can be reacted with a palladium catalyst such as Pd / C and a suitable reducing agent such as a hydrogen source in the presence of hydrogen gas to form compound A-3.

[0168] Step A-1-a: Ring Ar 1 Preparation of the compound:

Chemical formula

[0169] Step A-1-b: Ring Ar 1 Preparation of the compound:

Chemical formula

[0170] Step A-1-c: Ring Ar 1 Preparation of the compound: [ka] Further alternatively, the compound A-1 (wherein W 1 is halo, e.g., fluoro, chloro, or bromo, can be reacted with the appropriate R 2 After reaction with the reagent, it can be reacted with a suitable reducing agent, such as a palladium catalyst, such as Pd / C, and a hydrogen source in the presence of hydrogen gas to form compound A-3.

[0171] Compound A-1 is commercially available or can be synthesized by known methods by one of ordinary skill in the art.

[0172] Examples of compound A-1 include, but are not limited to, 1-fluoro-3-nitrobenzene, 1,3-difluoro-5-nitrobenzene, 1-fluoro-3-methylbenzene, or 2-bromo-1-fluoro-4-nitrobenzene.

[0173] 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, (4) 3,3,3-trifluoropropan-1-ol, (5) 2-aminoethan-1-ol, or (6) 2-amino-3-methylpropan-1-ol.

[0174] Examples of bases include, but are not limited to, diisopropylethylamine, potassium carbonate, or sodium hydride.

[0175] Step A-1-d: Ring Ar 1 Preparation of compounds: [ka] Furthermore, or alternatively, the compound A-1 defined in step A-1-c (wherein W 1(which is halo) can be preferably R in the amination reaction in the presence of a metal catalyst such as palladium or copper catalyst and in the presence of a base such as cesium carbonate, potassium carbonate or potassium phosphate. 2 After being reacted with the reagent, it can be reacted with a palladium catalyst such as Pd / C and a suitable reducing agent such as a hydrogen source in the presence of hydrogen gas to form Compound A-3.

[0176] Step A-2: Ring Ar 2 Preparation of compounds: [Chemical formula] In Step A-2, Compound A-6 (wherein each of W 2 and W 3 is independently halo, for example, fluoro, chloro, bromo, or iodo) can be reacted with MeOH in the presence of H2SO4 to form methyl ester A-7, which can then be reacted with R such as 2-amino-2-methyl-1-propanol, (1-aminocyclopropyl)methanol, or 2-aminoethan-1-ol in a suitable organic solvent such as NMP, acetonitrile, tetrahydrofuran, DMF, DMSO, etc. to form Compound A-8. Next, Compound A-8 is hydrolyzed using lithium chloride and neutralized with HCl to form A-9. 1

[0177] Step A-3a: Ring Ar 2 Attachment of Ring Ar to the compound 1 Coupling of the compound, followed by introduction of R 1 [Chemical formula] In Step A-3a, Compound A-5 obtained from Step A-9 can be reacted with an activator such as acid chloride (COCl)2 or SOCl2 in a suitable organic solvent such as tetrahydrofuran, methylene chloride, etc. to form an acid chloride derivative, which can then be reacted with Compound A-3 to form Compound A-10.

[0178] Alternatively, compound A-3 may be directly coupled with compound A-9 obtained from step A-2 in the presence of a coupling reagent such as N,N'-diisopropylcarbodiimide, N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide, benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate, O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, thionyl chloride, carbonyldiimidazole, and polyphosphonic anhydride in a suitable organic solvent such as acetonitrile, tetrahydrofuran, DMF, methylene chloride, etc.

[0179] One skilled in synthetic chemistry will readily understand that other coupling agents can be used. In a suitable organic solvent such as NMP, acetonitrile, tetrahydrofuran, DMF, methylene chloride, DMSO, etc., (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, etc., of R X By reacting with a reagent, the halogen group W 2 Further operations can be used to form compound (Ia). One skilled in the art will readily understand that coupling reactions such as those shown in step A-3 can be carried out under various known conditions. Alternatively, the halogen group W 2 Can be reacted with a boronic acid and a palladium catalyst in a palladium-catalyzed coupling reaction to form compound (Ia).

[0180] Scheme B: Alternative preparation of compound (I-A): Step B-1: Preparation of the ring Ar 1 Preparation of the compound: Refer to steps A-1 to A-1d of Scheme A above. Step B-2: Preparation of the ring Ar 2 Preparation of the compound:

Chemical formula

[0181] Step B-3: Ring Ar 2 Compound with ring Ar 1Coupling of Compounds and Removal of Protecting Groups:

Chem.

[0182] Scheme C: Alternative Preparation of Compound (I-A): Step C-1: Ring Ar 1 Preparation of Compound: Refer to Steps A-1 to A-1d of Scheme A above. Step C-2: Ring Ar 2 Preparation of Compound:

Chem.

[0183] Step C-3 can be carried out under the same conditions as the coupling reaction described above in step A-3 of Scheme A.

[0184] Scheme D: Preparation of compound (I-B): In another embodiment, a compound of formula (I-B):

Chemical formula

[0185] Examples of the compound of formula (I-B) include, but are not limited to,

Chemical formula

[0186] Step D-1: Preparation of the ring Ar 1 compound: ring Ar 1 compound: Preparation of the ring Ar 1 One embodiment of the compound is of the formula:

Chemical formula

[0187] Step D-2: Ring Ar 2 Preparation of compound:

Chemical formula

[0188] Step D-3: Ring Ar 2 Ring Ar to compound 1 Coupling of compound:

Chemical formula

[0189] Those skilled in the art will readily understand that coupling reactions such as those shown in Step D-3 can be carried out under various known conditions. It will also be recognized by those skilled in the art that the above conversions can be carried out at an earlier stage in the synthetic process based on the feasibility of the conversion.

[0190] Preparation of Synthetic Intermediates Ring AR 1 Preparation of Intermediates Intermediate 1: 3-Amino-N-(tert-butyl)-5-methylbenzenesulfonamide

Chem.

[0191] Step 2: To an ice-cooled solution of 2-methylpropan-2-amine (1.09 g, 14.94 mmol) in DCM (50 mL), a solution of DIPEA (3.56 mL, 20.37 mmol) and 3-methyl-5-nitrobenzene-1-sulfonyl chloride (3.2 g, 13.58 mmol) in DCM (50 mL) was slowly added. After the addition, the reaction mixture was slowly warmed to ambient temperature and stirred for 2 hours. Next, water (100 mL) was added and stirred for 10 minutes, and the aqueous layer was extracted with DCM (100 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated under reduced pressure. The crude material was absorbed onto a plug of silica gel and purified by chromatography eluting with 10% EtOAc in petroleum ether to obtain N-(tert-butyl)-3-methyl-5-nitrobenzenesulfonamide (1.9 g, 6.98 mmol, 51% yield) as a pale yellow solid. 11H NMR (300 MHz, chloroform-d) δ 8.56 (d, J = 5.8 Hz, 1H), 8.27 - 8.16 (m, 1H), 8.10 - 7.99 (m, 1H), 4.86 (s, 1H), 2.57 (s, 3H), and 1.27 (s, 9H). MS (ESI negative ion) m / z: 271.2 (M - 1).

[0192] Step 3: A mixture of N-(tert-butyl)-3-methyl-5-nitrobenzenesulfonamide (1.9 g, 6.98 mmol) and 10% Pd / C (0.6 g, 0.56 mmol) in MeOH (50 mL) was stirred under a hydrogen atmosphere (balloon pressure) for 3 h and then filtered through a CELITE® pad. The pad was washed with MeOH (150 mL) and the filtrate was concentrated under reduced pressure to give a white residue. The crude material was absorbed onto a silica gel plug and purified by chromatography eluting with a gradient of 15 - 20% EtOAc in petroleum ether to give 3-amino-N-(tert-butyl)-5-methylbenzenesulfonamide (1.2 g, 4.95 mmol, 71% yield) as an off-white solid. 1 1H NMR (400 MHz, DMSO-d6) δ 7.33 - 7.13 (m, 1H), 6.83 (dq, J = 4.7, 2.2 Hz, 1H), 6.76 (dd, J = 3.8, 2.2 Hz, 1H), 6.52 (q, J = 3.5, 2.6 Hz, 1H), 5.55 - 5.29 (m, 2H), 2.16 (s, 3H), and 1.09 (s, 9H). m / z (ESI): 243.1 (M + 1).

[0193] Intermediate 2: 3-Amino-N-(tert-butyl)-5-fluorobenzenesulfonamide

Chemical Structure

[0194] Procedure 2: To a 100 mL round-bottom flask, add benzyl (3-fluoro-5-nitrophenyl)sulfane (1.63 g, 6.19 mmol), acetonitrile (12 mL), water (0.3 mL), and acetic acid (0.45 mL). Cool the reaction mixture to 0 °C and add 1,3-dichloro-5,5-dimethylhydantoin (1.14 mL, 8.67 mmol) portionwise. When the addition is complete, stir the reaction mixture at 0 °C for 20 minutes. Then dilute the reaction mixture with NaHCO3 and extract with EtOAc. Wash the organic extract with water and dry over Na2SO4. Filter the solution and concentrate under reduced pressure to obtain 3-fluoro-5-nitrobenzene-1-sulfonyl chloride (1.48 g, 6.19 mmol, 100% yield) as a white solid, which was used without further purification. m / z (ESI): 240.2 (M+1).

[0195] Step 3: To a 100 mL round-bottom flask were added 3-fluoro-5-nitrobenzene-1-sulfonyl chloride (1.66 g, 6.92 mmol), DCM (10 mL), tert-butylamine (0.51 mL, 6.92 mmol), and DIPEA (1.81 mL, 10.38 mmol). The reaction mixture was stirred at room temperature for 3 h, diluted with saturated NaHCO3, and extracted with DCM (6 mL). The organic extract was washed with water and dried over Na2SO4. The solution was filtered and concentrated under reduced pressure to obtain the crude material as a white oil. The crude material was absorbed onto a plug of silica gel and purified by chromatography on a silica gel column eluting with a gradient of 0 - 25% EtOAc in heptane to give N-(tert-butyl)-3-fluoro-5-nitrobenzenesulfonamide (1.02 g, 3.70 mmol, 54% yield) as a white solid. m / z (ESI): 290.2 (M+Na).

[0196] Step 4: To a 25 mL glass vial (red cap) were added iron (0.30 g, 5.43 mmol), ammonium chloride (48 mg, 0.91 mmol), followed by N-(tert-butyl)-3-fluoro-5-nitrobenzenesulfonamide (0.50 g, 1.81 mmol) in EtOH (6 mL) and water (0.5 mL). The reaction mixture was stirred at 80 °C for 1 h, cooled to room temperature, filtered through CELITE® and the solvent was removed under reduced pressure. The reaction mixture was diluted with saturated NH4Cl (10 mL) and extracted with EtOAc (10 mL). The organic extract was washed with water (10 mL) and dried over Na2SO4. The solution was filtered and concentrated under reduced pressure to obtain the crude material as a white oil. The crude material was absorbed onto a plug of silica gel and purified by chromatography on a silica gel column eluting with a gradient of 0 - 40% EtOAc in heptane to give 3-amino-N-(tert-butyl)-5-fluorobenzenesulfonamide (0.33 g, 1.36 mmol, 75% yield) as a white solid. 11H NMR (400 MHz, chloroform-d) δ 6.99 (t, J = 1.76 Hz, 1H), 6.93 (td, J = 1.91, 8.12 Hz, 1H), 6.51 (td, J = 2.35, 10.17 Hz, 1H), 4.52 (s, 1H), 1.26 (s, 9H). m / z (ESI): 247.1 (M+1).

[0197] Intermediate 3: 3-Amino-N-(tert-butyl)-2-fluorobenzenesulfonamide

Chemical Structure

[0198] Step 2: To a solution of 3-bromo-N-(tert-butyl)-2-fluorobenzenesulfonamide (3.2 g, 10.32 mmol) in ethylene glycol (5 mL) was added, under a nitrogen atmosphere, N 1 ,N 2-Dimethylethane-1,2-diamine (0.091 g, 1.032 mmol), potassium carbonate (0.285 g, 2.06 mmol), copper(I) oxide (0.074 g, 0.516 mmol), and aqueous ammonia (7.5 mL) were added. The reaction vessel was added and stirred at 100 °C for 1 hour. The reaction mixture was cooled to room temperature, diluted with water (30 mL), and extracted with EtOAc (3 × 50 mL). The combined organic extracts were washed with aqueous saline solution (50 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude residue was absorbed onto a plug of silica gel and purified by silica gel chromatography eluting with 25 - 35% EtOAc in hexane to afford 3-amino-N-(tert-butyl)-2-fluorobenzenesulfonamide (0.9 g, 3.65 mmol, 35% yield) as an off-white solid. 1 1H NMR (400 MHz, DMSO-d6): δ 7.56 (s, 1H), 6.98 - 6.88 (m, 3H), 5.50 (s, 2H), 1.12 (s, 9H). m / z (ESI): 247.2 [M+1].

[0199] Intermediate 4: 3-((1-Methylcyclobutyl)sulfonyl)aniline

Chemical Structure

[0200] Step 2: To a solution of 3-((1-methylcyclobutyl)thio)aniline (3.0 g, 15.5 mmol) in THF (30 mL) was added Boc anhydride (7.21 mL, 31.0 mmol) and Et3N (3.24 mL, 23.28 mmol) at room temperature, and the mixture was stirred for 18 h. The reaction mixture was quenched with water (100 mL) and extracted with EtOAc (200 mL). The organic layer was washed with water (50 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude residue was absorbed onto a plug of silica gel and purified by flash chromatography through a silica gel column eluting with a gradient of 5 - 8% EtOAc in hexane to give tert-butyl (3-((1-methylcyclobutyl)thio)phenyl)carbamate (3.2 g, 70% yield) as a white solid. 1 1H NMR (300 MHz, chloroform-d): δ 7.44 - 7.33 (m, 2H), 7.30 - 7.20 (m, 1H), 7.10 (d, J = 7.6 Hz, 1H), 6.50 (s, 1H), 2.42 - 2.26 (m, 2H), 2.16 - 1.93 (m, 4H), 1.57 (s, 9H), 1.54 (s, 3H). m / z (ESI): 294.2 [M+1].

[0201] Procedure 3: To a solution of tert-butyl (3-((1-methylcyclobutyl)thio)phenyl)carbamate (2.0 g, 6.82 mmol) in MeOH (40 mL) and water (20 mL) was added oxone (9.22 g, 15.00 mmol), and the reaction mixture was stirred at room temperature for 18 h. The reaction mixture was concentrated under reduced pressure, adjusted to pH 7 using 10% aqueous NaHCO3 (50 mL), and extracted with EtOAc (3 × 50 mL). The combined organic extracts were washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude residue was absorbed onto a plug of silica gel (60 - 120 mesh) and purified by flash chromatography through a silica gel column eluting with a gradient of 1 - 40% EtOAc in hexanes to afford tert-butyl (3-((1-methylcyclobutyl)sulfonyl)phenyl)carbamate (1.8 g, 81% yield) as a white solid. 1 1H NMR (400 MHz, DMSO-d6) δ 9.78 (s, 1H), 8.05 (s, 1H), 7.78 - 7.68 (m, 1H), 7.53 (d, J = 8.0 Hz, 1H), 7.39 (d, J = 7.8 Hz, 1H), 2.72 - 2.64 (m, 2H), 2.00 (dt, J = 10.8, 7.8 Hz, 1H), 1.91 - 1.68 (m, 3H), 1.49 (s, 9H), 1.35 (s, 3H). m / z (ESI): 326.1 [M+1].

[0202] Procedure 4: To a solution of tert-butyl (3-((1-methylcyclobutyl)sulfonyl)phenyl)carbamate (1.8 g, 5.53 mmol) in 1,4-dioxane (20 mL) was added HCl (6.91 mL, 27.7 mmol, 4 M in dioxane) at 0 °C, and the mixture was stirred at ambient temperature for 18 h. The reaction mixture was concentrated under reduced pressure and adjusted to pH 7 - 8 using 10% aqueous NaHCO3. The reaction mixture was diluted with water (50 mL) and extracted with EtOAc (3 × 100 mL). The combined organic extracts were washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was triturated with diethyl ether (100 mL) to afford 3-((1-methylcyclobutyl)sulfonyl)aniline (1.05 g, 84% yield) as an off-white solid.1 1H NMR (400 MHz, DMSO-d6) δ 7.34 - 7.21 (m, 1H), 7.00 (s, 1H), 6.93 - 6.79 (m, 2H), 5.66 (s, 2H), 2.73 - 2.59 (m, 2H), 2.07 - 1.93 (m, 1H), 1.85 - 1.76 (m, 3H), 1.35 (s, 3H). m / z (ESI): 226.1 [M+1].

[0203] Intermediate 5: 3 - ((1,1,1-Trifluoro-2-methylpropan-2-yl)sulfonyl)aniline

Chemical formula

[0204] Step 2: To a solution of 3-aminobenzenethiol (4.43 g, 35.4 mmol) in DMF (225 mL) was added sodium hydride (1.77 g, 44.3 mmol) at 0 °C. After stirring for 30 minutes, 2,2,2-trifluoroethyl 4-methylbenzenesulfonate (9.0 g, 35.4 mmol) was added and the mixture was stirred for 18 hours. The reaction mixture was quenched with water (20 mL) and extracted with EtOAc (2 × 50 mL). The combined organic extracts were washed with water, dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude residue was purified by flash chromatography through a silica gel column eluting with a gradient of 5 - 10% EtOAc in hexane to afford 3-((2,2,2-trifluoroethyl)thio)aniline (5.0 g, 68% yield) as a pale brown oil. 1 1H NMR (300 MHz, DMSO-d6): δ 6.99 (d, J = 7.7 Hz, 1H), 6.66 - 6.58 (m, 2H), 6.47 (d, J = 8.0 Hz, 1H), 5.22 (s, 2H), 3.87 (q, J = 10.2 Hz, 2H). m / z (ESI): 208.2 [M+1].

[0205] Step 3: To a solution of 3-((2,2,2-trifluoroethyl)thio)aniline (5.0 g, 24.13 mmol) in THF (50 mL) were added Et3N (6.73 mL, 48.3 mmol) and Boc2O (8.40 mL, 36.2 mmol), and the mixture was stirred at room temperature for 18 hours. The reaction mixture was quenched with water (100 mL) and extracted with EtOAc (200 mL). The organic layer was washed with water (100 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude material was absorbed onto a plug of silica gel (60 - 120 mesh) and purified by flash chromatography through a silica gel column (50 g) eluting with a gradient of 5% - 8% EtOAc in hexane to afford tert-butyl (3-((2,2,2-trifluoroethyl)thio)phenyl)carbamate (4.2 g, 57% yield) as a white solid. 11H NMR (400 MHz, DMSO-d6): δ 9.43 (s, 1H), 7.62 (s, 1H), 7.33 (d, J = 8.4 Hz, 1H), 7.27 - 7.21 (m, 2H), 3.93 (q, J = 10.0 Hz, 2H), 1.48 (s, 9H). m / z (ESI): 308.1 [M+1].

[0206] Procedure 4: To a solution of tert-butyl (3-((2,2,2-trifluoroethyl)thio)phenyl)carbamate (4.2 g, 13.67 mmol) in MeOH (40 mL) and water (20 mL) was added oxone (9.24 g, 30.1 mmol), and the mixture was stirred at room temperature for 18 h. The reaction mixture was concentrated under reduced pressure. The reaction mixture was neutralized with 10% aqueous NaHCO3 and extracted with EtOAc (3 × 200 mL). The combined organic extracts were washed with water (2 × 100 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to afford tert-butyl (3-((2,2,2-trifluoroethyl)sulfonyl)phenyl)carbamate (3.2 g, 69% yield) as a white solid. 1 1H NMR (300 MHz, DMSO-d6): δ 9.86 (s, 1H), 8.22 (s, 1H), 7.70 (d, J = 7.2 Hz, 1H), 7.64 - 7.56 (m, 2H), 4.92 (q, J = 10.0 Hz, 2H), 1.49 (s, 9H). m / z (ESI): 338.0 [M-1].

[0207] Procedure 5: To a solution of tert-butyl (3-((2,2,2-trifluoroethyl)sulfonyl)phenyl)carbamate (8.5 g, 25.05 mmol) in dry THF (50 mL) were added methyl iodide (15.66 mL, 250 mmol) and HMPA (43.6 mL, 250 mmol) at 0 °C. The reaction mixture was cooled to -78 °C and LDA (31.3 mL, 62.6 mmol, 2 M solution in THF) was added under a nitrogen atmosphere. The reaction mixture was stirred at -78 °C for 30 minutes and then warmed to room temperature. The reaction mixture was diluted with saturated aqueous NH4Cl (100 mL) and extracted with diethyl ether (3 × 250 mL). The organic layer was washed with brine (250 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude residue was absorbed onto a plug of silica gel (60 - 120 mesh) and purified by silica gel chromatography eluting with 5 - 7% EtOAc in hexanes to afford tert-butyl (3-((1,1,1-trifluoro-2-methylpropan-2-yl)sulfonyl)phenyl)carbamate (1.9 g, 21% yield) as a pale yellow solid. 1 1H NMR (400 MHz, DMSO-d6) δ 9.86 (s, 1H), 8.15 (s, 1H), 7.80 (d, J = 8.7 Hz, 1H), 7.62 - 7.58 (m, 1H), 7.50 (d, J = 7.9 Hz, 1H), 1.58 - 1.43 (m, 15H). m / z (ESI): 366.2 [M - 1].

[0208] Procedure 6: To a solution of tert-butyl (3-((1,1,1-trifluoro-2-methylpropan-2-yl)sulfonyl)phenyl)carbamate (1.8 g, 4.90 mmol) in 1,4-dioxane (20.0 mL) was added hydrochloric acid (12.3 mL, 49.0 mmol, 4 M in dioxane) at 0 °C, and the mixture was stirred at room temperature for 18 h. The reaction mixture was concentrated under reduced pressure and adjusted to pH 7 - 8 using 10% aqueous NaHCO3. The reaction mixture was diluted with water (50 mL) and extracted with EtOAc (3 × 100 mL). The combined organic extracts were washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude residue was absorbed onto a plug of silica gel (60 - 120 mesh) and purified by silica gel chromatography eluting with 17 - 19% EtOAc in hexane to afford 3-((1,1,1-trifluoro-2-methylpropan-2-yl)sulfonyl)aniline (0.9 g, 69% yield) as an off-white solid. 1 1H NMR (400 MHz, DMSO-d6) δ 7.30 (d, J = 7.9 Hz, 1H), 7.08 (s, 1H), 7.00 - 6.89 (m, 2H), 5.75 (s, 2H), 1.49 (s, 6H). m / z (ESI): 268.1 [M + 1].

[0209] Intermediate 6: 3-((3,3-difluorocyclopentyl)sulfonyl)aniline

Chemical Structure

[0210] Procedure 2: A mixture of tert-butyl (3-mercaptophenyl)carbamate (10.0 g, 44.4 mmol) and cyclopenta-2-enone (14.58 g, 178 mmol) was stirred at 120 °C for 24 h. The reaction mixture was directly absorbed onto a plug of silica gel (60 - 120 mesh) and purified by silica gel chromatography eluting with 9 - 12% EtOAc in hexane to give tert-butyl (3-((3-oxocyclopentyl)thio)phenyl)carbamate (10.0 g, 73% yield) as a colorless viscous oil. 1 H NMR (300 MHz, chloroform-d) δ 7.54 (s, 1H), 7.28 - 7.17 (m, 2H), 7.07 (d, J = 8.8 Hz, 1H), 6.55 (s, 1H), 3.94 (t, J = 8.0 Hz, 1H), 2.71 - 2.58 (m, 1H), 2.45 - 2.20 (m, 4H), 2.12 - 1.97 (m, 1H), 1.53 (s, 9H). m / z (ESI): 308.6 [M + 1].

[0211] Step 3: To a solution of tert-butyl (3-((3-oxocyclopentyl)thio)phenyl)carbamate (5.0 g, 16.27 mmol) in MeOH (100 mL) was added a solution of oxone (22 g, 35.8 mmol) in water (50 mL), and the mixture was stirred at room temperature for 18 h. The reaction mixture was concentrated under reduced pressure to remove MeOH, and the resulting aqueous solution was extracted with DCM (3 × 100 mL). The combined organic phases were washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give tert-butyl (3-((3-oxocyclopentyl)sulfonyl)phenyl)carbamate (4.5 g, crude) as a pale yellow oil, which was carried on directly to the next step. 1 H NMR (400 MHz, DMSO-d6) δ 9.84 (s, 1H), 8.14 (s, 1H), 7.71 (d, J = 7.92 Hz, 1H), 7.60 - 7.53 (m, 1H), 7.48 (d, J = 7.7 Hz, 1H), 4.08 (t, J = 7.4 Hz, 1H), 2.45 - 2.16 (m, 6H), 1.50 (s, 9H).

[0212] Step 4: To a solution of tert-butyl (3-((3-oxocyclopentyl)sulfonyl)phenyl)carbamate (1.65 g, 4.86 mmol) in DCM (20 mL) was added DAST (1.61 mL, 12.15 mmol) at -78 °C under a nitrogen atmosphere, and the mixture was stirred at -78 °C for 30 min. The reaction mixture was slowly warmed to room temperature and stirred for 18 h. The reaction mixture was quenched with 1 N aqueous NaOH solution (10 mL), diluted with water (25 mL), and extracted with DCM (3 × 50 mL). The combined organic phases were washed with brine (80 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude material was absorbed onto a plug of silica gel and purified by silica gel chromatography eluting with 6 - 8% EtOAc in hexane to give tert-butyl (3-((3,3-difluorocyclopentyl)sulfonyl)phenyl)carbamate (0.95 g, 54% yield) as a colorless viscous oil. 11H NMR (300 MHz, DMSO-d6): δ 9.84 (s, 1H), 8.14 (s, 1H), 7.70 (d, J = 7.8 Hz, 1H), 7.61 - 7.43 (m, 2H), 4.13 - 3.98 (m, 1H), 2.41 - 1.95 (m, 6H), 1.49 (s, 9H). m / z (ESI): 360.2 [M - 1].

[0213] Step 5: To a solution of tert-butyl (3-((3,3-difluorocyclopentyl)sulfonyl)phenyl)carbamate (1.0 g, 2.77 mmol) in 1,4-dioxane (10 mL) was added HCl (10 mL, 40 mmol, 4 M in dioxane) at 0 °C, and the mixture was stirred at room temperature for 24 h. The reaction mixture was concentrated under reduced pressure. The crude residue was dissolved in water (15 mL), the pH was adjusted to about 7 using 10% aqueous NaHCO3 solution, diluted with water (20 mL), and extracted with EtOAc (3 × 30 mL). The combined organic extracts were washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was absorbed onto a plug of silica gel (60 - 120 mesh) and purified by silica gel chromatography eluting with 20 - 25% EtOAc in hexane to afford 3-((3,3-difluorocyclopentyl)sulfonyl)aniline (250 mg, 35% yield) as a viscous oil. 1 1H NMR (400 MHz, MeOD): δ 7.27 (t, J = 8.0 Hz, 1H), 7.15 (d, J = 2.0 Hz, 1H), 7.09 (dd, J = 8.0, 2.0 Hz, 1H), 6.95 (dd, J = 8.0, 2.0 Hz, 1H), 3.90 - 3.84 (m, 1H), 2.51 - 2.05 (m, 6H), NH2 proton was not visible. m / z (ESI): 262.1 [M + 1].

[0214] Ring AR 2 Preparation of Intermediate Intermediate 7: 3-Chloro-5-((1-hydroxy-2-methylpropan-2-yl)amino)pyrazine-2-carboxylic acid

Chemical Structure

[0215] Procedure 2: To a solution of methyl 3,5-dichloropyrazine-2-carboxylate (2.02 g, 9.78 mmol) in DMSO (40 mL) were added DIPEA (3.41 mL, 19.56 mmol) and 2-amino-2-methyl-1-propanol (0.93 mL, 9.78 mmol, Aldrich). The solution was stirred at room temperature for 4 days, then treated with water (150 mL) and extracted with EtOAc (5 × 20 mL). The combined organic extracts were concentrated under reduced pressure, adsorbed onto a silica gel plug, and chromatographed through a silica gel column eluting with 0 - 75% EtOAc in heptane to afford methyl 3-chloro-5-((1-hydroxy-2-methylpropan-2-yl)amino)pyrazine-2-carboxylate (1.46 g, 5.63 mmol, 58% yield) as an orange solid. 1 1H NMR (400 MHz, DMSO-d6) δ ppm 7.94 (s, 1H), 7.80 (s, 1H), 4.84 (t, J = 5.9 Hz, 1H), 3.78 (s, 3H), 3.55 (d, J = 5.9 Hz, 2H), 1.31 (s, 6H). m / z (ESI): 260.0 (M + H) + .

[0216] Step 3: To a solution of methyl 3-chloro-5-((1-hydroxy-2-methylpropan-2-yl)amino)pyrazine-2-carboxylate (1.46 g, 5.62 mmol) in THF (40 mL) and MeOH (13 mL) was added LiOH (20 mL of 1 M aqueous solution, 20.00 mmol). The resulting dark brown solution was stirred at room temperature for 16 h and then concentrated under reduced pressure to remove the organic solvents. The aqueous solution was neutralized with HCl (2 N) to about pH 7 and extracted with EtOAc (2 × 10 mL). The combined organic extracts were concentrated under reduced pressure to give 3-chloro-5-((1-hydroxy-2-methylpropan-2-yl)amino)pyrazine-2-carboxylic acid (1.15 g, 4.68 mmol, 83% yield) as a yellow solid. m / z (ESI): 246.1 (M+H) + 。

[0217]

Table 6

[0218] Intermediate 8: 5-(4,4-dimethyl-2-oxooxazolidin-3-yl)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxylic acid.

Chem.

[0219] Step 2: 4,4-Dimethyloxazolidin-2-one (4.1 g, 35.6 mmol) was dissolved in DMF (80 mL) in a jacketed reactor. The temperature was set to 15 °C. Potassium tert-butoxide solution (35.5 mL, 35.5 mmol, 1 M in THF) was added and the mixture became a thick gel. A solution of benzyl 3,5-dichloropyrazine-2-carboxylate (10.0 g, 35.3 mmol) in DMF (50 mL) was added all at once and the mixture was stirred for 2 h. Water (100 mL), saturated ammonium chloride (50 mL) and EtOAc (100 mL) were added and the mixture was stirred for 5 min. The aqueous phase was allowed to settle and then drained. The organic phase was washed with water (100 mL), then dried over Na2SO4 and evaporated to dryness under reduced pressure. Purification on a silica gel column (0 - 100% EtOAc in heptane) afforded benzyl 3-chloro-5-(4,4-dimethyl-2-oxooxazolidin-3-yl)pyrazine-2-carboxylate (10.7 g, 29.6 mmol, 84% yield) as an off-white solid. m / z (ESI): 362.3 (M+H) + 。

[0220] Step 3: Benzyl 3-chloro-5-(4,4-dimethyl-2-oxooxazolidin-3-yl)pyrazine-2-carboxylate (10.7 g, 29.6 mmol), 6-azaspiro[2.5]octane (3.5 g, 31.5 mmol), and cesium carbonate (12 g, 36.8 mmol) were combined with DMF (75 mL) under nitrogen. The mixture was stirred at room temperature for 16 h. Water (200 mL) and EtOAc (200 mL) were added, the phases were mixed and separated. The organic phase was washed with water (200 mL) and then evaporated to dryness under reduced pressure. The crude material was suspended in methyl tert-butyl ether (50 mL) and stirred for 10 min. Heptane (50 mL) was slowly added and the mixture was stirred for an additional 10 min. It was filtered through a sintered glass frit and the solid was washed with 2:1 heptane:methyl tert-butyl ether (10 mL). The solid was dried under reduced pressure to give benzyl 5-(4,4-dimethyl-2-oxooxazolidin-3-yl)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxylate (12.2 g, 27.9 mmol, 95% yield) as a pale yellow solid. The filtrate was evaporated to dryness under reduced pressure to give a yellow oil (0.7 g). Purification using ISCO (gradient from heptane to EtOAc) gave an additional 0.5 g. m / z (ESI): 437.2 (M+H) + 。

[0221] Step 4: Benzyl 5-(4,4-dimethyl-2-oxooxazolidin-3-yl)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxylate (12.2 g, 27.9 mmol) was dissolved in DCM:EtOH (1:1, 120 mL). Palladium on carbon (5 wt% Pd) (0.350 g, 0.082 mmol) was added and the suspension was hydrogenated at 40 psi for 1 hour. The suspension was filtered through a pad of CELITE® and the solid was washed with DCM (10 mL). The filtrate was evaporated to dryness under reduced pressure to afford 5-(4,4-dimethyl-2-oxooxazolidin-3-yl)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxylic acid (9.17 g, 26.5 mmol, 95% yield) as a pale yellow solid. 1 H NMR (400 MHz, chloroform-d) δ ppm 10.1 (br s,1H) 8.60 - 8.78 (m,1H) 4.06 - 4.26 (m,2H) 3.49 - 3.63 (m,4H) 1.65 - 1.81 (m,6H) 1.49 - 1.61 (m,4H) 0.34 - 0.50 (m,4H). m / z (ESI): 347.1 (M+H) + .

[0222]

Table 7

[0223] Intermediate 9: 3-(6-azaspiro[2.5]octan-6-yl)-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)pyrazine-2-carboxylic acid

Chem.

[0224] Procedure 2: A mixture of benzyl 3-chloro-5-(1-ethoxyvinyl)pyrazine-2-carboxylate and benzyl 5-chloro-3-(1-ethoxyvinyl)pyrazine-2-carboxylate (2.81 g, 8.82 mmol), DIPEA (4.62 mL, 26.4 mmol), and 6-azaspiro[2.5]octane (1.18 g, 10.58 mmol, Wuxi Apptec) in DMSO (10 mL) was heated at 70 °C for 3 h. The reaction mixture was cooled to room temperature, diluted with water (10 mL), and extracted with EtOAc (3×20 mL). The combined organic extracts were concentrated, and the residue was purified by ISCO column (0-20% EtOAc in heptane) to give benzyl 5-(1-ethoxyvinyl)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxylate (1.33 g, 3.40 mmol, 38% yield): 11H NMR (chloroform-d) δ: 8.30 (s, 1H), 7.45 - 7.51 (m, 2H), 7.31 - 7.40 (m, 3H), 5.42 (s, 2H), 5.40 (d, J = 1.8 Hz, 1H), 4.41 (d, J = 2.0 Hz, 1H), 3.96 (q, J = 7.0 Hz, 2H), 3.41 - 3.47 (m, 4H), 1.43 (t, J = 6.9 Hz, 3H), 1.38 (dd, J = 6.5, 4.7 Hz, 4H), 0.33 (s, 4H). m / z (ESI): 394.1 (M + H) + .

[0225] Step 3: To a stirred solution of benzyl 5-(1-ethoxyvinyl)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxylate (1.33 g, 3.38 mmol) in THF (15 mL) was added 5N HCl (3.38 mL, 16.90 mmol). After the addition, the mixture was stirred at room temperature for 3 hours. The mixture was diluted with EtOAc and slowly neutralized with saturated aqueous NaHCO3. The organic layer was separated, dried over MgSO4, and concentrated to give benzyl 5-acetyl-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxylate. 1 1H NMR (chloroform-d) δ: 8.51 (s, 1H), 7.46 - 7.50 (m, 2H), 7.31 - 7.41 (m, 3H), 5.44 (s, 2H), 3.42 - 3.52 (m, 4H), 2.62 (s, 3H), 1.33 - 1.41 (m, 4H), 0.36 (s, 4H). m / z (ESI): 366.1 (M + H) + .

[0226] Step 4: To a stirred mixture of benzyl 5-acetyl-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxylate (1.21 g, 3.31 mmol) and cesium fluoride (2.01 g, 13.23 mmol) in THF (2 mL) was added trimethyl(trifluoromethyl)silane (0.59 mL, 3.97 mmol, Aldrich). After the addition, the reaction mixture was stirred for 16 h. The reaction mixture was partitioned between EtOAc and water. The organic layer was concentrated and the residue was purified by silica gel column (0 - 30% EtOAc in heptane) to afford benzyl 3-(6-azaspiro[2.5]octan-6-yl)-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)pyrazine-2-carboxylate (1.21 g, 2.78 mmol, 84% yield). 1 1H NMR (chloroform-d) δ: 8.10 (s, 1H), 7.49 (dd, J = 7.8, 1.4 Hz, 2H), 7.37 (dd, J = 7.5, 1.9 Hz, 3H), 5.44 (s, 2H), 5.21 (s, 1H), 3.40 - 3.47 (m, 4H), 1.74 (s, 3H), 1.39 (dd, J = 6.5, 4.7 Hz, 4H), 0.36 (s, 4H). m / z (ESI): 436.3 (M + H) + .

[0227] Step 5: A solution of benzyl 3-(6-azaspiro[2.5]octan-6-yl)-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)pyrazine-2-carboxylate (1.2 g, 2.76 mmol) in ethanol (15 mL) was hydrogenated at 50 psi for 2 h in the presence of Pd / C (0.293 g of 10 wt%, 0.276 mmol). The catalyst was removed by filtration through a pad of CELITE® and the solid was washed with EtOH. The filtrate was concentrated to afford 3-(6-azaspiro[2.5]octan-6-yl)-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)pyrazine-2-carboxylic acid (0.90 g, 95% yield) as a yellow solid. m / z (ESI): 346.3 (M + H) + 。

[0228] Ar1 and Ar 2 Coupling of intermediates Intermediate 10: N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-3,5-dichloropyrazine-2-carboxamide

Chemical formula

[0229]

Table 8

[0230] Intermediate 11: 3-Chloro-N-(3-(cyclopentylsulfonyl)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)pyrazine-2-carboxamide

Chemical formula

[0231]

Table 9

[0232] Example 100: N-(3-(N-(tert-butyl)sulfamoyl)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide.

Chem.

[0233] Step 2: To a solution of N-(3-(N-(tert-butyl)sulfamoyl)phenyl)-3-chloro-5-((1-hydroxy-2-methylpropan-2-yl)amino)pyrazine-2-carboxamide (60 mg, 0.13 mmol) in DMSO (1.5 mL) were added DIPEA (0.046 mL, 0.26 mmol) and 6-azaspiro[2.5]octane (26 mg, 0.23 mmol). The solution was stirred at room temperature for 16 h. The reaction mixture was diluted with water (50 mL) and extracted with EtOAc (2 × 15 mL). The combined organic extracts were concentrated under reduced pressure, adsorbed onto a silica gel plug, and chromatographed through a silica gel column eluting with 0 - 60% EtOAc in heptane to afford N-(3-(N-(tert-butyl)sulfamoyl)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide (39 mg, 0.073 mmol, 55% yield) as a pale yellow solid. 1 H NMR (DMSO-d6) δ: 10.29 (s, 1H), 8.42 (s, 1H), 7.87 (d, J = 5.9 Hz, 1H), 7.40 - 7.51 (m, 4H), 6.99 (s, 1H), 4.74 (t, J = 4.8 Hz, 1H), 3.58 (d, J = 4.5 Hz, 2H), 3.36 - 3.45 (m, 4H), 1.42 (br s, 4H), 1.34 (s, 6H), 1.11 (s, 9H), 0.32 (s, 4H). m / z (ESI): 531.2 (M + H) + .

[0234]

Table 10

[0235]

Table 11

[0236]

Table 12

[0237] Example 101: N-(3-(N-(tert-Butyl)sulfamoyl)-5-methylphenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide. [Chemical formula] Step 1: To a 50 mL round-bottom flask, 5-(4,4-dimethyl-2-oxooxazolidin-3-yl)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxylic acid (111 mg, 0.320 mmol, Intermediate 8) and DCM (4 mL) were added. Next, oxalyl chloride (0.24 mL, 0.48 mmol, 2 M in DCM) was added, followed by a few drops of DMF. The reaction mixture was stirred at room temperature for 30 minutes and the solvent was removed under reduced pressure. The residue was redissolved in DCM (4 mL) and treated with 3-amino-N-(tert-butyl)-5-methylbenzenesulfonamide (78 mg, 0.32 mmol, Intermediate 1) and DIPEA (0.17 mL, 0.96 mmol). The reaction mixture was stirred at room temperature for 18 hours and the solvent was removed under reduced pressure. The crude material was absorbed onto a plug of silica gel and purified by chromatography through a silica gel column eluting with a gradient of 0 - 50% EtOAc in heptane to give N-(3-(N-(tert-butyl)sulfamoyl)-5-methylphenyl)-5-(4,4-dimethyl-2-oxooxazolidin-3-yl)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide (117 mg, 0.205 mmol, 64% yield) as a pale yellow solid. m / z (ESI): 571.3 (M+H) + 。

[0238] Procedure 2: To a 15 mL reaction vial were added N-(3-(N-(tert-butyl)sulfamoyl)-5-methylphenyl)-5-(4,4-dimethyl-2-oxooxazolidin-3-yl)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide (117 mg, 0.205 mmol), sodium hydroxide (0.410 mL, 2.050 mmol, 1 N), and MeOH (3 mL). The reaction mixture was heated at 70 °C for 1.5 h, cooled to room temperature, and the solvent was removed under reduced pressure. HCl (2 N) was added and the mixture was extracted with EtOAc. The organic extract was washed with saturated NaHCO3 and water, and dried over Na2SO4. N-(3-(N-(tert-butyl)sulfamoyl)-5-methylphenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide (86 mg, 0.16 mmol, 77% yield) was obtained as a pale yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ 10.20 (s, 1H), 8.20 (s, 1H), 7.73 (s, 1H), 7.41 (s, 1H), 7.29 (s, 1H), 6.98 (s, 1H), 4.79 (t, J = 5.77 Hz, 1H), 3.59 (d, J = 5.67 Hz, 2H), 3.35 - 3.48 (m, 4H), 2.35 (s, 3H), 1.38 - 1.47 (m, 4H), 1.34 (s, 6H), 1.13 (s, 9H), 0.33 (s, 4H). m / z (ESI): 545.3 (M+H) + .

[0239]

Table 13

[0240]

Table 14

[0241] Examples 102-1 and 102-2: (S)-N-(3-((3,3-Difluorocyclopentyl)sulfonyl)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide and (R)-N-(3-((3,3-Difluorocyclopentyl)sulfonyl)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide. [Chemical formula] Step 1: To a solution of 5-(4,4-dimethyl-2-oxooxazolidin-3-yl)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxylic acid (71 mg, 0.21 mmol, Intermediate 8) and DMF (2 mL) was added 3-((3,3-difluorocyclopentyl)sulfonyl)aniline (54 mg, 0.21 mmol, Intermediate 6) and 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholin-4-ium chloride (86 mg, 0.31 mmol). The solution was stirred at room temperature for 16 h. The reaction was treated with additional 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholin-4-ium chloride (50 mg). After an additional 24 h, the reaction was diluted with water (40 mL) and stirred for 20 min. The aqueous solution was filtered and the yellow solid was dried in the filter under reduced pressure. This material was adsorbed onto a plug of silica gel and chromatographed through a silica gel column eluting with 0-50% EtOAc in heptane to afford N-(3-((3,3-difluorocyclopentyl)sulfonyl)phenyl)-5-(4,4-dimethyl-2-oxooxazolidin-3-yl)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide (88 mg, 0.15 mmol, 72% yield) as a pale yellow solid.

[0242] Procedure 2: To a solution of N-(3-((3,3-difluorocyclopentyl)sulfonyl)phenyl)-5-(4,4-dimethyl-2-oxooxazolidin-3-yl)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide (88 mg, 0.15 mmol) in THF (2 mL):MeOH (0.5 mL) was added 1 M aqueous LiOH (0.45 mL, 0.45 mmol). The solution was stirred at room temperature. After 2 h, the reaction was diluted with water and extracted with EtOAc (2×15 mL). The combined organic extracts were concentrated under reduced pressure, adsorbed onto a plug of silica gel, and chromatographed through a silica gel column eluting with 0-100% EtOAc in heptane to afford N-(3-((3,3-difluorocyclopentyl)sulfonyl)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide (61 mg, 0.11 mmol, 73% yield) as an off-white solid. m / z (ESI): 590.2 (M+H) + This material was separated by chiral preparative SFC using an OX column (250×21 mm, 5 mic) with a mobile phase of 60% liquid CO2 and 40% MeOH and a flow rate of 75 mL / min to afford the following.

[0243] Example 102-1: (S)-N-(3-((3,3-difluorocyclopentyl)sulfonyl)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide. First eluting peak; ee > 99%, 11H NMR (400 MHz, DMSO-d6) δ ppm 10.43 (s, 1H), 8.43 (t, J = 1.8 Hz, 1H), 8.13 (d, J = 7.7 Hz, 1H), 7.59 (t, J = 7.9 Hz, 1H), 7.51 (d, J = 8.0 Hz, 1H), 7.45 (s, 1H), 7.03 (s, 1H), 4.72 - 4.89 (m, 1H), 4.01 - 4.11 (m, 1H), 3.56 - 3.60 (m, 2H), 3.38 - 3.44 (m, 4H), 2.37 - 2.46 (m, 1H), 1.99 - 2.24 (m, 4H), 1.76 (dt, J = 6.8, 3.2 Hz, 1H), 1.37 - 1.47 (m, 4H), 1.34 (s, 6H), 0.33 (s, 4H). 19 19F NMR (376 MHz, DMSO-d6) δ ppm -92.52--90.43 (m, 2 F). m / z (ESI): 564.2 (M + H) + .

[0244] Example 102-2: (R)-N-(3-((3,3-Difluorocyclopentyl)sulfonyl)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide. Second elution peak; ee 98%. 1 1H NMR (400 MHz, DMSO-d6) δ ppm 10.42 (s, 1H), 8.43 (t, J = 1.8 Hz, 1H), 8.13 (d, J = 8.4 Hz, 1H), 7.58 (t, J = 7.9 Hz, 1H), 7.50 (d, J = 7.8 Hz, 1H), 7.44 (s, 1H), 7.03 (s, 1H), 4.79 (br s, 1H), 4.05 (br t, J = 8.2 Hz, 1H), 3.58 (br s, 2H), 3.38 - 3.45 (m, 4H), 2.35 - 2.47 (m, 2H), 1.98 - 2.24 (m, 4H), 1.37 - 1.45 (m, 4H), 1.33 (s, 6H), 0.32 (s, 4H). m / z (ESI): 564.2 (M + H) + .

[0245] The stereochemistry was arbitrarily assigned.

[0246] Example 103: N-(3-(Cyclopentylsulfonyl)phenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide

Chemical Structure

[0247]

Table 15

[0248] Example 104: N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-3-(4-chlorophenyl)-5-((1-hydroxy-2-methylpropan-2-yl)amino)pyrazine-2-carboxamide.

Chemical formula

[0249]

Table 16

[0250] Examples 105-1 and 105-2: (R)-N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-3-(6-azaspiro[2.5]octan-6-yl)-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)pyrazine-2-carboxamide and (S)-N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-3-(6-azaspiro[2.5]octan-6-yl)-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)pyrazine-2-carboxamide. [Chemical formula] To a stirred mixture of 3-(6-azaspiro[2.5]octan-6-yl)-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)pyrazine-2-carboxylic acid (150 mg, 0.434 mmol, Intermediate 9), 3-amino-N-(tert-butyl)benzenesulfonamide (119 mg, 0.52 mmol), and DIPEA (0.19 mL, 1.09 mmol) in DMF (2 mL) was added HATU (0.21 g, 0.56 mmol). The reaction mixture was stirred at room temperature for 18 h and diluted with water. The precipitated solid was collected by filtration, washed with water, and dried. The crude material was purified by silica gel chromatography (0 - 30% EtOAc:EtOH (3:1) in heptane) to give N-(3-(N-(tert-butyl)sulfamoyl)phenyl)-3-(6-azaspiro[2.5]octan-6-yl)-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)pyrazine-2-carboxamide (192 mg, 0.346 mmol, 80% yield) as a white solid. 1 1H NMR (DMSO-d6) δ: 10.98 (s, 1H), 8.37 (s, 1H), 8.26 (s, 1H), 7.87 (dt, J = 6.9, 2.1 Hz, 1H), 7.51 - 7.61 (m, 3H), 6.94 (s, 1H), 3.50 - 3.59 (m, 4H), 1.69 (s, 3H), 1.34 - 1.41 (m, 4H), 1.11 (s, 9H), 0.32 (s, 4H). m / z (ESI): 556.1 (M + H) +The racemic material was separated by chiral preparative SFC using a mobile phase of 0.75% liquid CO2 and 25% MeOH and a flow rate of 60 mL / min using a Chiralpack AD column (250×20 mm, 5 mic) to obtain the following.

[0251] Example 105-1: (R)-N-(3-(N-(tert-butyl)sulfamoyl)phenyl)-3-(6-azaspiro[2.5]octan-6-yl)-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)pyrazine-2-carboxamide. First elution peak; ee>99%, 1 H NMR (DMSO-d6) δ: 10.98 (s, 1H), 8.37 (s, 1H), 8.26 (s, 1H), 7.87 (dt, J = 6.9, 2.1 Hz, 1H), 7.51 - 7.61 (m, 3H), 6.94 (s, 1H), 3.50 - 3.59 (m, 4H), 1.69 (s, 3H), 1.34 - 1.41 (m, 4H), 1.11 (s, 9H), 0.32 (s, 4H). m / z (ESI): 556.1 (M+H) + .

[0252] Example 105-2: (S)-N-(3-(N-(tert-butyl)sulfamoyl)phenyl)-3-(6-azaspiro[2.5]octan-6-yl)-5-(1,1,1-trifluoro-2-hydroxypropan-2-yl)pyrazine-2-carboxamide. Second elution peak; ee>99%, 1 H NMR (DMSO-d6) δ: 10.98 (s, 1H), 8.37 (s, 1H), 8.26 (s, 1H), 7.87 (dt, J = 6.9, 2.1 Hz, 1H), 7.51 - 7.61 (m, 3H), 6.94 (s, 1H), 3.50 - 3.59 (m, 4H), 1.69 (s, 3H), 1.34 - 1.41 (m, 4H), 1.11 (s, 9H), 0.32 (s, 4H). m / z (ESI): 556.1 (M+H) + .

[0253] The stereochemistry was arbitrarily assigned.

[0254] Example 106: N-(3-(N-(tert-Butyl)sulfamoyl)phenyl)-5-((1,3-dihydroxy-2-methylpropan-2-yl)amino)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide.

Chemical Structure

[0255] Procedure 2: To a solution of N-(3-(N-(tert-butyl)sulfamoyl)phenyl)-5-((2,2,3,3,6,9,9,10,10-nonamethyl-4,8-dioxa-3,9-disiloundecan-6-yl)amino)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide (266 mg, 0.257 mmol) in THF (5 mL) was added TBAF (1.6 mL, 1.6 mmol, 1 M in THF). The reaction mixture was stirred at room temperature for 16 h. Next, the crude mixture was adsorbed onto a plug of silica gel and chromatographed through a Redi-Sep® prepacked silica gel column eluting with 0–100% EtOAc in heptane to afford N-(3-(N-(tert-butyl)sulfamoyl)phenyl)-5-((1,3-dihydroxy-2-methylpropan-2-yl)amino)-3-(6-azaspiro[2.5]octan-6-yl)pyrazine-2-carboxamide (23.5 mg, 0.043 mmol, 17% yield) as an off-white solid. 1 H NMR (400 MHz, DMSO-d6) δ ppm 10.30 (s, 1H), 8.43 (s, 1H), 7.84–7.90 (m, 1H), 7.43–7.50 (m, 4H), 6.79 (s, 1H), 4.67 (t, J = 5.7 Hz, 2H), 3.59–3.71 (m, 4H), 3.35–3.43 (m, 4H), 1.38–1.46 (m, 4H), 1.30 (s, 3H), 1.11 (s, 9H), 0.33 (s, 4H). m / z (ESI): 547.1 (M+H) + .

[0256] Further Examples The following compounds can be made following the same procedures as in the above examples or following procedures well known to those skilled in the art, using commercially available starting materials.

[0257]

Table 17

[0258]

Table 18

[0259] Biological Examples For the compounds in Table A, the following assay conditions were used: KIF18A Enzyme Assay: The microtubule-stimulated ATPase activity assay is used to measure the KIF18A enzyme activity after treatment with the compound. The compound was serially diluted two-fold in DMSO (Sigma Inc) over a 22-point concentration range. Recombinant human KIF18A (tagged with 1-467 His) protein was expressed using a baculovirus system and purified by affinity chromatography by Amgen Inc. The concentrations of KIF18A protein, microtubules (MT), and ATP during the reaction were optimized for a standardized homogeneous enzyme assay using the ADP-Glo™ Kinase / ATPase Assay Kit (Promega Inc). The assay measures the ADP formed from the ATPase reaction. Prepare the 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 Taxol (Cytoskeleton Inc), and 30 μg / mL porcine microtubules (Cytoskeleton Inc)]. Add the compound and KIF18A protein (30 nM) to the prepared reaction buffer, incubate at room temperature for 15 minutes, and then add ATP (K mThen, 75 μM is added to the reaction mixture and incubated at room temperature for an additional 15 minutes. Mix 5 μl of the ADP-Glo™ reagent with 2.5 μl of the reaction mixture and incubate at room temperature for 40 minutes. Add 10 μl of the ADP-Glo™ detection reagent and incubate at room temperature for 40 minutes. Read the luminescence using an EnVision microplate reader (Perkin Elmer Inc) equipped with an ultra-luminescence module. Use Genedata Screener Software (Standard 15.0.1, Genedata Inc) with a 4-parameter logistic regression fitting model to fit the concentration-response curve and determine the IC 50 .

[0260]

Table 19

[0261]

Table 20

[0262]

Table 21

[0263] The above invention has been described in some detail by way of illustration and examples for the purpose of clarity and understanding. Those skilled in the art will understand that changes and modifications can be made within the scope of the appended claims. Therefore, it should be understood that the above description is intended to be illustrative and not limiting. Accordingly, the scope of the present invention should not be determined with reference to the above description, but with reference to the appended claims, along with the full scope of equivalents to which such claims are entitled.

[0264] 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 individually set forth.

Claims

1. Formula I: 【Chemical 1】 a compound of formula I, or any pharmaceutically acceptable salt thereof [wherein, L is -NR ８ -(C=O)-, and R １ is base-Z-R ９ where Z is -NR １１ -. R ２ is a group -Y-R １０ wherein Y is -NH-, -NHSO ２ -, SO ２ NH-, or SO ２ -; R ３ is H, halo, C １～４ alk, or C １～４ haloalk, and R ４ is H, halo, C １～４ alk, or C １～４ haloalk, and R ５ is H, halo, C １～８ alk, or C １～４ haloalk, and R ６ is H, halo, C １～８ alk, or C １～４ haloalk, and R ７ is H, halo, C １～４ alk, or C １～４ haloalk, and R ８ is H or C １～４ alk, and R ９ is H, R ９ａ or R ９ｂ and is R １０ is a halo, R １０ａ or R １０ｂ and R Ｘ is 【Chemical 2】 is, R Ｘａ 、R Ｘｂ 、R Ｘｃ 、R Ｘｄ 、R Ｘｅ 、R Ｘｆ 、R Ｘｇ 、R Ｘｈ 、R Ｘｉ 、and R Ｘｊ each of which is H, halo, or R Ｘｏ is, Alternatively, R Ｘａ and R Ｘｂ each of the pair independently combines with a carbon atom bonded thereto to form a saturated monocyclic 3-membered ring that is spiro to the piperidinyl ring, where the 3-membered monocyclic ring contains 0, 1 or 2 N atoms and 0 or 1 atom selected from O and S, and further, the 3-membered monocyclic ring is substituted with 0, 1, 2 or 3 groups selected from F, Cl, Br, C １～６ alk, C １～４ haloalk, -OR ａ , -OC １～４ haloalk, CN, -NR ａ R ａ , or oxo R Ｘｏ is, independently in each case, F, Cl, Br, -CH ２ F, -CHF ２ , -CF ３ , -C(=O)OR ａ , -OR ａ , -OC １～４ haloalk, CN, NH ２ , NH(CH ３ ), or N(CH ３ ) ２ selected from the group consisting of C １～８ alk substituted with 0, 1, 2, 3, 4, or 5 groups selected from the group consisting of: R ９ａ and R １０ａ each independently contains, in each case, 0, 1, 2 or 3 N atoms and 0 or 1 atom selected from O and S, and F, Cl, Br, C １～６ alk, C １～４ haloalk, -OR ａ , -OC １～４ haloalk, 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 alkNR ａ R ａ , -OC ２～６ alkOR ａ , -SR ａ , -S(=O)R ｂ , -S(=O) ２ R ｂ , -S(=O) ２ NR 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) ２ R ｂ , -N(R ａ )S(=O) ２ NR ａ R ａ , -NR ａ C ２～６ alkNR ａ R ａ , -NR ａ C ２～６ alkOR ａ 、 -C １～６ alkNR ａ R ａ 、 -C １～６ alkOR ａ 、 -C １～６ alkN(R ａ )C(=O)R ｂ 、 -C １～６ alkOC(=O)R ｂ 、 -C １～６ alkC(=O)NR ａ R ａ 、 -C １～６ alkC(=O)OR ａ 、 R １１ 、 and selected from 0, 1, 2 or 3 groups selected from oxo, a saturated, partially saturated or unsaturated 3-membered, 4-membered, 5-membered, 6-membered or 7-membered monocyclic ring or an 8-membered, 9-membered, 10-membered, 11-membered or 12-membered bicyclic ring, selected from the group consisting of R ９ｂ and R １０ｂ are each, independently, in each case, F, Cl, Br, -CH ２ F, -CHF ２ , -CF ３ , -C(=O)OR ａ , -OR ａ , -OC １～４ haloalk, CN, NH ２ , NH(CH ３ ), or N(CH ３ ), ２ and are selected from the group consisting of C １～６ alk which is substituted with 0, 1, 2, 3, 4, or 5 groups selected from R １１ is H or C １～８ alk, and R ａ is, independently in each case, H or R ｂ and R ｂ is, independently in each case, C １～６ alk, phenyl, or benzyl, where said C １～６ alk is substituted with 0, 1, 2 or 3 substituents selected from halo, -OH, -OC １～４ alk, -NH ２ , -NHC １～４ alk, -OC(=O)C １～４ alk, or -N(C １～４ alk)C １～４ alk, and said phenyl or benzyl is substituted with 0, 1, 2 or 3 substituents selected from halo, C １～４ alk, C １－３ haloalk, -OH, -OC １～４ alk, -NH ２ , -NHC １～４ alk, -OC(=O)C １～４ alk, or -N(C １～４ alk)C １～４ alk].

2. L is -NR ８ -(C=O), and the formula (Ia): 【Chemical 3】 having, wherein said R Ｘａ and R Ｘｂ pairs, in combination with the carbon atoms to which they are each attached, form a saturated monocyclic 3-membered ring that is spiro to the piperidinyl ring, where the 3-membered monocyclic ring contains 0 N, O, and S atoms, and further, the 3-membered monocyclic ring is substituted with 0, 1, 2, or 3 groups selected from F, Cl, Br, C １～６ alk, C １～４ haloalk, -OR ａ , -OC １～４ haloalk, CN, -NR ａ R ａ , or oxo, the compound according to claim 1.

3. R ８ The compound according to claim 1 or 2, wherein R is H or methyl.

4. R Ｘｃ 、R Ｘｄ 、R Ｘｅ 、R Ｘｆ 、R Ｘｇ 、R Ｘｈ 、R Ｘｉ 、and R Ｘｊ each of which is H, halo, C １～６ alk, or C １～４ haloalk, and each pair of R Ｘａ and R Ｘｂ combines with the carbon atom to which each of them is attached to form a saturated monocyclic 3-membered ring that is spiro to the piperidinyl ring, where the ring contains 0, 1 or 2 N atoms and 0 or 1 atom selected from O and S, a compound according to any one of claims 1 to 3.

5. R Ｘｃ 、R Ｘｄ 、R Ｘｅ 、R Ｘｆ 、R Ｘｇ 、R Ｘｈ 、R Ｘｉ 、and R Ｘｊ each of which is H, methyl, or ethyl, and each pair of R Ｘａ and R Ｘｂ forms a cyclopropyl ring that is spiro to the piperidinyl ring in combination with the carbon atom to which each of them is attached, the compound according to claim 1.

6. R Ｘ is 【Chemical Formula 4】 a compound according to any one of claims 1 to 5, selected from

7. R Ｘ is [Chemical Formula 5] a compound according to any one of claims 1 to 6, wherein

8. R ９ is C substituted with 0, 1, 2 or 3 groups selected from a) H, b) F, Cl, Br, -CF ３ , or -OH, or c) a saturated, partially saturated or unsaturated 3-membered, 4-membered, 5-membered, 6-membered or 7-membered monocyclic ring containing 0, 1, 2 or 3 N atoms and 0 or 1 atom selected from O and S, and substituted with 0, 1, 2 or 3 groups selected from F, Cl, Br, methyl, ethyl, -CF １～６ , -CH ３ OH, -OH, -OCH ２ , -NH ３ , or oxo, and is a compound according to any one of claims 1 to 7 ２、 selected from monocyclic rings

9. R ９ The compound according to any one of claims 1 to 8, wherein R is selected from cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl, azetidinyl, diazinyl, oxazolidinyl, isothiazolidinyl, pyrazolyl, or pyrrolidinyl.

10. R １ is a group-Z-R ９ wherein R ９ is 0, 1, 2 or 3 OH, -CH ２ OH, methyl, or cyclopropyl or oxazolinyl substituted with an oxo group, or R ９ is 0, 1, 2 or 3 OH or CF ３ substituted C １～６ alk, a compound according to any one of claims 1 to 9.

11. R １ is base-Z-R ９ where Z is -NHSO ２ -, and R ９ is -CH ２ -CH ２ -OH, the compound according to any one of claims 1 to 8 and 10.

12. R ２ is a group -Y-R １０ where Y is -NHSO ２ - and R １０ contains 0, 1, 2 or 3 N atoms and 0 or 1 atom selected from O and S, and is a saturated, partially saturated or unsaturated 3-membered, 4-membered, 5-membered, 6-membered or 7-membered monocyclic ring substituted with 0, 1, 2 or 3 groups selected from F, Cl, Br, methyl, ethyl, CF ３ , CH ２ OH, -OH, -OCH ３ , -NH ２ , -NH(CH ３ ), and oxo, or R １０ is C alk substituted with 0, 1, 2, 3, 4, or 5 groups selected from F, Cl, Br, -OH, or -CF ３ The compound according to any one of claims 1 to 11, which is １～６ alk.

13. R ２ is a morpholinyl ring, a thiomorpholinyl ring, a cyclobutyl ring, a cyclopentyl ring, or a piperidinyl ring, wherein each of said rings is substituted with 0, 1, 2 or 3 groups selected from F, Cl, Br, methyl, CF ３ , -OH, -OCHF ２ , or oxo, a compound according to any one of claims 1 to 12.

14. R ２ The compound according to any one of claims 1 to 13, wherein R is morpholinyl or piperidinyl substituted with 0, 1, 2 or 3 selected from F or methyl.

15. R ３ The compound according to any one of claims 1 to 14, wherein R is H or F.

16. R ４ The compound according to any one of claims 1 to 15, wherein R is H.

17. R ５ The compound according to any one of claims 1 to 16, wherein R is H.

18. R ６ The compound according to any one of claims 1 to 17, wherein R is H, F, or methyl.

19. R ７ The compound according to any one of claims 1 to 18, wherein R is H.

20. as follows: 【Table 1】 【Table 2】 【Table 3】 a compound selected from the group consisting of

21. a pharmaceutical composition comprising a compound according to any one of claims 1 to 20 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable diluent or carrier.

22. A pharmaceutical composition for use in a method for treating a condition that can be treated with a KIF18A inhibitor, comprising a therapeutically effective amount of a compound according to any one of claims 1 to 20 or a pharmaceutically acceptable salt thereof, wherein the method comprises administering the pharmaceutical composition to a patient in need thereof.

23. wherein the condition is selected from cancer, psoriasis, atopic dermatitis, autoimmune diseases or inflammatory bowel diseases, the cancer is melanoma, prostate cancer, cervical cancer, breast cancer, colon cancer, sarcoma or leukemia, the autoimmune disease is rheumatoid arthritis, systemic lupus erythematosus, Sjogren's syndrome, scleroderma, mixed connective tissue disease, dermatomyositis, polymyositis, Reiter's syndrome, autoimmune lymphoproliferative syndrome (ALPS) also known as Canale-Smith syndrome, or a central nervous system autoimmune disease such as multiple sclerosis, myasthenia gravis and encephalomyelitis, and the inflammatory bowel disease can be ulcerative colitis or Crohn's disease, the pharmaceutical composition according to claim 22.

24. A pharmaceutical composition for use in a method for reducing the size of a solid tumor in a subject, comprising a therapeutically effective amount of a compound according to any one of claims 1 to 20 or a pharmaceutically acceptable salt thereof, wherein the method comprises administering the pharmaceutical composition to the subject in need thereof.

25. A pharmaceutical composition for use in a method of treating a cell growth disorder in a subject, the composition comprising a therapeutically effective amount of a compound according to any one of claims 1 to 20 or a pharmaceutically acceptable salt thereof, the method comprising administering the pharmaceutical composition to the subject in need thereof.

26. A pharmaceutical composition for use in a method of inhibiting KIF18A in a cell, the composition comprising a compound according to any one of claims 1 to 20 or a pharmaceutically acceptable salt thereof, the method comprising contacting the cell with the pharmaceutical composition.

27. The pharmaceutical composition according to claim 22, wherein the disease state is (a) a solid tumor selected from 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, or a blood-derived tumor; (b) a lymphoid hematopoietic tumor selected from leukemia, acute lymphoblastic leukemia, acute lymphoblastic leukemia, B cell lymphoma, T cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, hairy cell lymphoma and Burkitt lymphoma; (c) a myeloid hematopoietic tumor selected from acute and chronic myeloid leukemia, myelodysplastic syndrome and promyelocytic leukemia; (d) a mesenchymal-derived tumor selected from fibrosarcoma and rhabdomyosarcoma; (e) a central and peripheral nervous system tumor selected from astrocytoma, neuroblastoma, glioma and schwannoma; or (f) a cancer selected from the group consisting of melanoma, seminoma, teratoma, osteosarcoma, xeroderma pigmentosum, keratoacanthoma, follicular thyroid cancer or Kaposi sarcoma.

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