Substituted piperazine derivatives useful as T cell activators

Abstract

Formula (I): TIFF2023508375000147.tif45150 [wherein R1, R2, R3, R4, R5, R6, and m are as defined herein] or a salt thereof. Also disclosed are methods of using such compounds, and pharmaceutical compositions containing such compounds, to inhibit the activity of one or both of diacylglycerol kinase alpha (DGKα) and diacylglycerol kinase zeta (DGKζ). These compounds are useful in the treatment of proliferative disorders such as viral infections and cancer.

Description

Technical Field

[0001] (Related Application) This application claims the priority of Indian Provisional Application No. 201911053559, filed on December 23, 2019, the entire content of which is incorporated herein by reference.

[0002] (Description of the Invention) The present invention generally relates to substituted compounds that activate T cells, promote T cell proliferation, and / or exhibit anti-tumor activity. Provided herein are substituted compounds, compositions containing such compounds, and methods of using them. The present invention further relates to pharmaceutical compositions comprising at least one compound of the present invention useful for the treatment of proliferative disorders (e.g., cancer and viral infections).

Background Art

[0003] Human cancers encompass numerous genetic and epigenetic changes and produce neoantigens that may be recognized by the immune system (Sjoblom et al., (2006) Science 314:268-74). The adaptive immune system, consisting of T and B lymphocytes, has the potential to exhibit potent anti-cancer effects and is equipped with a broad range of capabilities and high specificity to respond to diverse tumor antigens. Furthermore, this immune system has considerable flexibility and a memory component. By successfully harnessing all of these characteristics of the adaptive immune system, immunotherapy becomes distinctive among all cancer treatment methods. However, although an endogenous immune response to cancer has been observed in preclinical models and patients, this response is ineffective, and established cancers are regarded as "self" and show tolerance to the immune system. The resulting state of tolerance allows tumors to utilize multiple different mechanisms to actively break anti-tumor immunity. These mechanisms include dysfunctional T cell signaling (Mizoguchi et al., (1992) Science 258:1795-98), suppressive regulatory cells (Facciabene et al., (2012) Cancer Res. 72:2162-71), and the use of endogenous "immune checkpoints", which have the function of reducing the intensity of the adaptive immune response and protecting normal tissues from secondary damage by tumors to escape immune destruction (Topalian et al., (2012) Curr. Opin. Immunol. 24:1-6; Mellman et al., (2011) Nature 480:480-489).

[0004] Diacylglycerol kinase (DGK) is a lipid kinase that mediates the conversion of diacylglycerol to phosphatidic acid, thereby arresting the function of T cells propagated through the TCR signaling pathway. Therefore, DGK functions as an intracellular checkpoint, and inhibiting DGK is expected to promote T cell signaling pathways and T cell activation. As evidence to support this, knockout mouse models of either DGKα or DGKζ show an overreactive T cell phenotype and improved antitumor immune activity (Riese M.J. et al., Journal of Biological Chemistry, (2011) 7: 5254-5265; Zha Y et al., Nature Immunology, (2006) 12:1343; Olenchock B.A. et al., (2006) 11: 1174-81). Furthermore, tumor-infiltrating lymphocytes isolated from human renal cell carcinoma patients overexpress DGKα, and it has been confirmed that they inhibit the function of T cells (Prinz, P.U. et al., J Immunology (2012) 12:5990-6000). Therefore, DGKα and DGKζ are regarded as targets in cancer immunotherapy (Riese M.J. et al., Front Cell Dev Biol. (2016) 4: 108; Chen, S.S. et al., Front Cell Dev Biol. (2016) 4: 130; Avila-Flores, A. et al., Immunology and Cell Biology (2017) 95: 549-563; Noessner, E., Front Cell Dev Biol. (2017) 5: 16; Krishna, S., et al., Front Immunology (2013) 4:178; Jing, W. et al., Cancer Research (2017) 77: 5676-5686).

[0005] There is a need for compounds useful as inhibitors of one or both of DGKα and DGKζ. Further, there is a need for compounds having selectivity for other diacylglycerol kinases, protein kinases, and / or other lipid kinases that are useful as inhibitors of one or both of DGKα and DGKζ.

[0006] Accordingly, a safe and effective agent for restoring T cell activity, lowering the antigen threshold, enhancing antitumor function, and / or overcoming the inhibitory effects of one or more endogenous immune checkpoints (e.g., PD-1, LAG-3 and TGFβ) is an important addition to the treatment of patients with proliferative disorders (e.g., cancer, as well as viral infections). SUMMARY OF THE INVENTION

[0007] The inventors have found compounds having activity as inhibitors of one or both of DGKα and DGKζ. Further, the inventors have found compounds having activity as inhibitors of one or both of DGKα and DGKζ and having selectivity for other diacylglycerol kinases, protein kinases, and / or other lipid kinases. These compounds provide useful pharmaceuticals having desirable stability, bioavailability, therapeutic index, and toxicity values important for their drugability.

[0008] The present invention provides substituted compounds of formula (I), or salts and prodrugs thereof, useful as inhibitors of DGKα, DGKζ, or both DGKα and DGKζ.

[0009] The present invention also provides a pharmaceutical composition comprising a compound of formula (I) and / or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0010] The present invention also provides a method for treating a disease or disorder related to the activity of DGKα, DGKζ, or both DGKα and DGKζ, which comprises administering to a mammalian patient a compound of formula (I) and / or a pharmaceutically acceptable salt thereof.

[0011] The present invention also provides a method and intermediates for producing a compound of formula (I) and / or a salt thereof.

[0012] The present invention also provides a compound of formula (I) and / or a pharmaceutically acceptable salt thereof for use in therapy.

[0013] The present invention also provides the use of a compound of formula (I) and / or a pharmaceutically acceptable salt thereof for the manufacture of a therapeutic agent for proliferative disorders (such as cancer and viral infections).

[0014] The compounds of formula (I) and the compositions containing the compounds of formula (I) can be used for the treatment, prevention, or cure of viral infections and various proliferative disorders (such as cancer). The pharmaceutical compositions containing these compounds are useful for the treatment, prevention, or suppression of the progression of diseases or disorders in various therapeutic fields (such as viral infections and cancer).

[0015] The above and other features of the present invention will be described in a broader scope along with the disclosure.

DETAILED DESCRIPTION OF THE INVENTION

[0016] In a first aspect, the present invention relates to a compound of formula (I):

CHEMICAL FORMULA

Chemical formula

[0017] In a second aspect, the present invention provides a compound of formula (I): [Chemical formula: [wherein: X is CR6 or N; Y is CR3 or N; R1 is H, F, Cl, Br, -CN, C 1-3 alkyl (substituted with 0 to 4 Rs 1a ), C 3-4 cycloalkyl (substituted with 0 to 4 Rs 1a ), C 1-3 alkoxy (substituted with 0 to 4 Rs 1a ), -C(O)NR a R a , -NR a R a , -S(O) n R f , or -P(O)R f R f ; Each R 1a is independently F, Cl, -CN, -OH, -OCH3, or -NR a R a ; Each R a is independently H or C 1-3 alkyl; R2 is H, C 1-3 alkyl (substituted with 0 to 4 Rs 2a ), C 2-3 alkenyl (substituted with 0 to 4 Rs 2a ), or C 3-4 cycloalkyl (substituted with 0 to 4 Rs 2a ); Each R 2a is independently F, Cl, -CN, -OH, -O(C 1-2 alkyl), C 3-4 cycloalkyl, C 3-4 alkenyl, or C 3-4 alkynyl; R3 is H, F, Cl, Br, -CN, C 1-3 alkyl, C 1-2 fluoroalkyl, C 3-4 cycloalkyl, C 3-4 fluorocycloalkyl, -NO2, or pyridinyl (substituted with 0 to 2 Rs 3a ); Each R3a is halo, -CN, C 1-3 alkyl, or C 1-3 alkoxy; R4 is -CR 4a R 4c -L-R 4b ; L is L1 or -CH2L1-; L1 is -C(O)-, -C(O)O-, -C(O)NR d -, -NR d C(O)-, -NR d C(O)NR d -, -S(O)2-, -S(O)2NR d -, -NR d S(O)2-, or -P(O)R e ; R 4a is (i) H or C 1-6 alkyl (substituted with 0 to 4 substituents independently selected from F, Cl, -CN, -OH, -OCH3, -SCH3, C 1-3 fluoroalkoxy, -NR a R a , -S(O)2R f , or -NR a S(O)2R f ); or (ii) C 3-6 cycloalkyl, C 5-14 heterocyclyl, C 6-10 aryl, or C 5-14 heteroaryl, each substituted with 0 to 4 R b ; each R b is independently F, Cl, Br, -CN, -OH, C 1-6 alkyl, C 1-3 fluoroalkyl, C 1-4 hydroxyalkyl, -(CH2) 1-2 O(C 1-3 alkyl), C 1-4 alkoxy, -O(C 1-4 hydroxyalkyl), -O(CH) 1-3 O(C 1-3 alkyl), C 1-3Fluoroalkoxy, -O(CH) 1-3 NR c R c , -OCH2CH=CH2, -OCH2C≡CH, -C(O)(C 1-4 alkyl), -C(O)OH, -C(O)O(C 1-4 alkyl), -C(O)NH2, -C(O)NH(C 1-4 alkyl), -C(O)N(C 1-4 alkyl)2, -NR c R c , -NR a S(O)2(C 1-3 alkyl), -NR a C(O)(C 1-3 alkyl), -NR a C(O)O(C 1-4 alkyl), -P(O)(C 1-3 alkyl)2, -S(O)2(C 1-3 alkyl), -O(CH2) 1-2 (C 3-6 cycloalkyl), -O(CH2) 1-2 (morpholinyl), C 3-6 cycloalkyl, cyanocyclopropyl, methylazetidinyl, acetylazetidinyl, triazolyl, tetrahydropyranyl, morpholinyl, thiophenyl, methylpiperidinyl, or -CR c R c (phenyl); R 4b is (i) H, C 1-6 alkyl (F, Cl, -CN, -OH, -OCH3, -SCH3, C 1-3 fluoroalkoxy, -NR a R a , -S(O)2R f , -NR a S(O)2R f ,

Chemical formula

[0018] In a third aspect, the present invention provides a compound of formula (I): [Chemical formula] [wherein: X is CR6 or N; Y is CR3 or N; R1 is H, F, Cl, Br, -CN, C 1-3 alkyl (0 to 4 Rs1a substituted with), C 3-4 cycloalkyl (0 to 4 R 1a substituted with), C 1-3 alkoxy (0 to 4 R 1a substituted with), -C(O)NR a R a , -NR a R a , -S(O) n R f , or -P(O)R f R f ; each R 1a is independently F, Cl, -CN, -OH, -OCH3, or -NR a R a ; each R a is independently H or C 1-3 alkyl; R2 is H, C 1-3 alkyl (0 to 4 R 2a substituted with), C 2-3 alkenyl (0 to 4 R 2a substituted with), or C 3-4 cycloalkyl (0 to 4 R 2a substituted with); each R 2a is independently F, Cl, -CN, -OH, -O(C 1-2 alkyl), C 3-4 cycloalkyl, C 3-4 alkenyl, or C 3-4 alkynyl; R3 is H, F, Cl, Br, -CN, C 1-3 alkyl, C 1-2 fluoroalkyl, C 3-4 cycloalkyl, C 3-4 fluorocycloalkyl, -NO2, or pyridinyl (0 to 2 R 3a substituted with); each R 3a is halo, -CN, C 1-3 alkyl, or C 1-3 alkoxy; R4 is -CR 4a R4c -L-R 4b wherein; L is L1 or -(CH2) 1-3 L1-; L1 is -C(O)-, -C(O)O-, -C(O)NR d -, -C(O)NR d O-, -C(O)NR d S(O)2-, -NR d -, -NR d C(O)-, -NR d C(O)O-, -NR d C(O)NR d -, O-, -S(O)2-, -S(O)2NR d -, -NR d S(O)2-, or -P(O)R e -; R 4a is (i) H or C 1-6 alkyl (substituted with 0 to 4 substituents independently selected from F, Cl, -CN, -OH, -OCH3, -SCH3, C 1-3 fluoroalkoxy, -NR a R a , -S(O)2R f , or -NR a S(O)2R f ); or (ii) C 3-6 cycloalkyl, C 5-14 heterocyclyl, C 6-10 aryl, or C 5-14 heteroaryl, each of which is substituted with 0 to 4 R b ; each R b is independently F, Cl, Br, -CN, -OH, C 1-6 alkyl, C 1-3 fluoroalkyl, C 1-4 hydroxyalkyl, -(CH2) 1-2 O(C 1-3 alkyl), C 1-4 alkoxy, -O(C 1-4 hydroxyalkyl), -O(CH) 1-3 O(C 1-3 alkyl), C1-3 fluoroalkoxy, -O(CH) 1-3 NR c R c , -OCH2CH=CH2, -OCH2C≡CH, -C(O)(C 1-4 alkyl), -C(O)OH, -C(O)O(C 1-4 alkyl), -C(O)NH2, -C(O)NH(C 1-4 alkyl), -C(O)N(C 1-4 alkyl)2, -NR c R c , -NR a S(O)2(C 1-3 alkyl), -NR a C(O)(C 1-3 alkyl), -NR a C(O)O(C 1-4 alkyl), -P(O)(C 1-3 alkyl)2, -S(O)2(C 1-3 alkyl), -O(CH2) 1-2 (C 3-6 cycloalkyl), -O(CH2) 1-2 (morpholinyl), C 3-6 cycloalkyl, cyanocyclopropyl, methylazetidinyl, acetylazetidinyl, triazolyl, tetrahydropyranyl, morpholinyl, thiophenyl, methylpiperidinyl, or -CR c R c (phenyl); R 4b is (i) -CH2CH2OCH2CH2OCH2CH2NH2; or (ii) -CH2CH2OCH2CH2OCH2CH2NH2C(O)OC(CH3)3; or (iii)

Chemical formula

[0019] The present invention, in a fourth aspect, provides a compound of formula (I):

Chemical formula

Chemical formula

[0020] In one embodiment, a compound of formula (I) or a salt thereof, wherein R1 is H, F, Cl, Br, -CN, C 1-3 alkyl (substituted with 0 to 4 R 1a ), cyclopropyl (substituted with 0 to 3 R 1a ), C 1-3 alkoxy (substituted with 0 to 3 R 1a ), -C(O)NR a R a , -NR a R a , -S(O) n CH3, or -P(O)(CH3)2; each R 1a is independently F, Cl, or -CN; R2 is H, C 1-2 alkyl (substituted with 0 to 2 R 2a ), or C 2-3 alkenyl (substituted with 0 to 2 R 2a ); each R 2a is independently F, Cl, -CN, -OH, -O(C 1-2 alkyl), cyclopropyl, C 3-4 alkenyl, or C 3-4 alkynyl; R3 is H, F, Cl, Br, -CN, C 1-2 alkyl, C 1-2 fluoroalkyl, C 3-4 cycloalkyl, -NO2, or pyridinyl (substituted with 0 to 2 R 3a ); each R 3a is F, Cl, Br, -CN, C 1-3 alkyl, or C 1-2 alkoxy; R 4a is: (i) H or C 1-4 alkyl (F, Cl, -CN, -OH, -OCH3, -SCH3, C 1-2 fluoroalkoxy, or -NR a R aselected independently, and substituted with 0 to 4 substituents); or (ii) C 3-6 cycloalkyl, C 5-10 heterocyclyl, phenyl, or C 5-10 heteroaryl, each substituted with 0 to 4 R b ; each R b is independently F, Cl, Br, -CN, -OH, C 1-4 alkyl, C 1-2 fluoroalkyl, C 1-3 hydroxyalkyl, -(CH2) 1-2 O(C 1-3 alkyl), C 1-3 alkoxy, -O(C 1-3 hydroxyalkyl), -O(CH) 1-3 O(C 1-3 alkyl), C 1-2 fluoroalkoxy, -O(CH) 1-2 NR c R c 、-OCH2CH=CH2、-OCH2C≡CH、-C(O)(C 1-3 alkyl), -C(O)OH, -C(O)O(C 1-3 alkyl), -C(O)NH2, -C(O)NH(C 1-3 alkyl), -C(O)N(C 1-3 alkyl)2, -NR c R c 、-NR a S(O)2(C 1-2 alkyl), -NR a C(O)(C 1-2 alkyl), -NR a C(O)O(C 1-3 alkyl), -P(O)(C 1-2 alkyl)2, -S(O)2(C 1-2 alkyl), -O(CH2) 1-2 (C 3-6 cycloalkyl), -O(CH2) 1-2 (morpholinyl), C 3-6 cycloalkyl, or -CR c R c (phenyl); R 4b is: (i) H, -CN, or C 1-4Alkyl (F, Cl, -CN, -OH, -OCH3, -SCH3, C 1-3 fluoroalkoxy, -NR a R a 、-S(O)2(C 1-3 alkyl), cyclopropyl, hydroxycyclopropyl,

Chem.

[0021] In one embodiment, a compound of formula (I) or a salt thereof, wherein R1 is H, F, Cl, Br, -CN, C 1-3 (alkyl (substituted with 0 - 4 R 1a (substituted with 0 - 3 R 1a (), C 1-3 (alkoxy (substituted with 0 - 3 R 1a (substituted with 0 - 3 R a R a , -NR a R a , -S(O) n CH3, or -P(O)(CH3)2; each R 1a is independently F, Cl, or -CN; R2 is H, C 1-2 (alkyl (substituted with 0 - 2 R 2a (substituted with 0 - 2 R 2-3 (), or C 2a (alkenyl (substituted with 0 - 2 R 2a (substituted with 0 - 2 R 1-2 (alkyl), cycloalkyl, C 3-4 (alkenyl, or C 3-4 (alkynyl); R3 is H, F, Cl, Br, -CN, C 1-2 (alkyl, C 1-2 (fluoroalkyl, C 3-4Cycloalkyl, -NO2, or pyridinyl (substituted with 0 to 2 Rs 3a ); each R 3a is F, Cl, Br, -CN, C 1-3 alkyl, or C 1-2 alkoxy; L1 is -C(O)-, -C(O)O-, -C(O)NR d -, -NR d C(O)-, -S(O)2NR d -, -NR d S(O)2-, or -P(O)R e -; R 4a is (i) H or C 1-4 alkyl (substituted with 0 to 4 substituents independently selected from F, Cl, -CN, -OH, -OCH3, -SCH3, C 1-2 fluoroalkoxy, or -NR a R a ) or; (ii) C 3-6 cycloalkyl, C 5-10 heterocyclyl, phenyl, or C 5-10 heteroaryl, each substituted with 0 to 4 Rs b ; each R b is, independently, F, Cl, Br, -CN, -OH, C 1-4 alkyl, C 1-2 fluoroalkyl, C 1-3 hydroxyalkyl, -(CH2) 1-2 O(C 1-3 alkyl), C 1-3 alkoxy, -O(C 1-3 hydroxyalkyl), -O(CH) 1-3 O(C 1-3 alkyl), C 1-2 fluoroalkoxy, -O(CH) 1-2 NR c R c , -OCH2CH=CH2, -OCH2C≡CH, -C(O)(C 1-3 alkyl), -C(O)OH, -C(O)O(C 1-3 alkyl), -C(O)NH2, -C(O)NH(C 1-3 alkyl), -C(O)N(C 1-3 alkyl)2, -NR c Rc ,-NR a S(O)2(C 1-2 alkyl), -NR a C(O)(C 1-2 alkyl), -NR a C(O)O(C 1-3 alkyl), -P(O)(C 1-2 alkyl)2, -S(O)2(C 1-2 alkyl), -O(CH2) 1-2 (C 3-6 cycloalkyl), -O(CH2) 1-2 (morpholinyl), C 3-6 cycloalkyl, or -CR c R c (phenyl); R 4b is (i) H, C 1-4 alkyl (F, Cl, -CN, -OH, -OCH3, -SCH3, C 1-3 fluoroalkoxy, and -NR a R a , independently selected from 0 to 4 substituents); [Chemical formula] or (ii) C 3-6 cycloalkyl, C 5-10 heterocyclyl, phenyl, naphthalenyl, or C 5-10 heteroaryl, each substituted with 0 to 3 R b ; R 4c is H or C 1-3 alkyl; each R d is, independently, H or C 1-4 alkyl; R e is C 1-4 alkyl; each R5 is, independently, F, -CN, -OH, C 1-5 alkyl (substituted with 0 to 4 R g ); C 1-2 alkoxy (substituted with 0 to 3 R g ); C 2-3 alkenyl (substituted with 0 to 4 R g ); C 2-3 alkynyl (substituted with 0 to 4 R greplaced by), C 3-4 cycloalkyl (0 to 4 R g replaced by), phenyl (0 to 3 R g replaced by), oxadiazolyl (0 to 3 R g replaced by), pyridinyl (0 to 3 R g replaced by), -(CH2) 1-2 (0 to 4 R g replaced heterocyclyl), -(CH2) 1-2 NR c C(O)(C 1-4 alkyl), -(CH2) 1-2 NR c C(O)O(C 1-4 alkyl), -(CH2) 1-2 NR c S(O)2(C 1-4 alkyl), -C(O)(C 1-4 alkyl), -C(O)OH, -C(O)O(C 1-4 alkyl), -C(O)O(C 3-4 cycloalkyl), -C(O)NR a R a or -C(O)NR a (C 3-4 cycloalkyl), or two R5 are attached to the same carbon atom to form =O; each R6 is H, F, Cl, -CN, or -CH3; and m is 0, 1, 2, or 3, a compound or a salt thereof is provided.

[0022] In one embodiment, a compound or a salt thereof of formula (I), wherein X is N; R1 is -CN; R2 is -CH3; R3 is H; L is -C(O)-, -C(O)O-, -C(O)NH-, -C(O)N(CH3)-, -CH2C(O)O-, -CH2C(O)NH-, -CH2C(O)N(CH3)-, or -P(O)(CH3)-; R 4a is (i) -CH3 or; (ii) phenyl or pyridinyl, each being substituted with 0 to 2 substituents independently selected from F, -CH3, -CF3, and -OCH3; R 4b is (i) H, C 1-4alkyl, -CH2CF3, -CH2H2OH, -CH2CH2OCH3,

Chem.

[0023] In one embodiment, a compound of formula (I) or a salt thereof, wherein X is N; Y is N; R1 is -CN; R2 is -CH3; R3 is H; L is -C(O)-, -C(O)O-, -C(O)NR d -, -C(O)NHO-, -C(O)NHS(O)2-, -CH2C(O)-, -CH2NR d -, -CH2NHC(O)-, -CH2NHC(O)O-, -CH2NHS(O)2-, -CH2C(O)O-, -CH2C(O)NR d -, -CH2CH2NHC(O)O-, or -CH2CH2NHS(O)2-; R 4a is phenyl substituted with -CF3 or -OCH3; R 4b is (i) H, -CN, C 1-4 alkyl, -CH2CN, -CH2CF3, -CH2CH2CN, -CH2CH2CF3, -CH2CH2OCH3, -CH2CH2S(O)2CH3, -CH2C(CH3)2OH, -CH2(cyclopropyl), -CH2(hydroxycyclopropyl), or

Chem.

[0024] In one embodiment, a compound or a salt thereof of formula (I), wherein R1 is H, F, Cl, Br, -CN, C 1-3 alkyl (substituted with 0 - 4 R 1a ), cyclopropyl (substituted with 0 - 3 R 1a ), C 1-3 alkoxy (substituted with 0 - 3 R 1a ), -C(O)NR a R a , -NR a R a , -S(O) n CH3, or -P(O)(CH3)2. This embodiment includes compounds wherein R1 is H, F, Cl, Br, -CN, -CH3, -CHF2, -CF3, cyclopropyl, or -OCH3. This embodiment also includes compounds wherein R1 is -CN.

[0025] In one embodiment, a compound or a salt thereof of formula (I), wherein R2 is H, C 1-2 alkyl (substituted with 0 - 2 R 2a ), or C 2-3 alkenyl (substituted with 0 - 2 R 2a ). This embodiment includes compounds wherein R2 is H, -CH3, -CH2CN, or -CH2CH2F. This embodiment also includes compounds wherein R2 is -CH3.

[0026] In one embodiment, a compound or a salt thereof of formula (I), wherein R3 is H, F, Cl, Br, -CN, C 1-2 alkyl, C 1-2 fluoroalkyl, C 3-4Cycloalkyl, -NO2, or pyridinyl (substituted with 0 to 2 Rs 3a ), a compound or a salt thereof is provided. This embodiment includes a compound wherein R3 is H, F, Cl, -CN, -CH3, -CHF2, -CF3, cyclopropyl, or -NO2. This embodiment also includes a compound wherein R3 is H.

[0027] In one embodiment, a compound or a salt thereof of formula (I), wherein R1 is H, F, Cl, Br, -CN, -CH3, -CHF2, -CF3, cyclopropyl, or -OCH3; R2 is H, -CH3, -CH2CN, or -CH2CH2F; and R3 is H, F, Cl, -CN, -CH3, -CHF2, -CF3, cyclopropyl, or -NO2, is provided. This embodiment includes a compound wherein R1 is -CN; R2 is -CH3; and R3 is H. is included.

[0028] In one embodiment, a compound or a salt thereof of formula (I), wherein L is L1, is provided. In one embodiment, a compound or a salt thereof of formula (I), wherein L is -CH2L1-, is provided. This embodiment includes a compound wherein L is -CD2L1-.

[0029] In one embodiment, a compound or a salt thereof of formula (I), wherein L1 is -C(O)-, -C(O)O-, -C(O)NR d -, -C(O)NR d O-, -C(O)NR d S(O)2-, -NR d -, -NR d C(O)-, -NR d C(O)O-, -NR d C(O)NR d -, -C(O)NR d O-, -S(O)2-, -S(O)2NR d -, -NR d S(O)2-, or -P(O)R e- provided is a compound or a salt thereof. In this embodiment, L1 is -C(O)-, -C(O)O-, -C(O)NR d -, -C(O)NHO-, -C(O)NHS(O)2-, -NR d -, -NHC(O)-, -NHC(O)O-, -NHS(O)2-, or -P(O)(CH3)-; when L is -CH2NR d -, then R 4b is H or a group other than unsubstituted C 1-6 alkyl. This embodiment includes compounds where L1 is -C(O)-, -C(O)O-, -C(O)ND-, -C(O)NDO-, -C(O)NDS(O)2-, -ND-, -NDC(O)-, -NDC(O)O-, -NDS(O)2-, or -P(O)(CH3)-.

[0030] In one embodiment, provided is a compound or a salt thereof of formula (I) wherein L1 is -C(O)-, -C(O)O-, -C(O)NR d -, -NR d C(O)-, -S(O)2NR d -, -NR d S(O)2-, or -P(O)R e -. This embodiment includes compounds where L1 is -C(O)-, -C(O)O-, -C(O)NH-, -C(O)N(CH3)-, -CH2C(O)O-, -CH2C(O)NH-, -CH2C(O)N(CH3)-, or -P(O)(CH3)-. This embodiment also includes compounds where L is -C(O)-, -C(O)O-, -C(O)ND-, -C(O)N(CD3)-, -CD2C(O)O-, -CH2C(O)ND-, -CH2C(O)N(CD3)-, or -P(O)(CD3)-.

[0031] In one embodiment, provided is a compound or a salt thereof of formula (I) wherein L1 is -C(O)- or -C(O)O-. In one embodiment, provided is a compound or a salt thereof of formula (I) wherein L1 is -C(O)NR 4d -, -NR4d C(O)-, or -NR 4d C(O)NR 4d -, there is provided a compound or a salt thereof. In this embodiment, each R 4d is, independently, H, D, -CH3, or -CD3, and the compound is also included.

[0032] In one embodiment, a compound or a salt thereof of formula (I), wherein L1 is -C(O)NR d O- or -C(O)NR d S(O)2-, there is provided a compound or a salt thereof. In this embodiment, each R 4d is, independently, H, D, -CH3, or -CD3, and the compound is included. In one embodiment, a compound or a salt thereof of formula (I), wherein L1 is -NR d -, -NR d C(O)-, -NR d C(O)O-, or -NR d C(O)NR d -; when L is -CH2NR d -, then R 4b is H or a group other than unsubstituted C 1-6 alkyl, there is provided a compound or a salt thereof.

[0033] In one embodiment, a compound or a salt thereof of formula (I), wherein L1 is -S(O)2-, -S(O)2NR 4d -, or -NR 4d S(O)2-, there is provided a compound or a salt thereof. In this embodiment, each R 4d is, independently, H, D, -CH3, or -CD3, and the compound is included. In one embodiment, a compound or a salt thereof of formula (I), wherein L1 is -P(O)R 4e -, there is provided a compound or a salt thereof. In this embodiment, each R 4d is, independently, H, D, -CH3, or -CD3, and the compound is included.

[0034] In one embodiment, a compound of formula (I) or a salt thereof, wherein R 4a is (i) H or C 1-4 alkyl (substituted with 0 to 4 substituents independently selected from F, Cl, -CN, -OH, -OCH3, -SCH3, C 1-2 fluoroalkoxy, or -NR a R a ); or (ii) C 3-6 cycloalkyl, C 5-10 heterocyclyl, phenyl, or C 5-10 heteroaryl, each substituted with 0 to 4 R b . There is provided a compound or a salt thereof. In this embodiment, R 4a is (i) -CH3; or (ii) phenyl or pyridinyl, each substituted with 0 to 2 substituents independently selected from F, -CH3, -CF3, and -OCH3. Also included in this embodiment are compounds wherein (i) is -CH3; or (ii) is phenyl or pyridinyl, each substituted with 0 to 2 substituents independently selected from F, -CH3, -CF3, and -OCH3.

[0035] In one embodiment, a compound of formula (I) or a salt thereof, wherein R 4a is H or C 1-6 alkyl (substituted with 0 to 4 substituents independently selected from F, Cl, -CN, -OH, -OCH3, -SCH3, C 1-3 fluoroalkoxy, -NR a R a , -S(O)2R f , or -NR a S(O)2R f ). There is provided a compound or a salt thereof. In this embodiment, R 4a is H or C 1-4 alkyl (substituted with 0 to 4 substituents independently selected from F, Cl, -CN, -OH, -OCH3, -SCH3, C 1-2 fluoroalkoxy, or -NR a R aCompounds are included which are substituted with 0 to 4 substituents selected independently thereof. In this embodiment, R 4a Compounds where it is -CH3 are also included.

[0036] In one embodiment, a compound of formula (I) or a salt thereof, wherein R 4a is C 3-6 cycloalkyl, C 5-14 heterocyclyl, C 6-10 aryl, or C 5-14 heteroaryl, each being substituted with 0 to 4 R b is provided. In this embodiment, R 4a is C 3-6 cycloalkyl, C 5-10 heterocyclyl, phenyl, naphthalenyl, or C 5-10 heteroaryl, each being substituted with 0 to 3 R b Compounds are included. In this embodiment, R 4a is phenyl or pyridinyl, each being substituted with 0 to 2 substituents independently selected from F, -CH3, -CF3, and -OCH3 are also included.

[0037] In one embodiment, a compound of formula (I) or a salt thereof, wherein R 4b is (i) H, -CN, or C 1-4 alkyl (F, Cl, -CN, -OH, -OCH3, -SCH3, C 1-3 fluoroalkoxy, -NR a R a -S(O)2(C 1-3 alkyl), cyclopropyl, or hydroxycyclopropyl,

Chemical formula

[0038] In one embodiment, the compound of formula (I) or a salt thereof, wherein R 4b is (i) H, C 1-4 alkyl (substituted with 0 to 4 substituents independently selected from F, Cl, -CN, -OH, -OCH3, -SCH3, C 1-3 fluoroalkoxy, and -NR a R a ); or (ii) C

Chemical formula

Chemical formula

[0039] In one embodiment, the compound of formula (I) or a salt thereof, wherein R 4b is H, C 1-6 alkyl (substituted with 0 to 4 substituents independently selected from F, Cl, -CN, -OH, -OCH3, -SCH3, C 1-3 fluoroalkoxy, -NR a R a , -S(O)2R f , -NR a S(O)2R f );

Chemical formula

Chemical formula

Chemical formula

[0040] In one embodiment, a compound or a salt thereof of formula (I), wherein R 4b is (i) H, -CN, C 1-4 alkyl, -CH2CN, -CH2CHF2, -CH2CF3, -CH2CH2OH, -CH2CH2CN, -CH2CH2CF3, -CH2CH2OCH3, -CH2CH2S(O)2CH3, -CH2C(CH3)2OH, -CH2(cyclopropyl), -CH2(hydroxycyclopropyl),

Chemical formula

[0041] In one embodiment, it is a compound or a salt thereof of formula (I), wherein R 4b is (i) H, -CN, C 1-4 alkyl, -CH2CN, -CH2CHF2, -CH2CF3, -CH2CH2OH, -CH2CH2CN, -CH2CH2CF3, -CH2CH2OCH3, -CH2CH2S(O)2CH3, -CH2C(CH3)2OH, -CH2(cyclopropyl), -CH2(hydroxycyclopropyl),

Chemical formula

[0042] In one embodiment, it is a compound or a salt thereof of formula (I), wherein R 4b is azetidinyl, cyclopropyl, cyclopentyl, morpholinyl, pyrazolyl, isoxazolyl, piperidinyl, piperazinyl, pyrazolyl, pyrrolidinyl, dioxido thiomorpholinyl, or azaspiro[2.5]octanyl, and each is substituted with 0 to 2 R b and there is provided a compound or a salt thereof. In this embodiment, each R b is independently F, -OH, -CH3, -CH2OH, -C(CH3)2OH, -CH2OCH3, -OCH3, or -S(O)2CH3, and the compound is included.

[0043] In one embodiment, it is a compound or a salt thereof of formula (I), wherein R 4b is H, C 1-6 alkyl (F, Cl, -CN, -OH, -OCH3, -SCH3, C 1-3 fluoroalkoxy, -NR a R a , -S(O)2R f , or -NRa S(O)2R f A compound or a salt thereof is provided, which is substituted with 0 to 4 substituents independently selected from the group consisting of:). In this embodiment, R 4b is H, C 1-4 alkyl (F, Cl, -CN, -OH, -OCH3, -SCH3, C 1-3 fluoroalkoxy, and -NR a R a substituted with 0 to 4 substituents independently selected from the group consisting of:). The compound includes those where R 4b is H, C 1-4 alkyl-CH2CF3, -CH2H2OH, or -CH2CH2OCH3. In one embodiment, the compound of formula (I) or a salt thereof, wherein R 4b is

Chemical formula

[0044] In one embodiment, the compound of formula (I) or a salt thereof, wherein R 4b is C 3-6 cycloalkyl, C 5-14 heterocyclyl, C 6-10 aryl, or C 5-14 heteroaryl, each of which is substituted with 0 to 4 R b A compound or a salt thereof is provided. In this embodiment, R 4b is C 3-6 cycloalkyl, C 5-10 heterocyclyl, phenyl, naphthalenyl, or C 5-10 heteroaryl, each of which is substituted with 0 to 3 R b The compound includes those where R 4b is cyclopropyl, cyclopentyl, morpholinyl, pyrrolyl, methylpyrazolyl, isoxazolyl, piperidinyl, or methylpiperazinyl.

[0045] In one embodiment, a compound of formula (I) or a salt thereof, wherein R 4c is H or C 1-3 alkyl, a compound or a salt thereof is provided. This embodiment includes compounds wherein R 4c is H. This embodiment also includes compounds wherein R 4c is D.

[0046] In one embodiment, a compound of formula (I) or a salt thereof, wherein R 4a is (i) -CH3; or (ii) phenyl or pyridinyl, each substituted with 0 to 2 substituents independently selected from F, -CH3, -CF3, and -OCH3; R 4b is (i) H, C 1-4 alkyl, -CH2CF3, -CH2H2OH, -CH2CH2OCH3,

Chemical formula

[0047] In one embodiment, a compound of formula (I) or a salt thereof, wherein m is 1, 2, or 3; each R5 is independently F, -CN, -OH, C 1-5 alkyl (substituted with 0 to 4 R g ), C 1-2 alkoxy (substituted with 0 to 3 R g ), C 2-3 alkenyl (substituted with 0 to 4 R g ), C 2-3 alkynyl (substituted with 0 to 4 R g ), C 3-4 cycloalkyl (substituted with 0 to 4 R g ), phenyl (substituted with 0 to 3 R g ), oxadiazolyl (substituted with 0 to 3 R greplaced), pyridinyl (0 to 3 R g replaced), -(CH2) 1-2 (0 to 4 R g replaced heterocyclyl), -(CH2) 1-2 NR c C(O)(C 1-4 alkyl), -(CH2) 1-2 NR c C(O)O(C 1-4 alkyl), -(CH2) 1-2 NR c S(O)2(C 1-4 alkyl), -C(O)(C 1-4 alkyl), -C(O)OH, -C(O)O(C 1-4 alkyl), -C(O)O(C 3-4 cycloalkyl), -C(O)NR a R a or -C(O)NR a (C 3-4 cycloalkyl), or two R5 are bonded to the same carbon atom to form =O, a compound or a salt thereof is provided. In this embodiment, each R5 is, independently, F, -CN, -OH, C 1-2 alkyl (0 to 3 R g replaced), C 1-2 alkoxy (0 to 3 R g replaced), C 3-4 cycloalkyl (0 to 2 R g replaced), -C(O)(C 1-4 alkyl), -C(O)OH, -C(O)O(C 1-2 alkyl), or -C(O)NR a R a or two R5 are bonded to the same carbon atom to form =O, a compound is included. Also, in this embodiment, a compound is included wherein each R5 is -CH3 or two R5 are bonded to the same carbon atom to form =O.

[0048] In one embodiment, a compound or a salt thereof of formula (I) wherein m is 2 and two R5 are bonded to the same carbon atom to form =O is provided. In one embodiment, there is provided a compound of formula (I) or a salt thereof, wherein m is 0.

[0049] In one embodiment, there is provided a compound of formula (I) or a salt thereof, wherein m is 1, 2, or 3. This embodiment includes compounds wherein m is 1 or 2. This embodiment also includes compounds wherein m is 1. In one embodiment, there is provided a compound of formula (I) or a salt thereof, wherein m is 2 or 3. This embodiment includes compounds wherein m is 2. In one embodiment, there is provided a compound of formula (I) or a salt thereof, wherein m is 3.

[0050] In one embodiment, a compound of formula (I) or a salt thereof, (II):

Chemical formula

[0051] In one embodiment, a compound of formula (I) or a salt thereof, having the structural formula (III):

Chemical formula

[0052] In one embodiment, the compound of formula (III) or a salt thereof is provided where R 5a is -CH3 and R 5c is -CH3. In one embodiment, the compound of formula (III) or a salt thereof is provided where R 5a is -CH3 and R 5c is -CH2CH3. In one embodiment, the compound of formula (III) or a salt thereof is provided where R 5a is -CH2CH3 and R 5c is -CH3.

[0053] In one embodiment, the compound of formula (III) or a salt thereof is provided where R 5a is -CH2CH3 and R 5c is -CH2CH3. In one embodiment, the compound of formula (III) or a salt thereof is provided where R 5a is -CH3 and R 5c is -CH2OH. In one embodiment, the compound of formula (III) or a salt thereof is provided where R 5a is -CH3 and R 5c is -CH2OCH3.

[0054] In one embodiment, a compound of formula (I) or a salt thereof, having the structural formula:

Chemical formula

[0055] In one embodiment, a compound of formula (I) or a salt thereof, isopropyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate (1-2); methyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(5-fluoropyridin-2-yl)acetate (3-4); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-N-(2,2-difluoroethyl)-2-(4-fluorophenyl)acetamide (5-6); methyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate (7-8); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetic acid (9-10); (8-((2S,5R)-4-(1-(4-fluorophenyl)-2-oxo-2-(piperidin-1-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (11-12); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetamide (13-14); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)-N-(1-methyl-1H-pyrazol-3-yl)acetamide (15-16);8-((2S,5R)-4-(1-(4-Fluorophenyl)-2-morpholino-2-oxoethyl)-2,5-dimethylpiperazin-1-yl)-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (17-18); 8-((2S,5R)-4-(1-(4-Fluorophenyl)-2-(4-methylpiperazin-1-yl)-2-oxoethyl)-2,5-dimethylpiperazin-1-yl)-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (19-20); 8-((2S,5R)-4-(1-(4-Fluorophenyl)-2-oxo-2-(pyrrolidin-1-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (21-22); 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)-N,N-dimethylacetamide (23-24); 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)-N-methylacetamide (25-26); 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)-N-(2-hydroxyethyl)acetamide (27-28); 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)-N-(isoxazol-3-yl)acetamide (29-30); Methyl 3-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-3-(4-fluorophenyl)propanoate (31-32);N-(tert-Butyl)-3-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-3-(4-fluorophenyl)propanamide (33 - 34); 3-[(2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl]-3-(4-fluorophenyl)-N-(propan-2-yl)propanamide (35); 3-[(2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl]-3-(4-fluorophenyl)-N-(propan-2-yl)propanamide (36); 3-[(2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl]-3-(4-fluorophenyl)propanamide (37); 3-[(2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl]-3-(4-fluorophenyl)propanamide (38); 3-[(2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl]-3-(4-fluorophenyl)-N-methylpropanamide (39); 3-[(2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl]-3-(4-fluorophenyl)-N-methylpropanamide (40); 3-[(2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl]-3-(4-fluorophenyl)-N-methyl-N-(propan-2-yl)propanamide (41);3-[(2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl]-3-(4-fluorophenyl)-N-methyl-N-(propan-2-yl)propanamide (42); 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-N-cyclopentylpropanamide (43-44); 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-N-(1-methyl-1H-pyrazol-4-yl)propanamide (45); 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-N-(1-methyl-1H-pyrazol-4-yl)propanamide (46); 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-N-cyclopropylpropanamide (47); 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-N-cyclopropylpropanamide (48); 8-((2S,5R)-4-((Dimethylphosphoryl)(4-fluorophenyl)methyl)-2,5-dimethylpiperazin-1-yl)-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (49-50); 3-(3-(But-3-yn-1-yl)-3H-diazirin-3-yl)propyl 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate (57);or 2-(3-methyl-3H-diazirin-3-yl)ethyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate (62), provided as a compound.;

[0056] In one embodiment, a compound of formula (I) or a salt thereof, ethyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetate (51); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetic acid (52); 4-((2S,5R)-2,5-diethyl-4-(2-morpholino-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (53-54); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-N-(2-methoxyethyl)-2-(4-(trifluoromethyl)phenyl)acetamide (55-56); methyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-methoxyphenyl)acetate (58); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-methoxyphenyl)acetic acid (59); 3-(3-(but-3-yn-1-yl)-3H-diazirin-3-yl)propyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-methoxyphenyl)acetate (60-61); ethyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetate (69-70);2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetic acid (71-72); 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-N-((1-hydroxycyclopropyl)methyl)-2-(4-(trifluoromethyl)phenyl)acetamide (73-74); 4-((2S,5R)-2,5-diethyl-4-(2-morpholino-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-6-(12-methyl)-1-methylpyrido[3,2-d]pyrimidin-2(1H)-one (75-76); 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-N-(2-methoxyethyl)-2-(4-(trifluoromethyl)phenyl)acetamide (77-78); 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-N-(2-hydroxy-2-methylpropyl)-2-(4-(trifluoromethyl)phenyl)acetamide (79-80); 4-((2S,5R)-2,5-diethyl-4-(2-(3-hydroxy-3-methylazetidin-1-yl)-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (81-82); 4-((2S,5R)-2,5-diethyl-4-(2-oxo-2-(6-azaspiro[2.5]octan-6-yl)-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (83-84);2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-N-(cyclopropylmethyl)-N-methyl-2-(4-(trifluoromethyl)phenyl)acetamide (85 - 86); 4-((2S,5R)-2,5-diethyl-4-(2-(3-(hydroxymethyl)azetidin-1-yl)-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (87 - 88); 4-((2S,5R)-2,5-diethyl-4-(2-(3-(2-hydroxypropan-2-yl)azetidin-1-yl)-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (89 - 90); 4-((2S,5R)-2,5-diethyl-4-(2-(3-(methylsulfonyl)azetidin-1-yl)-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (91 - 92); 4-((2S,5R)-2,5-diethyl-4-(2-(4-(methylsulfonyl)piperidin-1-yl)-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (93 - 94); 4-((2S,5R)-4-(2-(3,3-difluoroazetidin-1-yl)-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)-2,5-diethylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (95 - 96);4-((2S,5R)-4-(2-(1,1-Dioxidothiomorpholino)-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)-2,5-diethylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (97 - 98); 4-((2S,5R)-2,5-diethyl-4-(2-(3-methoxy-3-methylazetidin-1-yl)-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (99 - 100); 4-((2S,5R)-2,5-diethyl-4-(2-((S)-3-hydroxypyrrolidin-1-yl)-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (101 - 102); 4-((2S,5R)-2,5-diethyl-4-(2-((S)-2-(hydroxymethyl)pyrrolidin-1-yl)-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (103 - 104); 4-((2S,5R)-2,5-diethyl-4-(2-((R)-2-(hydroxymethyl)pyrrolidin-1-yl)-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (105 - 106); 4-((2S,5R)-2,5-diethyl-4-(2-((R)-3-hydroxypyrrolidin-1-yl)-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (107 - 108);4-((2S,5R)-2,5-Diethyl-4-(2-((S)-2-(methoxymethyl)pyrrolidin-1-yl)-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (109-110); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-4-yl)-2,5-diethylpiperazin-1-yl)-N-isopropyl-N-methyl-2-(4-(trifluoromethyl)phenyl)acetamide (111-112); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-4-yl)-2,5-diethylpiperazin-1-yl)-N-isopropyl-2-(4-(trifluoromethyl)phenyl)acetamide (113-114); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-4-yl)-2,5-diethylpiperazin-1-yl)-N-(2-methoxyethyl)-N-methyl-2-(4-(trifluoromethyl)phenyl)acetamide (115-116); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-4-yl)-2,5-diethylpiperazin-1-yl)-N-methoxy-2-(4-(trifluoromethyl)phenyl)acetamide (117-118); N-cyano-2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetamide (119-120); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-4-yl)-2,5-diethylpiperazin-1-yl)-N-(cyanomethyl)-2-(4-(trifluoromethyl)phenyl)acetamide (121-122);2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-N-(2,2,2-trifluoroethyl)-2-(4-(trifluoromethyl)phenyl)acetamide (123-124); 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-N-(2-cyanoethyl)-2-(4-(trifluoromethyl)phenyl)acetamide (125-126); 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)-N-(3,3,3-trifluoropropyl)acetamide (127-128); 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-N-(2-(methylsulfonyl)ethyl)-2-(4-(trifluoromethyl)phenyl)acetamide (129-130); 2-((2R,5S)-4-(6-Cyano-1-methyl; -2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-N-methoxy-2-(4-(trifluoromethyl)phenyl)acetamide (131 - 132); ethyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetate (133); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetic acid (134); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-N-methoxy-2-(4-(trifluoromethyl)phenyl)acetamide (135 - 136); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-N-(cyanomethyl)-2-(4-(trifluoromethyl)phenyl)acetamide (137 - 138); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-N-(2,2,2-trifluoroethyl)-2-(4-(trifluoromethyl)phenyl)acetamide (139 - 140); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-N-(2-cyanoethyl)-2-(4-(trifluoromethyl)phenyl)acetamide (141 - 142);2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)-N-(3,3,3-trifluoropropyl)acetamide (143-144); 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-N-(2-(methylsulfonyl)ethyl)-2-(4-(trifluoromethyl)phenyl)acetamide (145-146); N-Cyano-2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetamide (147-148); 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-N-(methylsulfonyl)-2-(4-(trifluoromethyl)phenyl)acetamide (149-150); 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-N-(cyclopropylsulfonyl)-2-(4-(trifluoromethyl)phenyl)acetamide (151-152); Methyl 3-((2R,5S)-4-(6-chloro-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-3-(4-(trifluoromethyl)phenyl)propanoate (153); Methyl 3-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-3-(4-(trifluoromethyl)phenyl)propanoate (154-155);3-((2R,5S)-4-(6-chloro-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-3-(4-(trifluoromethyl)phenyl)propanoic acid (156); 3-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-3-(4-(trifluoromethyl)phenyl)propanoic acid (157-158); 4-((2S,5R)-2,5-diethyl-4-(3-oxo-3-(6-azaspiro[2.5]octan-6-yl)-1-(4-(trifluoromethyl)phenyl)propyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (159-160); 3-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-N-methyl-N-(1-methylpiperidin-4-yl)-3-(4-(trifluoromethyl)phenyl)propanamide (161-162); 3-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-N-(2-methoxyethyl)-N-methyl-3-(4-(trifluoromethyl)phenyl)propanamide (163-164); 4-((2S,5R)-2,5-diethyl-4-(3-((R)-2-(methoxymethyl)pyrrolidin-1-yl)-3-oxo-1-(4-(trifluoromethyl)phenyl)propyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (165-166); methyl (3-((2R,5S)-4-(6-chloro-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-3-(4-(trifluoromethyl)phenyl)propyl)carbamate (167);3-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-3-(4-(trifluoromethyl)phenyl)propanoic acid (168-169); N-(3-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-3-(4-(trifluoromethyl)phenyl)propyl)methanesulfonamide (170-171); 4-((2S,5R)-4-(2-((Cyanomethyl)(methyl)amino)-1-(4-(trifluoromethyl)phenyl)ethyl)-5-ethyl-2-methylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (172-173); 4-((2S,5R)-4-(2-((Cyanomethyl)amino)-1-(4-(trifluoromethyl)phenyl)ethyl)-5-ethyl-2-methylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (174-175); 4-((2S,5R)-5-Ethyl-2-methyl-4-(2-((2,2,2-trifluoroethyl)amino)-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (176-177); 4-((2S,5R)-4-(2-((2-Cyanoethyl)amino)-1-(4-(trifluoromethyl)phenyl)ethyl)-5-ethyl-2-methylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (178-179); 4-((2S,5R)-5-Ethyl-2-methyl-4-(1-(4-(trifluoromethyl)phenyl)-2-((3,3,3-trifluoropropyl)amino)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (180-181);4-((2S,5R)-4-(2-((cyanomethyl)(methyl)amino)-1-(4-(trifluoromethyl)phenyl)ethyl)-2,5-diethylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (182 - 183); 4-((2S,5R)-4-(2-((cyanomethyl)(methyl)amino)-1-(4-(trifluoromethyl)phenyl)ethyl)-2,5-diethylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (184 - 185); 4-((2S,5R)-2,5-diethyl-4-(2-((2,2,2-trifluoroethyl)amino)-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (186 - 187); 4-((2S,5R)-4-(2-((2-cyanoethyl)amino)-1-(4-(trifluoromethyl)phenyl)ethyl)-2,5-diethylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (188 - 189); 4-((2S,5R)-2,5-diethyl-4-(1-(4-(trifluoromethyl)phenyl)-2-((3,3,3-trifluoropropyl)amino)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (190 - 191); N-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl)acetamide (192 - 193); N-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl)isobutyramide (194 - 195);Methyl (2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl)carbamate (196-197);isopropyl (2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl)carbamate (198-199);N-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1 -yl)-2-(4-(trifluoromethyl)phenyl)ethyl)methanesulfonamide (200-201);N-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl)acetamide (202-203);N-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl)acetamide (202-203); (2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl)isobutyramide (204-205); Ethyl (2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl)carbamate (206-207); Isopropyl (2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl)carbamate (208-209); or N-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl)methanesulfonamide (210-211), a compound is provided.

[0057] In one embodiment, a compound of formula (I) or a salt thereof, tert-butyl (2-(2-(2-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetamido)ethoxy)ethoxy)ethyl)carbamate (63); N-(2-(2-(2-aminoethoxy)ethoxy)ethyl)-2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetamide·TFA (64); N-(2-(2-(2-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetamido)ethoxy)ethoxy)ethyl)-3’,6’-dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9’-xanthene]-5-carboxamide (65); 2,2-dimethyl-4-oxo-3,8,11-trioxa-5-azatridecan-13-yl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate (66); 2-(2-(2-aminoethoxy)ethoxy)ethyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate·TFA (67); or 2-(2-(2-(3’,6’-dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9’-xanthene]-5-carboxamide)ethoxy)ethoxy)ethyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate (68) is provided.

[0058] The present invention may be embodied in other specific forms without departing from its essence or essential characteristics. The present invention includes any combination of aspects and / or embodiments of the present invention described herein. It is understood that any and all embodiments of the present invention may be combined with any other embodiment for the purpose of explaining further embodiments. It is also understood that the individual elements of the embodiments are meant to be combined with any and all other elements from any embodiment for the purpose of explaining further embodiments.

[0059] Definitions The features and advantages of the present invention can be more readily understood by those skilled in the art by reading the following detailed description. For clarity, it is understood that certain features of the present invention described in the context of separate embodiments may be combined to form one embodiment. Conversely, for brevity, the various features of the present invention described in the context of a single embodiment may be combined to form sub-combinations thereof. The embodiments illustrated or described as preferred herein are intended as examples and not for purposes of limitation.

[0060] Unless otherwise specified herein, references in the singular may also include the plural. For example, "a" and "an" may refer to one, or one or more.

[0061] As used herein, the phrase "compound and / or a salt thereof" refers to at least one compound, at least one salt of a compound, or a combination thereof. For example, the compound of formula (I) and / or a salt thereof includes the compound of formula (I); two compounds of formula (I); a salt of the compound of formula (I); the compound of formula (I), and one or more salts of the compound of formula (I); and salts of two or more compounds of formula (I).

[0062] Unless otherwise specified, any atom with an unsatisfied valence is considered to include sufficient hydrogen atoms to satisfy the valence.

[0063] The definitions set forth herein supersede any definitions set forth in any patent, patent application, and / or patent application publication incorporated herein by reference.

[0064] Definitions of various terms used to describe the present invention are listed below. These definitions apply to the terms used throughout the specification, individually or as part of a larger group, unless otherwise limited in a particular case.

[0065] Throughout this specification, the groups and substituents may be selected by one of ordinary skill in the art to provide stable moieties and compounds.

[0066] In accordance with the convention used in the art,

Chem.

[0067] As used herein, the terms "halo" and "halogen" refer to F, Cl, Br, and I.

[0068] The term "cyano" refers to the group -CN.

[0069] The term "amino" refers to the group -NH2. The term "oxo" refers to the group =O.

[0070] As used herein, the term "alkyl" refers to both branched and straight-chain saturated aliphatic hydrocarbon groups having, for example, 1 to 12 carbon atoms, 1 to 6 carbon atoms, and 1 to 4 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (e.g., n-propyl and i-propyl), butyl (e.g., n-butyl, i-butyl, sec-butyl, and t-butyl), and pentyl (e.g., n-pentyl, isopentyl, neopentyl), n-hexyl, 2-methylpentyl, 2-ethylbutyl, 3-methylpentyl, and 4-methylpentyl. When a number is shown subscripted after the symbol "C", the subscripted letter more specifically limits the number of carbon atoms that a particular group may contain. For example, "C 1-4 alkyl" means straight-chain and branched-chain alkyl groups having 1 to 4 carbon atoms.

[0071] As used herein, the term "fluoroalkyl" means encompassing both branched and straight-chain saturated aliphatic hydrocarbon groups substituted with one or more fluorine atoms. For example, "C 1-4 fluoroalkyl" means encompassing C1, C2, C3, and C4 alkyl groups substituted with one or more fluorine atoms. Representative examples of fluoroalkyl groups include, but are not limited to, -CF3 and -CH2CF3.

[0072] The term "hydroxyalkyl" includes both branched and straight-chain saturated alkyl groups substituted with one or more hydroxyl groups. For example, "hydroxyalkyl" includes -CH2OH, -CH2CH2OH, and C 1-4 hydroxyalkyl.

[0073] The term "alkenyl" refers to a straight-chain or branched-chain hydrocarbon group containing 2 to 12 carbon atoms and at least one carbon-carbon double bond. Examples of such groups include ethenyl or allyl. For example, "C 2-6 alkenyl" represents a straight-chain or branched-chain alkenyl group having 2 to 6 carbon atoms.

[0074] The term "alkynyl" refers to a straight-chain or branched-chain hydrocarbon group containing 2 to 12 carbon atoms and at least one carbon-carbon triple bond. Examples of such groups include ethynyl. For example, "C 2-6 alkynyl" represents a straight-chain or branched-chain alkynyl group having 2 to 6 carbon atoms.

[0075] As used herein, the term "cycloalkyl" refers to a group derived from a non-aromatic monocyclic hydrocarbon molecule or a non-aromatic polycyclic hydrocarbon molecule by removing one hydrogen atom from a saturated ring carbon atom. Representative examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclopentyl, and cyclohexyl. When a number is shown subscripted after the symbol "C", the subscript more specifically limits the number of carbon atoms that a particular cycloalkyl group may contain. For example, "C 3-6 cycloalkyl" means a cycloalkyl group having 3 to 6 carbon atoms.

[0076] As used herein, the term "fluorocycloalkyl" means encompassing a cycloalkyl group substituted with one or more fluorine atoms.

[0077] As used herein, the term "alkoxy" is an alkyl group that is connected to a part of the parent molecule via an oxygen atom, for example, a methoxy group (-OCH3). For example, "C 1-3 alkoxy" means an alkoxy group having 1 to 3 carbon atoms.

[0078] The terms "fluoroalkoxy" and "-O(fluoroalkyl)" represent the fluoroalkyl groups described above that are bonded via an oxygen bond (-O-). For example, "C 1-4 fluoroalkoxy" is intended to encompass C1, C2, C3, and C4 fluoroalkoxy groups.

[0079] The terms "carbocyclo", "carbocyclic", or "carbocyclyl" may be used synonymously and refer to a cyclic group having at least one saturated or partially saturated non-aromatic ring in which all atoms of all rings are carbon. The carbocyclyl ring may be unsubstituted or may have one or more substituents within the valence range. Therefore, this term includes non-aromatic rings (e.g., cycloalkyl, cycloalkenyl, and cycloalkynyl rings). Examples of bicyclic carbocyclyl groups include indanyl, indenyl, dihydronaphthalenyl, tetrahydronaphthalenyl, hexahydronaphthalenyl, octahydronaphthalenyl, decahydronaphthalenyl, bicycloheptanyl, bicyclooctanyl, and bicyclononanyl.

[0080] As used herein, the term "aryl" refers to a group of atoms obtained by removing one hydrogen bonded to an aromatic ring from a molecule containing an aromatic carbon ring. Representative examples of aryl groups include, but are not limited to, phenyl and naphthyl. The aryl ring may be unsubstituted or may have one or more substituents as permitted by valence.

[0081] As used herein, the term "benzyl" refers to a methyl group in which one of the hydrogen atoms is replaced by a phenyl group. The phenyl ring may be unsubstituted or may have one or more substituents as permitted by valence.

[0082] The term "heteroatom" refers to oxygen (O), sulfur (S), and nitrogen (N).

[0083] The terms "heterocyclo", "heterocycle", or "heterocyclyl" may be used interchangeably and refer to a cyclic group having at least one saturated or partially saturated non-aromatic ring, where one or more of the rings have at least one heteroatom (O, S, or N), and the heteroatom-containing ring preferably has 1 to 3 heteroatoms independently selected from O, S, and / or N. The rings of such groups having heteroatoms may contain 1 or 2 oxygen or sulfur atoms, and / or 1 to 4 nitrogen atoms, provided that the total number of heteroatoms in each ring is 4 or less and further provided that the ring contains at least one carbon atom. The nitrogen and sulfur atoms may be optionally oxidized, and the nitrogen atoms may be optionally quaternized. The heterocyclo group may be attached by any available nitrogen or carbon atom. The heterocyclo ring may be unsubstituted or may have one or more substituents as permitted by valence.

[0084] Examples of monocyclic heterocyclyl groups include pyrrolidinyl, imidazolinyl, oxazolidinyl, isoxazolinyl, thiazolidinyl, isothiazolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, 2-oxazepinyl, azepinyl, 4-piperidonyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, thiomorpholinyl sulfoxide, thiomorpholinyl sulfone, 1,3-dioxolane, tetrahydro-1,1-dioxothienyl, dihydroisoindolyl, and tetrahydroquinolinyl.

[0085] The term "heteroaryl" refers to substituted and unsubstituted aromatic 5- or 6-membered monocyclic groups and 9- or 10-membered bicyclic groups having at least one heteroatom (O, S, or N) in at least one ring, said heteroatom-containing ring preferably having 1, 2, or 3 heteroatoms independently selected from O, S, and / or N. The heteroaryl group of each ring containing heteroatoms may contain 1 or 2 oxygen or sulfur atoms and / or 1 to 4 nitrogen atoms, provided that the total number of heteroatoms in each ring is 4 or less and each ring contains at least one carbon atom. The fused rings forming the bicyclic group may be aromatic and may contain only carbon atoms. The nitrogen and sulfur atoms may be oxidized as appropriate, and the nitrogen atoms may be quaternized as appropriate. The bicyclic heteroaryl group must contain only aromatic rings. The heteroaryl group may be bonded at any available nitrogen or carbon atom. The heteroaryl ring system may be unsubstituted or may have one or more substituents.

[0086] Examples of monocyclic heteroaryl groups include pyrrolyl, pyrazolyl, pyrazolinyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, thiadiazolyl, isothiazolyl, furanyl, thiophenyl, oxadiazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, and triazinyl.

[0087] Examples of bicyclic heteroaryl groups include indolyl, benzothiazolyl, benzodioxolyl, benzoxazolyl, benzothienyl, quinolinyl, tetrahydroisoquinolinyl, isoquinolinyl, benzimidazolyl, benzopyranyl, indolizinyl, benzofuranyl, chromonyl, coumarinyl, benzopyranyl, cinnolinyl, quinoxalinyl, indazolyl, and pyrrolopyridyl.

[0088] As used herein, the term "pharmaceutically acceptable" means that within the scope of ordinary medical judgment, it is suitable for contact with human and animal tissues without causing excessive toxicity, irritation, allergic reaction, or other problems or complications, and provides a reasonable benefit / risk ratio, referring to compounds, substances, compositions, and / or dosage forms.

[0089] The compounds of formula (I) can form salts, and such salts are also within the scope of the present invention. Unless otherwise specified, references to the compounds of the invention are understood to include references to one or more of their salts. The term "salt" refers to acid salts and / or base salts formed by inorganic and / or organic acids and bases. Further, the term "salt" can include zwitterions (inner salts) when, for example, the compounds of formula (I) have both a basic moiety (such as an amine or pyridine or imidazole ring) and an acidic moiety (such as a carboxylic acid). Pharmaceutically acceptable (i.e., non-toxic and physiologically acceptable) salts are preferably, for example, acceptable metal salts and amine salts such that the cation does not significantly contribute to the toxicity or biological activity of the salt. However, other salts may also be useful, for example, in isolation or purification steps used in the manufacturing process, and therefore, other salts are also considered to be within the scope of the present invention. The salts of the compounds of formula (I) can be formed, for example, by reacting the compounds of formula (I) with a certain amount of an acid or base (such as 1 equivalent) in a solvent, for example, by precipitating the salt or by subsequently lyophilizing an aqueous solution.

[0090] Examples of acid addition salts include acetates (e.g., acetates prepared from acetic acid or trihaloacetic acids (e.g., trifluoroacetic acid)), adipates, alginates, ascorbates, aspartates, benzoates, benzenesulfonates, bisulfates, borates, butyrates, citrates, camphorates, camphorsulfonates, cyclopentanepropionates, digluconates, dodecyl sulfates, ethanesulfonates, fumarates, glucoheptanoates, glycerophosphates, hemisulfates, heptanoates, hexanoates, hydrochlorides (prepared from hydrochloric acid), hydrobromides (prepared from hydrobromic acid), hydroiodides, maleates (prepared from maleic acid), 2-hydroxyethanesulfonates, lactates, methanesulfonates (prepared from methanesulfonic acid), 2-naphthalenesulfonates, nicotinates, nitrates, oxalates, pectates, persulfates, 3-phenylpropionates, phosphates, picrates, pivalates, propionates, salicylates, succinates, sulfates (e.g., prepared from sulfuric acid), sulfonates (e.g., those described herein), tartrates, thiocyanates, toluenesulfonates (e.g., tosylates), undecanoates, and the like.

[0091] Examples of base salts include ammonium salts, alkali metal salts (e.g., sodium, lithium, and potassium salts), alkaline earth metal salts (e.g., calcium and magnesium salts), barium, zinc, and aluminum salts, organic base salts, such as organic amines (e.g., trialkylamines (e.g., triethylamine), procaine, dibenzylamine, N-benzyl-β-phenethylamine, 1-ephenamine, N,N'-dibenzylethylene-diamine, dehydroabietylamine, N-ethylpiperidine, benzylamine, dicyclohexylamine), or similar pharmaceutically acceptable amines, and amino acid salts (e.g., arginine, lysine), etc. The basic nitrogen-containing group may be quaternized by reagents (e.g., lower alkyl halides (e.g., methyl, ethyl, propyl, and butyl chlorides, bromides and iodides), dialkyl sulfates (e.g., dimethyl, diethyl, dibutyl, and diamyl sulfates), long-chain halides (e.g., decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides), aralkyl halides (e.g., benzyl and phenethyl bromides), and other groups). Preferred salts include the monohydrochloride, bisulfate, methanesulfonate, phosphate or nitrate.

[0092] The compounds of formula (I) can be provided as amorphous solids or crystalline solids. The compounds of formula (I) can be provided as solids by lyophilization.

[0093] Furthermore, solvates (e.g., hydrates) of the compounds of formula (I) should also be considered within the scope of the present invention. The term "solvate" means a physical association of a compound of formula (I) with one or more organic or inorganic solvent molecules. This physical association includes hydrogen bonding. In some cases, it is possible to isolate the solvate, for example when one or more solvent molecules are incorporated into the crystal lattice of the crystalline solid. "Solvate" includes both the solution phase and separable solvates. Examples of solvates include hydrates, ethanolates, methanolates, isopropanolates, acetonitrile solvates, and ethyl acetate solvates. Methods for solvates are known in the art.

[0094] Various forms of prodrugs are well known in the art and are described in Rautio, J. et al., Nature Review Drug Discovery, 17, 559-587 (2018).

[0095] In addition, the compounds of formula (I) are isolated and purified after their preparation to obtain a composition containing the compound of formula (I) in an amount of 99% or more (a "substantially pure") and then used or formulated as described herein. Such "substantially pure" compounds of formula (I) are also considered to be part of the present invention here.

[0096] "Stable compounds" and "stable structures" are intended to be sufficiently robust compounds that do not decompose even when isolated from the reaction mixture to useful purity or formulated into effective therapeutic agents. The present invention is intended to embody stable compounds.

[0097] "Therapeutically effective amount" means an amount of the compound of the present invention alone, or an amount of the compounds claimed in combination, or an amount of the compound of the present invention in combination with other active ingredients, effective to act as an inhibitor of DGKα and / or DGKζ, or effective for the treatment or prevention of viral infections and proliferative disorders (e.g., cancer).

[0098] As used herein, the terms "treating" or "treatment" include treating a disorder in a mammal, particularly a human, and (a) preventing, in particular, a mammal from contracting a disorder when the mammal is predisposed to the disorder but has not yet been diagnosed as having it; (b) inhibiting the disorder, i.e., arresting the progress of the disorder; and / or (c) alleviating the disorder, i.e., causing regression of the disorder.

[0099] The compounds of the present invention are intended to contain all isotopes of atoms contained in the compounds of the present invention. Isotopes include atoms having the same atomic number but different mass numbers. General examples include, but are not limited to, deuterium (D) and tritium (T) as isotopes of hydrogen. Isotopes of carbon include 13 C and 14 C. The compounds of the present invention labeled with isotopes can be prepared by using appropriate isotope-labeled reagents in place of the unlabeled reagents used otherwise, by conventional techniques generally known to those skilled in the art or by methods similar to those described herein.

[0100] The compounds described by formula (I) and / or their pharmaceutically acceptable salts can be administered by any suitable means for the condition to be treated, which may depend on the need for site-specific treatment or the amount of the compound of formula (I) to be delivered.

[0101] Also, the present invention includes compounds of formula (I) and / or their pharmaceutically acceptable salts; and non-toxic, pharmaceutically acceptable one or more carriers and / or diluents and / or adjuvants (substances collectively referred to herein as "carriers") and, optionally, other active ingredients. The compounds of formula (I) may be administered by any suitable route, preferably in the form of a pharmaceutical composition adapted to such a route and in a dosage effective for the intended treatment. The compounds and compositions of the present invention may be administered, for example, orally, transmucosally, or parenterally, including intravenous, intraperitoneal, subcutaneous, intramuscular, and intrasternal, in dosage unit formulations containing conventional pharmaceutically acceptable carriers, adjuvants, and vehicles. For example, pharmaceutical carriers may include a mixture of mannitol or lactose and microcrystalline cellulose. The mixture may include additional ingredients such as lubricants (e.g., magnesium stearate) and disintegrants (e.g., crospovidone). The carrier mixture may be filled into gelatin capsules or compressed into tablets. The pharmaceutical composition may be administered, for example, in oral dosage form or by infusion.

[0102] For oral administration, the pharmaceutical composition may be, for example, in the form of tablets, capsules, liquid capsules, suspensions, or liquids. The pharmaceutical composition is preferably formulated in dosage unit form having a specific amount of the active ingredient. For example, the pharmaceutical composition may be provided as tablets or capsules containing an amount of the active ingredient in the range of about 0.1 to 1000 mg, preferably about 0.25 to 250 mg, more preferably about 0.5 to 100 mg. The appropriate daily dosage for administration to humans or other mammals may vary widely depending on the patient's condition and other factors, but can be determined using conventional methods.

[0103] Any of the pharmaceutical compositions contemplated herein can be delivered orally, for example, via any acceptable and suitable oral formulation. Examples of oral formulations include, but are not limited to, tablets, troches, lozenges, aqueous and oily suspensions, dispersible powders or granules, emulsions, hard and soft capsules, liquid capsules, syrups, and elixirs. Pharmaceutical compositions for oral administration can be manufactured according to any method known in the art of manufacturing pharmaceutical compositions for oral administration. To provide a pharmaceutically palatable formulation, the pharmaceutical compositions described herein can include at least one substance selected from sweetening agents, flavoring agents, coloring agents, glidants, antioxidants, and preservatives.

[0104] Tablets can be manufactured, for example, by mixing at least one compound of formula (I) and / or at least one pharmaceutically acceptable salt with at least one non-toxic and pharmaceutically acceptable additive suitable for tablet manufacture. Examples of additives include, but are not limited to, inert diluents (e.g., calcium carbonate, sodium carbonate, lactose, calcium phosphate, and sodium phosphate), granulating and disintegrating agents (e.g., microcrystalline cellulose, croscarmellose sodium, corn starch, and alginic acid), binding agents (e.g., starch, gelatin, polyvinylpyrrolidone, and gum arabic), and lubricants (e.g., magnesium stearate, stearic acid, and talc). Furthermore, tablets can be uncoated or can be coated by known techniques to mask the unpleasant taste of the drug, or to delay disintegration and absorption of the active ingredient in the gastrointestinal tract and to sustain the effect of the active ingredient over a longer period. Examples of water-soluble taste masking materials include, but are not limited to, hydroxypropylmethylcellulose and hydroxypropylcellulose. Examples of time delay materials include, but are not limited to, ethylcellulose and cellulose acetate butyrate.

[0105] Hard gelatin capsules can be produced, for example, by mixing at least one compound of formula (I) and / or at least one salt thereof with at least one inert solid diluent (such as calcium carbonate, calcium phosphate, and kaolin).

[0106] Soft gelatin capsules can be produced, for example, by mixing at least one compound of formula (I) and / or at least one pharmaceutically acceptable salt thereof with at least one water-soluble carrier (such as polyethylene glycol), and at least one oily medium (such as peanut oil, liquid paraffin, and olive oil).

[0107] An aqueous suspension can be prepared, for example, by mixing at least one compound of formula (I) and / or at least one pharmaceutically acceptable salt thereof with at least one additive suitable for the preparation of an aqueous suspension. Examples of additives suitable for the preparation of an aqueous suspension include, but are not limited to, for example, suspending agents (such as sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, sodium alginate, alginic acid, polyvinyl pyrrolidone, tragacanth gum, and gum arabic), dispersing agents or wetting agents (such as naturally occurring phosphatides (such as lecithin), condensation products of alkylene oxides and fatty acids (such as polyoxyethylene stearate), condensation products of ethylene oxide and long-chain aliphatic alcohols (such as heptadecaethyleneoxy cetanol), condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol (such as polyoxyethylene sorbitol monooleate), and condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides (such as polyethylene sorbitan monooleate)). Further, the aqueous suspension may include at least one preservative (such as ethyl p-hydroxybenzoate and n-propyl p-hydroxybenzoate), at least one coloring agent, at least one flavoring agent, and / or at least one sweetening agent (including, but not limited to, for example, sucrose, saccharin, and aspartame).

[0108] An oily suspension can be prepared, for example, by suspending at least one compound of formula (I) and / or at least one pharmaceutically acceptable salt thereof in either a vegetable oil (such as peanut oil, olive oil, sesame oil, and coconut oil) or a mineral oil (such as liquid paraffin). The oily suspension may also contain at least one thickening agent (such as beeswax, solid paraffin, and cetyl alcohol). To provide a palatable oily suspension, at least one sweetening agent already described above and / or at least one flavoring agent can be added to the oily suspension. The oily suspension may further contain at least one preservative (including, but not limited to, for example, antioxidants such as butylhydroxyanisole and α-tocopherol).

[0109] Dispersible powders and granules can be prepared, for example, by mixing at least one compound of formula (I) and / or at least one pharmaceutically acceptable salt thereof with at least one dispersing agent and / or wetting agent, at least one suspending agent, and / or at least one preservative. Suitable dispersing agents, wetting agents, and suspending agents have already been described above. Examples of preservatives include, but are not limited to, for example, antioxidants such as ascorbic acid. Further, the dispersible powders and granules may also contain at least one excipient (including, but not limited to, for example, sweetening agents, flavoring agents, and coloring agents).

[0110] An emulsion of at least one compound of formula (I) and / or at least one pharmaceutically acceptable salt can be produced, for example, as an oil-in-water emulsion. The oil phase of the emulsion containing the compound of formula (I) may be composed of known components by known methods. The oil phase can be provided, for example, but not limited to, vegetable oils (such as olive oil and peanut oil), mineral oils (such as liquid paraffin), and mixtures thereof. The oil phase may contain only an emulsifier, or may contain at least one emulsifier and a fat or an oil, or a mixture of both a fat and an oil. Suitable emulsifiers include, but are not limited to, for example, naturally occurring phosphatides (such as soy lecithin), esters or partial esters derived from fatty acids and hexitol anhydrides (such as sorbitan monooleate), and condensation products of partial esters and ethylene oxide (such as polyoxyethylene sorbitan monooleate). Preferably, a hydrophilic emulsifier is included together with a lipophilic emulsifier that acts as a stabilizer. It is also preferred to include both an oil and a fat. Additionally, the emulsifier, with or without a stabilizer, forms a so-called emulsifying wax, and the wax, together with the oil and the fat, forms a so-called emulsifying ointment base that forms the oily dispersed phase of the cream formulation. The emulsion may also include a sweetening agent, a flavoring agent, a preservative, and / or an antioxidant. Suitable emulsifiers and emulsion stabilizers for use in the formulations of the present invention include Tween 60, Span 80, cetostearyl alcohol, myristyl alcohol, glyceryl monostearate, sodium lauryl sulfate, glyceryl distearate, either alone or together with a wax; or other substances known in the art.

[0111] In addition, the compound of formula (I) and / or at least one of its pharmaceutically acceptable salts can be administered, for example, intravenously, subcutaneously, and / or intramuscularly via any pharmaceutically acceptable and suitable injection form. Examples of injection forms include, but are not limited to, for example, sterile aqueous solutions containing acceptable vehicles and solvents (such as water, Ringer's solution, and sodium chloride isotonic solution), sterile water-in-oil microemulsions, and aqueous or oily suspensions.

[0112] Formulations for parenteral administration may be in the form of aqueous or non-aqueous isotonic sterile injection solutions or suspensions. These solutions and suspensions may be prepared from sterile powders or granules by using one or more carriers or diluents described for use in formulations for oral administration or by using other suitable dispersing or wetting agents and suspending agents. The compound may be dissolved in water, polyethylene glycol, propylene glycol, ethanol, corn oil, cottonseed oil, peanut oil, sesame oil, benzyl alcohol, sodium chloride, tragacanth gum, and / or various buffer solutions. Other adjuvants and administration methods are known and widely known in the pharmaceutical field. Also, the active ingredient may be administered by injection as a composition with a suitable carrier (such as physiological saline, dextrose, or water), or cyclodextrin (i.e., Captisol), solubilizing co-solvent (i.e., propylene glycol), or solubilizing micelle (i.e., Tween 80).

[0113] Also, a sterile injection preparation may be a sterile injection solution or suspension (such as a solution in 1,3-butanediol) in a non-toxic and parenterally acceptable diluent or solvent. Among the acceptable vehicles and solvents that may be used are water, Ringer's solution, and sodium chloride isotonic solution. Furthermore, sterile fixed oils are conventionally used as solvents or suspension media. For this purpose, any sterile fixed oil containing synthetic monoglycerides or diglycerides may be used. Additionally, fatty acids such as oleic acid are used in injection preparations.

[0114] An oil-in-water microemulsion in sterile water for injection can be produced, for example, as follows. 1) Dissolve at least one compound of formula (I) in an oil phase (for example, a mixture of soybean oil and lecithin), 2) combine the oil phase containing formula (I) with a mixture of water and glycerol, and 3) treat the combination to form a microemulsion.

[0115] A sterile aqueous suspension or a sterile oily suspension can be produced according to methods known to those skilled in the art. For example, a sterile aqueous solution or a sterile aqueous suspension can be prepared using a non-toxic, parenterally acceptable diluent or solvent (for example, 1,3-butanediol), and a sterile oily suspension can be produced using a sterile, non-toxic, acceptable solvent or suspension medium (for example, a sterile, non-volatile oil (for example, synthetic monoglycerides or diglycerides), and a fatty acid (for example, oleic acid)).

[0116] Pharmaceutically acceptable carriers, adjuvants, and vehicles that can be used in the pharmaceutical compositions of the present invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, self-emulsifying drug delivery systems (SEDDS) (e.g., d-α-tocopherol polyethylene glycol 1000 succinate), surfactants used in pharmaceutical dosage forms (e.g., Tween, polyethoxylated castor oil (e.g., CREMOPHOR surfactants (BASF), or other similar polymeric delivery matrices), serum proteins (e.g., human serum albumin), buffer substances (e.g., phosphate, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes (e.g., protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate)), polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol, and lanolin). Also, cyclodextrins (e.g., α-, β-, and γ-cyclodextrins, or chemically modified derivatives (e.g., hydroxyalkyl cyclodextrins including 2- and 3-hydroxypropyl cyclodextrin, or other solubilizing derivatives)) may also be advantageously used to enhance the delivery of the compounds of the formulas described herein.

[0117] The pharmaceutically active compounds of the present invention can be processed according to conventional pharmaceutical methods to prepare agents for administration to patients (e.g., humans and other mammals). The pharmaceutical compositions may be subjected to conventional pharmaceutical operations (e.g., sterilization) and / or may include conventional adjuvants (e.g., preservatives, stabilizers, wetting agents, emulsifying agents, buffers, etc.). Tablets and pills can additionally be prepared using enteric coating agents. Such compositions may also include adjuvants (e.g., wetting agents, sweeteners, flavoring agents, and fragrances).

[0118] The amount of the compound and / or composition of the present invention to be administered for treating a medical condition, and the dosing schedule, depend on various factors (e.g., age, weight, gender, the medical condition of the patient, the type of disease, the severity of the disease, the route and frequency of administration, and the particular compound utilized). Therefore, the dosing schedule may vary significantly but can be determined in accordance with the standard method as prescribed. The daily dose can be appropriately between about 0.001 and 100 mg / kg of body weight, preferably between about 0.0025 and about 50 mg / kg of body weight, and most preferably between about 0.005 and 10 mg / kg of body weight. The daily dose can be administered 1 to 4 times a day. Other dosing schedules include once a week and once every two days cycles.

[0119] For therapeutic purposes, the active compounds of the present invention are combined with one or more adjuvants appropriate for the intended route of administration. When administered orally, the compound may be mixed with lactose, sucrose, starch powder, cellulose esters of alkanoic acids, cellulose alkyl esters, talc, stearic acid, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphoric and sulfuric acids, gelatin, gum arabic, sodium alginate, polyvinylpyrrolidone, and / or polyvinyl alcohol, and then may be tableted or encapsulated for convenient administration. Such capsules or tablets may include controlled-release formulations and may be provided by dispersing the active compound in hydroxypropylmethylcellulose.

[0120] The pharmaceutical composition of the present invention appropriately includes at least one compound of formula (I) and / or at least one pharmaceutically acceptable salt thereof, and additives selected from any pharmaceutically acceptable carrier, adjuvant, and vehicle. Another composition of the present invention includes the compound of formula (I) described herein, or a prodrug thereof, and a pharmaceutically acceptable carrier, adjuvant, or vehicle.

[0121] Usefulness The compound of formula (I) is useful for the treatment of cancer.

[0122] In another embodiment, the present invention provides a combination medicament of a compound of formula (I) and / or a pharmaceutically acceptable salt thereof, a stereoisomer or a tautomer thereof, and another therapeutic agent, which is used simultaneously, separately or sequentially in the treatment and / or prevention of a plurality of diseases or disorders associated with the targeted inhibition of DGK in T cells.

[0123] In another aspect, the present invention provides a method for treating a patient suffering from or susceptible to a medical condition associated with the targeted inhibition of DGK in T cells. A number of medical conditions can be treated. This method is characterized by administering to the patient a composition comprising a therapeutically effective amount of a compound of formula (I) and / or a pharmaceutically acceptable salt thereof, a stereoisomer or a tautomer thereof. For example, the compounds described herein may be used to treat or prevent viral infections and proliferative diseases (such as cancer).

[0124] The compound of formula (I) and a pharmaceutical composition comprising at least one compound of formula (I) are useful for the treatment or prevention of any disease or medical condition associated with the targeted inhibition of DGK in T cells. Such diseases or medical conditions include viruses and other infections (such as skin infections, GI infections, urinary tract infections, urogenital infections, systemic infections), and proliferative diseases (such as cancer). The compound of formula (I) and a pharmaceutical composition comprising at least one compound of formula (I) can be administered to animals, preferably mammals (such as domestic animals, cats, dogs, mice, rats), and more preferably humans. Any method of administration may be used to deliver this compound or pharmaceutical composition to a patient. In certain embodiments, the compound of formula (I) or a pharmaceutical composition comprising at least the compound of formula (I) is administered orally. In other embodiments, a pharmaceutical composition comprising the compound of formula (I) or at least the compound of formula (I) is administered parenterally.

[0125] The compound of formula (I) can inhibit the activities of diacylglycerol kinases α and ζ (DGKα / ζ). For example, the compound of formula (I) can be used to inhibit the activities of DGKα and DGKζ by administering an inhibitory amount of the compound of formula (I) or a salt thereof to a cell or an individual in which the regulation of DGKα and DGKζ is required.

[0126] The present invention further provides a method for treating a disease related to the activity or expression (e.g., abnormal activity and / or overexpression of DGKα and DGKζ in an individual (e.g., a patient)) by administering a therapeutically effective amount or dose of the compound of formula (I) or a pharmaceutical composition thereof to an individual in need of such treatment. Examples of diseases may include any disease, disorder or condition directly or indirectly involving the expression or activity (e.g., overexpression or abnormal activity) of DGKα and DGKζ enzymes. Also, DGKα- and DGKζ-related diseases may include any disease, disorder or condition that can be prevented, ameliorated, or recovered by regulating the activities of DGKα and DGKζ enzymes. Examples of DGKα- and DGKζ-related diseases include cancer and viral infections (e.g., HIV infection, hepatitis B, and hepatitis C).

[0127] In one aspect, the compound of formula (I) is administered continuously before the administration of an immuno-oncology agent. In another aspect, the compound of formula (I) is administered simultaneously with the immuno-oncology agent. In yet another aspect, the compound of formula (I) is administered subsequently after the administration of the immuno-oncology agent.

[0128] In another aspect, the compound of formula (I) may be formulated with an immuno-oncology agent.

[0129] Immuno-oncology agents include, for example, small molecule drugs, antibodies, or other biological molecules or small molecules. Examples of biological immuno-oncology agents include, but are not limited to, cancer vaccines, antibodies, and cytokines. In one aspect, the antibody is a monoclonal antibody. In another aspect, the monoclonal antibody is a humanized antibody or a human antibody.

[0130] In one aspect, an immuno-oncology agent is an agonist of a stimulatory receptor (including co-stimulation) on T cells or an antagonist of an inhibitory signal (including co-inhibition), both of which result in amplification of antigen-specific T cell responses (often referred to as immune checkpoint regulators).

[0131] Certain stimulatory and inhibitory molecules belong to the immunoglobulin superfamily (IgSF). An important family of membrane-bound ligands that bind to co-stimulatory or co-inhibitory receptors is the B7 family, which includes B7-1, B7-2, B7-H1 (PD-L1), B7-DC (PD-L2), B7-H2 (ICOS-L), B7-H3, B7-H4, B7-H5 (VISTA), and B7-H6. Another family of membrane-bound ligands that bind to co-stimulatory or co-inhibitory receptors is the TNF family molecules that bind to the homologous TNF receptor family, which includes CD40 and CD40L, OX-40, OX-40L, CD70, CD27L, CD30, CD30L, 4-1BBL, CD137 (4-1BB), TRAIL / Apo2-L, TRAILR1 / DR4, TRAILR2 / DR5, TRAILR3, TRAILR4, OPG, RANK, RANKL, TWEAKR / Fn14, TWEAK, BAFFR, EDAR, XEDAR, TACI, APRIL, BCMA, LTβR, LIGHT, DcR3, HVEM, VEGI / TL1A, TRAMP / DR3, EDAR, EDA1, XEDAR, EDA2, TNFR1, lymphotoxin α / TNFβ, TNFR2, TNFα, LTβR, lymphotoxin α1β2, FAS, FASL, RELT, DR6, TROY, NGFR.

[0132] In certain embodiments, the T cell response can be stimulated by a combination of a compound of formula (I) with one or more (i) protein antagonists that inhibit T cell activation (e.g., immune checkpoint inhibitors), such as CTLA-4, PD-1, PD-L1, PD-L2, LAG-3, TIM-3, galectin 9, CEACAM-1, BTLA, CD69, galectin 1, TIGIT, CD113, GPR56, VISTA, 2B4, CD48, GARP, PD1H, LAIR1, TIM-1, and TIM-4, and (ii) agonists of proteins that stimulate T cell activation, such as B7-1, B7-2, CD28, 4-1BB (CD137), 4-1BBL, ICOS, ICOS-L, OX40, OX40L, GITR, GITRL, CD70, CD27, CD40, DR3 and CD28H.

[0133] For cancer treatment, other agents that can be combined with a compound of formula (I) can include antagonists of inhibitory receptors on NK cells or agonists of activating receptors on NK cells. For example, a compound of formula (I) can be combined with an antagonist of KIR, such as lirilumab.

[0134] Still other agents for use in combination therapy include agents that inhibit or ablate macrophages or monocytes, including, but not limited to, CSF-1R antagonists, such as CSF-1R antagonist antibodies including RG7155 (WO11 / 70024, WO11 / 107553, WO11 / 131407, WO13 / 87699, WO13 / 119716, WO13 / 132044) or FPA-008 (WO11 / 140249; WO13169264; WO14 / 036357).

[0135] In another aspect, the compound of formula (I) is used with an agonistic agent that binds to a positive costimulatory receptor, a blocking agent that attenuates signal transduction via an inhibitory receptor, an antagonist, and one or more agents that systemically increase the frequency of anti-tumor T cells, agents that overcome different immunosuppressive pathways in the tumor microenvironment (e.g., blocking the involvement of inhibitory receptors (e.g., PD-L1 / PD-1 interaction), drastic reduction or inhibition of regulatory (Treg) cells (e.g., use of anti-CD25 monoclonal antibodies (e.g., daclizumab) or depletion of anti-CD25 beads in vitro), inhibition of metabolic enzymes such as IDO, or restoration / prevention of T cell anergy or T cell depletion), and one or more agents that cause innate immune activation and / or inflammation of the tumor portion.

[0136] In one aspect, the immuno-oncology agent is a CTLA-4 antagonist, e.g., an antagonistic CTLA-4 antibody. Suitable CTLA-4 antibodies include, for example, Yervoy (ipilimumab) or tremelimumab.

[0137] In another aspect, the immuno-oncology agent is a PD-1 antagonist, e.g., an antagonistic PD-1 antibody. Suitable PD-1 antibodies include, for example, Opdivo (nivolumab), Keytruda (pembrolizumab), or MEDI-0680 (AMP-514; WO2012 / 145493). The immuno-oncology agent may also include pidilizumab (CT-011), although its specificity for PD-1 binding has been questioned. Another approach targeting the PD-1 receptor is a recombinant protein consisting of the extracellular domain of PD-L2 (B7-DC) fused to the Fc portion of IgG1, designated AMP-224.

[0138] In another aspect, the immuno-oncology agent is a PD-L1 antagonist, such as an antagonistic PD-L1 antibody. Suitable PD-L1 antibodies include, for example, MPDL3280A (RG7446; WO2010 / 077634), durvalumab (MEDI4736), BMS-936559 (WO2007 / 005874), and MSB0010718C (WO2013 / 79174).

[0139] In another aspect, the immuno-oncology agent is a LAG-3 antagonist, such as an antagonistic LAG-3 antibody. Suitable LAG3 antibodies include, for example, BMS-986016 (WO10 / 19570, WO14 / 08218), or IMP-731 or IMP-321 (WO08 / 132601, WO09 / 44273).

[0140] In another aspect, the immuno-oncology agent is a CD137 (4-1BB) agonist, such as an agonistic CD137 antibody. Suitable CD137 antibodies include, for example, urelumab and PF-05082566 (WO12 / 32433).

[0141] In another aspect, the immuno-oncology agent is a GITR agonist, such as an agonistic GITR antibody. Suitable GITR antibodies include, for example, BMS-986153, BMS-986156, TRX-518 (WO06 / 105021, WO09 / 009116) and MK-4166 (WO11 / 028683).

[0142] In another aspect, the immuno-oncology agent is an IDO antagonist. Suitable IDO antagonists include, for example, INCB-024360 (WO2006 / 122150, WO07 / 75598, WO08 / 36653, WO08 / 36642), indoximod, BMS-986205, or NLG-919 (WO09 / 73620, WO09 / 1156652, WO11 / 56652, WO12 / 142237).

[0143] In another aspect, the immuno-oncology agent is an OX40 agonist, such as an agonistic OX40 antibody. Suitable OX40 antibodies include, for example, MEDI-6383 or MEDI-6469.

[0144] In another aspect, the immuno-oncology agent is an OX40L antagonist, such as an antagonistic OX40 antibody. Suitable OX40L antagonists include, for example, RG-7888 (WO06 / 029879).

[0145] In another aspect, the immuno-oncology agent is a CD40 agonist, such as an agonistic CD40 antibody. In yet another embodiment, the immuno-oncology agent is a CD40 antagonist, such as an antagonistic CD40 antibody. Suitable CD40 antibodies include, for example, lucatumumab or dacetuzumab.

[0146] In another aspect, the immuno-oncology agent is a CD27 agonist, such as an antagonistic CD27 antibody. Suitable CD27 antibodies include, for example, varlilumab.

[0147] In another aspect, the immuno-oncology agent against B7H3 is MGA271 (WO11 / 109400).

[0148] Combination therapy is intended to include administration of these therapeutic agents in a sequential manner, i.e., each therapeutic agent is administered at various different time points, as well as these therapeutic agents or at least two of the therapeutic agents being administered in a substantially simultaneous manner. Substantially simultaneous administration can be achieved, for example, by administering to a patient a single dosage form at a fixed ratio of each therapeutic agent, or multiple single dosage forms for each of the therapeutic agents. The sequential or substantially simultaneous administration of each therapeutic agent can be carried out, for example, by any suitable route such as the oral route, intravenous route, intramuscular route, and direct absorption through mucosal membrane tissues, but is not limited thereto. The therapeutic agents can be administered by the same route or different routes. For example, the first therapeutic agent in a selected combination can be administered by intravenous injection, while another therapeutic agent within this combination can be administered orally. Alternatively, for example, all of the therapeutic agents can be administered orally, or all of the therapeutic agents can be administered by intravenous injection. The above administration of the therapeutic agents can be combined with further other biologically active ingredients and non-drug therapies (e.g., surgery or radiation therapy) to perform combination therapy. When this combination therapy further includes non-pharmacological treatment, the non-pharmacological treatment can be carried out at any appropriate time as long as a useful effect resulting from the combined action of the therapeutic agent and the non-pharmacological treatment is achieved. For example, in a preferred case, this useful effect is achieved even when the non-drug treatment is temporarily suspended, probably for several days or weeks, from the administration of the therapeutic agent.

[0149] As used herein, the term "cell" is intended to refer to cells in vitro, ex vivo, or in vivo. In some embodiments, ex vivo cells can be a part of a tissue sample excised from an organism (e.g., a mammal). In some embodiments, in vitro cells can be cells in cell culture. In some embodiments, in vivo cells are living cells in an organism (e.g., a mammal).

[0150] As used herein, the term "contact" refers to the joining of a designated portion of an in vitro or in vivo system. For example, "contacting" a DGKα and DGKζ enzyme with a compound of formula (I) includes administering the compound of the invention to an individual or patient (e.g., a human) having DGKα and DGKζ, and introducing the compound of formula (I) into, for example, a sample containing cells or a preparation containing DGKα and DGKζ enzymes.

[0151] The term "DGKα and DGKζ inhibitor" refers to an agent that can inhibit the activity of diacylglycerol kinase α and / or diacylglycerol kinase ζ (DGKα and DGKζ) in T cells and that provides a stimulus to T cells. A DGKα and DGKζ inhibitor can be a reversible or irreversible DGKα and DGKζ inhibitor. A "reversible DGKα and DGKζ inhibitor" is a compound that reversibly inhibits DGKα and DGKζ enzyme activity at either the catalytic or non-catalytic site, and an "irreversible DGKα and DGKζ inhibitor" is a compound that irreversibly impairs DGKα and DGKζ enzyme activity by forming a covalent bond with the enzyme.

[0152] The types of cancer that may be treated with the compound of formula (I) include, but are not limited to, brain cancer, skin cancer, bladder cancer, ovarian cancer, breast cancer, stomach cancer, pancreatic cancer, prostate cancer, colon cancer, blood cancer, lung cancer, and bone cancer. Examples of such types of cancer include neuroblastoma, intestinal cancer (e.g., rectal cancer, colon cancer, familial adenomatous polyposis, and hereditary non-polyposis colorectal cancer), esophageal cancer, lip cancer, laryngeal cancer, hypopharyngeal cancer, tongue cancer, salivary gland cancer, stomach cancer, adenocarcinoma, medullary thyroid cancer, papillary thyroid cancer, kidney cancer, renal parenchymal cancer, ovarian cancer, cervical cancer, endometrial cancer, choriocarcinoma, pancreatic cancer, prostate cancer, testicular cancer, breast cancer, urinary tract cancer, melanoma, brain tumors such as glioblastoma, astrocytoma, meningioma, medulloblastoma, and peripheral primitive neuroectodermal tumor, Hodgkin lymphoma, non-Hodgkin lymphoma, Burkitt lymphoma, acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), adult T-cell leukemia lymphoma, diffuse large B-cell lymphoma (DLBCL), hepatocellular cancer, gallbladder cancer, bronchial cancer, small cell lung cancer, non-small cell lung cancer, multiple myeloma, basal cell carcinoma, teratoma, retinoblastoma, choroidal melanoma, seminoma, rhabdomyosarcoma, craniopharyngioma, osteosarcoma, chondrosarcoma, sarcoma, liposarcoma, fibrosarcoma, Ewing sarcoma, and plasmacytoma.

[0153] For the treatment of diseases, disorders or conditions related to DGKα and DGKζ, one or more other pharmaceuticals or treatment methods (e.g., antiviral agents, chemotherapeutic agents or other anti-cancer agents, immune enhancers, immunosuppressants, radiation, anti-tumor and anti-viral vaccines, cytokine therapy (e.g., IL2 and GM-CSF), and / or tyrosine kinase inhibitors) may be used in combination with the compound of formula (I) as appropriate. The above agents may be combined with the present compound in a single dosage form, or the agents may be administered simultaneously or sequentially in different dosage forms.

[0154] Suitable chemotherapeutic agents or other anticancer agents include, for example, alkylating agents (including, but not limited to, nitrogen mustards, ethyleneimine derivatives, alkyl sulfonic acids, nitrosoureas, and triazenes), such as uracil mustard, chloromethine, cyclophosphamide (Cytoxan®), ifosfamide, melphalan, chlorambucil, pipobroman, triethylenemelamine, triethylenethiophosphoramide, busulfan, carmustine, lomustine, streptozocin, dacarbazine, and temozolomide.

[0155] In the treatment of melanoma, suitable agents for use in combination with the compound of formula (I) include dacarbazine (DTIC), and optionally other chemotherapeutic agents (such as carmustine (BCNU) and cisplatin); the "Dartmouth regimen" consisting of DTIC, BCNU, cisplatin, and tamoxifen; combinations of cisplatin, vinblastine, and DTIC, temozolomide, or Yervoy®. The compound of formula (I) can also be combined with immunotherapeutic agents (such as cytokines (interferon α, interleukin 2, and tumor necrosis factor (TNF), etc.)) in the treatment of melanoma.

[0156] The compound of formula (I) can also be used in combination with vaccine therapy in the treatment of melanoma. Antimelanoma vaccines are somewhat similar in some respects to antiviral vaccines used to prevent virus-caused diseases (such as polio, measles, and mumps). Weakened melanoma cells or a part of melanoma cells called antigens can be injected into a patient to stimulate the body's immune system and destroy melanoma cells.

[0157] Melanomas of the wrist or leg can also be treated using thermoperfusion therapy in combination with an agent comprising one or more compounds of formula (I). In this treatment protocol, the circulation of the limb at the disease site is temporarily isolated from the rest of the body's circulation, and a high concentration of chemotherapeutic agent is injected into the artery of the corresponding limb to administer a high dose that can cause serious side effects if exposed to the internal organs to the tumor site. Usually, in this treatment, the body fluid is heated to 38.9 °C to 40 °C. Melphalan is the most frequently used drug in this chemotherapy. Another agent called tumor necrosis factor (TNF) can also be used.

[0158] Suitable chemotherapeutic agents or other anticancer agents include, for example, antimetabolites (including, but not limited to, folic acid antagonists, pyrimidine analogs, purine analogs, and adenosine deaminase inhibitors, etc.), such as methotrexate, 5-fluorouracil, floxuridine, cytarabine, 6-mercaptopurine, 6-thioguanine, fludarabine phosphate, pentostatin, and gemcitabine.

[0159] Suitable chemotherapeutic agents or other anticancer agents further include, for example, certain natural products and their derivatives (such as vinca alkaloids, antitumor antibiotics, enzymes, lymphokines, and epipodophyllotoxins), such as vinblastine, vincristine, vindesine, bleomycin, dactinomycin, daunorubicin, doxorubicin, epirubicin, idarubicin, cytarabine, paclitaxel (Taxol), mitomycin, deoxycoformycin, mitomycin C, L-asparaginase, interferon (especially IFNα), etoposide, and teniposide.

[0160] Other cytotoxic agents include navelbine, CPT-11, anastrozole, letrozole, capecitabine, raloxifene, and droloxifene.

[0161] In addition, suitable cytotoxic agents include, for example, epipodophyllotoxin; anti-cancer enzymes; topoisomerase inhibitors; procarbazine; mitoxantrone; platinum coordination complexes (such as cisplatin and carboplatin); biological response modifiers; growth inhibitors; anti-hormone therapeutic agents; leucovorin; tegafur; and hematopoietic growth factors.

[0162] Other anti-cancer agents include antibody drugs, such as trastuzumab (Herceptin®), antibodies against co-stimulatory molecules (such as CTLA-4, 4-1BB, and PD-1), or antibodies against cytokines (IL-10 or TGF-β).

[0163] Still other anti-cancer agents include anti-cancer agents that block immune cell migration, such as antagonists against chemokine receptors (such as CCR2 and CCR4).

[0164] Still other anti-cancer agents include anti-cancer agents that enhance the immune system, such as adjuvants or adoptive T cell transfer.

[0165] Anti-cancer vaccines include dendritic cells, synthetic peptides, DNA vaccines, and recombinant viruses.

[0166] The pharmaceutical composition of the present invention may optionally contain at least one signal transduction inhibitor (STI). A "signal transduction inhibitor" is an agent that selectively inhibits one or more important steps in a signal transduction pathway in the normal function of cancer cells, thereby causing apoptosis. Suitable STIs include, but are not limited to, (i) bcr / abl kinase inhibitors (e.g., STI 571 (Gleevec®)); (ii) epidermal growth factor (EGF) receptor inhibitors (e.g., kinase inhibitors (Iressa®, SSI-774) and antibodies (Imclone: C225 [Goldstein et al., Clin. Cancer Res., 1: 1311-1318 (1995)], and Abgenix: ABX-EGF)); (iii) her-2 / neu receptor inhibitors (e.g., farnesyl transferase inhibitors (FTIs) (e.g., L-744,832 (Kohl et al., Nat. Med., 1(8):792-797 (1995))); (iv) inhibitors of Akt family kinases or the Akt pathway (e.g., rapamycin (see, e.g., Sekulic et al., Cancer Res., 60: 3504-3513 (2000))); (v) cell cycle kinase inhibitors (e.g., flavopiridol and UCN-O1 (see, e.g., Sausville, Curr. Med. Chem. Anti-Canc. Agents, 3:47-56(2003)); and (vi) phosphatidylinositol kinase inhibitors (e.g., LY294002 (see, e.g., Vlahos et al., J. Biol. Chem., 269: 5241-5248 (1994))). Alternatively, at least one STI and at least one compound of formula (I) may be formulated in separate pharmaceutical compositions. In certain embodiments of the invention, at least one compound of formula (I) and at least one STI may be administered to a patient simultaneously or sequentially. In other words, at least one compound of formula (I) may be administered first, at least one STI may be administered first, or at least one compound of formula (I) and at least one STI may be administered simultaneously.Furthermore, when one or more compounds of formula (I) and / or an STI are used, the compounds can be administered in any order.

[0167] Furthermore, the present invention provides a pharmaceutical composition comprising at least one compound of formula (I), optionally at least one chemotherapeutic agent, and optionally at least one antiviral agent, in a pharmaceutically acceptable carrier, for treating a chronic viral infection in a patient.

[0168] Also provided is a method of treating a chronic viral infection in a patient by administering an effective amount of the above pharmaceutical composition.

[0169] In certain embodiments of the invention, at least one compound of formula (I) and at least one chemotherapeutic agent are administered to the patient simultaneously or sequentially. In other words, at least one compound of formula (I) can be administered first, at least one chemotherapeutic agent can be administered first, or at least one compound of formula (I) and at least one chemotherapeutic agent can be administered simultaneously. Furthermore, when one or more compounds of formula (I) and / or chemotherapeutic agents are used, the compounds can be administered in any order. Similarly, any antiviral agent or STI can be administered at any point in time compared to the administration of the compound of formula (I).

[0170] Chronic viral infections that can be treated using the present combination therapy include, but are not limited to, diseases caused by hepatitis C virus (HCV), human papillomavirus (HPV), cytomegalovirus (CMV), herpes simplex virus (HSV), Epstein - Barr virus (EBV), varicella - zoster virus, coxsackievirus, and human immunodeficiency virus (HIV). In particular, parasitic infections (e.g., malaria) can be treated by the above - mentioned method by appropriately adding compounds known in the treatment of parasitosis in place of the antiviral agent.

[0171] Suitable antiviral drugs for which use in combination with a compound of formula (I) is contemplated may include nucleoside and nucleotide reverse transcriptase inhibitors (NRTIs), non-nucleoside reverse transcriptase inhibitors (NNRTIs), protease inhibitors and other antiviral drugs.

[0172] Examples of suitable NRTIs include zidovudine (AZT); didanosine (ddI); zalcitabine (ddC); stavudine (d4T); lamivudine (3TC); abacavir (1592U89); adefovir pivoxil [bis(POM)-PMEA]; lobucavir; BCH-I0652; emtricitabine [(-)-FTC]; β-L-FD4 (also referred to as β-L-D4C, the name being β-L-2’,3’-dideoxy-5-fluorocytidine); DAPD, ((-)-β-D-2,6-diaminopurine dioxolane); and rodenosine (FddA). Representative and suitable NNRTIs include nevirapine (BI-RG-587); delavirdine (BHAP, U-90152); efavirenz (DMP-266); PNU-142721; AG-1549; MKC-442 (1-(ethoxymethyl)-5-(1-methylethyl)-6-(phenylmethyl)-(2,4(1H,3H)-pyrimidinedione); and (+)-calanolide A (NSC-675451) and B. Representative and suitable protease inhibitors include saquinavir (Ro 31-8959); ritonavir (ABT-538); indinavir (MK-639); nelfinavir (AG-1343); amprenavir (141W94); lasinavir; DMP-450; BMS-2322623; ABT-378; and AG-1549. Other antiviral drugs include hydroxyurea, ribavirin, IL-2, IL-12, pentafuside and Yissum Project No.11607.

[0173] The present invention also includes a pharmaceutical kit useful, for example, in the treatment or prevention of DGKα- and DGKζ-related diseases or disorders, and other diseases described herein, and includes a pharmaceutical composition containing a therapeutically effective amount of a compound of formula (I) in one or more containers. Such a kit may further optionally include one or more of the various conventional components of a pharmaceutical kit (e.g., a container containing one or more pharmaceutically acceptable carriers, another container), which will be readily apparent to those skilled in the art. Instructions in either an accompanying document or label indicating the amount of components to be administered, dosing guidelines, and / or mixing guidelines for the components may also be included in the kit.

[0174] Combination therapy is intended to include administration of these therapeutic agents in a sequential manner, i.e., each therapeutic agent is administered at various different times, and also that these therapeutic agents or at least two of them are administered in a substantially simultaneous manner. Substantially simultaneous administration can be achieved, for example, by administering to a patient in a single dosage form at a fixed ratio of each therapeutic agent, or in multiple single dosage forms for each of the therapeutic agents. The sequential or substantially simultaneous administration of each therapeutic agent can be carried out, for example, by any suitable route such as oral route, intravenous route, intramuscular route, and direct absorption through mucosal membrane tissues, but is not limited thereto. The therapeutic agents can be administered by the same route or different routes. For example, the first therapeutic agent in a selected combination can be administered by intravenous injection, while another therapeutic agent in this combination can be administered orally. Alternatively, for example, all therapeutic agents can be administered orally, or all therapeutic agents can be administered by intravenous injection. Combination therapy can also include the administration in combination with the above-described therapeutic agents of further other biologically active components and non-drug therapies (e.g., surgery or radiation therapy). If this combination therapy further includes non-pharmacological treatment, the non-pharmacological treatment can be carried out at any appropriate time as long as a useful effect resulting from the combined action of the therapeutic agent and the non-pharmacological treatment is achieved. For example, in a preferred case, this useful effect is achieved even when the non-drug treatment is temporarily suspended, perhaps for several days or weeks, from the administration of the therapeutic agent.

[0175] The present invention also provides a pharmaceutically acceptable composition comprising a therapeutically effective amount of one or more compounds of formula (I), one or more pharmaceutically acceptable carriers (additives) and / or diluents, and optionally one or more of the above-mentioned other therapeutic agents formulated together.

[0176] The compounds of the present invention can be administered by any suitable means (e.g., oral administration (e.g., tablets, capsules (including sustained-release formulations or timed-release formulations), pills, powders, granules, elixirs, tinctures, suspensions (including nanosuspensions, microsuspensions, spray-dried dispersions), syrups, and emulsions); sublingual administration; buccal administration; parenteral administration (e.g., subcutaneous, intravenous, intramuscular, or intracardiac injection, or infusion techniques (e.g., sterile aqueous or non-aqueous solutions or suspensions)); nasal administration including administration to the nasal membrane (e.g., inhalation sprays); topical administration (e.g., in the form of cream formulations or ointments); or rectal administration (e.g., in the form of suppositories)) for any of the uses described herein. These may be administered alone, but are generally administered with a pharmaceutical carrier selected based on the chosen route of administration and standard pharmaceutical criteria.

[0177] As used herein, the term "pharmaceutically acceptable carrier" means a pharmaceutically acceptable substance, composition or vehicle, for example, a liquid or solid bulking agent, diluent, excipient, processing aid (e.g., lubricant, magnesium talc, calcium or zinc stearate, or stearic acid), or solvent encapsulant, which is involved in the transport or delivery of a particular compound from one organ or part of the body to a different organ or part of the body. Each carrier must be "acceptable" in the sense that it is compatible with the other ingredients in the formulation (i.e., including adjuvants, excipients or vehicles (e.g., diluents, preservatives, bulking agents, flow regulators, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, fragrances, antibacterial agents, antifungal agents, lubricants and pharmaceutical agents for compounding) which vary depending on the method of administration and the nature of the dosage form) and not harmful to the patient.

[0178] The term "pharmaceutical composition" means a composition comprising a combination of a compound of the present invention and at least one other pharmaceutically acceptable carrier.

[0179] Pharmaceutically acceptable carriers are formulated according to many factors that are within the expertise of a person of ordinary skill in the art. Pharmaceutically acceptable carriers are formulated according to many factors that are within the expertise of a person of ordinary skill in the art. These factors include, but are not limited to, the type and nature of the active agent being formulated, the patient to whom the composition containing the active agent is administered, the intended route of administration of the composition, and the targeted therapeutic indication. Pharmaceutically acceptable carriers include both aqueous and non-aqueous liquid media, as well as various solid and semi-solid dosage forms. Such carriers can include, in addition to the active agent, many different components and additives, and such additional components are included in the formulation for various reasons, for example, for stabilization reasons such as active agents, binders, etc. well-known to those skilled in the art. Explanation of suitable pharmaceutically acceptable carriers and the factors in selecting them are described in various readily available literature, for example, Allen, L. V. Jr. et al. Remington: The Science and Practice of Pharmacy (2 volumes), 22nd edition (2012), Pharmaceutical Press.

[0180] The dosing regimen of the compounds of the present invention will of course vary according to known factors (such as the pharmacodynamic properties of a particular drug and its method and route of administration; the species, age, sex, health status, medical condition, and body weight of the recipient; the nature and extent of the symptoms; the type of concomitant therapy; the frequency of treatment; the route of administration, the kidney and liver function of the patient, and the desired effect).

[0181] In general guidance, the daily oral dose of each active ingredient, when used to obtain the intended effect, ranges from about 0.001 to about 5000 mg / day, preferably from about 0.01 to about 1000 mg / day, and most preferably from about 0.1 to about 250 mg / day. The most preferred dose for a constant rate infusion in intravenous administration ranges from about 0.01 to about 10 mg / kg / min. The compounds of the present invention can be administered in a once-daily dose or in doses that divide the total daily dose into two, three, or four times.

[0182] This compound is generally appropriately selected for the intended dosage form (e.g., oral tablets, capsules, elixirs, and syrups) and is administered as a mixture with a suitable pharmaceutical diluent, excipient, or carrier (collectively referred to herein as a pharmaceutical carrier), which is in accordance with conventional pharmaceutical standards.

[0183] Dosage forms (pharmaceutical compositions) suitable for administration may contain from about 1 mg to about 2000 mg of the active ingredient per dosage unit. In these pharmaceutical compositions, the active ingredient is usually present in a weight of about 0.1 to 95% of the total weight of the composition.

[0184] Typical capsules for oral administration contain at least one compound of the present invention (250 mg), lactose (75 mg), and magnesium stearate (15 mg). This mixture is sieved through a 60-mesh sieve and filled into No. 1 gelatin capsules.

[0185] A typical injection preparation is produced by aseptically adding at least one compound of the present invention (250 mg) to a vial, freeze-drying, and sealing it aseptically. When in use, the contents of the vial are mixed with physiological saline (2 mL) to prepare an injection preparation.

[0186] The present invention encompasses within its scope pharmaceutical compositions comprising, as an active ingredient, a therapeutically effective amount of at least one compound of the present invention, either alone or in combination with a pharmaceutical carrier. The compounds of the present invention can be used as appropriate alone, in combination with other compounds of the present invention, or in combination with one or more other therapeutic agents (e.g., anti-cancer agents or other pharmaceutically active substances).

[0187] Regardless of the selected route of administration, the compounds of the present invention and / or the pharmaceutical compositions of the present invention, which can be used in an appropriate hydrated form, are formulated into pharmaceutically acceptable dosage forms by conventional methods known to those skilled in the art.

[0188] The actual dosage level of the active ingredient in the pharmaceutical compositions of the present invention can be varied so as to be non-toxic to the patient and to include an amount of the active ingredient effective to obtain a therapeutic effect for a particular patient, composition, and mode of administration.

[0189] The selected dosage level depends on a variety of factors including the activity of the particular compound or its ester, salt, or amide of the present invention being used, the route of administration of the particular compound being used, the time of administration, the rate of excretion or metabolism, the rate and extent of absorption, the duration of the treatment, other drugs, compounds, and / or substances being used in combination with the particular compound being used, the age, sex, weight, symptoms, health status, and medical history of the patient being treated, as well as factors known in the medical arts.

[0190] A physician or veterinarian having ordinary skill in the art can readily determine and prescribe the effective amount of the required pharmaceutical composition. For example, a physician or veterinarian can start the dosage of the compound of the present invention used in the pharmaceutical composition at a level lower than the required amount and gradually increase the dosage until the desired effect is obtained in order to achieve a therapeutic effect.

[0191] Generally, the appropriate daily dosage of the compounds of the present invention is the amount of the compound at the lowest dosage effective to obtain a therapeutic effect. Such effective dosages are generally determined by the above factors. Generally, the dosage of the compounds of the present invention to a patient is about 0.01 to about 50 mg / kg body weight / day for oral, intravenous, intraventricular and subcutaneous administration.

[0192] Optionally, the effective daily dosage of the active compound can be administered in divided dosages two, three, four, five, six or more times, at appropriate intervals throughout the day, in suitable unit dosage forms. In certain embodiments of the present invention, the dosing is once a day.

[0193] It is possible to administer the compounds of the present invention alone, but it is preferred to administer the compounds as pharmaceutical formulations (compositions).

[0194] When the other therapeutic agents described above are used in combination with the compounds of the present invention, they may be used, for example, in the amounts described in the Physicians' Desk Reference (PDR) or otherwise as specifically determined by those skilled in the art. In the methods of the present invention, the other therapeutic agents may be administered before, simultaneously with, or after the administration of the compounds of the present invention.

[0195] Manufacturing Method The compounds of the present invention can be synthesized by many methods available to those skilled in organic chemistry. General synthetic schemes for producing the compounds of the present invention are described below. This scheme is for illustration only and is not intended to limit the possible techniques that those skilled in the art can use to produce the compounds described herein. Various methods for producing the compounds of the present invention will be apparent to those skilled in the art. Examples of the compounds of the present invention produced by the methods described in the general scheme are shown in the Examples section below. The production of homochiral examples can be carried out by techniques known to those skilled in the art. For example, homochiral compounds can be produced by separating racemic products or diastereomers by chiral phase preparative HPLC. Alternatively, the compounds can be produced by known methods that give enantiomerically enriched or diastereomerically enriched products.

[0196] The reactions and techniques described in this section are carried out in a solvent suitable for the reagents and substances used and are appropriate for the transformations effected. Also, in the description of the synthetic methods set forth below, it is to be understood that all of the reaction conditions presented (including solvent selection, reaction atmosphere, reaction temperature, experimental time, and workup method) are chosen to be those standard for the reaction and should be readily recognizable to one skilled in the art. It is understood by one skilled in the art of organic synthesis that the functional groups present on various portions of the molecule must be compatible with the reagents and reactions presented. It is readily apparent to one skilled in the art that the substituents compatible with the reaction conditions are so limited, and alternative schemes are required when the substituents present are not suitable. The reaction may also require a judgment to change the order of synthetic steps or select a different particular reaction process in order to obtain the compounds of the present invention. Also, in any synthetic route planning in this field, it is recognized that another important consideration is the judicious selection of protecting groups to be used for the protection of the reactive functional groups present in the desired compounds described in the present invention. For the skilled experimenter, an authoritative literature describing many alternatives of protecting groups is Greene's Protective Groups in Organic Synthesis by Wuts and Greene (Fourth Edition, Wiley & Sons, 2007).

[0197] Examples The following examples illustrate certain and preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Chemical abbreviations and symbols, as well as scientific abbreviations and symbols, have their general and customary meanings unless otherwise noted. Further abbreviations utilized in the examples and throughout this specification are as defined above. Common intermediates are generally useful in the preparation of one or more of the examples and are identified in order (e.g., Intermediate 1, Intermediate 2, etc.) and abbreviated as Int.1 or I1, Int.2 or I2, etc. The compounds of the examples are identified either by the example and step in which they are prepared (e.g., "1-A" represents Step A of Example 1) or, only when the compound is the title compound of the example, by the example (e.g., "1" represents the title compound of Example 1). In some cases, alternative methods of preparing the intermediates or examples are described. Those skilled in the art of synthesis will frequently be able to devise desirable alternative methods of preparation based on one or more considerations (e.g., shorter reaction times, less expensive starting materials, ease of operation and isolation, improved yields, handling of catalysts, avoidance of toxic reagents, availability of special equipment, and reduction in the number of linear steps, etc.). The intention of describing alternative methods of preparation is to further enable the preparation of the examples of the present invention. In some cases, some of the functional groups in the schematic examples and claims may be replaced by well-known biological equivalents known to those skilled in the art (e.g., substitution of a carboxylic acid group with a tetrazole or phosphate moiety). Collected in deuterated dimethyl sulfoxide 1 The 1H NMR data were collected using water suppression in data processing. In the reporting of the spectra, no correction was made for the effect of water suppression. Protons adjacent to the water suppression frequency of 3.35 ppm show a decrease in signal intensity.

[0198] Abbreviations

Table 1

Table 2

[0199] The retention times reported for each example and intermediate by analytical LC-MS / HPLC were obtained using one of the following general analytical LC-MS / HPLC conditions:

[0200] Method A: Column: XBridge BEH XP C18 (2.1x50mm), 2.5μm; Mobile phase A: 10 mM ammonium acetate: acetonitrile (95:5), Mobile phase B: 10 mM ammonium acetate: acetonitrile (5:95), gradient = 0 - 100% B over 3 minutes; Temperature: 50°C; Flow rate: 1.1 mL / min; Detection: UV (220 nm)

[0201] Method B: Column: Waters Acquity UPLC BEH C18, 2.1x50mm, 1.7μm particles; Mobile phase A: 5:95 acetonitrile: water + 10 mM NH4OAc; Mobile phase B: 95:5 acetonitrile: water + 10 mM NH4OAc; Temperature: 50°C; Gradient: 0 - 100% B over 3 minutes, then hold at 100% B for 0.75 minutes; Flow: 1.0 mL / min; Detection: UV (220 nm)

[0202] Method C: Column: Waters Acquity UPLC BEH C18, 2.1x50mm, 1.7μm particles; Mobile phase A: 5:95 acetonitrile: water + 0.1% TFA; Mobile phase B: 95:5 acetonitrile: water + 0.1% TFA; Temperature: 50°C; Gradient: 0 - 100% B over 3 minutes, then hold at 100% B for 0.75 minutes; Flow: 1.0 mL / min; Detection: UV (220 nm)

[0203] Method D: Column: Waters XBridge C18, 2.1mmx50mm, 1.7μm particles; Mobile phase A: 5:95 acetonitrile: water + 10 mM NH4OAc; Mobile phase B: 95:5 acetonitrile: water + 10 mM NH4OAc; Temperature: 50°C; Gradient: 0% B - 100% B over 3 minutes, then hold at 100% B for 0.50 minutes; Flow: 1 mL / min; Detection: UV (220 nm)

[0204] Method E: Column: Waters Acquity UPLC BEH C18, 2.1 mm x 50 mm, 1.7 μm particles; Mobile phase A: 5:95 acetonitrile: water + 0.1% TFA; Mobile phase B: 95:5 acetonitrile: water + 0.1% TFA; Temperature: 50 °C; Gradient: 0% B to 100% B over 3 minutes, then hold at 100% B for 0.50 minutes; Flow rate: 1 mL / min; Detection: UV (220 nm)

[0205] Intermediate 2 tert-Butyl (2S,5R)-4-(1-(4-fluorophenyl)-2-isopropoxy-2-oxoethyl)-2,5-dimethylpiperazine-1-carboxylate

Chem.

[0206] To a stirred solution of 2-((2R,5S)-4-(tert-butoxycarbonyl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetic acid (500 mg, 1.36 mmol) in DMF (10 mL) was added K2CO3 (566 mg, 4.09 mmol) and 2-iodopropane (0.82 mL, 8.19 mmol). The reaction mixture was stirred at room temperature for 16 h. The reaction was quenched with water (50 mL). The reaction mixture was extracted with ethyl acetate (3 x 100 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and evaporated under reduced pressure to give the crude product (320 mg, 64%). LCMS: m / z = 409.0 (M+H); Retention time: 1.48, 1.65 minutes (LCMS method: Column: Waters Acquity UPLC BEH C18 (2.1 x 50 mm), 1.7 μm; Mobile phase A: 0.1% TFA in water; Mobile phase B: 0.1% TFA in acetonitrile; Gradient = 20 - 90% B over 1.1 minutes, then hold at 90% B for 0.6 minutes; Temperature: 50 °C; Flow rate: 0.7 mL / min; Detection: UV (220 nm)

[0207] Intermediate 3 Isopropyl 2-((2R,5S)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate

Chem.

[0208] To a stirred solution of tert-butyl (2S,5R)-4-(1-(4-fluorophenyl)-2-isopropoxy-2-oxoethyl)-2,5-dimethylpiperazine-1-carboxylate (300 mg, 0.73 mmol) in DCM (5 mL) was added HCl in dioxane (2 mL, 65.8 mmol). The reaction mixture was stirred at room temperature for 1 h. The reaction mixture was concentrated under reduced pressure to afford the crude product. The crude compound was triturated with DCM and hexane (1:4) and filtered to give isopropyl 2-((2R,5S)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate (180 mg, 71%). LCMS: m / z = 309.5 (M+H); retention time: 0.73 min (LCMS method: column: Waters Acquity UPLC BEH C18 (2.1x50 mm) 1.7 μm, mobile phase A: 10 mM NH4OAc: acetonitrile (95:5); mobile phase B: 10 mM NH4OAc: acetonitrile (5:95), gradient = 20 - 90% B over 1.1 min then hold at 90% B for 0.6 min; temperature: 50 °C; flow rate: 0.7 mL / min; detection: UV (220 nm)

[0209] Examples 1 and 2 Isopropyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate

Chem.

[0210] To a stirred solution of 6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl trifluoromethanesulfonate·HCl (200 mg, 0.54 mmol) in acetonitrile (8 mL) was added DIPEA (0.283 mL, 1.62 mmol). The reaction mixture was stirred for 5 minutes and isopropyl 2-((2R,5S)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate (167 mg, 0.54 mmol) was added. The reaction mixture was heated at 85 °C for 3 hours. The reaction mixture was cooled to room temperature and evaporated under reduced pressure to give the crude product, which was purified by preparative HPLC (method: column: Sunfire C18 (150x19) mm - 5 μm; mobile phase A: 10 mM ammonium acetate in water (pH: 4.5); mobile phase B: acetonitrile; flow rate: 20 mL / min) to give Examples 1 and 2.

[0211] Example 1: 13.0 mg, yield 4.9%; LCMS: m / z = 492.2 (M+H); retention time: 2.25 minutes (LCMS method: column: Ascentis Express C18 (2.1x50 mm), 2.7 μm; mobile phase A: 10 mM NH4OAc: acetonitrile (95:5), mobile phase B: 10 mM NH4OAc: acetonitrile (5:95), gradient = 0 - 100% B over 3 minutes; temperature: 50 °C; flow rate: 1.1 mL / min; detection: UV (220 nm)); 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.12 - 8.18 (m, 1H), 8.04 - 8.10 (m, 1H), 7.53 - 7.62 (m, 2H) 7.22 (t, J = 8.80 Hz, 2H), 6.04 (s, 1H), 4.92 (quin, J = 6.17 Hz, 1H), 4.35 - 4.45 (m, 1H), 4.32 (s, 1H), 3.61 - 3.71 (m, 1H), 3.49 - 3.61 (m, 4H), 3.37 (brs, 1H), 2.76 (dd, J = 11.37, 3.30 Hz, 1H), 2.06 - 2.14 (m, 1H), 1.17 - 1.23 (m, 6H), 1.11 (t, J = 6.60 Hz, 6H)

[0212] Example 2: 22.0 mg, yield 8.1%; LCMS: m / z = 492.2 (M+H); retention time: 2.23 minutes (LCMS method: column: Ascentis Express C18 (2.1x50 mm), 2.7 μm; mobile phase A: 10 mM NH4OAc: acetonitrile (95:5), mobile phase B: 10 mM NH4OAc: acetonitrile (5:95), gradient = 0-100% B over 3 minutes; temperature: 50 °C; flow rate: 1.1 mL / min; detection: UV (220 nm)); 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.13 - 8.18 (m, 1H), 8.03 - 8.11 (m, 1H), 7.60 (dd, J = 8.56, 5.62 Hz, 2H), 7.24 (t, J = 8.80 Hz, 2H), 5.99 (s, 1H), 4.90 (quin, J = 6.24 Hz, 1H), 4.52 - 4.65 (m, 1H), 4.29 (s, 1H), 3.55 - 3.63 (m, 1H), 3.52 (s, 3H), 3.42 (dd, J = 2.59, 3.30 Hz, 1H), 3.13 (dd, J = 11.25, 3.42 Hz, 1H), 2.77 - 2.88 (m, 1H), 2.44 (brd, J = 10.52 Hz, 1H), 1.28 (d, J = 6.36 Hz, 3H), 1.16 (d, J = 6.11 Hz, 3H), 1.11 (d, J = 6.36 Hz, 3H), 1.00 (d, J = 6.36 Hz, 3H)

[0213] Examples 3 and 4 Methyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(5-fluoropyridin-2-yl)acetate [Chemical formula]

[0214] To a stirred solution of 8-((2S,5R)-2,5-dimethylpiperazin-1-yl)-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile, HCl (300 mg, 0.9 mmol) in acetonitrile (8 mL) was added DIPEA (0.471 mL, 2.70 mmol), followed by methyl 2-bromo-2-(5-fluoropyridin-2-yl)acetate (268 mg, 1.08 mmol). The reaction mixture was heated at 85 °C for 16 h. The reaction mixture was filtered through a pad of celite and concentrated under reduced pressure to give the crude product. The crude product was purified by preparative SFC (method: column / dimension: Lux Amylose-2 (250x4.6) mm, 5 μm; %CO2: 65%; % co-solvent: 35% ACN:MeOH (50:50); total flow: 4.0 g / min; back pressure: 100 bar; temperature: 30 °C; UV: 225 nm) to give Examples 3 and 4.

[0215] Example 3: 20.3 mg, yield 39.4%; LCMS: m / z = 465.2 (M+H); retention time: 1.67 min (LCMS method: column: Ascentis Express C8 (50x2.1 mm) 2.7 μm; mobile phase A: 10 mM ammonium formate:acetonitrile (98:2), mobile phase B: 10 mM ammonium formate:acetonitrile (2:98), gradient = 0-100% B over 1.5 min, then hold at 100% B for 0.6 min; temperature: 27 °C; flow rate: 1.0 mL / min; detection: UV (220 nm)); 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.53 (d, J = 2.9 Hz, 1H), 8.21 - 8.13 (m, 1H), 8.10 - 8.03 (m, 1H), 7.87 - 7.79 (m, 1H), 7.78 - 7.71 (m, 1H), 6.08 (s, 1H), 4.68 (s, 1H), 4.28 - 4.15 (m, 1H), 3.71 - 3.58 (m, 4H), 3.53 (s, 4H), 3.32 - 3.23 (m, 1H), 2.82 (dd, J = 3.5, 11.9 Hz, 1H), 2.31 (dd, J = 4.4, 12.0 Hz, 1H), 1.14 (dd, J = 6.4, 16.6 Hz, 6H)

[0216] Example 4: 21.7 mg, yield 41.2%; LCMS: m / z = 465.2 (M+H); retention time: 1.67 minutes (LCMS method: column: Ascentis Express C8 (50x2.1 mm) 2.7 μm; mobile phase A: 10 mM ammonium formate: acetonitrile (98:2), mobile phase B: 10 mM ammonium formate: acetonitrile (2:98), gradient = 0 - 100% B over 1.5 minutes, then hold at 100% B for 0.6 minutes; temperature: 27 °C; flow rate: 1.0 mL / min; detection: UV (220 nm)); 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.53 (d, J = 2.7 Hz, 1H), 8.19 - 8.13 (m, 1H), 8.09 - 8.03 (m, 1H), 7.86 - 7.79 (m, 1H), 7.78 - 7.73 (m, 1H), 6.03 (s, 1H), 4.73 (s, 1H), 4.53 - 4.39 (m, 1H), 3.66 (s, 3H), 3.59 - 3.49 (m, 5H), 3.32 (brd, J = 3.2 Hz, 1H), 3.11 - 3.02 (m, 1H), 2.40 (dd, J = 2.6, 11.9 Hz, 1H), 1.17 (d, J = 6.6 Hz, 3H), 1.06 (d, J = 6.6 Hz, 3H)

[0217] Intermediate 4 tert-Butyl (2S,5R)-4-(1-(4-fluorophenyl)-2-methoxy-2-oxoethyl)-2,5-dimethylpiperazine-1-carboxylate

Chemical Structure

[0218] To a solution of methyl 2-bromo-2-(4-fluorophenyl)acetate (1.0 g, 4.05 mmol) in acetonitrile (10 mL) was added DIPEA (2.12 mL, 12.14 mmol), followed by tert-butyl (2S,5R)-2,5-dimethylpiperazine-1-carboxylate (1.04 g, 4.86 mmol). The reaction mixture was stirred at 85 °C overnight. The reaction mixture was concentrated under reduced pressure to remove volatile substances, and the crude residue was dissolved in ethyl acetate (150 mL) and washed with water. The aqueous layer was back-extracted with ethyl acetate (100 mL x 2). The combined organic layers were flashed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product. The crude residue was purified by silica gel chromatography (10-20% ethyl acetate / petroleum ether; 40 g column) to give tert-butyl (2S,5R)-4-(1-(4-fluorophenyl)-2-methoxy-2-oxoethyl)-2,5-dimethylpiperazine-1-carboxylate (1.201 g, 3.16 mmol, 78% yield). LCMS: m / z = 381.4 (M+H); rt 2.18 minutes (LCMS method: column: Waters Acquity UPLC BEH C18 (2.1 x 50 mm) 1.7 μm, mobile phase A: 10 mM NH4OAc: acetonitrile (95:5); mobile phase B: 10 mM NH4OAc: acetonitrile (5:95), gradient = 20-90% B over 1.1 minutes, then hold at 90% B for 0.6 minutes; temperature: 50 °C; flow rate: 0.7 mL / min; detection: UV (220 nm))

[0219] Intermediate 5 2-((2R,5S)-4-(tert-butoxycarbonyl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetic acid

Chemical Structure

[0220] To a solution of tert-butyl (2S,5R)-4-(1-(4-fluorophenyl)-2-methoxy-2-oxoethyl)-2,5-dimethylpiperazine-1-carboxylate (500 mg, 1.31 mmol) in THF (12 mL) / water (4 mL) was added LiOH (315 mg, 13.14 mmol). The reaction mixture was stirred at 60 °C overnight. The reaction mixture was partially concentrated under reduced pressure to remove volatiles and extracted with ethyl acetate. The organic layers were combined, washed with brine, dried over Na2SO4, concentrated under reduced pressure to give 2-((2R,5S)-4-(tert-butoxycarbonyl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetic acid (451 mg, 94% yield). LCMS: m / z = 367.2 (M+H); rt 0.97 and 1.01 minutes (LCMS method: column: Waters Acquity UPLC BEH C18 (2.1x50 mm) 1.7 μm, mobile phase A: 10 mM NH4OAc: acetonitrile (95:5); mobile phase B: 10 mM NH4OAc: acetonitrile (5:95), gradient = 20 - 90% B over 1.1 minutes then hold at 90% B for 0.6 minutes; temperature: 50 °C; flow rate: 0.7 mL / min; detection: UV (220 nm))

[0221] Intermediate 6 tert-butyl (2S,5R)-4-(2-((2,2-difluoroethyl)amino)-1-(4-fluorophenyl)-2-oxoethyl)-2,5-dimethylpiperazine-1-carboxylate [Chemical formula]

[0222] To a solution of 2-((2R,5S)-4-(tert-butoxycarbonyl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetic acid (100 mg, 0.27 mmol) in DMF (5 mL), 1-propanephosphonic anhydride (0.55 mL, 0.55 mmol) was added, followed by DIPEA (0.05 mL, 0.27 mmol). The reaction mixture was stirred at room temperature for 30 minutes. Next, 2,2-difluoroethan-1-amine (22.12 mg, 0.27 mmol) was added and the reaction mixture was stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure to remove volatile substances, and the crude residue was dissolved in ethyl acetate (20 mL) and washed with water. The aqueous layer was back-extracted with ethyl acetate (15 mL x 2). The organic layers were combined, flashed with brine, dried over Na2SO4, and concentrated under reduced pressure to obtain the crude product. The crude residue was purified by silica gel chromatography (10 - 20% MeOH / CHCl3; 12 g column) to give tert-butyl (2S,5R)-4-(2-((2,2-difluoroethyl)amino)-1-(4-fluorophenyl)-2-oxoethyl)-2,5-dimethylpiperazine-1-carboxylate (65 mg, yield 55.5%). LCMS: m / z = 430.4 (M+H); rt 1.86 minutes (LCMS method: column: Waters Acquity UPLC BEH C18 (2.1 x 50 mm) 1.7 μm, mobile phase A: 10 mM NH4OAc: acetonitrile (95:5); mobile phase B: 10 mM NH4OAc: acetonitrile (5:95), gradient = 20 - 90% B over 1.1 minutes, then hold at 90% B for 0.6 minutes; temperature: 50 °C; flow rate: 0.7 mL / min; detection: UV (220 nm))

[0223] Intermediate 7 N-(2,2-Difluoroethyl)-2-((2R,5S)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetamide [Chemical Structure]

[0224] To a solution of tert-butyl (2S,5R)-4-(2-((2,2-difluoroethyl)amino)-1-(4-fluorophenyl)-2-oxoethyl)-2,5-dimethylpiperazine-1-carboxylate (60 mg, 0.14 mmol) in DCM (10 mL) was added HCl (0.04 mL, 1.39 mmol) in dioxane. The reaction mixture was stirred for 3 h. The reaction mixture was evaporated under reduced pressure and the crude residue was triturated with hexane. The solid was filtered through a sintered funnel and dried under reduced pressure to give N-(2,2-difluoroethyl)-2-((2R,5S)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetamide (42 mg, 91% yield). LCMS: m / z = 330.3 (M+H); rt 0.81 and 0.91 min (LCMS method: column: Waters Acquity UPLC BEH C18 (2.1x50 mm) 1.7 μm, mobile phase A: 10 mM NH4OAc: acetonitrile (95:5); mobile phase B: 10 mM NH4OAc: acetonitrile (5:95), gradient = 20 - 90% B over 1.1 min then hold at 90% B for 0.6 min; temperature: 50 °C; flow rate: 0.7 mL / min; detection: UV (220 nm))

[0225] Examples 5 and 6 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-N-(2,2-difluoroethyl)-2-(4-fluorophenyl)acetamide

Chemical formula

[0226] To a solution of 6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl trifluoromethanesulfonate (47.4 mg, 0.14 mmol) in acetonitrile (3 mL) was added DIPEA (0.06 mL, 0.33 mmol), followed by N-(2,2-difluoroethyl)-2-((2R,5S)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetamide·HCl (40 mg, 0.11 mmol). The reaction mixture was stirred at 85 °C overnight. The reaction mixture was concentrated under reduced pressure to remove volatile substances, and the crude residue was dissolved in ethyl acetate and washed with water. The aqueous layer was back-extracted with ethyl acetate (15 mL x 2). The combined organic layers were flashed with brine, dried over Na2SO4, concentrated under reduced pressure to obtain the crude product. The crude residue was purified by preparative HPLC (method: column: DAD-1-cellulose-2 (250 x 4.6 mm), 5 micron; DAD-2-cellulose-4 (250 x 4.6 mm), 5 micron; mobile phase: 0.1% DEA in acetonitrile; flow: 2.0 mL / min) to obtain Examples 5 and 6.

[0227] Example 5: 1.2 mg, yield 2.1%; LCMS: m / z = 513.2 (M+H); rt 1.731 minutes (LCMS method: column: XBridge BEH XP C18 (2.1 x 50 mm), 2.5 μm; mobile phase A: 10 mM ammonium acetate:acetonitrile (95:5), mobile phase B: 10 mM ammonium acetate:acetonitrile (5:95), gradient = 0 - 100% B over 3 minutes; temperature: 50 °C; flow rate: 1.1 mL / min; detection: UV (220 nm); 1 1H NMR (400 MHz, DMSO-d6): δ 8.30 - 8.44 (m, 1H), 8.12 - 8.20 (m, 1H), 8.01 - 8.10 (m, 1H), 7.55 - 7.69 (m, 2H), 7.15 - 7.29 (m, 2H), 5.84 - 6.19 (m, 1H), 4.58 - 4.69 (m, 1H), 4.12 (s, 1H), 3.42 - 3.59 (m, 6H), 3.03 - 3.08 (m, 1H), 2.81 - 2.93 (m, 3H), 2.59 (brd, J = 0.73 Hz, 1H), 1.20 - 1.34 (m, 3H), 0.94 (d, J = 6.60 Hz, 3H) ppm

[0228] Example 6: (0.5 mg, yield 0.9%; LCMS: m / z = 513.2 (M+H); rt 1.734 minutes (LCMS method: column: XBridge BEH XP C18 (2.1x50 mm), 2.5 μm; mobile phase A: 10 mM ammonium acetate: acetonitrile (95:5), mobile phase B: 10 mM ammonium acetate: acetonitrile (5:95), gradient = 0 - 100% B over 3 minutes; temperature: 50 °C; flow rate: 1.1 mL / min; detection: UV (220 nm)); 1 H NMR (400 MHz, DMSO-d6): δ 8.55 - 8.74 (m, 1H), 8.12 - 8.22 (m, 1H), 7.98 - 8.09 (m, 1H), 7.45 - 7.64 (m, 2H), 7.06 - 7.25 (m, 2H), 5.83 - 6.18 (m, 2H), 4.39 - 4.49 (m, 1H), 4.18 - 4.28 (m, 1H), 3.66 - 3.77 (m, 1H), 3.48 - 3.63 (m, 4H), 3.36 (brs, 1H), 2.85 - 2.94 (m, 1H), 2.67 - 2.75 (m, 2H), 2.07 (brd, J = 11.74 Hz, 1H), 0.97 - 1.33 (m, 6H)

[0229] Examples 7 and 8 Methyl 2 - ((2R,5S)-4-(6 - cyano - 1 - methyl - 2 - oxo - 1,2 - dihydro - 1,5 - naphthyridin - 4 - yl)-2,5 - dimethylpiperazin - 1 - yl)-2-(4 - fluorophenyl)acetate

Chemical Structure

[0230] To a solution of 8 - ((2S,5R)-2,5 - dimethylpiperazin - 1 - yl)-5 - methyl - 6 - oxo - 5,6 - dihydro - 1,5 - naphthyridine - 2 - carbonitrile·HCl (200 mg, 0.59 mmol) in acetonitrile (8 mL) was added DIPEA (0.31 mL, 1.8 mmol). The reaction mixture was stirred for 5 minutes and methyl 2 - bromo - 2-(4 - fluorophenyl)acetate (148 mg, 0.6 mmol) was added. The reaction mixture was heated at 85 °C for 16 hours. The reaction mixture was filtered through a cartridge and concentrated under reduced pressure to obtain a solid residue. The crude compound was purified by silica gel column (12 g silica gel column; using 60 - 67% ethyl acetate / petroleum ether) to give methyl 2 - ((2R,5S)-4-(6 - cyano - 1 - methyl - 2 - oxo - 1,2 - dihydro - 1,5 - naphthyridin - 4 - yl)-2,5 - dimethylpiperazin - 1 - yl)-2-(4 - fluorophenyl)acetate (250 mg, yield 90%) as a brown solid. The purified compound (30 mg) was further purified by preparative HPLC (method: column: Inersil ODS (250 mm x 19 mm ID, 5 μm); mobile phase A = 0.1% TFA in water; mobile phase B = acetonitrile; gradient: 40 - 100% B over 20 minutes; flow rate 17 mL / min) to obtain Examples 7 and 8.

[0231] Example 7: Methyl 2 - ((2R,5S)-4-(6 - cyano - 1 - methyl - 2 - oxo - 1,2 - dihydro - 1,5 - naphthyridin - 4 - yl)-2,5 - dimethylpiperazin - 1 - yl)-2-(4 - fluorophenyl)acetate·TFA (12.2 mg, yield 32.4%); LCMS: m / z = 464.2 (M + H); rt 2.07 minutes (LCMS method: column: Ascentis Express C8 (50 x 2.1 mm) 2.7 μm; mobile phase A: 10 mM ammonium formate:acetonitrile (98:2), mobile phase B: 10 mM ammonium formate:acetonitrile (2:98), gradient = 0 - 100% B over 1.5 minutes then hold at 100% B for 0.6 minutes; temperature: 27 °C; flow rate: 1.0 mL / min; detection: UV (220 nm)); 11H NMR (400 MHz, DMSO-d6) δ 8.20 - 8.15 (m, 1H), 8.10 - 8.03 (m, 1H), 7.62 (dd, J = 5.6, 8.3 Hz, 2H), 7.27 (t, J = 8.9 Hz, 2H), 7.24 - 6.95 (m, 1H), 6.03 (s, 1H), 4.71 - 4.51 (m, 2H), 3.65 (s, 4H), 3.53 (s, 4H), 3.23 - 3.13 (m, 1H), 2.92 (brd, J = 6.6 Hz, 1H), 2.55 (s, 1H), 1.32 - 1.19 (m, 3H), 1.11 - 0.95 (m, 3H)

[0232] Example 8: Methyl 2 - ((2R,5S)-4-(6 - cyano - 1 - methyl - 2 - oxo - 1,2 - dihydro - 1,5 - naphthyridin - 4 - yl)-2,5 - dimethylpiperazin - 1 - yl)-2-(4 - fluorophenyl)acetate·TFA (17.5 mg, yield 46.8%); LCMS: m / z = 464.2 (M + H); rt 2.08 minutes (LCMS method: column: Ascentis Express C8 (50 x 2.1 mm) 2.7 μm; mobile phase A: 10 mM ammonium formate: acetonitrile (98:2), mobile phase B: 10 mM ammonium formate: acetonitrile (2:98), gradient = 0 - 100% B over 1.5 minutes, then hold at 100% B for 0.6 minutes; temperature: 27 °C; flow rate: 1.0 mL / min; detection: UV (220 nm)); 1 1H NMR (400 MHz, DMSO-d6) δ 8.19 - 8.13 (m, 1H), 8.11 - 8.05 (m, 1H), 7.62 - 7.53 (m, 2H), 7.28 - 7.18 (m, 2H), 6.05 (s, 1H), 4.52 - 4.37 (m, 2H), 3.66 (s, 3H), 3.53 (s, 3H), 3.39 - 3.32 (m, 1H), 2.85 - 2.76 (m, 1H), 2.55 (s, 2H), 2.20 - 2.08 (m, 1H), 1.19 (d, J = 6.6 Hz, 3H), 1.12 (d, J = 6.4 Hz, 3H)

[0233] Examples 9 and 10 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetic acid

Chem.

[0234] To a solution of methyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate (0.9 g, 1.94 mmol) in 1,4-dioxane (15 mL) was added trimethylstannyl hydroxide (1.76 g, 9.71 mmol). The reaction mixture was heated at 100 °C for 28 h. The reaction mixture was cooled in an ice bath and diluted with ethyl acetate (50 mL). The precipitate was filtered off through a Buchner funnel. The filtrate was concentrated under reduced pressure to give crude 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetic acid (crude product = 1.11 g). LCMS: m / z = 450.4 and 450.3 (M+H); rt 0.88 and 0.93 min (LCMS method: column: Waters Acquity UPLC BEH C18 (2.1x50 mm) 1.7 μm, mobile phase A: 10 mM NH4OAc: acetonitrile (95:5); mobile phase B: 10 mM NH4OAc: acetonitrile (5:95), gradient = 20 - 90% B over 1.1 min, then hold at 90% B for 0.6 min; temperature: 50 °C; flow rate: 0.7 mL / min; detection: UV (220 nm)). The crude compound (0.08 g) was purified by preparative HPLC (method column: Sapphire C18 (150 mm x 19 mm ID, 5 μm); mobile phase A = 10 mM AA in water; mobile phase B = acetonitrile; gradient: 10 - 55% B over 25 min; flow 19 mL / min) to give diastereomers 1 and 2.

[0235] Example 9: 2.0 mg, yield 2.5%. LCMS: m / z = 450.1 (M+H); rt 1.07 minutes (LCMS method: column: Ascentis Express C8 (50x2.1 mm) 2.7 μm; mobile phase A: 10 mM ammonium formate: acetonitrile (98:2), mobile phase B: 10 mM ammonium formate: acetonitrile (2:98), gradient = 0 - 100% B over 1.5 minutes, then hold at 100% B for 0.6 minutes; temperature: 27 °C; flow rate: 1.0 mL / min; detection: UV (220 nm)); 1 H NMR (400 MHz, DMSO-d6) δ 8.13 - 8.19 (m, 1H), 8.02 - 8.09 (m, 1H), 7.60 (dd, J = 8.56, 5.62 Hz, 2H), 7.23 (t, J = 8.80 Hz, 2H), 6.00 (s, 1H), 4.51 - 4.64 (m, 1H), 4.22 (s, 1H), 3.57 - 3.66 (m, 1H), 3.52 (s, 4H), 3.42 - 3.46 (m, 1H), 3.11 (dd, J = 11.25, 4.16 Hz, 1H), 2.78 - 2.86 (m, 1H), 1.28 (d, J = 6.36 Hz, 3H), 1.00 (d, J = 6.60 Hz, 3H)

[0236] Example 10: 4.5 mg, yield 5.6%; LCMS: m / z = 450.1 (M+H); rt 1.15 minutes (LCMS method: column: Ascentis Express C8 (50x2.1 mm) 2.7 μm; mobile phase A: 10 mM ammonium formate: acetonitrile (98:2), mobile phase B: 10 mM ammonium formate: acetonitrile (2:98), gradient = 0 - 100% B over 1.5 minutes, then hold at 100% B for 0.6 minutes; temperature: 27 °C; flow rate: 1.0 mL / min; detection: UV (220 nm)); 11H NMR (400 MHz, DMSO-d6) δ 8.11 - 8.18 (m, 1H), 8.02 - 8.09 (m, 1H), 7.57 (dd, J = 8.68, 5.50 Hz, 2H), 7.21 (t, J = 8.93 Hz, 2H), 6.04 (s, 1H), 4.38 - 4.48 (m, 1H), 4.25 (s, 1H), 3.65 (brs, 1H), 3.55 - 3.61 (m, 1H), 3.53 (s, 4H), 3.43 - 3.49 (m, 1H), 2.75 (dd, J = 11.49, 3.42 Hz, 1H), 1.20 (d, J = 6.60 Hz, 3H), 1.12 (d, J = 6.60 Hz, 3H)

[0237] Examples 11 and 12 (8 - ((2S,5R)-4-(1-(4-Fluorophenyl)-2-oxo-2-(piperidin-1-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile [Chemical formula]

[0238] A stirred solution of 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetic acid (70 mg, 0.16 mmol) in DCM (5 mL) was added with EDC (35.8 mg, 0.19 mmol), HOBt (28.6 mg, 0.19 mmol), and DIPEA (0.05 mL, 0.31 mmol) at room temperature, followed by the addition of piperidine (0.02 mL, 0.19 mmol). The reaction mixture was stirred for 12 h. The reaction mixture was extracted with DCM (2 x 30 mL), washed with water and brine, dried over sodium sulfate, and the solvent was removed under reduced pressure to obtain a crude residue, which was purified by preparative HPLC [Method: Column: Waters XBridge C18, 150 mm x 19 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; Gradient: held at 15% B for 2 min, changed to 15 - 52% B over 25 min, then held at 100% B for 5 min; Flow rate: 15 mL / min; Column temperature: 25 °C]. The fractions containing the product were combined and dried via centrifugal evaporation to obtain Examples 11 and 12.

[0239] Example 11: 1 mg, yield 1%; LCMS: m / z = 517.2 (M + H); rt 1.98 min (LCMS method: Column: XBridge BEH XP C18 (2.1 x 50 mm), 2.5 μm; Mobile phase A: 10 mM ammonium acetate: acetonitrile (95:5), Mobile phase B: 10 mM ammonium acetate: acetonitrile (5:95), Gradient = 0 - 100% B over 3 min; Temperature: 50 °C; Flow rate: 1.1 mL / min; Detection: UV (220 nm)); 11H NMR (400 MHz, DMSO-d6) δ ppm 8.09 - 8.17 (m, 1H), 7.99 - 8.07 (m, 1H), 7.59 (dd, J = 8.44, 5.75 Hz, 2H), 7.20 (t, J = 8.80 Hz, 2H), 5.99 (s, 1H), 4.81 (s, 1H), 4.47 (brdd, J = 7.70, 4.28 Hz, 1H), 3.61 - 3.71 (m, 1H), 3.47 - 3.58 (m, 6H), 3.38 - 3.47 (m, 4H), 3.07 - 3.18 (m, 1H), 2.83 - 2.92 (m, 1H), 1.46 - 1.61 (m, 2H), 1.26 - 1.44 (m, 4H), 1.20 (d, J = 6.36 Hz, 4H), 1.02 (d, J = 6.36 Hz, 3H)

[0240] Example 12: 1 mg, yield 1%; LCMS: m / z = 517.2 (M + H); rt 1.98 minutes (LCMS method: column: XBridge BEH XP C18 (2.1 x 50 mm), 2.5 μm; mobile phase A: 10 mM ammonium acetate: acetonitrile (95:5), mobile phase B: 10 mM ammonium acetate: acetonitrile (5:95), gradient = 0 - 100% B over 3 minutes; temperature: 50 °C; flow rate: 1.1 mL / min; detection: UV (220 nm)); 1 1H NMR (400 MHz, DMSO-d6) δ ppm 8.11 - 8.16 (m, 1H), 8.01 - 8.10 (m, 1H), 7.58 (dd, J = 8.44, 5.50 Hz, 2H), 7.11 - 7.23 (m, 2H), 6.02 (s, 1H), 4.81 (s, 1H), 4.29 - 4.41 (m, 1H), 3.69 - 3.83 (m, 1H), 3.50 - 3.63 (m, 6H), 2.73 - 2.82 (m, 1H), 2.19 - 2.26 (m, 2H), 1.48 - 1.61 (m, 3H), 1.24 - 1.41 (m, 4H), 1.13 (d, J = 6.60 Hz, 3H), 1.09 (d, J = 6.36 Hz, 3H)

[0241] Table 1

Table 3

Table 4

Table 5

Table 6

Table 7

[0242] Intermediate 8: tert-Butyl (2S,5R)-4-(1-(4-fluorophenyl)-3-methoxy-3-oxopropyl)-2,5-dimethylpiperazine-1-carboxylate

Chem.

[0243] To a stirred solution of tert-butyl (2S,5R)-2,5-dimethylpiperazine-1-carboxylate (1 g, 4.67 mmol) in DMSO (10 mL) was added 4-fluorobenzaldehyde (0.87 g, 7.00 mmol) and trimethyl borate (0.52 mL, 4.67 mmol). The reaction mixture was stirred at room temperature for 15 minutes. The resulting solution was treated by dropwise addition of 1-(tert-butyldimethylsilyloxy)-1-methoxyethane (1.5 mL, 7.00 mmol) and stirred at room temperature for 16 hours. The reaction mixture was extracted with EtOAc (2 x 100 mL), washed with water, brine, and dried over sodium sulfate. The solvent was removed under reduced pressure to give a crude residue, which was purified by silica gel flash column chromatography (30% EtOAc in n-hexane; 24 g column) to give tert-butyl (2S,5R)-4-(1-(4-fluorophenyl)-3-methoxy-3-oxopropyl)-2,5-dimethylpiperazine-1-carboxylate (1.2 g, 65% yield) as a mixture of diastereomers. LCMS: m / z, 453.2 (M+59); rt 2.17 minutes (LCMS method: column: Waters Acquity UPLC BEH C18 (2.1 x 50 mm) 1.7 μm, mobile phase A: 10 mM NH4OAc: acetonitrile (95:5); mobile phase B: 10 mM NH4OAc: acetonitrile (5:95), gradient = 20 - 100% B over 2 minutes, then hold at 100% B for 0.3 minutes; temperature: 50 °C; flow rate: 0.7 mL / min; detection: UV (220 nm))

[0244] Intermediate 9: Methyl 3-((2R,5S)-2,5-dimethylpiperazin-1-yl)-3-(4-fluorophenyl)propanoate hydrochloride

Chemical formula

[0245] To a stirred solution of tert-butyl (2S,5R)-4-(1-(4-fluorophenyl)-3-methoxy-3-oxopropyl)-2,5-dimethylpiperazine-1-carboxylate (125 mg, 0.32 mmol) in DCM (5 mL) was added 4N HCl (0.8 mL, 3.17 mmol) in 1,4-dioxane at room temperature. The reaction mixture was stirred for 2 h. The reaction mixture was concentrated under reduced pressure to afford a mixture of diastereomers of methyl 3-((2R,5S)-2,5-dimethylpiperazine-1-yl)-3-(4-fluorophenyl)propanoate (120 mg, 64% yield). LCMS: m / z, 295.4 (M+H); rt 1.08 and 1.11 min (LCMS method: column: Acquity UPLC BEH C18 (3.0x50 mm) 1.7 μm; mobile phase A: 10 mM ammonium acetate: acetonitrile (95:5), mobile phase B: 10 mM ammonium acetate: acetonitrile (5:95), method: %B: 0 min - 20: 2 min - 100: 2.3 min - 100, temperature: 27 °C; flow rate: 0.7 mL / min; detection: UV (220 nm))

[0246] Examples 31 and 32 Methyl 3-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazine-1-yl)-3-(4-fluorophenyl)propanoate

Chemical Structure

[0247] To a stirred solution of methyl 3-((2R,5S)-2,5-dimethylpiperazin-1-yl)-3-(4-fluorophenyl)propanoate (100 mg, 0.34 mmol) in acetonitrile (5 mL) were added DIPEA (0.3 mL, 1.70 mmol) and 6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl trifluoromethanesulfonate (113 mg, 0.34 mmol) at room temperature. The reaction mixture was heated at 80 °C for 3 hours. The reaction mixture was cooled to room temperature and the solvent was removed under reduced pressure to give a mixture of diastereomers of the product, and the crude mixture was purified by preparative HPLC (chiral separation method: column: Lux cellulose C2 (250*21) mm, 5 micron, mobile phase: 0.1% DEA in MeOH, flow rate: 17 mL / min) to give Peak 1 and Peak 2.

[0248] Example 31: 10 mg, yield 6%; LCMS: m / z, 478.1 (M+H); rt 1.99 minutes; column: X Bridge BEH XP C18 (2.1x50 mm), 2.5 μm; mobile phase A: 10 mM ammonium acetate: acetonitrile (95:5), mobile phase B: 10 mM ammonium acetate: acetonitrile (5:95), gradient = 0-100% B over 3 minutes; temperature: 50 °C; flow rate: 1.1 mL / min; detection: UV (220 nm); 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.13 - 8.17 (m, 1H), 8.05 - 8.08 (m, 1H), 7.39 - 7.46 (m, 2H), 7.15 - 7.22 (m, 2H), 6.05 (s, 1H), 4.24 - 4.34 (m, 1H), 4.13 - 4.18 (m, 1H), 3.53 (s, 3H), 3.50 (s, 3H), 3.39 - 3.44 (m, 1H), 3.19 - 3.25 (m, 1H), 3.04 - 3.12 (m, 2H), 2.68 - 2.77 (m, 1H), 2.54 - 2.63 (m, 1H), 2.36 - 2.44 (m, 1H), 1.14 (d, J = 6.4 Hz, 3H), 1.01 (d, J = 6.4 Hz, 3H)

[0249] Example 32: 15 mg, yield 9%; LCMS: m / z, 478.2 (M+H); rt 2.0 minutes; column: XBridge BEH XP C18 (2.1x50 mm), 2.5 μm; mobile phase A: 10 mM ammonium acetate: acetonitrile (95:5), mobile phase B: 10 mM ammonium acetate: acetonitrile (5:95), gradient = 0 - 100% B over 3 minutes; temperature: 50 °C; flow rate: 1.1 mL / min; detection: UV (220 nm); 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.13 - 8.17 (m, 1H), 8.05 - 8.09 (m, 1H), 7.40 (dd, J = 8.4, 5.7 Hz, 2H), 7.15 (t, J = 8.8 Hz, 2H), 6.01 (s, 1H), 4.28 - 4.36 (m, 1H), 3.97 (dd, J = 9.4, 5.3 Hz, 1H), 3.62 - 3.69 (m, 1H), 3.48 - 3.55 (m, 5H, 3.45 (s, 3H), 3.40 - 3.44 (m, 1H), 3.04 - 3.13 (m, 1H), 2.70 - 2.76 (m, 1H), 2.13 - 2.20 (m, 1H), 1.12 (d, J = 6.4 Hz, 3H), 1.05 (d, J = 6.6 Hz, 3H)

[0250] Intermediate 11: 3 - ((2R,5S)-4-(tert-butoxycarbonyl)-2,5-dimethylpiperazin-1-yl)-3-(4-fluorophenyl)propanoic acid

Chemical formula

[0251] To a stirred solution of tert-butyl (2S,5R)-4-(1-(4-fluorophenyl)-3-methoxy-3-oxopropyl)-2,5-dimethylpiperazine-1-carboxylate (1.0 g, 2.53 mmol) in THF (20 mL) and water (10 mL) was added lithium hydroxide monohydrate (0.53 g, 12.67 mmol) at room temperature. The reaction mixture was stirred for 16 h. The reaction mixture was neutralized with aqueous 1.5 N HCl and extracted with EtOAc (2 x 100 mL). The combined organic extracts were washed with water, brine, dried over sodium sulfate, concentrated, and 3-((2R,5S)-4-(tert-butoxycarbonyl)-2,5-dimethylpiperazine-1-yl)-3-(4-fluorophenyl)propanoic acid (0.85 g, 88% yield) was obtained as a mixture of diastereomers. LCMS: m / z, 381.3 (M+H); rt 1.11 min (LCMS method: column: Acquity UPLC BEH C18 (3.0 x 50 mm) 1.7 μm; mobile phase A: 10 mM ammonium acetate:acetonitrile (95:5), mobile phase B: 10 mM ammonium acetate:acetonitrile (5:95), method: %B: 0 min - 20: 2 min - 100: 2.3 min - 100, temperature: 27 °C; flow rate: 0.7 mL / min; detection: UV (220 nm))

[0252] Intermediate 12: tert-butyl (2S,5R)-4-(3-(tert-butylamino)-1-(4-fluorophenyl)-3-oxopropyl)-2,5-dimethylpiperazine-1-carboxylate [Chemical formula]

[0253] To a stirred solution of 3-((2R,5S)-4-(tert-butoxycarbonyl)-2,5-dimethylpiperazin-1-yl)-3-(4-fluorophenyl)propanoic acid (350 mg, 0.92 mmol) in DMF (5 mL) were added DIPEA (0.32 mL, 1.84 mmol), BOP (450 mg, 1.01 mmol) and 2-methylpropan-2-amine (135 mg, 1.84 mmol) at room temperature. The reaction mixture was stirred for 3 h. The reaction mixture was extracted with EtOAc (2 x 50 mL) and washed with water. The combined organic extracts were washed with brine, dried over sodium sulfate and concentrated under reduced pressure. The crude residue was purified by flash column chromatography (column: 12 g silica; solvent run: 40 - 50% EtOAc in n-hexane) to give a mixture of diastereomers of tert-butyl (2S,5R)-4-(3-(tert-butylamino)-1-(4-fluorophenyl)-3-oxopropyl)-2,5-dimethylpiperazine-1-carboxylate (250 mg, 62% yield). LCMS: m / z, 436.2 (M+H); rt 3.61 min (LC-MS method information: column - Kinetex - XB-C18 (75 x 3 mm - 2.6 μm); mobile phase A: 10 mM ammonium formate in water: acetonitrile (98:2); mobile phase B: 10 mM ammonium formate in water: acetonitrile (2:98); gradient: 0 - 100% B over 24 min, flow rate 1.0 mL / min, then hold at 100% B for 0.6 min, flow rate 1.5 mL / min; then gradient: 100 - 20% B over 0.1 min, flow rate 1.5 mL / min)

[0254] Intermediate 13: N-(tert-butyl)-3-((2R,5S)-2,5-dimethylpiperazin-1-yl)-3-(4-fluorophenyl)propanamide

Chemical Structure

[0255] A stirred solution of tert-butyl (2S,5R)-4-(3-(tert-butylamino)-1-(4-fluorophenyl)-3-oxopropyl)-2,5-dimethylpiperazine-1-carboxylate (0.2 g, 0.46 mmol) in DCM (5 mL) was treated with 2,6-lutidine (0.32 mL, 2.75 mmol) and TMS-OTf (0.58 mL, 3.21 mmol) at 0 °C. The reaction mixture was allowed to reach room temperature and stirred for 2 h. The reaction mixture was concentrated under reduced pressure and co-distilled with acetonitrile (2 x 10 mL) to afford a mixture of diastereomers of N-(tert-butyl)-3-((2R,5S)-2,5-dimethylpiperazin-1-yl)-3-(4-fluorophenyl)propanamide (150 mg, 97%) as a brown solid. LCMS: m / z, 336.2 (M+H); rt 0.95 and 1.05 min (LCMS method: column: Waters Acquity UPLC BEH C18 (2.1 x 50 mm) 1.7 μm, mobile phase A: 10 mM NH4OAc: acetonitrile (95:5); mobile phase B: 10 mM NH4OAc: acetonitrile (5:95), gradient = 20-100% B over 2 min then hold at 100% B for 0.3 min; temperature: 50 °C; flow rate: 0.7 mL / min; detection: UV (220 nm))

[0256] Examples 33 and 34 N-(tert-butyl)-3-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-3-(4-fluorophenyl)propanamide

Chemical Structure

[0257] To a stirred solution of N-(tert-butyl)-3-((2R,5S)-2,5-dimethylpiperazin-1-yl)-3-(4-fluorophenyl)propanamide (0.10 g, 0.30 mmol) in acetonitrile (2 mL) were added sodium bicarbonate (0.13 g, 1.50 mmol) and 6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl trifluoromethanesulfonate (0.1 g, 0.30 mmol) at room temperature. The reaction mixture was heated at 80 °C for 12 h. The reaction mixture was cooled to room temperature and the solvent was removed under reduced pressure to afford a mixture of diastereomers of the product. The crude mixture was purified by preparative HPLC (chiral separation method: column: cellulose-4 (250x4.6) mm, 5 micron, mobile phase A: 0.1% DEA in ACN, mobile phase B: 0.1% DEA in MeOH, gradient: 0 - 100% B over 12 min) to give Peak-1 and Peak-2.

[0258] Example 33: LCMS: m / z, 519.3 (M+H); rt 1.92 min; column: X Bridge BEH XP C18 (2.1x50 mm), 2.5 μm; mobile phase A: 10 mM ammonium acetate: acetonitrile (95:5), mobile phase B: 10 mM ammonium acetate: acetonitrile (5:95), gradient = 0 - 100% B over 3 min; temperature: 50 °C; flow rate: 1.1 mL / min; detection: UV (220 nm); 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.15 (d, J = 8.8 Hz, 1H), 8.05 - 8.09 (m, 1H), 7.36 (dd, J = 8.6, 5.9 Hz, 2H), 7.22 (s, 1H), 7.12 (t, J = 8.8 Hz, 2H), 6.03 (s, 1H), 4.23 - 4.34 (m, 1H), 3.87 - 3.95 (m, 1H), 3.62 - 3.70 (m, 1H), 3.48 - 3.55 (m, 4H), 3.38 - 3.45 (m, 1H), 2.62 - 2.75 (m, 2H), 2.27 - 2.35 (m, 1H), 2.08 - 2.17 (m, 1H), 1.15 (d, J = 6.4 Hz, 3H), 1.02 - 1.08 (m, 12H)

[0259] Example 34: LCMS: m / z, 519.2 (M+H); rt 1.89 minutes; column: XBridge BEH XP C18 (2.1x50 mm), 2.5 μm; mobile phase A: 10 mM ammonium acetate: acetonitrile (95:5), mobile phase B: 10 mM ammonium acetate: acetonitrile (5:95), gradient = 0-100% B over 3 minutes; temperature: 50 °C; flow rate: 1.1 mL / min; detection: UV (220 nm) 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.14 (d, J = 8.8 Hz, 1H), 8.04 - 8.08 (m, 1H), 7.33 - 7.40 (m, 3H), 7.16 (t, J = 8.8 Hz, 2H), 6.03 (s, 1H), 4.31 - 4.53 (m, 1H), 4.04 - 4.15 (m, 1H), 3.52 (s, 3H), 3.38 - 3.43 (m, 2H), 2.98 - 3.11 (m, 1H), 2.68 - 2.81 (m, 2H), 2.42 - 2.47 (m, 1H), 2.13 - 2.27 (m, 1H), 1.17 (d, J = 6.4 Hz, 3H), 1.09 (s, 9H), 1.03 (d, J = 6.1 Hz, 3H)

[0260] The examples shown in Table 35 were prepared according to the general procedures of Examples 33 and 34 by replacing 2-methylpropan-2-amine with the appropriate amine in the synthetic route. When a mixture of diastereomers was obtained by the reaction, the mixture was separated using either preparative chromatography or preparative chiral chromatography at the final stage. The absolute stereochemistry was not assigned to the newly formed carbon-nitrogen bond.

[0261] Table 2

Table 8

Table 9

[0262] Intermediate 14 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)propanoic acid

Chem.

[0263] To a solution of 2-bromopropanoic acid (0.77 g, 5.04 mmol) in dry acetonitrile (3.0 mL) were added 8-((2S,5R)-2,5-dimethylpiperazin-1-yl)-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (1.0 g, 3.36 mmol) and DIPEA (0.59 mL, 3.36 mmol) at room temperature. The reaction mixture was stirred at room temperature for 10 minutes, then heated to 85 °C and stirred for 16 hours. The reaction mixture was cooled to room temperature and concentrated under reduced pressure to give 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)propanoic acid (1.20 g, yield 58%) as a brown gummy liquid. LCMS: m / z, 370.3 (M+H); rt 0.60 minutes; LCMS method: column: Waters Acquity UPLC BEH C18 (2.1x50 mm) 1.7 μm, mobile phase A: 10 mM ammonium acetate: acetonitrile (95:5); mobile phase B: 10 mM ammonium acetate: acetonitrile (5:95), gradient = 20 - 100% B over 2 minutes, then hold at 100% B for 0.2 minutes; temperature: 50 °C; flow rate: 0.7 mL / min; detection: UV (220 nm)

[0264] Examples 43 and 44 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-N-cyclopentylpropanamide

Chem.

[0265] To a solution of 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)propanoic acid (0.05 g, 0.14 mmol) in DMF (2 mL), HATU (0.15 g, 0.41 mmol) and DIPEA (0.07 mL, 0.41 mmol) were added at room temperature. The reaction mixture was stirred for 5 minutes, and then cyclopentanamine (0.023 g, 0.27 mmol) was added. The mixture was stirred at room temperature for 3 hours. The reaction was quenched with saturated NaHCO3 solution (5 mL). The reaction mixture was dissolved in DCM (100 mL). The organic layer was washed with water (20 mL) and brine (20 mL), dried over anhydrous sodium sulfate, filtered, and evaporated under reduced pressure. The crude product was purified by preparative HPLC followed by chiral HPLC (chiral separation method: column: cellulose-2 (250x4.6 mm), 5 μm; mobile phase: 10 mM ammonium acetate in MeOH, flow rate: 2.0 mL / min) to obtain Examples 43 and 44.

[0266] Example 43 (9.3 mg, yield 16%); LCMS: m / z 437.2 (M+H); rt 1.61 minutes (LCMS method: column: XBridge BEH XP C18 (50x2.1 mm) 2.5 μm; mobile phase A: 5:95 acetonitrile: water + 10 mM NH4OAc; mobile phase B: 95:5 acetonitrile: water + 10 mM NH4OAc; gradient: 0-100% B over 3 minutes; flow rate: 1.1 mL / min; temperature: 50°C; 11H NMR (400 MHz, DMSO-d6) δ ppm 8.17 - 8.15 (m, 1H), 8.08 - 8.06 (m, 1H), 7.42 (d, J = 7.8 Hz, 1H), 6.06 (s, 1H), 4.43 (brd, J = 1.7 Hz, 1H), 4.03 - 4.00 (m, 1H), 3.58 - 3.56 (m, 1H), 3.50 (s, 3H), 3.48 - 3.40 (m, 1H), 3.35 - 3.20 (m, 1H), 3.10 - 3.04 (m, 2H), 1.94 - 1.73 (m, 3H), 1.70 - 1.50 (m, 4H), 1.49 - 1.35 (m, 2H), 1.18 (t, J = 6.1 Hz, 6H), 1.05 (d, J = 6.6 Hz, 3H)

[0267] Example 44 (7.1 mg, yield 12%); LCMS: m / z, 437.2 (M + H); rt 1.6 minutes (LCMS method: column: XBridge BEH XP C18 (2.1 x 50 mm), 2.5 μm; mobile phase A: 10 mM ammonium acetate: acetonitrile (95:5), mobile phase B: 10 mM ammonium acetate: acetonitrile (5:95), gradient = 0 - 100% B over 3 minutes; temperature: 50 °C; flow rate: 1.1 mL / min; detection: UV (220 nm); 1 1H NMR (400 MHz, DMSO-d6) δ ppm 8.17 - 8.15 (m, 1H), 8.09 - 8.06 (m, 1H), 7.67 (d, J = 7.8 Hz, 1H), 6.06 (s, 1H), 4.43 (brd, J = 1.7 Hz, 1H), 4.03 - 4.00 (m, 1H), 3.58 - 3.56 (m, 1H), 3.50 (s, 3H), 3.48 - 3.40 (m, 1H), 3.35 - 3.20 (m, 1H), 3.10 - 3.04 (m, 2H), 1.94 - 1.73 (m, 3H), 1.70 - 1.50 (m, 4H), 1.49 - 1.35 (m, 2H), 1.18 (t, J = 6.1 Hz, 6H), 1.05 (d, J = 6.6 Hz, 3H)

[0268] The compounds shown in Table 2 were prepared from the acid intermediates using the corresponding amines as described in Examples 42 and 43. When a mixture of diastereomers was obtained by the reaction, the mixture was separated using either preparative chromatography or preparative chiral chromatography at the final stage. The absolute stereochemistry was not assigned to the newly formed carbon-nitrogen bond.

[0269] Table 3

Table 10

[0270] Intermediate 17 tert-butyl (2S,5R)-4-((dimethylphosphoryl)(4-fluorophenyl)methyl)-2,5-dimethylpiperazine-1-carboxylate

Chem.

[0271] To a solution of tert-butyl (2S,5R)-2,5-dimethylpiperazine-1-carboxylate (700 mg, 3.27 mmol) in dry toluene (15 mL) was added 4-fluorobenzaldehyde (486 mg, 3.92 mmol) at room temperature. The reaction mixture was stirred at the same temperature for 1 h, and then dimethylphosphine oxide (306 mg, 3.92 mmol) was added. A Dean Stark apparatus was attached to the reaction mixture, and the mixture was heated under reflux for 16 h. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was triturated with diethyl ether (2 x 20 mL) to obtain the crude compound. The crude compound was purified by silica gel chromatography (12 g) using 0 - 10% MeOH / CHCl3 as the eluent to give tert-butyl (2S,5R)-4-((dimethylphosphoryl)(4-fluorophenyl)methyl)-2,5-dimethylpiperazine-1-carboxylate (1.1 g, 85% yield). LCMS: m / z, 399.5 (M+H); rt 1.52 and 1.57 min; LCMS method: column: Waters Acquity UPLC BEH C18 (2.1 x 50 mm) 1.7 μm, mobile phase A: 10 mM ammonium acetate: acetonitrile (95:5); mobile phase B: 10 mM ammonium acetate: acetonitrile (5:95), gradient = 20 - 100% B over 2 min, then hold at 100% B for 0.3 min; flow rate: 0.7 mL / min; detection: UV (220 nm)

[0272] Intermediate 18 (((2R,5S)-2,5-Dimethylpiperazin-1-yl)(4-fluorophenyl)methyl)dimethylphosphine oxide·TFA [Chemical formula]

[0273] To a stirred solution of tert-butyl (2S,5R)-4-((dimethylphosphoryl)(4-fluorophenyl)methyl)-2,5-dimethylpiperazine-1-carboxylate (200 mg, 0.5 mmol) in DCM (3 mL) was added TFA (0.387 mL, 5.02 mmol). The reaction mixture was stirred at room temperature for 2 h. The solvent was removed under reduced pressure to afford the TFA salt of (((2R,5S)-2,5-dimethylpiperazin-1-yl)(4-fluorophenyl)methyl)dimethylphosphine oxide as a brown semi-solid. LCMS: m / z, 299.0 (M+H); rt 0.51 and 0.53 min; LCMS method: column: Waters Acquity UPLC BEH C18 (2.1x50 mm) 1.7 μm, mobile phase A: 10 mM ammonium acetate:acetonitrile (95:5); mobile phase B: 10 mM ammonium acetate:acetonitrile (5:95), gradient = 20 - 100% B over 2 min then hold at 100% B for 0.3 min; flow rate: 0.7 mL / min; detection: UV (220 nm)

[0274] Examples 49 and 50 8-((2S,5R)-4-((dimethylphosphoryl)(4-fluorophenyl)methyl)-2,5-dimethylpiperazin-1-yl)-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile

Chemical Structure

[0275] A solution of ((2R,5S)-2,5-dimethylpiperazin-1-yl)(4-fluorophenyl)methyl)dimethylphosphine oxide (59.1 mg, 0.2 mmol) in dry acetonitrile (3 mL) was added with 6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl trifluoromethanesulfonate (60 mg, 0.18 mmol) and sodium hydrogen carbonate (45.4 mg, 0.54 mmol) under an argon atmosphere at room temperature, and the mixture was stirred at the same temperature for 10 minutes. The reaction mixture was heated to 85 °C and maintained at the same temperature for 16 hours. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with 10% MeOH in DCM to give 8-((2S,5R)-4-((dimethylphosphoryl)(4-fluorophenyl)methyl)-2,5-dimethylpiperazin-1-yl)-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile. The crude material was further purified via preparative HPLC followed by chiral HPLC (Chiral HPLC method: column: cellulose-4 (250 mm x 21.2 mm) 5 μm, mobile phase A: 0.1% DEA in acetonitrile, mobile phase B: 0.1% DEA in MeOH; gradient = 0 - 100% B over 12 minutes then 100 - 0% B over 2 minutes and held at 0% B for 6 minutes) to give Examples 51 and 52.

[0276] Example 49 (7 mg, 8% yield); LCMS: m / z, 482.1 (M + H); rt 1.48 minutes (LCMS method: column: XBridge BEH XP C18 (50 x 2.1 mm) 2.5 μm; mobile phase A: 95% water: 5% acetonitrile; 10 mM ammonium acetate; mobile phase B: 5% water: 95% acetonitrile; 10 mM ammonium acetate; flow: 1.1 mL / minute; temperature: 50 °C; time (minutes): 0 - 3; %B: 0 - 100%); 11H NMR (400 MHz, DMSO-d6) δ ppm 8.18 - 8.11 (m, 1H), 8.10 - 8.03 (m, 1H), 7.61 (dd, J = 6.0, 7.9 Hz, 2H), 7.28 (t, J = 8.8 Hz, 2H), 6.18 (s, 1H), 4.14 (d, J = 12.2 Hz, 1H), 4.08 - 3.95 (m, 1H), 3.69 - 3.57 (m, 2H), 3.54 (s, 3H), 3.15 - 3.02 (m, 1H), 2.78 - 2.69 (m, 1H), 2.55 (s, 1H), 1.46 (d, J = 12.7 Hz, 3H), 1.23 (d, J = 12.7 Hz, 3H), 1.07 (dd, J = 6.1, 14.2 Hz, 6H)

[0277] Example 50 (15 mg, yield 16%); LCMS: m / z, 482.1 (M + H); rt 1.47 minutes (LCMS method: column: XBridge BEH XP C18 (50x2.1 mm) 2.5 μm; mobile phase A: 95% water: 5% acetonitrile; 10 mM ammonium acetate; mobile phase B: 5% water: 95% acetonitrile; 10 mM ammonium acetate; flow rate: 1.1 mL / min; temperature: 50 °C; time (min): 0 - 3; %B: 0 - 100%); 1 1H NMR (400 MHz, DMSO-d6) δ ppm 8.17 - 8.11 (m, 1H), 8.08 - 8.02 (m, 1H), 7.63 - 7.50 (m, 2H), 7.23 (t, J = 8.8 Hz, 2H), 6.01 (s, 1H), 4.67 - 4.50 (m, 1H), 4.13 (d, J = 10.8 Hz, 1H), 3.67 - 3.47 (m, 4H), 3.26 (brs, 1H), 3.09 - 3.02 (m, 1H), 3.00 - 2.91 (m, 1H), 2.58 - 2.54 (m, 1H), 1.52 (d, J = 12.5 Hz, 3H), 1.26 (d, J = 6.4 Hz, 3H), 1.13 - 0.96 (m, 6H)

[0278] Intermediate 36 Ethyl 2-bromo-2-(4-(trifluoromethyl)phenyl)acetate

Chemical formula

[0279] To a stirred solution of ethyl 2-(4-(trifluoromethyl)phenyl)acetate (2 g, 8.61 mmol) in CCl4 (50 mL) was added NBS (1.84 g, 10.34 mmol) and HBr (0.1 mL, 0.86 mmol). The reaction mixture was heated at reflux temperature for 16 h. The reaction mixture was cooled to room temperature, the resulting solid was filtered off, the filtrate was washed with water, the organic layer was dried over anhydrous Na2SO4, filtered, evaporated under reduced pressure to give ethyl 2-bromo-2-(4-(trifluoromethyl)phenyl)acetate (2.3 g, 7.32 mmol, 85% yield) as a liquid. LCMS: m / z = 309 (M-H); rt 3.19 min (LCMS method: column: Kinetex XB-C18 (75x3mm-2.6μm) mobile phase A: 10 mM ammonium formate: acetonitrile (98:2); mobile phase B: 10 mM ammonium acetate: acetonitrile (2:98), gradient = 20% B over, then hold at 100% B for 0.6 min; flow rate: 1.0 mL / min; detection: UV (220 nm))

[0280] Example 51 Ethyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetate

Chemical formula

[0281] To a stirred solution of 4-((2S,5R)-2,5-diethylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile, HCl (0.5 g, 1.38 mmol) in acetonitrile (15 mL), DIPEA (0.722 mL, 4.13 mmol) was added, followed by ethyl 2-bromo-2-(4-(trifluoromethyl)phenyl)acetate (0.857 g, 2.76 mmol). The reaction mixture was heated at 85 °C for 16 h. The reaction mixture was concentrated under reduced pressure to remove volatile materials and a crude compound was obtained. The crude residue was purified by silica gel chromatography on ISCO® (50 - 70% EtOAc / petroleum ether; 40 g column) to give ethyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetate (0.5 g, 0.862 mmol, 62.6% yield) as an off-white solid. LCMS: m / z = 557.2 (M+H); rt 3.411 min (LCMS method: column: Kinetex XB-C18 (75x3mm - 2.6μm) mobile phase A: 10 mM ammonium formate:acetonitrile (98:2); mobile phase B: 10 mM ammonium acetate:acetonitrile (2:98), gradient = 20% B over 2 min then hold at 100% B for 0.6 min; flow rate: 1.0 mL / min; detection: UV (220 nm))

[0282] Example 52 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetic acid

Chemical Structure

[0283] A stirred solution of ethyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetate (500 mg, 0.898 mmol) in 1,4-dioxane (10 mL) was treated with trimethylstannanol (1624 mg, 8.98 mmol). The reaction mixture was heated at 100 °C for 16 h. The reaction mixture was filtered through a pad of celite, washed with ethyl acetate, and the filtrate was removed under reduced pressure to afford the crude product, which was purified by preparative HPLC (HPLC method: column: ACE C18 PFP (250 mm x 21.2 mm ID, 5 μm), mobile phase A: A = 10 mM ammonium acetate in water (pH 4.5); mobile phase B: acetonitrile:MeOH (1:1) acetonitrile; gradient: 80% B over 2 min then hold at 100% B for 16 min; flow: 19 mL / min). The fractions were concentrated under reduced pressure, the residue was diluted (EtOH / H2O, 1:5), lyophilized to afford 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetic acid (150 mg, yield 31.6%). LCMS: m / z = 529.2 (M+H); rt 1.602 min (LCMS method: column: Kinetex XB-C18 (75 x 3 mm - 2.6 μm) mobile phase A: 10 mM ammonium formate:acetonitrile (98:2); mobile phase B: 10 mM ammonium acetate:acetonitrile (2:98), gradient = 20% B over 2 min then hold at 100% B for 0.6 min; flow rate: 1.0 mL / min; detection: UV (220 nm))

[0284] Examples 53 and 54 4-((2S,5R)-2,5-Diethyl-4-(2-morpholino-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile [Chemical Structure]

[0285] To a stirred solution of 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetic acid (50 mg, 0.095 mmol) in DMF (2 mL) was added TEA (0.04 mL, 0.28 mmol) and HATU (71.9 mg, 0.189 mmol). The reaction mixture was stirred at room temperature for 5 minutes and then morpholine (8.24 mg, 0.095 mmol) was added. The reaction mixture was stirred at the same temperature for 16 hours. The reaction mixture was concentrated under reduced pressure to remove volatile materials and a crude compound was obtained. The crude compound was purified by preparative HPLC (HPLC method: column: ACE C18 PFP (250 mm x 21.2 mm ID, 5 μm), mobile phase A: A = 10 mM ammonium acetate in water (pH 4.5); mobile phase B: acetonitrile:MeOH (1:1) acetonitrile; gradient: 80% B over 2 minutes then hold at 100% B for 16 minutes; flow: 19 mL / min) to give Examples 53 and 54.

[0286] Example 53 was dried via centrifugal evaporation (4.7 mg, 8.2% yield); LCMS: m / z = 598.3 (M+H); rt 1.847 minutes ((LCMS method: column: XBridge BEH XP C18 (50 x 2.1) mm, 2.5 μm; mobile phase A: 95% water: 5% acetonitrile; 10 mM NH4OAc; mobile phase B: 5% water: 95% acetonitrile; 10 mM NH4OAc; flow: 1.1 mL / min; temperature: 50°C; time (min)); 11H NMR (400 MHz, DMSO-d6) δ 8.30 - 8.13 (m, 1H), 8.06 - 7.91 (m, 1H), 7.86 - 7.66 (m, 4H), 6.07 - 5.58 (m, 1H), 5.58 - 5.17 (m, 1H), 5.07 - 4.64 (m, 1H), 4.02 - 3.86 (m, 1H), 3.70 - 3.55 (m, 3H), 3.54 - 3.45 (m, 4H), 3.43 (s, 3H), 3.09 - 2.89 (m, 1H), 2.65 - 2.55 (m, 1H), 2.10 - 1.67 (m, 2H), 1.56 - 1.34 (m, 2H), 1.00 - 0.80 (m, 3H), 0.68 (t, J = 7.3 Hz, 3H) (2 protons were integrated with the DMSO peak).

[0287] Example 54 was dried via centrifugal evaporation (7.4 mg, yield 13%); LCMS: m / z = 598.3 (M + H); rt 1.885 minutes ((LCMS method: column: XBridge BEH XP C18 (50 x 2.1) mm, 2.5 μm; mobile phase A: 95% water: 5% acetonitrile; 10 mM NH4OAc; mobile phase B: 5% water: 95% acetonitrile; 10 mM NH4OAc; flow rate: 1.1 mL / min; temperature: 50 °C; time (min)); 1 1H NMR (400 MHz, DMSO-d6) δ 8.23 (d, J = 8.8 Hz, 1H), 7.98 (d, J = 8.6 Hz, 1H), 7.76 (s, 4H), 5.89 - 5.31 (m, 1H), 5.01 (s, 1H), 4.95 - 4.81 (m, 1H), 3.81 - 3.71 (m, 1H), 3.57 - 3.35 (m, 9H), 3.30 - 3.14 (m, 2H), 3.01 - 2.87 (m, 1H), 2.73 - 2.61 (m, 1H), 2.45 - 2.35 (m, 1H), 2.21 - 1.89 (m, 1H), 1.85 - 1.71 (m, 1H), 1.55 - 1.33 (m, 2H), 0.89 - 0.77 (m, 3H), 0.76 - 0.55 (m, 3H)

[0288] The examples shown in Table 6 were prepared in the synthetic route using appropriate amines according to the general procedures described in the preparation of Examples 54 and 55. When a mixture of diastereomers was obtained by the reaction, the mixture was separated using either preparative chromatography or preparative chiral chromatography at the final step. The absolute stereochemistry was not assigned to the newly formed carbon-nitrogen bond.

[0289] Table 6 [Table 11]

[0290] Intermediate 45 3-(3-(But-3-yn-1-yl)-3H-diazirin-3-yl)propan-1-ol [Chemical formula]

[0291] 3-(3-(But-3-yn-1-yl)-3H-diazirin-3-yl)propanoic acid (0.53 g, 3.17 mmol) (synthesized from 4-oxoocta-7-ynoic acid according to the procedure in the literature: Parker, C. G. et al., Cell, 168(3), 527-541, 2017) in THF (10 mL) at -10 °C was treated with N-methylmorpholine (0.53 mL, 4.75 mmol), followed by isobutyl chloroformate (0.55 mL, 4.11 mmol). The reaction mixture was stirred at -5 °C for about 10 minutes, and then NaBH4 (0.305 g, 7.91 mmol) was added. The resulting mixture was stirred at room temperature for 16 hours. The reaction was quenched with water. The reaction mixture was extracted with EtOAc (2 x 25 mL). The combined organic layers were washed with brine, dried over anhydrous MgSO4, filtered, and the filtrate was evaporated to give a pale yellow oil, which was purified by silica gel column chromatography (24 g RediSep® column, eluting with a gradient of 0 - 50% hexane in EtOAc). The fractions containing the product were combined and evaporated to give 3-(3-(but-3-yn-1-yl)-3H-diazirin-3-yl)propan-1-ol as a pale yellow oil (0.37 g, 77% yield). 1 H NMR (500 MHz, chloroform-d) δ 3.65 - 3.57 (m, 2H), 2.07 - 1.97 (m, 3H), 1.69 - 1.62 (m, 2H), 1.58 - 1.47 (m, 3H), 1.44 - 1.34 (m, 2H)

[0292] Example 57 3-(3-(But-3-yn-1-yl)-3H-diazirin-3-yl)propyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate

Chemical Structure

[0293] To a solution of 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetic acid (0.045 g, 0.1 mmol) in DCM (2 mL) were added DCC (0.041 g, 0.2 mmol) and DMAP (3 mg, 0.02 mmol). The reaction mixture was stirred at room temperature for 10 minutes, and then a solution of 3-(3-(but-3-yn-1-yl)-3H-diazirin-3-yl)propan-1-ol (0.030 g, 0.2 mmol) in DCM (ca. 0.5 mL) was added. Stirring of the resulting mixture was continued at room temperature for 1 hour, then the volatile materials were evaporated under reduced pressure to give a residue, which was dissolved in DMF and purified via preparative HPLC using the following conditions (column: XBridge C18, 200 mm x 19 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile: water + 10 mM ammonium acetate; gradient: held at 43% B for 0 minutes, brought to 43 - 83% B over 20 minutes, then held at 100% B for 6 minutes; flow rate: 20 mL / min; column temperature: 25 °C). The fractions containing the product were combined and dried under vacuum to give 3-(3-(but-3-yn-1-yl)-3H-diazirin-3-yl)propyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate. MS (ES), m / z = 584.4 [M+H]+; HPLC retention times 2.37 minutes and 2.09 minutes (for HPLC methods B and C respectively); 11H NMR (500 MHz, DMSO-d6) δ ppm 8.18 - 8.11 (m, 1H), 8.10 - 8.02 (m, 1H), 7.63 - 7.52 (m, 2H), 7.29 - 7.17 (m, 2H), 6.04 (s, 1H), 4.46 - 4.34 (m, 2H), 4.11 - 3.95 (m, 2H), 3.75 - 3.63 (m, 1H), 3.61 - 3.49 (m, 2H), 2.83 - 2.72 (m, 2H), 2.54 (s, 3H), 2.14 - 2.05 (m, 1H), 2.00 - 1.92 (m, 2H), 1.57 - 1.47 (m, 2H), 1.42 - 1.31 (m, 4H), 1.23 - 1.10 (m, 6H)

[0294] Intermediate 46 Methyl 2-(4-methoxyphenyl)acetate [Chemical Structure]

[0295] A solution of methyl 2-(4-hydroxyphenyl)acetate (2.5 g, 14.74 mmol) in THF (5 mL) was added dropwise to a suspension of NaH (0.590 g, 14.74 mmol) (60% in mineral oil) in THF (15 mL) at 0 °C. The reaction mixture was stirred at 0 °C for 90 minutes, and then a solution of MeI (2.95 mL, 47.2 mmol) in THF (5 mL) was added. The resulting mixture was stirred at room temperature for 16 hours. The reaction was quenched with saturated aqueous NH4Cl and brine. The solution was diluted with EtOAc and separated into two layers. The aqueous layer was back-extracted with EtOAc (2 x 25 mL). The organic layers were combined, washed with brine, dried over anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure to give an oil. The crude oil was purified by silica gel column chromatography (80 g RediSep® column, eluting with 35% hexane in EtOAc). The fractions containing the product were evaporated to give methyl 2-(4-methoxyphenyl)acetate as a colorless oil (2.39 g, 90% yield). MS (ES), m / z = 181.0 [M+H]+; HPLC retention time 0.84 minutes (HPLC method C)

[0296] Example 58 Methyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-methoxyphenyl)acetate

Chemical formula

[0297] Methyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-methoxyphenyl)acetate was obtained as a mixture of diastereomers using the general experimental conditions described for the synthesis of methyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate by reacting methyl 2-bromo-2-(4-methoxyphenyl)acetate (Intermediate 48, synthesized according to the procedures in the literature: Okano, K. et al., Angewandte Chemie, International Ed., 49(34), 5925-5929, 2010) with 4-((2S,5R)-2,5-dimethylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (Intermediate 49). MS(ES), m / z = 477.4[M+H]+; HPLC retention time 0.73 minutes (diastereomer 1) and 0.76 minutes (diastereomer 2) (HPLC method C)

[0298] Example 59 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-methoxyphenyl)acetic acid

Chemical formula

[0299] 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-methoxyphenyl)acetic acid was obtained as a mixture of diastereomers using experimental conditions similar to those for 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetic acid by reacting methyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-methoxyphenyl)acetate with trimethylstannanol. MS(ES), m / z = 463.2 [M+H]+; HPLC retention time 0.62 minutes (diastereomers 1 and 2) (HPLC method C)

[0300] Examples 60 and 61 3-(3-(But-3-yn-1-yl)-3H-diazirin-3-yl)propyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-methoxyphenyl)acetate

Chemical formula

[0301] 3-(3-(But-3-yn-1-yl)-3H-diazirin-3-yl)propyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-methoxyphenyl)acetate was synthesized according to the general procedure described in Example 57 by coupling a mixture of diastereomers of 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-methoxyphenyl)acetic acid with 3-(3-(but-3-yn-1-yl)-3H-diazirin-3-yl)propan-1-ol. MS(ES), m / z = 597.4 [M+H]+; HPLC retention time 0.87 min (diastereomer 1) and 0.91 min (diastereomer 2) (HPLC method C). The crude material was purified via preparative LC / MS using the following conditions (column: XBridge C18, 200 mm x 19 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + 0.05% trifluoroacetic acid; mobile phase B: 95:5 acetonitrile: water + 0.05% trifluoroacetic acid; gradient: hold at 20% B for 0 min, make 20 - 70% B over 25 min, then hold at 100% B for 0 min; flow rate: 20 mL / min; column temperature: 25 °C). The fractions containing the product were combined and evaporated to give Examples 60 and 61.

[0302] Example 60: (6.4 mg, yield 8.1%); MS(ES), m / z = 597.2 [M+H]+; HPLC retention time 2.21 min and 1.91 min (each, HPLC methods D and E); 11H NMR (500 MHz, DMSO-d6) δ 8.32 - 8.17 (m, 1H), 8.05 - 7.96 (m, 1H), 7.49 - 7.35 (m, 2H), 7.03 - 6.91 (m, 2H), 5.73 - 5.46 (m, 1H), 5.02 - 4.77 (m, 1H), 4.28 - 4.18 (m, 1H), 4.12 - 3.92 (m, 2H), 3.84 - 3.74 (m, 3H), 2.88 - 2.78 (m, 1H), 2.65 - 2.46 (m, 6H), 2.32 - 2.14 (m, 1H), 2.02 - 1.91 (m, 2H), 1.58 - 1.48 (m, 2H), 1.44 - 1.20 (m, 7H), 1.16 - 0.98 (m, 3H)

[0303] Example 61: (5.5 mg, yield 6.9%); MS (ES), m / z = 597.2 [M + H]+; HPLC retention times 2.29 minutes and 1.85 minutes (each, HPLC methods D and E); 1 1H NMR (500 MHz, DMSO-d6) δ ppm 8.29 - 8.15 (m, 1H), 8.04 - 7.91 (m, 1H), 7.54 - 7.37 (m, 2H), 7.04 - 6.91 (m, 2H), 5.75 - 5.37 (m, 1H), 5.14 - 4.58 (m, 1H), 4.23 - 4.13 (m, 1H), 4.09 - 3.90 (m, 2H), 3.84 - 3.72 (m, 3H), 3.84 - 3.72 (m, 2H), 3.53 - 3.40 (m, 1H), 3.00 - 2.86 (m, 2H), 2.85 - 2.77 (m, 1H), 2.60 - 2.54 (m, 2H), 2.50 - 2.41 (m, 1H), 2.02 - 1.89 (m, 2H), 1.64 - 1.38 (m, 5H), 1.37 - 1.27 (m, 3H), 0.96 - 0.78 (m, 3H)

[0304] Intermediate 52 2-(3-Methyl-3H-diazirin-3-yl)ethan-1-ol

Chemical Structure

[0305] 2-(3-Methyl-3H-diazirin-3-yl)ethan-1-ol was synthesized according to Wang et al., Angewandte Chemie, International Edition, 56(3), 870-873, 2017.

[0306] Example 62 2-(3-Methyl-3H-diazirin-3-yl)ethyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate [Chemical Structure]

[0307] 2-(3-Methyl-3H-diazirin-3-yl)ethyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate was synthesized according to the general procedure described in the preparation of 3-(3-(but-3-yn-1-yl)-3H-diazirin-3-yl)propyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate) by coupling 2-(3-methyl-3H-diazirin-3-yl)ethan-1-ol with 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetic acid. MS(ES), m / z = 532.4[M+H]+; HPLC retention times 2.28 minutes and 1.96 minutes (HPLC methods B and C, respectively); 11H NMR (500 MHz, DMSO-d6) δ 8.19 - 8.11 (m, 1H), 8.08 - 8.02 (m, 1H), 7.67 - 7.55 (m, 2H), 7.29 - 7.16 (m, 2H), 6.08 - 6.01 (m, 1H), 4.46 - 4.35 (m, 2H), 4.09 - 3.92 (m, 2H), 3.73 - 3.36 (m, 6H), 2.81 - 2.71 (m, 1H), 2.58 - 2.53 (m, 3H), 2.14 - 2.05 (m, 1H), 1.68 - 1.56 (m, 2H), 1.26 - 1.19 (m, 3H), 1.17 - 1.10 (m, 3H)

[0308] Intermediate 53 2,5-Dioxopyrrolidin-1-yl 3’,6’-dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9’-xanthene]-5-carboxylate [Chemical Structure]

[0309] To a solution of 3’,6’-dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9’-xanthene]-5-carboxylic acid (0.05 g, 0.133 mmol) in DMF (1 mL) was added EDC (0.032 g, 0.166 mmol), followed by 1-hydroxypyrrolidine-2,5-dione (0.02 g, 0.173 mmol). The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was diluted with EtOAc, and the organic layer was washed with phosphate buffer (pH = 7.4). The organic layer was separated, and the aqueous washings were subjected to back extraction with EtOAc (2 x 5 mL). The combined organic layers were washed with brine, dried over anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure to give crude 2,5-dioxopyrrolidin-1-yl 3’,6’-dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9’-xanthene]-5-carboxylate as an orange oil, which was then solidified (0.056 g, 89% yield). MS (ES), m / z = 474.0 [M+H]+; HPLC retention time 1.48 minutes (HPLC method C)

[0310] Example 63 tert-Butyl (2-(2-(2-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetamido)ethoxy)ethoxy)ethyl)carbamate

Chem.

[0311] A solution of 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetic acid (0.065 g, 0.145 mmol), tert-butyl (2-(2-(2-aminoethoxy)ethoxy)ethyl)carbamate (0.044 g, 0.174 mmol), HATU (0.066 g, 0.174 mmol) and DIPEA (0.05 mL, 0.289 mmol) in DMF (1.5 mL) was stirred at room temperature for 45 minutes. The reaction mixture was diluted with water and extracted with a 5% MeOH solution in DCM (3 x 5 mL). The organic layers were combined, washed with water and brine, dried over anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure to give a yellow oil. The crude product was purified by silica gel chromatography (12 g RediSep® column, eluting with a gradient of 0 - 5% MeOH in DCM). The fractions containing the product were combined and evaporated to give tert-butyl (2-(2-(2-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetamido)ethoxy)ethoxy)ethyl)carbamate as a yellow oil (0.067 g, 68% yield). MS(ES), m / z = 680.3[M+H]+; HPLC retention time 0.81 minutes (HPLC method C)

[0312] Example 64 N-(2-(2-(2-Aminoethoxy)ethoxy)ethyl)-2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetamide·TFA

Chemical formula

[0313] To a solution of tert-butyl (2-(2-(2-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetamide)ethoxy)ethoxy)ethyl)carbamate (0.067 g, 0.099 mmol) in DCM (1 mL) was added TFA (0.09 mL, 1.183 mmol). The reaction mixture was stirred at room temperature for 1 hour. The volatile materials were evaporated under reduced pressure to obtain N-(2-(2-(2-aminoethoxy)ethoxy)ethyl)-2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetamide as the TFA salt (0.067 g, yield >99%). MS(ES), m / z = 580.3 [M+H]+; HPLC retention time 0.63 minutes (HPLC method C)

[0314] Example 65 N-(2-(2-(2-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetamide)ethoxy)ethoxy)ethyl)-3’,6’-dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9’-xanthene]-5-carboxamide

Chemical formula

[0315] To a solution of N-(2-(2-(2-aminoethoxy)ethoxy)ethyl)-2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetamide·TFA (0.068 g, 0.098 mmol) in DMF (1 mL), DIPEA (0.09 mL, 0.490 mmol) was added, followed by 2,5-dioxopyrrolidin-1-yl 3’,6’-dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9’-xanthene]-5-carboxylate (0.056 g, 0.118 mmol). The reaction mixture was stirred at room temperature for 1 hour. The mixture was diluted with DMF and purified by preparative HPLC under the following conditions (column: X Bridge C18, 200 mm x 19 mm, 5 μm particles; mobile phase A: 5:95 acetonitrile: water + ammonium acetate; mobile phase B: 95:5 acetonitrile: water + ammonium acetate; gradient: held at 12% B for 0 minutes, changed to 12 - 52% B over 30 minutes, then held at 100% B for 0 minutes; flow rate: 20 mL / min; column temperature: 25 °C). The fractions containing the product were combined and dried under vacuum to give N-(2-(2-(2-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetamide)ethoxy)ethoxy)ethyl)-3’,6’-dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9’-xanthene]-5-carboxamide). MS(ES), m / z = 938.1 [M+H]+; HPLC retention times 1.42 minutes and 1.41 minutes (for HPLC methods B and C respectively)

[0316] Example 66 2,2-Dimethyl-4-oxo-3,8,11-trioxa-5-azatridecan-13-yl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate [Chemical formula]

[0317] To a solution of 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetic acid (0.07 g, 0.156 mmol) in DCM (1 mL) were added DCC (0.039 g, 0.187 mmol) and DMAP (1.9 mg, 0.016 mmol). The reaction mixture was stirred at room temperature for about 15 minutes, and then tert-butyl (2-(2-(2-hydroxyethoxy)ethoxy)ethyl)carbamate (0.049 g, 0.187 mmol) was added. The mixture was stirred at room temperature for 1 hour and then diluted with water and DCM. The two layers were separated, and the aqueous layer was back-extracted with DCM (2 x 5 mL). The organic layers were combined, washed with brine, dried over anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure to give 2,2-dimethyl-4-oxo-3,8,11-trioxa-5-azatridecan-13-yl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate as a pale yellow solid (0.1 g, yield 94%). MS(ES), m / z = 681.3 [M+H]+; HPLC retention time 0.88 minutes (HPLC method C)

[0318] Example 67 2-(2-(2-Aminoethoxy)ethoxy)ethyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate·TFA [Chemical formula]

[0319] To a solution of 2,2-dimethyl-4-oxo-3,8,11-trioxa-5-azatridecan-13-yl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate (0.1 g, 0.147 mmol) in DCM (1.5 mL) was added TFA (0.12 mL, 1.47 mmol). The reaction mixture was stirred at room temperature for 1 h. Volatiles were evaporated under reduced pressure to afford crude 2-(2-(2-aminoethoxy)ethoxy)ethyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate as the TFA salt (0.099 g, yield >99%). MS(ES), m / z = 581.3 [M+H]+; HPLC retention time 0.69 min (HPLC method C)

[0320] Example 68 2-(2-(2-(3’,6’-Dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9’-xanthene]-5-carboxamido)ethoxy)ethoxy)ethyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate

Chemical Structure

[0321] 2-(2-(2-(3’,6’-Dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9’-xanthene]-5-carboxamide)ethoxy)ethoxy)ethyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate was synthesized according to the general procedure described in the preparation of N-(2-(2-(2-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetamido)ethoxy)ethoxy)ethyl)-3’,6’-dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9’-xanthene]-5-carboxamide by reacting 2-(2-(2-aminoethoxy)ethoxy)ethyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate·TFA with 2,5-dioxopyrrolidin-1-yl 3’,6’-dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9’-xanthene]-5-carboxylate. MS(ES), m / z = 939.2 [M+H]+; HPLC retention times 1.722 minutes and 1.675 minutes (HPLC methods B and C respectively)

[0322] Additional LCMS conditions: Method FA: Column: Kinetex XB-C18 (75x3.0) mm, 2.6 μm, Mobile phase A: 5 mM ammonium formate (pH 3.3): ACN (98:2), Mobile phase B: ACN: Buffer (98:2), Flow rate: 1.0 mL / min, Detection: UV (220 nm) Method FB: Column: Kinetex biphenyl (100x4.6) mm, 2.6 μm, Mobile phase A: 0.05% TFA in water: ACN (95:5), Mobile phase B: ACN: 0.05% TFA in water (95:5), Flow: 1.0 mL / min, Detection: UV (220 nm) Method FC: Column: Kinetex EVO C18 (100x4.6) mm, 2.6 μm, Mobile Phase A: 0.05% TFA in water: ACN (95:5), Mobile Phase B: ACN: 0.05% TFA in water (95:5), Flow rate: 1.0 mL / min, Detection: UV (220 nm) Method FD: Column: X-Bridge C8 (50x4.6 mm) 5 μm, Mobile Phase A: 0.1% TFA in H2O, Mobile Phase B: 0.1% TFA in ACN, Flow rate: 1.5 mL / min, Detection: UV (220 nm)

[0323] Method FE: Column: X-Bridge C8 (50x4.6) mm, 3.5 μm, Mobile Phase: A: 10 mM NH4HCO3 in H2O, Mobile Phase: B: ACN, Flow rate: 1.5 mL / min, Detection: UV (220 nm) Method FF: Column: X-Bridge C8 (50x4.6 mm) 5 μm, Mobile Phase A: 10 mM ammonium acetate in H2O, Mobile Phase: B: ACN, Flow rate: 1.5 mL / min, Detection: UV (220 nm) Method FG: Column: X-Bridge C8 (50x4.6) mm, 3.5 μm, Mobile Phase B: 10 mM ammonium bicarbonate in water, Mobile Phase A: acetonitrile, Flow rate: 1.0 mL / min Method FH: Column: Kinetex biphenyl (100x4.6) mm, 2.6 μm, Mobile Phase A: 10 mM ammonium acetate in water, Mobile Phase B: acetonitrile, Flow rate: 1.0 mL / min Method FI: Column: Kinetex EVO C18 (100x4.6) mm, 2.6 μm, Mobile Phase: A: 10 mM ammonium acetate, Mobile Phase B: ACN, Flow rate: 1.0 mL / min

[0324] Examples 69 and 70 Ethyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetate

Chemical formula

[0325] To a stirred solution of 4-((2S,5R)-2,5-diethylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile, HCl (0.3 g, 0.83 mmol) in acetonitrile (10 mL) was added DIPEA (0.4 mL, 2.48 mmol), followed by ethyl 2-bromo-2-(4-(trifluoromethyl)phenyl)acetate (0.514 g, 1.65 mmol) and sodium iodide (0.124 g, 0.83 mmol), and the reaction mixture was heated at 85 °C for 16 h. The reaction mixture was cooled to room temperature, filtered through a pad of celite, washed with ethyl acetate, and the filtrate was evaporated under reduced pressure to give the crude compound, which was purified by silica gel column chromatography (80 g silica gel, eluting with 60 - 70% ethyl acetate / petroleum ether) to give ethyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetate (130 mg, yield 28%). The product was further purified by preparative HPLC (HPLC method: column: Sunfire C18 (150 x 19 mm ID, 5 μm); mobile phase A = 0.1% TFA in water; mobile phase B = acetonitrile; flow 19 mL / min; gradient: time: 0.01 - 22.0; % concentration of B: 10 - 75) to give Example 69 and Example 70.

[0326] Example 69: LCMS: m / z = 557.2 (M + H); retention time: 2.34 min ((LCMS method: column: XBridge BEH XP C18 (50 x 2.1) mm, 2.5 μm; mobile phase A: 95% water: 5% acetonitrile; 10 mM NH4OAc; mobile phase B: 5% water: 95% acetonitrile; 10 mM NH4OAc; flow: 1.1 mL / min; temperature: 50 °C; time (min); 11H NMR (400 MHz, DMSO-d6) δ 8.22 (s, 1H), 7.99 (d, J = 9.0 Hz, 1H), 7.85 - 7.67 (m, 4H), 5.91 - 5.37 (m, 1H), 4.90 - 4.92 (m, 1H), 4.54 (s, 1H), 4.19 - 3.97 (m, 2H), 3.44 (s, 3H), 3.24 - 2.80 (m, 2H), 2.32 - 2.38 (m, 2H), 2.11 - 1.72 (m, 2H), 1.55 - 1.29 (m, 2H), 1.18 - 1.11 (m, 3H), 0.97 - 0.80 (m, 3H), 0.72 - 0.45 (m, 3H)

[0327] Example 70; LCMS: m / z = 557.2 (M + H); retention time: 2.34 minutes (LCMS method: column: XBridge BEH XP C18 (50x2.1) mm, 2.5 μm; mobile phase A: 95% water: 5% acetonitrile; 10 mM NH4OAc; mobile phase B: 5% water: 95% acetonitrile; 10 mM NH4OAc; flow rate: 1.1 mL / min; temperature: 50 °C); 1 1H NMR (400 MHz, DMSO-d6) δ 8.32 - 8.14 (m, 1H), 7.98 - 8.02 (m, 1H), 7.77 - 7.75 (m, 4H), 6.06 - 5.33 (m, 1H), 5.05 - 4.78 (m, 1H), 4.60 - 4.48 (m, 1H), 4.24 - 4.13 (m, 1H), 4.13 - 4.00 (m, 1H), 3.41 - 3.46 (m, 4H), 3.04 - 2.87 (m, 1H), 2.64 - 2.54 (m, 1H), 2.30 - 1.71 (m, 3H), 1.66 - 1.43 (m, 2H), 1.16 (t, J = 7.0 Hz, 3H), 0.99 - 0.79 (m, 3H), 0.68 (t, J = 7.5 Hz, 3H)

[0328] Examples 71 and 72 2 - ((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetic acid

Chemical formula

[0329] To a stirred solution of ethyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetate (60 mg, 0.11 mmol) in 1,4-dioxane (10 mL) was added trimethylstannanol (100 mg, 0.55 mmol), and the reaction mixture was heated at 100 °C for 16 h. The reaction mixture was cooled to room temperature, filtered through a pad of celite, washed with ethyl acetate, and the filtrate was evaporated under reduced pressure to give the crude compound, which was purified by preparative HPLC (HPLC method: preparative column: C-5; mobile phase A: 10 mM ammonium acetate in MeOH; mobile phase B, flow rate: 20 mL / min) to give Example 71 and Example 72.

[0330] Example 71: (8.3 mg, yield 15%); LCMS: m / z = 529.3 (M+H); retention time: 1.41 min (LCMS method: column: XBridge BEH XP C18 (50x2.1) mm, 2.5 μm; mobile phase A: 95% water: 5% acetonitrile; 10 mM NH4OAc; mobile phase B: 5% water: 95% acetonitrile; 10 mM NH4OAc; flow rate: 1.1 mL / min; temperature: 50 °C); 1 H NMR (400 MHz, DMSO-d6) δ 12.85-12.57 (m, 1H), 8.22 (td, J = 1.6, 8.8 Hz, 1H), 7.97 (d, J = 8.8 Hz, 1H), 7.74-7.70 (m, 4H), 5.86-5.23 (m, 1H), 5.01-4.75 (m, 1H), 4.46-4.17 (m, 1H), 3.43 (s, 3H), 3.19-3.12 (m, 1H), 2.89-2.70 (m, 2H), 2.38-1.78 (m, 3H), 1.48-1.28 (m, 2H), 1.00-0.81 (m, 3H), 0.73-0.47 (m, 3H)

[0331] Example 72: (6 mg, yield 10%); LCMS: m / z = 529.3 (M+H); retention time: 1.42 minutes (LCMS method: column: XBridge BEH XP C18 (50x2.1) mm, 2.5 μm; mobile phase A: 95% water: 5% acetonitrile; 10 mM NH4OAc; mobile phase B: 5% water: 95% acetonitrile; 10 mM NH4OAc; flow rate: 1.1 mL / min; temperature: 50 °C); 1 H NMR (400 MHz, DMSO-d6) δ 8.31 - 8.12 (m, 1H), 8.06 - 7.89 (m, 1H), 7.81 - 7.59 (m, 4H), 6.06 - 5.28 (m, 1H), 5.09 - 4.73 (m, 1H), 4.41 - 4.20 (m, 1H), 3.68 - 3.59 (m, 1H), 3.43 (s, 3H), 3.19 - 3.11 (m, 1H), 2.51 - 2.56 (m, 1H), 2.30 - 1.75 (m, 3H), 1.63 - 1.40 (m, 2H), 0.99 - 0.80 (m, 3H), 0.66 - 0.68 (m, 3H), 0.43 - 0.17 (m, 1H)

[0332] Examples 73 and 74 2 - ((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-N-((1-hydroxycyclopropyl)methyl)-2-(4-(trifluoromethyl)phenyl)acetamide

Chemical Structure

[0333] To a stirred solution of 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetic acid (50 mg, 0.1 mmol) in DMF (3 mL) was added HATU (71.9 mg, 0.19 mmol), DIPEA (0.05 mL, 0.28 mmol) and 1-(aminomethyl)cyclopropan-1-ol (12.4 mg, 0.14 mmol), and the reaction mixture was stirred at room temperature for 16 h. The reaction was quenched with water (20 mL). The reaction mixture was extracted with ethyl acetate (3 x 20 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and evaporated under reduced pressure to give the crude product, which was purified by preparative SFC (SFC method: column / dimension: Chiralpak IC (250 x 30) mm, 5 μm; % CO2: 75%; co-solvent: 25% methanol; overall flow: 100.0 mL / min; back pressure: 100 bar; temperature: 35 °C; UV: 220 nm) to afford Example 73 and Example 74.

[0334] Example 73: LCMS: m / z = 598.3 (M+H); retention time: 1.71 min (LCMS method: column: XBridge BEH XP C18 (50 x 2.1) mm, 2.5 μm; mobile phase A: 95% water: 5% acetonitrile; 10 mM NH4OAc; mobile phase B: 5% water: 95% acetonitrile; 10 mM NH4OAc; flow: 1.1 mL / min; temperature: 50 °C); 1 1H NMR (400 MHz, DMSO-d6) δ 8.45 (brd, J = 4.0 Hz, 1H), 8.31 - 8.15 (m, 1H), 7.96 - 8.01 (m, 1H), 7.71 - 7.77 (m, 4H), 6.06 - 4.77 (m, 3H), 4.39 (brd, J = 7.8 Hz, 1H), 3.44 - 3.32 (m, 3H), 3.28 - 3.13 (m, 2H), 3.10 - 2.95 (m, 1H), 2.61 - 2.54 (m, 1H), 2.30 - 1.80 (m, 3H), 1.67 - 1.42 (m, 2H), 1.04 - 0.80 (m, 3H), 0.71 (t, J = 7.4 Hz, 3H), 0.55 - 0.33 (m, 4H)

[0335] Example 74: LCMS: m / z = 598.2 (M+H); retention time: 1.72 minutes (LCMS method: column: XBridge BEH XP C18 (50x2.1) mm, 2.5 μm; mobile phase A: 95% water: 5% acetonitrile; 10 mM NH4OAc; mobile phase B: 5% water: 95% acetonitrile; 10 mM NH4OAc; flow rate: 1.1 mL / min; temperature: 50 °C); 1 H NMR (400 MHz, DMSO-d6) δ (400 MHz, DMSO-d6) δ 8.30 - 8.12 (m, 2H), 7.97 - 7.80 (m, 1H), 7.77 - 7.71 (m, 4H), 5.93 - 4.83 (m, 3H), 4.35 (brs, 1H), 3.56 (brs, 1H), 3.43 - 3.32 (m, 3H), 3.23 - 3.17 (m, 3H), 2.87 - 2.71 (m, 2H), 2.45 - 2.36 (m, 2H), 2.11 - 1.91 (m, 2H), 1.80 (s, 3H), 1.48 - 1.28 (m, 2H), 1.23 - 1.25 (m, 1H), 0.99 - 0.32 (m, 4H)

[0336] The examples shown in Table 1 were prepared according to the general procedures described in the production of Examples 73 and 74 using appropriate amines in the synthetic route. When a mixture of diastereomers was obtained by the reaction, the mixture was separated using preparative HPLC at the final stage. The absolute stereochemistry was not assigned to the newly formed carbon-nitrogen bond.

[0337] Table 1 [Table 12] [Table 13] [Table 14] [Table 15] [Table 16]

Table 17

[0338] Examples 117 and 118 2 - ((2R,5S)-4-(6 - Cyano - 1 - methyl - 2 - oxo - 1,2 - dihydropyrido[3,2 - d]pyrimidin - 4 - yl)-2,5 - diethylpiperazin - 1 - yl)-N - methoxy - 2-(4 - (trifluoromethyl)phenyl)acetamide

Chemical Structure

[0339] To a stirred solution of 2 - ((2R,5S)-4-(6 - cyano - 1 - methyl - 2 - oxo - 1,2 - dihydropyrido[3,2 - d]pyrimidin - 4 - yl)-2,5 - diethylpiperazin - 1 - yl)-2-(4 - (trifluoromethyl)phenyl)acetic acid (200 mg, 0.378 mmol) in anhydrous DMF (3 mL), O - methylhydroxylamine·hydrochloride (37.9 mg, 0.454 mmol), DIPEA (0.13 mL, 0.76 mmol) and HATU (216 mg, 0.568 mmol) were added. The reaction mixture was stirred at ambient temperature. After 16 h, the reaction mixture was diluted with ethyl acetate (10 mL), washed with water (2 x 10 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the crude product, which was purified by preparative HPLC (HPLC method: column: Luna C18 (250 x 21) mm, 10 micron; mobile phase A: 0.1% TFA in water; mobile phase B: acetonitrile; diluent: acetonitrile: water (50:50); gradient: 30% - 70% B over 15 min; flow: 15 mL / min) to give Examples 117 and 118.

[0340] The fraction of Example 117 was concentrated under reduced pressure, diluted with ACN / H2O (1:1), and lyophilized to obtain 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-N-methoxy-2-(4-(trifluoromethyl)phenyl)acetamide·TFA (35 mg, yield 14%) as an off-white solid. LCMS: m / z = 558.2 (M+H) + ; Retention time: 2.56 minutes (LCMS method: Column: Kinetex XB-C18 (75x3.0) mm, 2.6 μm; Mobile phase: A: 5 mM ammonium formate (pH 3.3): ACN (98:2); Mobile phase: B: ACN: buffer (98:2); Flow rate: 1.0 mL / min; HPLC retention times 8.75 and 8.22 minutes (each, HPLC methods FB and FC); 1 H NMR (400 MHz, DMSO-d6): δ 11.56 (s, 1H), 8.24 (m, 1H), 8.00 (m, 1H), 7.78 (s, 4H), 5.87 - 5.40 (m, 1H), 4.96 - 4.89 (m, 1H), 4.16 (s, 1H), 3.57 (s, 3H), 3.43 (s, 3H), 3.24 - 3.21 (m, 1H), 2.82 (m, 1H), 2.70 (m, 2H), 2.45 - 2.30 (m, 2H), 2.15 - 1.88 (m, 2H), 1.42 - 1.30 (m, 2H), 0.96 - 0.87 (m, 3H), 0.68 - 0.52 (m, 3H)

[0341] The fraction of Example 118 was concentrated under reduced pressure, diluted with ACN / H2O (1:1), and lyophilized to obtain 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-N-methoxy-2-(4-(trifluoromethyl)phenyl)acetamide·TFA (30 mg, yield 11%). LCMS: m / z = 558.0 (M+H) +; HPLC retention time: 2.48 minutes (LCMS method: column: Kinetex XB-C18 (75x3.0) mm, 2.6 μm; mobile phase: A: 5 mM ammonium formate (pH 3.3): ACN (98:2); mobile phase: B: ACN: buffer (98:2); flow rate: 1.0 mL / min); HPLC retention times 9.38 and 8.97 minutes (each, HPLC methods FB and FC); 1 1H NMR (400 MHz, DMSO-d6): δ 11.76 (s, 1H), 8.29 - 8.21 (m, 1H), 8.03 - 7.97 (m, 1H), 7.78 - 7.72 (m, 4H), 6.04 - 5.35 (m, 1H), 5.07 - 4.82 (m, 1H), 4.10 (s, 1H), 3.60 (s, 3H), 3.44 (s, 3H), 3.06 - 2.98 (m, 1H), 2.6 - 2.85 (m, 1H), 2.27 - 2.08 (m, 2H), 1.92 - 1.83 (m, 2H), 1.53 - 1.51 (m, 2H), 0.98 - 0.91 (m, 3H), 0.71 - 0.69 (m, 3H)

[0342] The examples shown in Table 2 were prepared in the synthetic route according to the general procedures described in the preparation of Examples 117 and 118 using the appropriate amine. When a mixture of diastereomers was obtained by the reaction, the mixture was separated using preparative HPLC in the final step. The absolute stereochemistry was not assigned to the newly formed carbon - nitrogen bond.

[0343] Table 2

Table 18

Table 19

[0344] Example 133 Ethyl 2 - ((2R,5S)-4-(6 - cyano - 1 - methyl - 2 - oxo - 1,2 - dihydropyrido[3,2 - d]pyrimidin - 4 - yl)-2 - ethyl - 5 - methylpiperazin - 1 - yl)-2-(4 - (trifluoromethyl)phenyl)acetate [Chemical formula]

[0345] To a stirred solution of 4 - ((2S,5R)-5 - ethyl - 2 - methylpiperazin - 1 - yl)-1 - methyl - 2 - oxo - 1,2 - dihydropyrido[3,2 - d]pyrimidine - 6 - carbonitrile·TFA (2.00 g, 4.69 mmol) in anhydrous acetonitrile (20 mL) at 0 °C was added DIPEA (1.638 mL, 9.38 mmol), followed by ethyl 2 - bromo - 2-(4-(trifluoromethyl)phenyl)acetate (1.837 g, 5.63 mmol). Next, the reaction mixture was heated to 75 °C, and the progress of the reaction was monitored by TLC analysis. After 3 hours, the volatile materials were removed under reduced pressure, and the residue thus obtained was diluted with DCM (50 mL), washed with water (2 x 20 mL), saturated brine solution (20 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude material thus obtained was subjected to flash column chromatography on silica gel (230:400 mesh), eluting with 0 - 100% EtOAc in petroleum ether for purification to obtain ethyl 2 - ((2R,5S)-4-(6 - cyano - 1 - methyl - 2 - oxo - 1,2 - dihydropyrido[3,2 - d]pyrimidine - 4 - yl)-2 - ethyl - 5 - methylpiperazin - 1 - yl)-2-(4-(trifluoromethyl)phenyl)acetate (1.8 g, yield 67.9%) as a brown solid. LCMS: m / z = 543.2 (M + H); retention time 3.31 minutes (LCMS method: column: Kinetex XB - C18 (75 x 30) mm, 2.6 μm mobile phase: A: 5 mM ammonium formate (pH 3.3): ACN (98:2) mobile phase: B: ACN: buffer (98:2) flow rate: 1.0 mL / min)

[0346] Example 134 2 - ((2R,5S)-4-(6 - cyano - 1 - methyl - 2 - oxo - 1,2 - dihydropyrido[3,2 - d]pyrimidine - 4 - yl)-2 - ethyl - 5 - methylpiperazin - 1 - yl)-2-(4-(trifluoromethyl)phenyl)acetic acid [Chemical]

[0347] Ethyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetate (500 mg, 0.885 mmol) in anhydrous toluene (20 mL) was added to a stirred solution of trimethylstannanol (1440 mg, 7.96 mmol). The reaction mixture was then heated to 100 °C and the progress of the reaction was monitored using TLC analysis. After 36 h, the reaction mixture was cooled to room temperature and the volatile materials were removed under reduced pressure. The residue obtained was then dissolved in DCM (30 mL) and filtered through a celite bed. The filtrate was washed with water (3 x 20 mL), the organic layer was dried over Na2SO4 and concentrated under reduced pressure to give 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetic acid (400 mg, 71% yield) as a brown solid. LCMS: m / z = 515.2 (M+H); retention times: 1.49 and 1.78, (LCMS method column: Kinetex XB-C18 (75 x 3.0) mm, 2.6 μm, mobile phase: A: 5 mM ammonium formate (pH 3.3): ACN (98:2), mobile phase: B: ACN: buffer (98:2), flow rate: 1.0 mL / min)

[0348] Examples 135 and 136 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-N-methoxy-2-(4-(trifluoromethyl)phenyl)acetamide [Chemical]

[0349] To a stirred solution of 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetic acid (240 mg, 0.378 mmol) and O-methylhydroxylamine hydrochloride (47.3 mg, 0.567 mmol) in anhydrous DMF (2 mL) was added HATU (172 mg, 0.453 mmol) and DIPEA (0.13 mL, 0.756 mmol). The reaction mixture was then stirred at ambient temperature for 16 h. After completion of the reaction, the reaction mixture was diluted with ethyl acetate (15 mL) and washed with aqueous sodium bicarbonate solution (10 mL), water (10 mL), and saturated brine solution (10 mL). The organic layer was dried over sodium sulfate, filtered, concentrated under reduced pressure to obtain the crude product, which was purified by preparative HPLC (HPLC method: column: XSelect C18 (150x19) mm, 5 micron; mobile phase A: 0.1% TFA in water; mobile phase B: acetonitrile; diluent: acetonitrile: water (50:50); gradient: 20% - 80% B in 0 - 12 min; flow rate: 15 mL / min) to obtain Examples 135 and 136.

[0350] The fraction of Example 135 was concentrated under reduced pressure, diluted with ACN / H2O (1:1), freeze-dried to obtain 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-N-methoxy-2-(4-(trifluoromethyl)phenyl)acetamide·TFA (45 mg, yield 17%). LCMS: m / z = 544.2 (M+H); retention time: 2.24; (LCMS method: column: Kinetex XB-C18 (75x3.0) mm, 2.6 micron; mobile phase: A: 5 mM ammonium formate (pH 3.3): ACN (98:2); mobile phase: B: ACN: buffer (98:2); flow rate: 1.0 mL / min; HPLC retention times 8.31 and 7.63 min (for HPLC methods B and C respectively); 11H NMR (400 MHz, DMSO-d6): δ 11.55 (s, 1H), 8.25 - 8.23 (m, 1H), 8.01 - 7.99 (m, 1H), 7.79 (s, 4H), 5.84 (m, 1H), 4.93 (m, 1H), 4.22 (s, 1H), 3.62 (s, 3H), 3.42 (s, 3H), 2.94 (m, 1H), 1.60 (m, 1H), 1.42 (m, 4H), 0.64 (m, 3H). (-3H)

[0351] The fraction of Example 136 was concentrated under reduced pressure, diluted with ACN / H2O (1:1), freeze-dried to obtain 2 - ((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-N-methoxy-2-(4-(trifluoromethyl)phenyl)acetamide·TFA (29 mg, yield 12%) as an off-white solid. LCMS: m / z = 544.2 (M + H); retention time: 2.18; LCMS method: column: Kinetex XB-C18 (75x3.0) mm, 2.6 μm; mobile phase: A: 5 mM ammonium formate (pH 3.3): ACN (98:2); mobile phase: B: ACN: buffer (98:2); flow rate: 1.0 mL / min; HPLC retention times 8.75 and 8.16 minutes (each, HPLC methods FB and FC); 1 1H NMR (400 MHz, DMSO-d6): δ 11.76 (s, 1H), 8.26 (m, 1H), 8.01 (m, 1H), 7.78 - 7.72 (m, 4H), 5.99 (m, 1H), 4.94 (m, 2H), 4.13 (s, 1H), 3.61 (m, 4H), 3.45 (s, 3H), 3.1 - 2.99 (m, 1H), 2.23 - 2.21 (m, 1H), 1.58 - 1.46 (m, 3H), 1.30 (m, 2H), 0.99 (s, 3H)

[0352] The examples shown in Table 3 were produced in the synthetic route according to the general procedures described in the production of Examples 135 and 136 using appropriate amines. When a mixture of diastereomers was obtained by the reaction, the mixture was separated using preparative HPLC in the final step. The absolute stereochemistry was not assigned to the newly formed carbon-nitrogen bond.

[0353] Table 3

Table 20

Table 21

[0354] Examples 147 and 148 N-Cyano-2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetamide

Chem.

[0355] To a stirred solution of 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetic acid (250 mg, 0.394 mmol) in anhydrous DCM (2 mL) was added cyanamide (24.8 mg, 0.59 mmol), EDC·HCl (113 mg, 0.59 mmol) and DMAP (96 mg, 0.787 mmol). The reaction mixture was stirred at ambient temperature for 16 h. After completion of the reaction, the reaction mixture was diluted with DCM (15 mL) and washed with aqueous sodium bicarbonate solution (10 mL), water (10 mL), brine saturated solution (10 mL), dried over sodium sulfate, filtered, concentrated under reduced pressure to give the crude product, which was purified by preparative HPLC (HPLC method: column: Luna C18 (250x21.2) mm, 10 micron; mobile phase A: 0.1% TFA in water; mobile phase B: acetonitrile; diluent: acetonitrile: water (50:50); gradient: 40% - 80% B in 0 - 15 min; flow: 15 mL / min) to give Examples 147 and 148.

[0356] The fraction of Example 147 was concentrated under reduced pressure, diluted with ACN / H2O (1:1) and lyophilized to give N-cyano-2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetamide·TFA (16 mg, yield 6%) as an off-white solid. LCMS: m / z = 539.2 (M+H) + ; retention time: 1.617 min (LCMS method: column: Kinetex XB-C18 (75x3.0) mm, 2.6 micron; mobile phase A: 5 mM ammonium formate (pH 3.3): ACN (98:2); mobile phase: B: ACN: buffer (98:2); flow rate: 1.0 mL / min; HPLC retention times 7.58 and 7.16 min (each, HPLC methods FB and FC); 11H NMR (400 MHz, DMSO-d6): δ 9.94 (bs, 1H), 8.28 - 8.26 (m, 1H), 8.05 - 8.03 (m, 1H), 7.92 - 7.87 (m, 4H), 5.94 - 5.74 (m, 1H), 5.19 - 4.94 (m, 2H), 4.00 (m, 1H), 3.39 - 3.46 (m, 3H), 3.23 (m, 1H), 2.83 - 2.82 (m, 1H), 1.65 - 1.47 (m, 5H), 0.57 (s, 3H)

[0357] The fraction of Example 148 was concentrated under reduced pressure, diluted with ACN / H2O (1:1), lyophilized, and N-cyano-2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetamide·TFA (38 mg, yield 15%) was obtained as an off-white solid. LCMS: m / z = 539.2 (M+H) + ; Retention time: 1.794 minutes (LCMS method: Column: Kinetex XB-C18 (75x3.0) mm, 2.6 μm; Mobile phase: A: 5 mM ammonium formate (pH 3.3): ACN (98:2); Mobile phase: B: ACN: buffer (98:2); Flow rate: 1.0 mL / min; HPLC retention times 7.80 and 7.22 minutes (each, HPLC methods B and C); 1 1H NMR (400 MHz, DMSO-d6): δ 8.29 - 8.26 (m, 1H), 8.04 - 8.02 (m, 1H), 7.87 - 7.81 (m, 4H), 6.03 - 5.74 (m, 1H), 5.08 - 4.97 (m, 1H), 4.81 (s, 1H), 3.88 (m, 1H), 3.46 (s, 3H), 3.21 (m, 1H), 3.06 (m, 1H), 2.61 - 2.51 (m, 1H), 1.58 - 1.35 (m, 5H), 0.82 (s, 3H)

[0358] The examples shown in Table 4 were prepared in the synthetic route according to the general procedures described in the preparation of Examples 147 and 148 using the appropriate amine. When a mixture of diastereomers was obtained by the reaction, the mixture was separated using preparative HPLC at the final stage. The absolute stereochemistry was not assigned to the newly formed carbon-nitrogen bond.

[0359] Table 4

Table 22

[0360] Example 153 Methyl 3-((2R,5S)-4-(6-chloro-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-3-(4-(trifluoromethyl)phenyl)propanoate

Chem.

[0361] To a stirred solution of 6-chloro-4-((2S,5R)-2,5-diethylpiperazin-1-yl)-1-methylpyrido[3,2-d]pyrimidin-2(1H)-one (500 mg, 1.49 mmol) in DMSO (5 mL) was added 4-(trifluoromethyl)benzaldehyde (0.31 mL, 2.23 mmol) and trimethyl borate (0.17 mL, 1.49 mmol). The reaction mixture was stirred at room temperature for 15 minutes. The resulting solution was treated dropwise with 1-(tert-butyldimethylsilyloxy)-1-methoxyethane (0.5 mL, 2.23 mmol) and stirred at room temperature for 16 hours. The reaction mixture was extracted with EtOAc (2 x 100 mL), washed with water, brine, and dried over sodium sulfate. The solvent was removed under reduced pressure to give a crude residue, which was purified by silica gel flash column chromatography (10% MeOH in DCM; 24 g column) to afford methyl 3-((2R,5S)-4-(6-chloro-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-3-(4-(trifluoromethyl)phenyl)propanoate (700 mg, 83% yield) as a mixture of diastereomers. LCMS: m / z, 566.2 (M+1); retention time: 1.23 minutes (LCMS method: column: Waters Acquity UPLC BEH C18 (2.1 x 50 mm) 1.7 μm, mobile phase A: 10 mM NH4OAc: acetonitrile (95:5); mobile phase B: 10 mM NH4OAc: acetonitrile (5:95), gradient = 20 - 100% B over 2 minutes, then hold at 100% B for 0.3 minutes; temperature: 50 °C; flow rate: 0.7 mL / min; detection: UV (220 nm))

[0362] Examples 154 and 155 Methyl 3-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-3-(4-(trifluoromethyl)phenyl)propanoate

Chemical formula

[0363] To a stirred solution of methyl 3-((2R,5S)-4-(6-chloro-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-3-(4-(trifluoromethyl)phenyl)propanoate (150 mg, 0.26 mmol) in DMF (3.0 mL) was added zinc (26.0 mg, 0.40 mmol) and TEA (0.15 mL, 1.06 mmol). The reaction mixture was subjected to degassing with argon for 5 minutes, followed by the addition of zinc cyanide (93 mg, 0.79 mmol) and dichloro[9,9-dimethyl-4,5-bis(diphenylphosphino)xanthene]palladium(II) (80 mg, 0.11 mmol). The reaction mixture was heated at 90 °C for 16 hours. The reaction mixture was cooled to room temperature and concentrated under reduced pressure to give a residue, which was purified by preparative HPLC (column: cellulose-5 (250x21.2 mm), 5 μm; mobile phase: 10 mM NH4OAc in MeOH; flow rate: 20 mL / min).

[0364] Example 154: (7 mg, 5% yield): LCMS: m / z = 557.2 (M+H); retention time: 2.14 minutes (LCMS method: column: XBridge BEH XP C18 (50x2.1 mm), 2.5 μm; mobile phase A: 95% water: 5% acetonitrile; 10 mM ammonium acetate; mobile phase B: 5% water: 95% acetonitrile; 10 mM ammonium acetate; flow: 1.1 mL / min; temperature: 50 °C; time (min): 0-3; %B: 0-100; 11H NMR (400 MHz, DMSO-d6) δ ppm 8.24 (d, J = 8.8 Hz, 1H), 7.98 (d, J = 8.8 Hz, 1H), 7.74 (d, J = 8.1 Hz, 2H), 7.58 - 7.63 (m, 2H), 5.84 - 5.91 (m, 0.5H, 5.39 - 5.46 (m, 0.5H, 4.84 - 4.93 (m, 1H), 4.21 (brt, J = 6.2 Hz, 1H), 3.45 - 3.50 (m, 3H), 3.44 (s, 3H), 3.09 - 3.19 (m, 2H), 2.93 - 3.03 (m, 1H), 2.56 - 2.63 (m, 1H), 2.35 - 2.47 (m, 1H), 1.86 - 2.04 (m, 1H), 1.68 - 1.86 (m, 2H), 1.28 - 1.49 (m, 2H), 0.87 - 0.99 (m, 3H), 0.60 - 0.79 (m, 3H)

[0365] Example 155: (15 mg, yield 10%; LCMS: m / z = 557.2 (M + H); retention time: 2.12 minutes (LCMS method: column: XBridge BEH XP C18 (50 x 2.1 mm), 2.5 μm; mobile phase A: 95% water: 5% acetonitrile; 10 mM ammonium acetate; mobile phase B: 5% water: 95% acetonitrile; 10 mM ammonium acetate; flow rate: 1.1 mL / min; temperature: 50 °C; time (min): 0 - 3; %B: 0 - 100; 1 1H NMR (400 MHz, DMSO-d6) δ ppm 8.17 - 8.27 (m, 1H), 7.93 - 8.01 (m, 1H), 7.69 (d, J = 8.3 Hz, 2H), 7.59 (d, J = 8.3 Hz, 2H), 5.96 - 6.07 (m, 0.5H, 5.25 - 5.35 (m, 0.5H), 5.00 - 5.09 (m, 0.5H, 4.69 - 4.79 (m, 0.5H, 4.04 (dd, J = 9.2, 5.5 Hz, 1H), 3.56 - 3.68 (m, 1H), 3.45 (s, 3H), 3.42 (s, 3H), 2.98 - 3.13 (m, 2H), 2.53 - 2.64 (m, 2H), 2.24 - 2.38 (m, 1H), 1.67 - 2.01 (m, 2H), 1.30 - 1.52 (m, 2H), 0.82 - 0.99 (m, 3H), 0.63 (brt, J = 7.2 Hz, 3H)

[0366] Example 156 3-((2R,5S)-4-(6-Chloro-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-3-(4-(trifluoromethyl)phenyl)propanoic acid

Chem.

[0367] To a stirred solution of methyl 3-((2R,5S)-4-(6-chloro-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-3-(4-(trifluoromethyl)phenyl)propanoate (550 mg, 0.97 mmol) in THF (12 mL) and water (6 mL) was added lithium hydroxide monohydrate (408 mg, 9.72 mmol) at room temperature. The reaction mixture was stirred for 16 h. The reaction mixture was neutralized with aqueous 1.5 N HCl and extracted with EtOAc (2 x 100 mL). The combined organic extracts were washed with water, brine, dried over sodium sulfate and concentrated to give 3-((2R,5S)-4-(6-chloro-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-3-(4-(trifluoromethyl)phenyl)propanoic acid (500 mg, 93% yield) as a mixture of diastereomers. LCMS: m / z, 552.2 (M+H); retention times: 1.29 and 1.39 min (LCMS method: column: Acquity UPLC BEH C18 (3.0 x 50 mm) 1.7 μm; mobile phase A: 10 mM ammonium acetate:acetonitrile (95:5), mobile phase B: 10 mM ammonium acetate:acetonitrile (5:95), method: %B: 0 min - 20: 2 min - 100: 2.3 min - 100, temperature: 27 °C; flow rate: 0.7 mL / min; detection: UV (220 nm))

[0368] Examples 157 and 158 3-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-3-(4-(trifluoromethyl)phenyl)propanoic acid

Chem.

[0369] To a stirred solution of methyl 3-((2R,5S)-4-(6-chloro-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-3-(4-(trifluoromethyl)phenyl)propanoate (100 mg, 0.18 mmol) in DMF (3.0 mL) were added zinc (18 mg, 0.27 mmol) and TEA (0.1 mL, 0.72 mmol). The reaction mixture was degassed with argon for 5 minutes, followed by the addition of zinc cyanide (64 mg, 0.54 mmol) and dichloro[9,9-dimethyl-4,5-bis(diphenylphosphino)xanthene]palladium(II) (55 mg, 0.07 mmol). The reaction mixture was heated at 90 °C for 16 hours. The reaction mixture was cooled to room temperature and concentrated under reduced pressure to give a residue, which was purified by preparative HPLC (column: Waters XBridge C18, 19x150 mm, 5 μm; mobile phase A: 10 mM ammonium acetate; mobile phase B: acetonitrile; gradient: 15 - 55% B over 20 minutes, then held at 100% B for 5 minutes; flow rate: 15 mL / min).

[0370] Example 157: (22 mg, 22% yield): LCMS: m / z = 543.2 (M+H); retention time: 1.36 minutes (LCMS method: column: XBridge BEH XP C18 (50x2.1 mm), 2.5 μm; mobile phase A: 95% water: 5% acetonitrile; 10 mM ammonium acetate; mobile phase B: 5% water: 95% acetonitrile; 10 mM ammonium acetate; flow rate: 1.1 mL / min; temperature: 50 °C; time (min): 0 - 3; %B: 0 - 100

[0371] Example 158: (7 mg, yield 7%); LCMS: m / z = 543.2 (M+H); retention time: 1.47 minutes (LCMS method: column: XBridge BEH XP C18 (50x2.1 mm), 2.5 μm; mobile phase A: 95% water: 5% acetonitrile; 10 mM ammonium acetate; mobile phase B: 5% water: 95% acetonitrile; 10 mM ammonium acetate; flow rate: 1.1 mL / min; temperature: 50 °C; time (min): 0 - 3; %B: 0 - 100

[0372] Examples 159 and 160 4 - ((2S,5R)-2,5 - diethyl - 4 - (3 - oxo - 3 - (6 - azaspiro[2.5]octan - 6 - yl)-1-(4 - (trifluoromethyl)phenyl)propyl)piperazin - 1 - yl)-1 - methyl - 2 - oxo - 1,2 - dihydropyrido[3,2 - d]pyrimidine - 6 - carbonitrile

Chemical formula

[0373] To a stirred solution of 3 - ((2R,5S)-4 - (6 - cyano - 1 - methyl - 2 - oxo - 1,2 - dihydropyrido[3,2 - d]pyrimidin - 4 - yl)-2,5 - diethylpiperazin - 1 - yl)-3 - (4 - (trifluoromethyl)phenyl)propanoic acid (100 mg, 0.18 mmol) in EtOAc (2 mL), 6 - azaspiro[2.5]octane, HCl (43.5 mg, 0.29 mmol), TEA (0.05 mL, 0.37 mmol) were added, followed by 1 - propanephosphonic anhydride (50% in EtOAc) (0.18 mL, 0.29 mmol) at 0 °C. The reaction mixture was warmed to room temperature and stirred for 16 hours. The reaction mixture was extracted with EtOAc (2x30 mL), washed with water and brine, dried over sodium sulfate, concentrated under reduced pressure to obtain a crude residue, which was purified by preparative HPLC [column: Lux cellulose - 4 (250 mmx21 mm); mobile phase: 10 mM NH4OAc in MeOH; flow rate: 19 mL / min].

[0374] Example 159: (3.9 mg, yield 3%): LCMS: m / z = 636.3 (M+H); retention time: 2.25 minutes (LCMS method: column: XBridge BEH XP C18 (50x2.1 mm), 2.5 μm; mobile phase A: 95% water: 5% acetonitrile; 10 mM ammonium acetate; mobile phase B: 5% water: 95% acetonitrile; 10 mM ammonium acetate; flow rate: 1.1 mL / min; temperature: 50 °C; time (min): 0 - 3; %B: 0 - 100; 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.24 (brd, J = 8.8 Hz, 1H), 7.98 (d, J = 9.0 Hz, 1H), 7.71 (d, J = 8.0 Hz, 2H), 7.60 (brd, J = 8.3 Hz, 2H), 6.51 (s, 1H), 5.42 (brs, 1H), 4.97 - 4.81 (m, 1H), 4.36 - 4.21 (m, 1H), 3.49 - 3.38 (m, 6H), 3.11 - 2.91 (m, 2H), 2.77 - 2.68 (m, 1H), 2.61 - 2.55 (m, 1H), 2.45 - 2.37 (m, 1H), 2.05 - 1.68 (m, 2H), 1.54 - 1.42 (m, 1H), 1.40 - 1.19 (m, 2H), 1.15 - 1.02 (m, 3H), 0.99 - 0.85 (m, 3H), 0.81 - 0.60 (m, 3H), 0.15 - 0.30 (m, 4H)

[0375] Example 160: (2 mg, yield 2%); LCMS: m / z = 636.3 (M+H); retention time: 2.24 minutes (LCMS method: column: XBridge BEH XP C18 (50x2.1 mm), 2.5 μm; mobile phase A: 95% water: 5% acetonitrile; 10 mM ammonium acetate; mobile phase B: 5% water: 95% acetonitrile; 10 mM ammonium acetate; flow rate: 1.1 mL / min; temperature: 50 °C; time (min): 0 - 3; %B: 0 - 100; 11H NMR (400 MHz, DMSO-d6) δ ppm 8.17 - 8.30 (m, 1H), 7.92 - 8.04 (m, 1H), 7.65 - 7.71 (m, 2H), 7.57 - 7.63 (m, 2H), 6.00 - 6.13 (m, 1H), 5.25 - 5.34 (m, 1H), 5.03 - 5.13 (m, 1H), 4.69 - 4.79 (m, 1H), 4.03 - 4.13 (m, 1H), 3.58 - 3.73 (m, 1H), 3.48 - 3.56 (m, 1H), 3.43 (s, 3H), 3.11 - 3.20 (m, 2H), 2.73 - 2.87 (m, 2H), 1.68 - 2.04 (m, 3H), 1.41 - 1.55 (m, 2H), 1.20 - 1.27 (m, 1H), 0.74 - 1.10 (m, 7H), 0.64 (t, J = 7.5 Hz, 3H), 0.14 - 0.30 (m, 4H)

[0376] The examples shown in Table 5 were prepared according to the general procedures described in the preparation of Examples 159 and 160 using the appropriate amine in the synthetic route. When a mixture of diastereomers was obtained by the reaction, the mixture was separated using preparative HPLC at the final step. The absolute stereochemistry was not assigned to the newly formed carbon-nitrogen bond.

[0377] Table 5 [Table 23]

[0378] Intermediate 54 6-Chloro-4-((2S,5R)-2,5-diethyl-4-(3-hydroxy-1-(4-(trifluoromethyl)phenyl)propyl)piperazin-1-yl)-1-methylpyrido[3,2-d]pyrimidin-2(1H)-one [Chemical Structure]

[0379] 3-((2R,5S)-4-(6-chloro-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-3-(4-(trifluoromethyl)phenyl)propanoic acid (1.5 g, 2.72 mmol) and 4-methylmorpholine (0.6 mL, 5.43 mmol) in THF (15 mL) was stirred and isobutyl chloroformate (0.5 mL, 3.26 mmol) was added thereto at 0 °C. The reaction mixture was warmed to room temperature and stirred for 2 h. The reaction mixture was filtered through a Celite pad and the residue was washed with an excess amount of THF (10 mL). The filtrate was cooled to 0 °C and sodium borohydride (0.31 g, 8.15 mmol) was added. The reaction mixture was warmed to room temperature and stirred overnight. The reaction mixture was cooled to 0 °C. The reaction was quenched with saturated aqueous NH4Cl. The reaction mixture was extracted with EtOAc (3x100 mL). The combined organic extracts were washed with brine, dried over Na2SO4 and concentrated to give the crude material which was purified by flash chromatography (using a 40 g silica gel column, eluting with 8-10% MeOH in DCM) to give 6-chloro-4-((2S,5R)-2,5-diethyl-4-(3-hydroxy-1-(4-(trifluoromethyl)phenyl)propyl)piperazin-1-yl)-1-methylpyrido[3,2-d]pyrimidin-2(1H)-one (1.1 g, 2.045 mmol, 75% yield). LCMS: m / z, 538.2 (M+H); retention time: 2.27 min (LCMS method: column: Kinetex XB-C18 (3x75 mm) 2.6 μm; mobile phase A: 10 mM ammonium formate: acetonitrile (98:2), mobile phase B: 10 mM ammonium formate: acetonitrile (2:98), gradient = 20-100% B over 4 min then hold at 100% B for 0.6 min; temperature: 27 °C; flow rate: 1.0 mL / min; detection: UV (220 nm))

[0380] Intermediate 55 4-((2S,5R)-4-(3-azido-1-(4-(trifluoromethyl)phenyl)propyl)-2,5-diethylpiperazin-1-yl)-6-chloro-1-methylpyrido[3,2-d]pyrimidin-2(1H)-one

Chemical formula

[0381] A stirred solution of 6-chloro-4-((2S,5R)-2,5-diethyl-4-(3-hydroxy-1-(4-(trifluoromethyl)phenyl)propyl)piperazin-1-yl)-1-methylpyrido[3,2-d]pyrimidin-2(1H)-one (1.1 g, 2.04 mmol) in DCM (12 mL) was added with TEA (0.8 mL, 6.13 mmol) and mesyl-Cl (0.32 mL, 4.09 mmol) at 0 °C. The reaction mixture was stirred at room temperature for 2 h. The reaction mixture was extracted with DCM (3x50 mL), washed with water and brine, dried over sodium sulfate and concentrated under reduced pressure to give the crude mesylate derivative. The crude mesylate was dissolved in DMF (10 mL) and sodium azide (0.580 g, 8.93 mmol) was added at room temperature. The reaction mixture was heated at 50 °C for 14 h. The reaction mixture was cooled to room temperature, diluted with water and extracted with EtOAc (3x50 mL). The combined organic extracts were washed with brine, dried over Na2SO4 and concentrated to give the crude material, which was purified by flash chromatography (using a 40 g silica gel column, eluting with 5-7% MeOH in DCM) to give 4-((2S,5R)-4-(3-azido-1-(4-(trifluoromethyl)phenyl)propyl)-2,5-diethylpiperazin-1-yl)-6-chloro-1-methylpyrido[3,2-d]pyrimidin-2(1H)-one (900 mg, 90% yield). LCMS: m / z, 563.2 (M+1); retention times: 1.16 and 1.25 min (LCMS method: column: Kinetex XB-C18 (3x75 mm) 2.6 μm; mobile phase A: 10 mM ammonium formate: acetonitrile (98:2), mobile phase B: 10 mM ammonium formate: acetonitrile (2:98), gradient = 20-100% B over 4 min, then hold at 100% B for 0.6 min; temperature: 27 °C; flow rate: 1.0 mL / min; detection: UV (220 nm))

[0382] Intermediate 56 4-((2S,5R)-4-(3-amino-1-(4-(trifluoromethyl)phenyl)propyl)-2,5-diethylpiperazin-1-yl)-6-chloro-1-methylpyrido[3,2-d]pyrimidin-2(1H)-one [Chemistry]

[0383] To a stirred solution of 4 - ((2S,5R)-4-(3 - azido - 1-(4-(trifluoromethyl)phenyl)propyl)-2,5 - diethylpiperazin - 1 - yl)-6 - chloro - 1 - methylpyrido[3,2 - d]pyrimidin - 2(1H)-one (400 mg, 0.71 mmol) in MeOH (5 mL) were added zinc (464 mg, 7.10 mmol) and ammonium chloride (380 mg, 7.10 mmol) / water (1 mL) at room temperature. The reaction mixture was stirred for 1 hour. The reaction mixture was filtered through a Celite pad and washed with an excess of MeOH (10 mL). The filtrate was concentrated under reduced pressure to give a crude residue, which was extracted with EtOAc (3 x 50 mL). The combined organic extracts were washed with brine, dried over Na2SO4, and concentrated to give 4 - ((2S,5R)-4-(3 - amino - 1-(4-(trifluoromethyl)phenyl)propyl)-2,5 - diethylpiperazin - 1 - yl)-6 - chloro - 1 - methylpyrido[3,2 - d]pyrimidin - 2(1H)-one (320 mg, 84% yield). LCMS: m / z, 537.2 (M+1); retention times: 2.09 and 2.31 minutes (LCMS method: column: Kinetex XB - C18 (3 x 75 mm) 2.6 μm; mobile phase A: 10 mM ammonium formate:acetonitrile (98:2), mobile phase B: 10 mM ammonium formate:acetonitrile (2:98), gradient = 20 - 100% B over 4 minutes, then hold at 100% B for 0.6 minutes; temperature: 27 °C; flow rate: 1.0 mL / min; detection: UV (220 nm))

[0384] Example 167 Methyl (3 - ((2R,5S)-4-(6 - chloro - 1 - methyl - 2 - oxo - 1,2 - dihydropyrido[3,2 - d]pyrimidin - 4 - yl)-2,5 - diethylpiperazin - 1 - yl)-3-(4-(trifluoromethyl)phenyl)propyl)carbamate [Chemistry]

[0385] To a stirred solution of 4-((2S,5R)-4-(3-amino-1-(4-(trifluoromethyl)phenyl)propyl)-2,5-diethylpiperazin-1-yl)-6-chloro-1-methylpyrido[3,2-d]pyrimidin-2(1H)-one (120 mg, 0.22 mmol) in DCM (2 mL) were added TEA (0.1 mL, 0.67 mmol) and methyl chloroformate (0.04 mL, 0.45 mmol) at 0 °C. The reaction mixture was allowed to reach room temperature and stirred for 2 h. The reaction mixture was extracted with DCM (3 x 30 mL), washed with water, brine, dried over sodium sulfate and concentrated under reduced pressure to give methyl (3-((2R,5S)-4-(6-chloro-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-3-(4-(trifluoromethyl)phenyl)propyl)carbamate (110 mg, 0.185 mmol, 83% yield). LCMS: m / z, 595.2 (M+H); retention times: 2.01 and 2.02 min (LCMS method: column: Acquity UPLC BEH C18 (3.0 x 50 mm) 1.7 μm; mobile phase A: 10 mM ammonium acetate:acetonitrile (95:5), mobile phase B: 10 mM ammonium acetate:acetonitrile (5:95), method: %B: 0 min - 20: 2 min - 100: 2.3 min - 100, temperature: 27 °C; flow rate: 0.7 mL / min; detection: UV (220 nm))

[0386] Examples 168 and 169 3-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-3-(4-(trifluoromethyl)phenyl)propanoic acid

Chemical formula

[0387] To a stirred solution of methyl (3-((2R,5S)-4-(6-chloro-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-3-(4-(trifluoromethyl)phenyl)propyl)carbamate (80 mg, 0.13 mmol) in NMP (2 mL) was added dppf (5 mg, 8.07 μmol), zinc (2 mg, 0.027 mmol) and zinc cyanide (48 mg, 0.403 mmol). The reaction mixture was subjected to degassing for 5 minutes and Pd2(dba)3 (12 mg, 0.013 mmol) was added. The reaction mixture was heated at 90 °C overnight. The reaction mixture was cooled to room temperature, diluted with ethyl acetate and filtered through a pad of celite. The filtrate was washed with water, brine, the organic layer was dried over anhydrous Na2SO4, filtered and evaporated under reduced pressure to give a crude residue which was purified by preparative SFC [column / dimension: Chiralpak IC (250x21) mm, 5 μm, %CO2: 60%; % co-solvent: 40% ACN:MeOH (50:50); overall flow: 80.0 g / min; back pressure: 100 bar; temperature: 30 °C; UV: 270 nm].

[0388] Example 168: (5.3 mg, 7% yield): LCMS: m / z = 586.2 (M+H); retention time: 2.10 minutes (LCMS method: column: X Bridge BEH XP C18 (50x2.1 mm), 2.5 μm; mobile phase A: 95% water: 5% acetonitrile; 10 mM ammonium acetate; mobile phase B: 5% water: 95% acetonitrile; 10 mM ammonium acetate; flow: 1.1 mL / min; temperature: 50 °C; time (min): 0 - 3; %B: 0 - 100; 11H NMR (400 MHz, DMSO-d6) δ ppm 8.22 - 8.27 (m, 1H), 7.99 (d, J = 9.0 Hz, 1H), 7.75 (d, J = 8.3 Hz, 2H), 7.54 - 7.63 (m, 2H), 7.02 - 7.12 (m, 1H), 5.82 - 5.92 (m, 0.5H, 5.46 - 5.55 (m, 0.5H, 4.83 - 5.00 (m, 1H), 3.78 - 3.85 (m, 1H), 3.48 (s, 3H), 3.44 (s, 3H), 3.18 (d, J = 3.4 Hz, 1H), 2.85 - 2.91 (m, 1H), 2.76 - 2.84 (m, 2H), 2.30 - 2.44 (m, 2H), 1.97 - 2.16 (m, 2H), 1.70 - 1.93 (m, 3H), 1.40 - 1.52 (m, 1H), 1.27 - 1.37 (m, 1H), 0.88 - 1.04 (m, 3H), 0.57 - 0.76 (m, 3H)

[0389] Example 169: (7.7 mg, yield 9%): LCMS: m / z = 586.2 (M + H); retention time: 2.09 minutes (LCMS method: column: XBridge BEH XP C18 (50 x 2.1 mm), 2.5 μm; mobile phase A: 95% water: 5% acetonitrile; 10 mM ammonium acetate; mobile phase B: 5% water: 95% acetonitrile; 10 mM ammonium acetate; flow rate: 1.1 mL / min; temperature: 50 °C; time (min): 0 - 3; %B: 0 - 100; 1 1H NMR (400 MHz, DMSO-d6) δ ppm 8.15 - 8.31 (m, 1H), 7.92 - 8.08 (m, 1H), 7.69 - 7.75 (m, 2H), 7.58 - 7.64 (m, 2H), 7.09 - 7.15 (m, 1H), 6.01 - 6.08 (m, 0.5H, 5.24 - 5.35 (m, 0.5H, 5.06 - 5.15 (m, 0.5H, 4.71 - 4.82 (m, 0.5H, 3.60 - 3.68 (m, 1H), 3.49 (s, 3H), 3.44 (s, 3H), 3.26 - 3.31 (m, 1H), 3.09 - 3.23 (m, 1H), 2.62 - 2.82 (m, 2H), 2.15 - 2.29 (m, 1H), 1.96 - 2.08 (m, 1H), 1.62 - 1.88 (m, 3H), 1.35 - 1.56 (m, 2H), 0.89 - 1.02 (m, 3H), 0.62 (t, J = 7.3 Hz, 3H)

[0390] The examples shown in Table 6 were prepared in the synthetic route according to the general procedures described in the production of Examples 168 and 169 using appropriate amines. When a mixture of diastereomers was obtained by the reaction, the mixture was separated using preparative HPLC at the final stage. The absolute stereochemistry was not assigned to the newly formed carbon-nitrogen bond.

[0391] Table 6 [Table 24]

[0392] Intermediate 57 1-Benzyl 4-(tert-butyl) (2S,5R)-5-ethyl-2-methylpiperazine-1,4-dicarboxylate [Chemical formula]

[0393] To a stirred solution of tert-butyl (2R,5S)-2-ethyl-5-methylpiperazine-1-carboxylate (1.0 g, 4.38 mmol) in dioxane (10 mL) were added DIPEA (2.3 mL, 13.14 mmol) and Cbz-Cl (0.94 mL, 6.57 mmol). The reaction mixture was heated at 95 °C for 16 h. The reaction mixture was quenched with water (100 mL) and extracted with ethyl acetate (3 x 100 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, evaporated under reduced pressure to give the crude product, which was purified by silica gel chromatography (24 g silica gel column; using 10%-15% ethyl acetate / petroleum ether) to afford 1-benzyl 4-(tert-butyl) (2S,5R)-5-ethyl-2-methylpiperazine-1,4-dicarboxylate (1.5 g, 94% yield). LCMS: m / z = 363.2 (M+H); retention time: 2.05 min [LCMS method: column: Waters Acquity UPLC BEH C18 (2.1 x 50 mm) 1.7 μm, mobile phase A: 10 mM NH4OAc: acetonitrile (95:5); mobile phase B: 10 mM NH4OAc: acetonitrile (5:95), gradient = 20-90% B over 1.1 min then hold at 90% B for 0.6 min; temperature: 50 °C; flow rate: 0.7 mL / min; detection: UV (220 nm)]

[0394] Intermediate 58 Benzyl (2S,5R)-5-ethyl-2-methylpiperazine-1-carboxylate

Chemical formula

[0395] A stirred solution of 1-benzyl 4-(tert-butyl) (2S,5R)-5-ethyl-2-methylpiperazine-1,4-dicarboxylate (1.0 g, 2.76 mmol) in DCM (10 mL) was treated with 4N HCl in dioxane (4.2 mL, 16.55 mmol). The reaction mixture was stirred at room temperature for 16 h. The reaction mixture was concentrated under reduced pressure to give the crude product, which was triturated with DCM / hexane (1:4) to give a solid, filtered and dried under reduced pressure to give benzyl (2S,5R)-5-ethyl-2-methylpiperazine-1-carboxylate·HCl (0.8 g, 97% yield). LCMS: m / z = 263.2 (M+H); retention time: 1.02 min [LCMS method: column: Waters Acquity UPLC BEH C18 (2.1x50 mm) 1.7 μm, mobile phase A: 10 mM NH4OAc: acetonitrile (95:5); mobile phase B: 10 mM NH4OAc: acetonitrile (5:95), gradient = 20-90% B over 1.1 min then hold at 90% B for 0.6 min; temperature: 50 °C; flow rate: 0.7 mL / min; detection: UV (220 nm)]

[0396] Intermediate 59 Benzyl (2S,5R)-4-(2-ethoxy-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)-5-ethyl-2-methylpiperazine-1-carboxylate

Chemical formula

[0397] To a stirred solution of benzyl (2S,5R)-5-ethyl-2-methylpiperazine-1-carboxylate·HCl (0.85 g, 2.84 mmol) in acetonitrile (10 mL) was added DIPEA (1.5 mL, 8.53 mmol). The reaction mixture was stirred for 5 minutes, and ethyl 2-bromo-2-(4-(trifluoromethyl)phenyl)acetate (1.770 g, 5.69 mmol) and sodium iodide (0.213 g, 1.42 mmol) were added. The reaction mixture was heated at 85 °C for 16 hours. The reaction mixture was filtered through a celite bed and concentrated under reduced pressure to give the crude product, which was purified by silica gel chromatography (eluting with 10%-20% ethyl acetate / petroleum ether using a 24 g silica gel column) to give benzyl (2S,5R)-4-(2-ethoxy-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)-5-ethyl-2-methylpiperazine-1-carboxylate (0.8 g, 57.1% yield). LCMS: m / z = 493.0 (M+H); retention time: 1.50 minutes [LCMS method: column: Waters Acquity UPLC BEH C18 (2.1x50 mm) 1.7 μ, mobile phase A: 10 mM NH4OAc: acetonitrile (95:5); mobile phase B: 10 mM NH4OAc: acetonitrile (5:95), gradient = 20-90% B over 1.1 minutes then hold at 90% B for 0.6 minutes; temperature: 50 °C; flow rate: 0.7 mL / min; detection: UV (220 nm)]

[0398] Intermediate 60 Benzyl (2S,5R)-5-ethyl-4-(2-hydroxy-1-(4-(trifluoromethyl)phenyl)ethyl)-2-methylpiperazine-1-carboxylate

Chemical formula

[0399] A stirred solution of benzyl (2S,5R)-4-(2-ethoxy-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)-5-ethyl-2-methylpiperazine-1-carboxylate (0.3 g, 0.61 mmol) in tetrahydrofuran (5 mL) was treated with LiBH4 / THF (2 M, 1.5 mL, 3.05 mmol), followed by MeOH (0.3 mL). The reaction mixture was heated at 60 °C for 2 h. The reaction was quenched with saturated aqueous ammonium chloride (50 mL), and the reaction mixture was extracted with ethyl acetate (3 x 100 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and evaporated under reduced pressure to afford benzyl (2S,5R)-5-ethyl-4-(2-hydroxy-1-(4-(trifluoromethyl)phenyl)ethyl)-2-methylpiperazine-1-carboxylate (180 mg, 66%). LCMS: m / z = 451.0 (M+H); retention time: 2.09 min [LCMS method: column: Waters Acquity UPLC BEH C18 (2.1 x 50 mm) 1.7 μ, mobile phase A: 10 mM NH4OAc: acetonitrile (95:5); mobile phase B: 10 mM NH4OAc: acetonitrile (5:95), gradient = 20 - 90% B over 1.1 min, then hold at 90% B for 0.6 min; temperature: 50 °C; flow rate: 0.7 mL / min; detection: UV (220 nm)]

[0400] Examples 172 and 173 4-((2S,5R)-4-(2-((cyanomethyl)(methyl)amino)-1-(4-(trifluoromethyl)phenyl)ethyl)-5-ethyl-2-methylpiperazine-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile

Chemical formula

[0401] To a stirred solution of 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl methanesulfonate (500 mg, 0.864 mmol) in anhydrous acetonitrile (5 mL) was added DIPEA (0.3 mL, 1.728 mmol), followed by 2-(methylamino)acetonitrile (182 mg, 2.59 mmol). The resulting reaction mixture was heated at 85 °C for 6 hours. After completion of the reaction, the reaction mixture was diluted with water (100 mL) and extracted with EtOAc (100 mL x 2). The combined organic layers were washed with brine-saturated solution (150 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue thus obtained was purified by flash column chromatography on silica gel (230 - 400 mesh) using 70 - 80% EtOAc in hexane. The material thus obtained was further purified by preparative HPLC (HPLC method: Kinetex EVO C18 (250 x 21.2) mm, 5 micron; mobile phase A: 0.1% TFA in water; mobile phase B: acetonitrile; diluent: acetonitrile: water (50:50); gradient: 20% - 60% B over 0 - 12 minutes; flow rate: 15 mL / min) to obtain Examples 1 and 2 as a mixture of diastereomers. The mixture of diastereomers thus obtained was separated using chiral SFC purification (method: column: Chiralpak AS-H, co-solvent: 30% IPA, flow rate: 4 mL / min) to obtain Examples 172 and 173 as individual diastereomers.

[0402] The fraction of Example 172 was concentrated under reduced pressure, diluted with ACN / H2O (1:1), freeze-dried, and 4-((2S,5R)-4-(2-((cyanomethyl)(methyl)amino)-1-(4-(trifluoromethyl)phenyl)ethyl)-5-ethyl-2-methylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (47 mg, yield 9%) was obtained as an off-white solid. LCMS: m / z = 553.1 (M+H); RT - 1.889; (LCMS method: column: XBridge C8 (50x4.6 mm) 5 μm; mobile phase: A: 0.1% TFA in H2O; mobile phase: B: 0.1% TFA in ACN; flow rate: 1.5 mL / min; HPLC retention times 8.448 and 7.126 minutes (each, HPLC methods FB and FC); 1 H NMR (400 MHz, DMSO-d6): δ 8.24 (d, J = 8.80 Hz, 1H), 7.98 (d, J = 9.20 Hz, 1H), 7.71 (d, J = 8.00 Hz, 2H), 7.59 (d, J = 8.00 Hz, 2H), 6.20 - 5.35 (m, 1H), 5.10 - 4.70 (m, 1H), 3.94 (d, J = 17.20 Hz, 1H), 3.70 - 3.67 (m, 1H), 3.42 (s, 3H), 3.07 (dd, J = 6.40, 13.40 Hz, 1H), 2.81 - 2.77 (m, 2H), 2.40 - 2.30 (m, 1H), 2.28 (s, 3H), 1.34 - 1.12 (m, 6H), 0.89 (t, J = 6.80 Hz, 3H). (-2H)

[0403] The fraction of Example 173 was concentrated under reduced pressure, diluted with ACN / H2O (1:1), freeze-dried, and 4-((2S,5R)-4-(2-((cyanomethyl)(methyl)amino)-1-(4-(trifluoromethyl)phenyl)ethyl)-5-ethyl-2-methylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (53.96 mg, yield 11.09%) was obtained as an off-white solid. LCMS: m / z = 553.2 (M+H); retention time: 1.881; (LCMS method: column: XBridge C8 (50x4.6 mm) 5 μm; mobile phase: A: 0.1% TFA in H2O; mobile phase: B: 0.1% TFA in ACN; flow rate: 1.5 mL / min; HPLC retention times 8.453 and 7.171 minutes (each, HPLC method B and C); 1 H NMR (400 MHz, DMSO-d6): δ 8.23 (d, J = 8.80 Hz, 1H), 7.98 (d, J = 9.20 Hz, 1H), 7.73 (d, J = 8.40 Hz, 2H), 7.59 (d, J = 8.00 Hz, 2H), 6.95 - 5.45 (m, 1H), 5.15 - 4.70 (m, 1H), 3.97 (d, J = 17.20 Hz, 1H), 3.74 - 3.64 (m, 2H), 3.48 (s, 3H), 2.89 (dd, J = 6.00, 13.40 Hz, 1H), 2.80 - 2.76 (m, 2H), 2.70 - 2.60 (m, 1H), 2.28 (s, 3H), 1.40 - 1.15 (m, 6H), 0.91 (t, J = 7.60 Hz, 3H)

[0404] The examples shown in Table 9 were prepared in the synthetic route according to the general procedures described in the preparation of Examples 172 and 173 using appropriate amines. When a mixture of diastereomers was obtained by the reaction, the mixture was separated using a preparative chiral SFC purification method at the final stage. The absolute stereochemistry was not assigned to the newly formed carbon-nitrogen bond.

[0405] Table 9

Table 25

[0406] TIFF0007712274000118.tif58150

[0407] Examples 182 and 183 4-((2S,5R)-4-(2-((Cyanomethyl)(methyl)amino)-1-(4-(trifluoromethyl)phenyl)ethyl)-2,5-diethylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile [Chemical formula]

[0408] To a solution of 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl methanesulfonate (600 mg, 1.012 mmol) in anhydrous acetonitrile (10 mL) in a 20 mL sealed tube was added DIPEA (0.5 mL, 3.04 mmol), followed by 2-(methylamino)acetonitrile (213 mg, 3.04 mmol). The resulting reaction mixture was heated at 85 °C for 6 hours. After completion of the reaction, the reaction mixture was diluted with water (100 mL) and extracted with EtOAc (100 mL x 2). The organic layers were combined, washed with brine-saturated solution (150 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue thus obtained was purified by flash column chromatography on silica gel (230 - 400 mesh) using 70 - 80% EtOAc in hexane. The material obtained was further purified by preparative HPLC (HPLC method: Luna C18 (250 x 21.2) mm, 5 micron; mobile phase: 10 mM ammonium acetate in water / ACN) to obtain a mixture of diastereomers. The mixture of diastereomers was separated using chiral SFC (method: column: Chiralpak AS-H, co-solvent: 30% IPA, flow rate: 4 mL / minute) to obtain Examples 182 and 183 as individual diastereomers.

[0409] The fraction of Example 182 was concentrated under reduced pressure, diluted with ACN / H2O (1:1), lyophilized, and 4-((2S,5R)-4-(2-((cyanomethyl)(methyl)amino)-1-(4-(trifluoromethyl)phenyl)ethyl)-2,5-diethylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (17.5 mg, yield 2.75%) was obtained as an off-white solid. LCMS: m / z = 567.0 (M+H); rt 2.532; LCMS method: column: XBridge C8 (50x4.6) mm, 3.5 μm; mobile phase A: 10 mM NH4HCO3 in H2O; mobile phase B: ACN; flow rate: 1.5 mL / min; HPLC retention times 8.955 and 8.453 minutes (each, HPLC methods FB and FC); 1 H NMR (400 MHz, DMSO-d6): δ 8.22 (d, J = 9.20 Hz, 1H), 7.96 (d, J = 8.80 Hz, 1H), 7.71 (d, J = 8.00 Hz, 2H), 7.60 (d, J = 8.00 Hz, 2H), 6.00 - 5.10 (m, 1H), 5.00 - 4.70 (m, 1H), 3.92 (d, J = 17.20 Hz, 1H), 3.66 - 3.63 (m, 2H), 3.42 (s, 3H), 3.13 - 3.02 (m, 2H), 2.80 - 2.68 (m, 2H), 2.27 (s, 3H), 1.90 - 1.70 (m, 1H), 1.70 - 1.20 (m, 4H), 0.93 - 0.83 (m, 3H), 0.71 - 0.67 (m, 3H)

[0410] The fraction of Example 183 was concentrated under reduced pressure, diluted with ACN / H2O (1:1), lyophilized, and 4-((2S,5R)-4-(2-((cyanomethyl)(methyl)amino)-1-(4-(trifluoromethyl)phenyl)ethyl)-2,5-diethylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (87.22 mg, yield 15.07%) was obtained as an off-white solid. LCMS: m / z = 567.2 (M+H); rt 2.979; LCMS method: column: Kinetex XB-C18 (75x3.0) mm, 2.6 μm; mobile phase: A: 5 mM ammonium formate (pH 3.3): ACN (98:2); mobile phase: B: ACN: buffer (98:2); flow rate: 1.0 mL / min; HPLC retention times 6.193 and 5.753 minutes (for HPLC methods B and C, respectively); 1 H NMR (400 MHz, DMSO-d6): δ 8.24 (d, J = 8.80 Hz, 1H), 7.98 (d, J = 8.80 Hz, 1H), 7.73 (d, J = 8.00 Hz, 2H), 7.57 (d, J = 8.00 Hz, 2H), 6.00 - 5.30 (m, 1H), 4.88 - 4.84 (m, 1H), 3.95 (d, J = 17.20 Hz, 1H), 3.74 - 3.65 (m, 2H), 3.43 (s, 3H), 2.98 - 2.87 (m, 2H), 2.79 (dd, J = 8.00, 13.20 Hz, 1H), 2.70 - 2.63 (m, 2H), 2.23 (s, 3H), 1.85 - 1.50 (m, 2H), 1.45 - 1.25 (m, 2H), 1.00 - 0.70 (m, 6H)

[0411] The examples shown in Table 10 were prepared according to the general procedures described in the preparation of Examples 182 and 183 using the appropriate amine in the synthetic route. When a mixture of diastereomers was obtained by the reaction, the mixture was separated using chiral SFC in the final step. The absolute stereochemistry was not assigned to the newly formed carbon-nitrogen bond.

[0412] Table 10

Table 26

[0413] Intermediates 61 and 62 4-((2S,5R)-4-(2-Amino-1-(4-(trifluoromethyl)phenyl)ethyl)-5-ethyl-2-methylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile

Chemical Structure

[0414] A stirred solution of 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl methanesulfonate (1.3 g, 2.247 mmol) in anhydrous THF (10 mL) was added to 0.5 M ammonia in THF (44.9 mL, 22.47 mmol). The resulting reaction mixture was heated at 70 °C for 16 h. After completion of the reaction, the volatile materials were removed under reduced pressure and the resulting crude reaction mixture was purified by flash column chromatography on silica gel (230 - 400 mesh) eluting with 5 - 10% MeOH / DCM to afford 4-((2S,5R)-4-(2-amino-1-(4-(trifluoromethyl)phenyl)ethyl)-5-ethyl-2-methylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (750 mg, 45.4% yield) as a brown solid. 300 mg of the resulting product was purified again by preparative HPLC (HPLC method: column: XBridge C18 (19x250), 5 micron; mobile phase A: 0.1% FA in water; mobile phase B: 0.1% FA in acetonitrile) to afford Examples 86 and 87 (150 mg, 95% LCMS purity) as a mixture of diastereomers. The resulting mixture of diastereomers was purified by chiral SFC (SFC method: column: Chiralpak AD-H, co-solvent: 0.5% IPA in MeOH, flow rate: 4 mL / min) to afford Intermediate 61 and Intermediate 62 as individual diastereomers.

[0415] The fraction of Intermediate 61 was concentrated and further purified by preparative HPLC (HPLC method: column: XBridge C18 (19x250), 5 μm; mobile phase A: 0.1% formic acid in water; mobile phase B: 0.1% formic acid in acetonitrile). The fraction was evaporated under reduced pressure, diluted with ACN / H2O (1:1), freeze-dried, and 4-((2S,5R)-4-(2-amino-1-(4-(trifluoromethyl)phenyl)ethyl)-5-ethyl-2-methylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile·HCOOH (30 mg, yield 2.036%) was obtained as an off-white solid. LCMS: m / z = 500.0 (M+H); retention time: 1.837; (LCMS method: column: Kinetex XB-C18 (75x3.0) mm, 2.6 μm; mobile phase: A: 5 mM ammonium formate (pH 3.3): ACN (98:2); mobile phase: B: ACN: buffer (98:2); flow rate: 1.0 mL / min; HPLC retention times 6.076 and 7.178 minutes (for HPLC methods FG and FC respectively); 1 H NMR (400 MHz, DMSO-d6): δ 8.56 (s, 1H, formate peak), 8.10 (d, J = 8.80 Hz, 1H), 8.02 (d, J = 8.80 Hz, 1H), 7.74 (d, J = 8.40 Hz, 2H), 7.66 (d, J = 8.40 Hz, 2H), 6.50 - 5.60 (m, 1H), 5.50 - 5.00 (m, 1H), 4.39 (dd, J = 5.20, 8.60 Hz, 1H), 3.60 (s, 3H), 3.09 (dd, J = 4.80, 12.00 Hz, 1H), 2.92 (dd, J = 5.60, 13.40 Hz, 1H), 2.79 - 2.70 (m, 2H), 2.54 (dd, J = 3.60, 12.20 Hz, 1H), 1.80 - 1.30 (m, 5H), 1.05 (t, J = 7.60 Hz, 3H)

[0416] The fraction of intermediate 62 was concentrated under reduced pressure, diluted with ACN / H2O (1:1), lyophilized, and 4 - ((2S,5R)-4-(2 - amino - 1-(4-(trifluoromethyl)phenyl)ethyl)-5 - ethyl - 2 - methylpiperazin - 1 - yl)-1 - methyl - 2 - oxo - 1,2 - dihydropyrido[3,2 - d]pyrimidine - 6 - carbonitrile (15.22 mg, yield 1.311%) was obtained as an off - white solid. LCMS: m / z = 500.3 (M + H); retention time: 2.088; (LCMS method: column: XBridge C8 (50x4.6 mm) 5 μm; mobile phase: A: 10 mM ammonium acetate in H2O; mobile phase: B: ACN; flow rate: 1.5 mL / min; HPLC retention times 5.902 and 8.113 minutes (each, HPLC methods FH and FC); 1 H NMR (400 MHz, DMSO - d6): δ 8.10 (d, J = 8.80 Hz, 1H), 8.02 (d, J = 8.80 Hz, 1H), 7.72 (d, J = 8.00 Hz, 2H), 7.65 (d, J = 8.00 Hz, 2H), 5.50 - 5.00 (m, 1H), 6.50 - 5.60 (m, 1H), 4.28 - 4.24 (m, 1H), 3.60 (s, 3H), 3.01 (dd, J = 4.00, 12.20 Hz, 1H), 2.81 - 2.70 (m, 3H), 2.67 (d, J = 1.60 Hz, 1H), 1.60 - 1.42 (m, 5H), 1.02 (t, J = 7.20 Hz, 3H)

[0417] Examples 192 and 193 N-(2 - ((2R,5S)-4-(6 - cyano - 1 - methyl - 2 - oxo - 1,2 - dihydropyrido[3,2 - d]pyrimidine - 4 - yl)-2 - ethyl - 5 - methylpiperazin - 1 - yl)-2-(4-(trifluoromethyl)phenyl)ethyl)acetamide

Chemical Structure

[0418] To a stirred solution of 4-((2S,5R)-4-(2-amino-1-(4-(trifluoromethyl)phenyl)ethyl)-5-ethyl-2-methylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (350 mg, 0.7 mmol) in anhydrous dichloromethane (10 mL) was added DIPEA (0.61 mL, 0.35 mmol), followed by acetyl chloride (0.1 mL, 1.4 mmol) at 0 °C. The resulting reaction mixture was warmed to room temperature and stirred for 2 h. After completion of the reaction, the reaction mixture was diluted with water (100 mL) and extracted with EtOAc (100 mL x 2). The combined organic layers were washed with brine-saturated solution (150 mL) and dried over sodium sulfate. The organic layer was filtered and concentrated under reduced pressure. The crude product was subjected to flash column chromatography on silica gel (230 - 400 mesh) eluting with 5 - 10% MeOH / DCM, followed by preparative HPLC (method: column: ZORBAX C18 (50 x 21.5) mm, 5 micron, mobile phase A: 10 mM ammonium acetate in water, mobile phase B: acetonitrile, gradient: 90 - 40% A and 10 - 60% B, flow rate: 15 mL / min) to afford N-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl)acetamide. The resulting mixture of diastereomers was subjected to chiral SFC (method: column: Chiralpak RACES C4, co-solvent: 0.5% isopropylamine in IPA, flow rate: 3 mL / min) to afford Example 192 and Example 193 as individual diastereomers.

[0419] The fraction of Example 192 was concentrated under reduced pressure, diluted with ACN / H2O (1:1), freeze-dried, and N-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl)acetamide (32.27 mg, yield 8.39%) was obtained as an off-white solid. LCMS: m / z = 542.0 (M+H); retention time: 1.731; (Method D); HPLC retention times 7.336 and 6.213 minutes (HPLC methods FB and FC, respectively); 1 H NMR (400 MHz, DMSO-d6): δ 8.34 (d, J = 8.40 Hz, 1H), 8.24 (d, J = 9.20 Hz, 1H), 7.99 (d, J = 8.80 Hz, 1H), 7.71 (d, J = 8.00 Hz, 2H), 7.59 (d, J = 8.00 Hz, 2H), 6.00 - 5.50 (m, 1H), 5.05 - 5.00 (m, 2H), 3.44 (s, 3H), 2.85 (dd, J = 4.40, 12.00 Hz, 1H), 2.76 (dd, J = 7.20, 12.80 Hz, 1H), 2.69 - 2.64 (m, 1H), 2.70 - 2.60 (m, 1H), 2.50 - 2.45 (m, 1H), 1.87 (s, 3H), 1.50 - 1.20 (m, 5H), 0.91 (t, J = 7.60 Hz, 3H)

[0420] The fraction of Example 193 was concentrated under reduced pressure, diluted with ACN / H2O (1:1), freeze-dried, and N-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl)acetamide (27.02 mg, yield 7.02%) was obtained as an off-white solid. LCMS: m / z = 544.2 (M+H); retention time: 1.746; (LCMS Method D). HPLC retention time 7.444 (HPLC Method B); 11H NMR (400 MHz, DMSO-d6): δ 8.32 (d, J = 8.00 Hz, 1H), 8.25 (d, J = 8.80 Hz, 1H), 7.99 (d, J = 9.20 Hz, 1H), 7.69 (d, J = 8.00 Hz, 2H), 7.57 (d, J = 8.00 Hz, 2H), 6.00 - 5.45 (m, 1H), 5.03 - 4.97 (m, 2H), 3.44 (s, 3H), 2.84 (dd, J = 4.00, 12.00 Hz, 1H), 2.73 - 2.63 (m, 3H), 1.90 (s, 3H), 1.42 - 1.08 (m, 6H), 0.88 (t, J = 7.20 Hz, 3H)

[0421] The examples shown in Table 12 were prepared according to the general procedures described in the preparation of Examples 192 and 193 using an appropriate amine in the synthetic route. When a mixture of diastereomers was obtained by the reaction, the mixture was separated using chiral SFC purification at the final stage. The absolute stereochemistry was not assigned to the newly formed carbon-nitrogen bond.

[0422] Table 12

Table 27

[0423] Intermediates 63 and 64 4 - ((2S,5R)-4-(2-Amino-1-(4-(trifluoromethyl)phenyl)ethyl)-2,5-diethylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile

Chemical formula

[0424] To a stirred solution of 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl methanesulfonate (3.5 g, 5.91 mmol) in anhydrous THF (50 mL) was added 0.5 M ammonia in THF (118 mL, 59.1 mmol). The resulting reaction mixture was heated at 70 °C for 16 h. After completion of the reaction, the volatile materials were removed under reduced pressure to afford crude 4-((2S,5R)-4-(2-amino-1-(4-(trifluoromethyl)phenyl)ethyl)-2,5-diethylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (3.0 g, 75% yield). 300 mg of the resulting product was purified again by preparative HPLC (HPLC method: column: XSelect C18 (150x19) 5.0 micron; mobile phase A: 10 mM ammonium acetate in water; mobile phase B: acetonitrile) to give a mixture of diastereomers. The resulting mixture of diastereomers was subjected to chiral SFC (method: column: Chiralpak-(R,R) Whelk-01, co-solvent: 0.5% IPA in MeOH, flow rate: 3 mL / min) for purification to obtain Intermediate 63 and Intermediate 64 as individual diastereomers.

[0425] The fraction of Intermediate 63 was concentrated, diluted with ACN / H2O (1:1), lyophilized, and ((2S,5R)-4-(2-amino-1-(4-(trifluoromethyl)phenyl)ethyl)-2,5-diethylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (26.09 mg, yield 8.6%) was obtained as a light brown solid. LCMS: m / z = 514.0 (M+H); retention time: 1.830; (method: column: Kinetex XB-C18 (75x3.0) mm, 2.6 μm; mobile phase: A: 5 mM ammonium formate (pH 3.3): ACN (98:2); mobile phase: B: ACN: buffer (98:2); flow rate: 1.0 mL / min); HPLC retention times 7.801 and 6.305 minutes (each, HPLC methods FH and FI); 1 H NMR (400 MHz, DMSO-d6): δ 8.25 (d, J = 8.80 Hz, 1H), 7.99 (d, J = 8.80 Hz, 1H), 7.72 - 7.58 (m, 4H), 6.00 - 5.30 (m, 1H), 5.05 - 4.80 (m, 1H), 4.08 (t, J = 6.40 Hz, 1H), 3.44 (s, 3H), 2.83 (dd, J = 4.00, 12.00 Hz, 1H), 2.70 - 2.60 (m, 3H), 2.20 - 1.90 (m, 2H), 1.88 - 1.70 (m, 2H), 1.50 - 1.30 (m, 2H), 1.00 - 0.70 (m, 6H)

[0426] The fraction of Intermediate 64 was concentrated, diluted with ACN / H2O (1:1), lyophilized, and ((2S,5R)-4-(2-amino-1-(4-(trifluoromethyl)phenyl)ethyl)-2,5-diethylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (13.6 mg, yield 4.53%) was obtained as a light brown solid. LCMS: m / z = 514.0 (M+H); retention time: 1.850; (LCMS method: column: Kinetex XB-C18 (75x3.0) mm, 2.6 μm; mobile phase: A: 5 mM ammonium formate (pH 3.3):ACN (98:2); mobile phase: B: ACN:buffer (98:2); flow rate: 1.0 mL / min; HPLC retention times 5.480 and 4.588 minutes (each, HPLC methods FB and FC); 1 H NMR (400 MHz, DMSO-d6): δ 8.25 (d, J = 8.80 Hz, 1H), 8.00 (d, J = 8.00 Hz, 1H), 7.68 - 7.59 (m, 4H), 6.00 - 5.30 (m, 1H), 5.00 - 4.90 (m, 1H), 4.09 (dd, J = 4.80, 9.00 Hz, 1H), 3.44 (s, 3H), 2.80 - 2.60 (m, 3H), 2.45.00 - 2.35 (m, 2H), 2.30 - 2.00 (m, 3H), 1.90 - 1.70 (m, 2H), 1.00 - 0.70 (m, 6H)

[0427] Examples 202 and 203 N-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl)acetamide

Chemical Structure

[0428] To a stirred solution of 4-((2S,5R)-4-(2-amino-1-(4-(trifluoromethyl)phenyl)ethyl)-2,5-diethylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (400 mg, 0.78 mmol) in anhydrous dichloromethane (5 mL) was added DIPEA (0.41 mL, 2.34 mmol), followed by acetyl chloride (0.09 mL, 1.17 mmol) at 0 °C. The resulting reaction mixture was warmed to room temperature and stirred for 2 hours. After completion of the reaction, the reaction mixture was diluted with DCM (50 mL) and washed with saturated sodium bicarbonate (50 mL) and then brine solution (50 mL), and dried over sodium sulfate. Then the organic layer was filtered and concentrated under reduced pressure. The crude product was purified by preparative HPLC (method: Kinetex EVO C18 (250x21.2) mm, 5 micron, mobile phase A: 10 mM ammonium acetate in water, mobile phase B: acetonitrile, gradient: 60-10% A and 40-90% B, flow rate: 15 mL / min) to give N-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl)acetamide as a mixture of diastereomers. The resulting mixture of diastereomers was purified by chiral SFC (method: column: Chiralpak Lux A1, co-solvent: IPA, flow rate: 3 mL / min) to give Example 202 and Example 203 as individual diastereomers.

[0429] The fraction of Example 202 was concentrated under reduced pressure, diluted with ACN / H2O (1:1), freeze-dried, and N-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl)acetamide (52.21 mg, yield 11.97%) was obtained as an off-white solid. LCMS: m / z = 556.0 (M+H); retention time: 2.115; (LCMS method A); HPLC retention times 7.190 and 6.840 minutes (HPLC methods FH and FI, respectively); 1 H NMR (400 MHz, DMSO-d6): δ 8.30 (d, J = 8.40 Hz, 1H), 8.23 (d, J = 9.20 Hz, 1H), 7.98 (d, J = 8.80 Hz, 1H), 7.71 (d, J = 8.40 Hz, 2H), 7.58 (d, J = 8.00 Hz, 2H), 6.00 - 5.20 (m, 1H), 5.04 - 4.99 (m, 1H), 4.95 - 4.80 (m, 1H), 3.44 (s, 3H), 3.15 - 3.00 (m, 1H), 2.80 - 2.71 (m, 2H), 2.69 - 2.61 (m, 2H), 2.20 - 1.90 (m, 1H), 1.88 (s, 3H), 1.80 - 1.65 (m, 1H), 1.41 - 1.37 (m, 2H), 1.00 - 0.70 (m, 6H)

[0430] The fraction of Example 203 was concentrated under reduced pressure, diluted with ACN / H2O (1:1), freeze-dried, and N-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl)acetamide (34.77 mg, yield 7.91%) was obtained as an off-white solid. LCMS: m / z = 556.2 (M+H); retention time: 2.131; (LCMS method A); HPLC retention times 7.218 and 6.846 (HPLC methods FH and FI, respectively); 11H NMR (400 MHz, DMSO-d6): δ 8.28 (d, J = 7.20 Hz, 1H), 8.24 (d, J = 8.80 Hz, 1H), 7.99 (d, J = 8.80 Hz, 1H), 7.69 (d, J = 8.00 Hz, 2H), 7.57 (d, J = 8.00 Hz, 2H), 6.00 - 5.30 (m, 1H), 5.10 - 4.30 (m, 2H), 3.44 (s, 3H), 2.90 - 2.60 (m, 4H), 1.90 (s, 3H), 1.80 - 1.60 (m, 1H), 1.45 - 1.25 (m, 3H), 1.00 - 0.70 (m, 6H)

[0431] The examples shown in Table 13 were produced in the synthetic route according to the general procedures described in the production of Examples 202 and 203 using appropriate amines. When a mixture of diastereomers was obtained by the reaction, the mixture was separated using chiral SFC purification at the final stage. The absolute stereochemistry was not assigned to the newly formed carbon-nitrogen bond.

[0432] Table 13

Table 28

[0433] Biological assay The pharmacological properties of the compounds of the present invention can be confirmed by a number of biological assays. The following biological assays exemplified were performed using the compounds of the present invention.

[0434] Assay 1: In vitro DGK inhibition assay (Method A) The reactions of DGKα and DGKζ were performed using either extruded liposomes (LIPGLO assay for DGKα and DGKζ) or surfactant / lipid micelle substrates (DGKα and DGKζ assays). The reactions were carried out in 50 mM MOPS pH 7.5, 100 mM NaCl, 10 mM MgCl2, 1 μM CaCl2, and 1 mM DTT (assay buffer). The reactions using surfactant / lipid micelle substrates also contained 50 mM octyl B-D-glucopyranoside. The lipid substrate concentrations were 11 mM PS and 1 mM DAG for the surfactant / lipid micelle reaction. The lipid substrate concentrations were 2 mM PS, 0.25 mM DAG, and 2.75 mM PC for the extruded liposome reaction. The reactions were conducted in 150 μM ATP. The enzyme concentration was 5 nM for both DGKα and DGKζ.

[0435] The inhibition experiments of the compounds were conducted as follows: 50 nL droplets of each test compound dissolved in DMSO (serial 3-fold dilutions from a maximum concentration of 10 mM for each compound at 11 points) were transferred to the wells of a white 1536-well plate (Corning 3725). A 5 mL enzyme / substrate solution was prepared at 2x the final reaction concentration by combining 2.5 mL of 4x enzyme solution (20 nM DGKα or DGKζ in assay buffer (prepared as described below)) and either 2.5 mL of 4x liposome solution or 4x surfactant / lipid micelle solution (composition described below), and incubated at room temperature for 10 minutes. Next, 1 μL of the 2x enzyme / substrate solution was added to the wells containing the test compound, and 1 μL of 300 μM ATP was added to initiate the reaction. The reaction was allowed to proceed for 1 hour, after which 2 μL of Glo reagent (Promega V9101) was added and incubated for 40 minutes. Then, 4 μL of kinase detection reagent was added and incubated for 30 minutes. Luminescence was recorded using an EnVision microplate reader. % Inhibition was calculated from the ATP conversion generated by setting the enzyme-free control reaction as 100% inhibition and the vehicle-only reaction as 0% inhibition. The compounds were evaluated at 11 concentrations, and the IC 50 was determined.

[0436] 4x surfactant / lipid micelle preparation Surfactant / lipid micelles were prepared by combining 15 g of phosphatidylserine (Avanti 840035P) and 1 g of diacylglycerol (800811O) in a 2 L round-bottom flask and dissolving them in 150 mL of chloroform. The chloroform was removed under high vacuum using a rotary evaporator. The resulting colorless viscous oil was resuspended by vigorous mixing in 400 mL of 50 mM MOPS pH 7.5, 100 mM NaCl, 20 mM NaF, 10 mM MgCl2, 1 μM CaCl2, 1 mM DTT, and 200 mM octyl glucoside. The lipid / surfactant solution was aliquoted into 5 mL portions and stored at -80 °C.

[0437] Preparation of 4x liposomes The lipid composition in the 4x liposome solution was 5 mol% DAG (Avanti 800811O), 40 mol% PS (Avanti 840035P), and 55 mol% PC (Avanti 850457), and the total lipid concentration was 15.2 mg / mL. PC, DAG, and PS were dissolved in chloroform, combined, and dried under vacuum to form a thin film. The lipids were neutralized to 20 mM in 50 mM MOPS pH 7.5, 100 mM NaCl, 5 mM MgCl2, and freeze-thaw cycles were repeated 5 times. The lipid suspension was extruded 11 times through a 100 nm polycarbonate filter. Dynamic light scattering was performed to confirm the size of the liposomes (radius 50 - 60 nm). The liposome preparation was stored at 4 °C for a long period of 4 weeks.

[0438] Baculovirus expression in human DGKα and DGKζ Baculovirus samples of human DGK-alpha-TVMV-His-pFB gate and human DGK-zeta-transcription variant-2-TVMV-His-pFB gate were prepared according to the manufacturer's protocol using the Bac-to-Bac® baculovirus expression system (Invitrogen). The DNAs used for the expression of DGK-alpha and DGK-zeta were SEQ ID NO: 1 and SEQ ID NO: 3, respectively. The amplification of baculovirus was performed using Sf9 cells infected at a virus / cell ratio of 1:1500 and grown at 27°C for 65 hours after gene introduction. The scale-up of the expression of each protein was performed in a Cellbag 50L WAVE bioreactor system 20 / 50 (manufactured by GE Healthcare Bioscience). 12 L of Sf9 cells (Expression System, Davis, CA) at 2x10 6 cells / mL were infected with a virus stock solution at a virus / cell ratio of 1:200 and grown at 27°C for 66 to 68 hours after infection. The infected cell culture was collected by centrifugation using a SORVALL® RC12BP centrifuge (2000 rpm, 20 minutes, 4°C). The cell pellet was stored at -70°C until purification.

[0439] Purification of Human DGK-alpha and DGK-zeta Each full-length human DGKα and DGKζ, which are capable of TVMV cleavage, contain a C-terminal Hex-His tag sequence (SEQ ID NO: 2 and 4 respectively), are expressed and produced as described above, and were purified from Sf9 baculovirus-infected insect cell paste. The cells were lysed with a nitrogen cavitation method using a nitrogen disruptor (Parr Instruments), and the lysate was clarified by centrifugation. The clarified lysate was purified using three consecutive column chromatography steps on an AKTA Purifier Plus system until a homogeneity of approximately 90% was achieved. The three-step column chromatography included nickel affinity resin capture (i.e., HisTrap FF crude, GE Healthcare), followed by size exclusion chromatography (i.e., HiLoad 26 / 600 Superdex 200 prep grade for DGK-α, GE Healthcare; HiPrep 26 / 600 Sephacryl S 300_HR for DGK-ζ, GE Healthcare). The third step was ion exchange chromatography, which differed for the two isoforms. DGKα was refined using Q-Sepharose anion exchange chromatography (GE Healthcare). DGKζ was refined using SP-Sepharose cation exchange chromatography (GE Healthcare). The protein was > delivered at a concentration of 2 mg / mL. The formulation buffer was the same for both proteins and was 50 mM Hepes, pH 7.2, 500 mM NaCl, 10% v / v glycerol, 1 mM TCEP, and 0.5 mM EDTA.

[0440] Assay 2: In Vitro DGK Inhibition Assay (Method B) The reactions of DGKα and DGKζ were carried out using either extruded liposomes (LIPGLO assay for DGKα and DGKζ) or a surfactant / lipid micelle substrate (DGKα and DGKζ assay). The reactions were performed in 50 mM MOPS pH 7.5, 100 mM NaCl, 10 mM MgCl2, 1 μM CaCl2, and 1 mM DTT (assay buffer). The reaction using the surfactant / lipid micelle substrate also contained 50 mM octyl B-D-glucopyranoside. The lipid substrate concentrations were 11 mM PS and 1 mM DAG for the surfactant / lipid micelle reaction. The lipid substrate concentrations for the extruded liposome reaction (total 5 mM lipid) were 2 mM PS, 0.25 mM DAG, and 2.75 mM PC. The reactions were carried out in 150 μM ATP. The enzyme concentration was 5 nM for both DGKα and DGKζ.

[0441] The inhibition experiments of the compounds were carried out as follows: 25 nL droplets of each test compound dissolved in DMSO (serial 3-fold dilutions from a maximum concentration of 10 mM for each compound at 11 points) were transferred to the wells of a white 1536-well plate (Corning 3725). A 5 mL enzyme / lipid substrate solution was prepared at 2x the final reaction concentration by combining 2.5 mL of 4x enzyme solution (20 nM DGKα or DGKζ in assay buffer, prepared as described below) and 2.5 mL of 4x liposome or 4x surfactant / lipid micelle solution (the following compositions), and incubated at room temperature for 10 minutes. Next, 1 μL of the 2x enzyme / lipid substrate solution was added to the wells containing the test compound, and 1 μL of 300 μM ATP was added to initiate the reaction. The reaction was allowed to proceed for 2 hours, after which 2 μL of Glo reagent (Promega V9101) was added and incubated for 40 minutes. Then, 4 μL of kinase detection reagent was added and incubated for 30 minutes. Luminescence was recorded using an Envision microplate reader. % Inhibition was calculated from the ATP conversion generated by setting the enzyme-free control reaction as 100% inhibition and the vehicle-only reaction as 0% inhibition. The compounds were evaluated at 11 concentrations and the IC 50 was determined.

[0442] 4x Surfactant / Lipid Micelle Preparation Surfactant / Lipid micelles were prepared by combining 15 g of phosphatidylserine (Avanti 840035P) and 1 g of diacylglycerol (800811O) in a 2 L round-bottom flask and dissolving them in 150 mL of chloroform. The chloroform was removed under high vacuum using a rotary evaporator. The resulting colorless viscous oil was resuspended by vigorous mixing in 400 mL of 50 mM MOPS pH 7.5, 100 mM NaCl, 20 mM NaF, 10 mM MgCl2, 1 μM CaCl2, 1 mM DTT, and 200 mM octyl glucoside. The lipid / surfactant solution was aliquoted into 5 mL portions and stored at -80 °C.

[0443] Preparation of 2x Liposomes The lipid composition in the liposome solution was 5 mol% DAG (Avanti 800811O), 40 mol% PS (Avanti 840035P), and 55 mol% PC (Avanti 850457), and the overall lipid concentration was 7 - 8 mg / mL. PC, DAG, and PS were dissolved in chloroform, combined, and dried under vacuum to form a thin film. The lipid was hydrated to 20 mM in 50 mM MOPS, pH 7.5, 100 mM NaCl, 5 mM MgCl2, and 5 freeze-thaw cycles were repeated. The lipid suspension was extruded 10 - 12 times through a 100 nm polycarbonate filter. Dynamic light scattering was performed to confirm the size of the liposomes (radius 50 - 60 nm). The liposome preparation was stored at 4 °C for a long period of 4 weeks.

[0444] Baculovirus Expression of Near-Full-Length Human DGKα and Full-Length Human DGKζ Baculovirus samples of the human MA-hDGKα-(S9-S727)-Ct-TVMV-His-pFB gate and the full-length human DGK-ζ-transcript variant-2-TVMV-His-pFB gate were prepared according to the manufacturer's protocol using the Bac-to-Bac baculovirus expression system (Invitrogen) (the "MA-" in the name of the DGKα reagent indicates that two additional amino acids were added before Ser-9). The DNA sequences used for the expression of DGKα(9-727) and DGKζ are SEQ ID NO: 5 and SEQ ID NO: 3, respectively. Amplification of the baculovirus was achieved using Sf9 cells infected at a virus / cell ratio of 1:1500 and grown at 27 °C for 65 hours after transfection.

[0445] Scaling up the expression for the near-full-length DGK-α(9-727) protein was performed in flasks (2 L), and for the full-length DGKζ, it was performed using a Cellbag 50L WAVE bioreactor system (GE Healthcare Bioscience). The proteins were expressed at different volumes using similar conditions. For th...

Claims

1. Formula (I): 【Chemical 1】 [wherein: X is N; Y is CR ３ or N; R １ is Cl or -CN; R ２ is CH ３ and; R ３ is H; R ４ is -CR ４ａ R ４ｃ -L-R ４ｂ and is; L is L １ or -(CH ２ ) １－２ L １ -; L １ is -C(O)-, -C(O)O-, -C(O)NR ｄ -, -C(O)NHO-, -C(O)NHSO ２ -, -CH ２ C(O)-, -CH ２ NR ｄ -, -CH ２ NHCO-, -CH ２ NHCOO-, -CH ２ NHSO ２ -, -CH ２ COO-, -CH ２ CONR ｄ -, -CH ２ CH ２ NHCOO-, -CH ２ CH ２ NHSO ２ -, or -P(O)(CH ３ )-; R ４ａ is (i) -CH ３ or; or (ii) phenyl or pyridinyl, each substituted with 0 to 2 substituents independently selected from F, -CH ３ , -CF ３ , and -OCH ３ ; R ４ｂ is (i) H, -CN, C １－４ alkyl, -CH ２ CN, -CH ２ CHF ２ 、 -CH ２ CF ３ 、 -CH ２ CH ２ OH, -CH ２ CH ２ CN, -CH ２ CH ２ CF ３ 、 -CH ２ CH ２ OCH ３ 、 -CH ２ CH ２ S(O) ２ CH ３ 、 -CH ２ C(CH ３ ) ２ OH, -CH ２ (cyclopropyl), -CH ２ (hydroxycyclopropyl), 【Chemical Formula 2】 or (ii) azetidinyl, cyclopropyl, cyclopentyl, morpholinyl, pyrazolyl, isoxazolyl, piperidinyl, piperazinyl, pyrazolyl, pyrrolidinyl, dioxido thiomorpholinyl, or azaspiro[2.5]octanyl, each being substituted with 0 to 2 R ｂ and substituted with; Each R ｂ is independently F, -OH, -CH ３ , -CH ２ OH, -C(CH ３ ) ２ OH, -CH ２ OCH ３ , -OCH ３ , or -S(O) ２ CH ３ ; R ４ｃ is H; Each R ６ is H; R ｄ is, independently, H or -CH ３ ; Each R ５ is independently H, -CH ３ , -CH ２ CH ３ , -CH ２ OH, or -CH ２ OCH ３ ; m is 0, 1, or 2] A compound represented thereby or a salt thereof.

2. L is L １ The compound or a salt thereof according to claim 1, which is represented by being.

3. L is -(CH ２ ) １－２ L １ - is, The compound or a salt thereof according to Claim 1.

4. Structural formula: 【Chemical 3】 [wherein, R ５ａ and R ５ｃ are each independently selected from R ５ ​ The compound or a salt thereof according to Claim 1, represented thereby.

5. Structural formula: [Chemical Formula 4] The compound or a salt thereof according to Claim 1, represented thereby.

6. L １ The compound or a salt thereof according to claim 1, wherein L is -C(O)- or -C(O)O-.

7. L １ is -C(O)-, -C(O)O-, or -C(O)NR ｄ -, the compound or a salt thereof according to claim 1.

8. L １ is -P(O)(CH ３ )-, the compound or a salt thereof according to claim 1.

9. Isopropyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate (Example 1-2); Methyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(5-fluoropyridin-2-yl)acetate (Example 3-4); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-N-(2,2-difluoroethyl)-2-(4-fluorophenyl)acetamide (Example 5-6); Methyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate (Example 7-8); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetic acid (Example 9-10); (8-((2S,5R)-4-(1-(4-fluorophenyl)-2-oxo-2-(piperidin-1-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (Example 11-12); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetamide (Example 13-14); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)-N-(1-methyl-1H-pyrazol-3-yl)acetamide (Examples 15-16); 8-((2S,5R)-4-(1-(4-fluorophenyl)-2-morpholino-2-oxoethyl)-2,5-dimethylpiperazin-1-yl)-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (Examples 17-18); 8-((2S,5R)-4-(1-(4-fluorophenyl)-2-(4-methylpiperazin-1-yl)-2-oxoethyl)-2,5-dimethylpiperazin-1-yl)-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (Examples 19-20); 8-((2S,5R)-4-(1-(4-fluorophenyl)-2-oxo-2-(pyrrolidin-1-yl)ethyl)-2,5-dimethylpiperazin-1-yl)-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (Examples 21-22); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)-N,N-dimethylacetamide (Examples 23-24); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)-N-methylacetamide (Examples 25-26); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)-N-(2-hydroxyethyl)acetamide (Examples 27-28); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)-N-(isoxazol-3-yl)acetamide (Examples 29-30); Methyl 3-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-3-(4-fluorophenyl)propanoate (Examples 31-32); N-(tert-Butyl)-3-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-3-(4-fluorophenyl)propanamide (Examples 33-34); 3-[(2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl]-3-(4-fluorophenyl)-N-(propan-2-yl)propanamide (Example 35); 3-[(2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl]-3-(4-fluorophenyl)-N-(propan-2-yl)propanamide (Example 36); 3-[(2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl]-3-(4-fluorophenyl)propanamide (Example 37); 3-[(2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl]-3-(4-fluorophenyl)propanamide (Example 38); 3-[(2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl]-3-(4-fluorophenyl)-N-methylpropanamide (Example 39); 3-[(2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl]-3-(4-fluorophenyl)-N-methylpropanamide (Example 40); 3-[(2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl]-3-(4-fluorophenyl)-N-methyl-N-(propan-2-yl)propanamide (Example 41); 3-[(2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl]-3-(4-fluorophenyl)-N-methyl-N-(propan-2-yl)propanamide (Example 42); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-N-cyclopentylpropanamide (Examples 43-44); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-N-(1-methyl-1H-pyrazol-4-yl)propanamide (Example 45); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-N-(1-methyl-1H-pyrazol-4-yl)propanamide (Example 46); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-N-cyclopropylpropanamide (Example 47); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-N-cyclopropylpropanamide (Example 48); 8-((2S,5R)-4-((dimethylphosphoryl)(4-fluorophenyl)methyl)-2,5-dimethylpiperazin-1-yl)-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (Examples 49-50); Ethyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetate (Example 51); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetic acid (Example 52); 4-((2S,5R)-2,5-Diethyl-4-(2-morpholino-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (Examples 53-54); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-N-(2-methoxyethyl)-2-(4-(trifluoromethyl)phenyl)acetamide (Examples 55-56); 3-(3-(But-3-yn-1-yl)-3H-diazirin-3-yl)propyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate (Example 57); Methyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-methoxyphenyl)acetate (Example 58); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-methoxyphenyl)acetic acid (Example 59); 3-(3-(But-3-yn-1-yl)-3H-diazirin-3-yl)propyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-methoxyphenyl)acetate (Examples 60 - 61); 2-(3-Methyl-3H-diazirin-3-yl)ethyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate (Example 62); Ethyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetate (Examples 69 - 70); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetic acid (Examples 71 - 72); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-N-((1-hydroxycyclopropyl)methyl)-2-(4-(trifluoromethyl)phenyl)acetamide (Examples 73 - 74); 4-((2S,5R)-2,5-diethyl-4-(2-morpholino-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-6-(λ ２ -methyl)-1-methylpyrido[3,2-d]pyrimidin-2(1H)-one (Examples 75-76); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-N-(2-methoxyethyl)-2-(4-(trifluoromethyl)phenyl)acetamide (Examples 77 - 78); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-N-(2-hydroxy-2-methylpropyl)-2-(4-(trifluoromethyl)phenyl)acetamide (Examples 79 - 80); 4-((2S,5R)-2,5-Diethyl-4-(2-(3-hydroxy-3-methylazetidin-1-yl)-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (Examples 81-82); 4-((2S,5R)-2,5-Diethyl-4-(2-oxo-2-(6-azaspiro[2.5]octan-6-yl)-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (Examples 83-84); 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-4-yl)-2,5-diethylpiperazin-1-yl)-N-(cyclopropylmethyl)-N-methyl-2-(4-(trifluoromethyl)phenyl)acetamide (Examples 85-86); 4-((2S,5R)-2,5-Diethyl-4-(2-(3-(hydroxymethyl)azetidin-1-yl)-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (Examples 87-88); 4-((2S,5R)-2,5-Diethyl-4-(2-(3-(2-hydroxypropan-2-yl)azetidin-1-yl)-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (Examples 89-90); 4-((2S,5R)-2,5-Diethyl-4-(2-(3-(methylsulfonyl)azetidin-1-yl)-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (Examples 91-92); 4-(((2S,5R)-2,5-Diethyl-4-(2-(4-(methylsulfonyl)piperidin-1-yl)-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (Examples 93-94); 4-(((2S,5R)-4-(2-(3,3-Difluoroazetidin-1-yl)-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)-2,5-diethylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (Examples 95-96); 4-(((2S,5R)-4-(2-(1,1-Dioxidothiomorpholino)-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)-2,5-diethylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (Examples 97-98); 4-(((2S,5R)-2,5-Diethyl-4-(2-(3-methoxy-3-methylazetidin-1-yl)-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (Examples 99-100); 4-(((2S,5R)-2,5-Diethyl-4-(2-((S)-3-Hydroxypyrrolidin-1-yl)-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (Examples 101-102); 4-(((2S,5R)-2,5-Diethyl-4-(2-((S)-2-(Hydroxymethyl)pyrrolidin-1-yl)-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (Examples 103-104); 4-((2S,5R)-2,5-Diethyl-4-(2-((R)-2-(hydroxymethyl)pyrrolidin-1-yl)-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (Examples 105-106); 4-((2S,5R)-2,5-Diethyl-4-(2-((R)-3-hydroxypyrrolidin-1-yl)-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (Examples 107-108); 4-((2S,5R)-2,5-Diethyl-4-(2-((S)-2-(methoxymethyl)pyrrolidin-1-yl)-2-oxo-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (Examples 109-110); 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-4-yl)-2,5-diethylpiperazin-1-yl)-N-isopropyl-N-methyl-2-(4-(trifluoromethyl)phenyl)acetamide (Examples 111-112); 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-4-yl)-2,5-diethylpiperazin-1-yl)-N-isopropyl-2-(4-(trifluoromethyl)phenyl)acetamide (Examples 113-114); 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-4-yl)-2,5-diethylpiperazin-1-yl)-N-(2-methoxyethyl)-N-methyl-2-(4-(trifluoromethyl)phenyl)acetamide (Examples 115-116); 2-((2R,5S)-4-(6-Cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-4-yl)-2,5-diethylpiperazin-1-yl)-N-methoxy-2-(4-(trifluoromethyl)phenyl)acetamide (Examples 117-118); N-Cyano-2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetamide (Examples 119 - 120); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-N-(cyanomethyl)-2-(4-(trifluoromethyl)phenyl)acetamide (Examples 121 - 122); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-N-(2,2,2-trifluoroethyl)-2-(4-(trifluoromethyl)phenyl)acetamide (Examples 123 - 124); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-N-(2-cyanoethyl)-2-(4-(trifluoromethyl)phenyl)acetamide (Examples 125 - 126); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)-N-(3,3,3-trifluoropropyl)acetamide (Examples 127 - 128); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-N-(2-(methylsulfonyl)ethyl)-2-(4-(trifluoromethyl)phenyl)acetamide (Examples 129 - 130); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-N-methoxy-2-(4-(trifluoromethyl)phenyl)acetamide (Examples 131 - 132); Ethyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetate (Example 133); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetic acid (Example 134); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-N-methoxy-2-(4-(trifluoromethyl)phenyl)acetamide (Examples 135 - 136); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-N-(cyanomethyl)-2-(4-(trifluoromethyl)phenyl)acetamide (Examples 137 - 138); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-N-(2,2,2-trifluoroethyl)-2-(4-(trifluoromethyl)phenyl)acetamide (Examples 139 - 140); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-N-(2-cyanoethyl)-2-(4-(trifluoromethyl)phenyl)acetamide (Examples 141 - 142); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)-N-(3,3,3-trifluoropropyl)acetamide (Examples 143 - 144); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-N-(2-(methylsulfonyl)ethyl)-2-(4-(trifluoromethyl)phenyl)acetamide (Examples 145-146); N-cyano-2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)acetamide (Examples 147-148); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-N-(methylsulfonyl)-2-(4-(trifluoromethyl)phenyl)acetamide (Examples 149-150); 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-N-(cyclopropylsulfonyl)-2-(4-(trifluoromethyl)phenyl)acetamide (Examples 151-152); Methyl 3-((2R,5S)-4-(6-chloro-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-3-(4-(trifluoromethyl)phenyl)propanoate (Example 153); Methyl 3-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-3-(4-(trifluoromethyl)phenyl)propanoate (Examples 154-155); 3-((2R,5S)-4-(6-chloro-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-3-(4-(trifluoromethyl)phenyl)propanoic acid (Example 156); 3-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-3-(4-(trifluoromethyl)phenyl)propanoic acid (Examples 157-158); 4-((2S,5R)-2,5-diethyl-4-(3-oxo-3-(6-azaspiro[2.5]octan-6-yl)-1-(4-(trifluoromethyl)phenyl)propyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (Examples 159-160); 3-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-N-methyl-N-(1-methylpiperidin-4-yl)-3-(4-(trifluoromethyl)phenyl)propanamide (Examples 161-162); 3-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-N-(2-methoxyethyl)-N-methyl-3-(4-(trifluoromethyl)phenyl)propanamide (Examples 163-164); 4-((2S,5R)-2,5-diethyl-4-(3-((R)-2-(methoxymethyl)pyrrolidin-1-yl)-3-oxo-1-(4-(trifluoromethyl)phenyl)propyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (Examples 165-166); Methyl (3-((2R,5S)-4-(6-chloro-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-3-(4-(trifluoromethyl)phenyl)propyl)carbamate (Example 167); 3-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-3-(4-(trifluoromethyl)phenyl)propanoic acid (Examples 168-169); N-(3-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-3-(4-(trifluoromethyl)phenyl)propyl)methanesulfonamide (Examples 170 - 171); 4-((2S,5R)-4-(2-((cyanomethyl)(methyl)amino)-1-(4-(trifluoromethyl)phenyl)ethyl)-5-ethyl-2-methylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (Examples 172 - 173); 4-((2S,5R)-4-(2-((cyanomethyl)amino)-1-(4-(trifluoromethyl)phenyl)ethyl)-5-ethyl-2-methylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (Examples 174 - 175); 4-((2S,5R)-5-ethyl-2-methyl-4-(2-((2,2,2-trifluoroethyl)amino)-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (Examples 176 - 177); 4-((2S,5R)-4-(2-((2-cyanoethyl)amino)-1-(4-(trifluoromethyl)phenyl)ethyl)-5-ethyl-2-methylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (Examples 178 - 179); 4-((2S,5R)-5-ethyl-2-methyl-4-(1-(4-(trifluoromethyl)phenyl)-2-((3,3,3-trifluoropropyl)amino)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (Examples 180 - 181); 4-((2S,5R)-4-(2-((cyanomethyl)(methyl)amino)-1-(4-(trifluoromethyl)phenyl)ethyl)-2,5-diethylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (Examples 182 - 183); 4-(((2S,5R)-4-(2-((cyanomethyl)(methyl)amino)-1-(4-(trifluoromethyl)phenyl)ethyl)-2,5-diethylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (Examples 184 - 185); 4-(((2S,5R)-2,5-diethyl-4-(2-(((2,2,2-trifluoroethyl)amino)-1-(4-(trifluoromethyl)phenyl)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (Examples 186 - 187); 4-(((2S,5R)-4-(2-((2-cyanoethyl)amino)-1-(4-(trifluoromethyl)phenyl)ethyl)-2,5-diethylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (Examples 188 - 189); 4-(((2S,5R)-2,5-diethyl-4-(1-(4-(trifluoromethyl)phenyl)-2-(((3,3,3-trifluoropropyl)amino)ethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-6-carbonitrile (Examples 190 - 191); N-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl)acetamide (Examples 192 - 193); N-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl)isobutyramide (Examples 194 - 195); Methyl (2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidine-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl)carbamate (Examples 196 - 197); Isopropyl (2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl)carbamate (Examples 198-199); N-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2-ethyl-5-methylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl)methanesulfonamide (Examples 200-201); N-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl)acetamide (Examples 202-203); N-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl)isobutyramide (Examples 204-205); Methyl (2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl)carbamate (Examples 206-207); Isopropyl (2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl)carbamate (Examples 208-209); or N-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydropyrido[3,2-d]pyrimidin-4-yl)-2,5-diethylpiperazin-1-yl)-2-(4-(trifluoromethyl)phenyl)ethyl)methanesulfonamide (Examples 210-211) A compound selected from the above or a salt thereof.

10. A pharmaceutical composition comprising the compound according to Claim 1 or a salt thereof and a pharmaceutically acceptable carrier.

11. A pharmaceutical composition for the treatment of cancer or viral infection, comprising the compound according to any one of claims 1 to 9 or a salt thereof.

12. The pharmaceutical composition according to claim 11, wherein the cancer is selected from colon cancer, pancreatic cancer, breast cancer, prostate cancer, lung cancer, ovarian cancer, cervical cancer, kidney cancer, head and neck cancer, lymphoma, leukemia and melanoma.

13. A pharmaceutical composition for inhibiting the activity of at least one diacylglycerol kinase selected from diacylglycerol kinase alpha (DGKα) and diacylglycerol kinase zeta (DGKζ), comprising the compound according to any one of claims 1 to 9 or a salt thereof.

14. Formula (I): 【Chemical Formula 5】 [wherein: X is CR ６ or N; Y is CR ３ or N; R １ is H, F, Cl, Br, -CN, C １－３ alkyl (substituted with 0 to 4 R １ａ groups), C ３－４ cycloalkyl (substituted with 0 to 4 R １ａ groups), C １－３ alkoxy (substituted with 0 to 4 R １ａ groups), -C(O)NR ａ R ａ , -NR ａ R ａ , -S(O) ｎ R ｆ , or -P(O)R ｆ R ｆ ; Each R １ａ is independently F, Cl, -CN, -OH, -OCH ３ or -NR ａ R ａ ; Each R ａ is independently H or C １－３ alkyl; R ２ is H, C １－３ alkyl (substituted with 0 to 4 R ２ａ groups), C ２－３ alkenyl (substituted with 0 to 4 R ２ａ groups), or C ３－４ cycloalkyl (substituted with 0 to 4 R ２ａ groups); Each R ２ａ is, independently, F, Cl, -CN, -OH, -O(C １－２ alkyl), C ３－４ cycloalkyl, C ３－４ alkenyl, or C ３－４ alkynyl; R ３ is H, F, Cl, Br, -CN, C １－３ alkyl, C １－２ fluoroalkyl, C ３－４ cycloalkyl, C ３－４ fluorocycloalkyl, -NO ２ , or pyridinyl (substituted with 0 to 2 R ３ａ ); Each R ３ａ is halo, -CN, C １－３ alkyl, or C １－３ alkoxy; R ４ is -CR ４ａ R ４ｃ -L-R ４ｂ and is; L is L １ or -(CH ２ ) １－３ L １ -; L １ is -C(O)-, -C(O)O-, -C(O)NR ｄ -, -C(O)NR ｄ O-, -NR ｄ -, -NR ｄ C(O)-, -NR ｄ C(O)O-, -NR ｄ C(O)NR ｄ -, -C(O)NR ｄ O-, -S(O) ２ -, -S(O) ２ NR ｄ -, -NR ｄ S(O) ２ -, or -P(O)R ｅ -; R ４ａ is (i) H or C １－６ alkyl (F, Cl, -CN, -OH, -OCH ３ , -SCH ３ , C １－３ fluoroalkoxy, -NR ａ R ａ , -S(O) ２ R ｆ , or -NR ａ S(O) ２ R ｆ substituted with 0 to 4 substituents independently selected therefrom); or (ii) C ３－６ cycloalkyl, C ５－１４ heterocyclyl, C ６－１０ aryl, or C ５－１４ heteroaryl, each being substituted with 0 to 4 R ｂ ; Each R ｂ is independently F, Cl, Br, -CN, -OH, C １－６ alkyl, C １－３ fluoroalkyl, C １－４ hydroxyalkyl, -(CH ２ ) １－２ O(C １－３ alkyl), C １－４ alkoxy, -O(C １－４ hydroxyalkyl), -O(CH) １－３ O(C １－３ alkyl), C １－３ fluoroalkoxy, -O(CH) １－３ NR ｃ R ｃ , -OCH ２ CH=CH ２ , -OCH ２ C≡CH, -C(O)(C １－４ alkyl), -C(O)OH, -C(O)O(C １－４ alkyl), -C(O)NH ２ , -C(O)NH(C １－４ alkyl), -C(O)N(C １－４ alkyl) ２ , -NR ｃ R ｃ , -NR ａ S(O) ２ (C １－３ alkyl), -NR ａ C(O)(C １－３ alkyl), -NR ａ C(O)O(C １－４ alkyl), -P(O)(C １－３ alkyl) ２ , -S(O) ２ (C １－３ alkyl), -O(CH ２ ) １－２ (C ３－６ cycloalkyl), -O(CH ２ ) １－２ (morpholinyl), C ３－６ cycloalkyl, cyanocyclopropyl, methylazetidinyl, acetylazetidinyl, triazolyl, tetrahydropyranyl, morpholinyl, thiophenyl, methylpiperidinyl, or -CR ｃ R ｃ is (phenyl); R ４ｂ is (i)-CH ２ CH ２ OCH ２ CH ２ OCH ２ CH ２ NH ２ ; (ii) -CH ２ CH ２ OCH ２ CH ２ OCH ２ CH ２ NHCOOC(CH ３ ) ３ ; or (iii) [Chemical Formula 6] is; R ４ｃ is H or C １－６ and is alkyl; Each R ｃ is independently H or C １－２ alkyl; Each R ｄ is, independently, H or C １－６ alkyl; R ｅ is C １－６ alkyl; Each R ｆ is, independently, C ３－４ cycloalkyl or C １－３ alkyl (substituted with 0 to 4 R １ａ groups); Each R ５ is independently F, Cl, -CN, -OH, C １－６ alkyl (substituted with 0 to 4 R ｇ ), C １－３ alkoxy (substituted with 0 to 4 R ｇ ), C ２－４ alkenyl (substituted with 0 to 4 R ｇ ), C ２－４ alkynyl (substituted with 0 to 4 R ｇ ), C ３－４ cycloalkyl (substituted with 0 to 4 R ｇ ), phenyl (substituted with 0 to 4 R ｇ ), oxadiazolyl (substituted with 0 to 3 R ｇ ), pyridinyl (substituted with 0 to 4 R ｇ ), -(CH ２ ), １－２ heterocyclyl (substituted with 0 to 4 R ｇ ) substituted with -(CH ２ ), １－２ NR ｃ C(O)(C １－４ alkyl), -(CH ２ ), １－２ NR ｃ C(O)O(C １－４ alkyl), -(CH ２ ), １－２ NR ｃ S(O) ２ (C １－４ alkyl), -C(O)(C １－４ alkyl), -C(O)OH, -C(O)O(C １－４ alkyl), -C(O)O(C ３－４ cycloalkyl), -C(O)NR ａ R ａ , or -C(O)NR ａ (C ３－４ cycloalkyl), or two R ５ are bonded to the same carbon atom to form =O; Each R ｇ is independently F, Cl, -CN, -OH, C １－３ alkoxy, C １－３ fluoroalkoxy, -O(CH ２ ) １－２ O(C １－２ alkyl), C ３－５ cycloalkyl, or -NR ｃ R ｃ ; Each R ６ is H, F, Cl, -CN, -CH ３ , -CH ２ F, -CHF ２ , -CF ３ , or -OCH ３ ; m is 0, 1, 2, or 3; and n is 0, 1, or 2] A compound represented by or a salt thereof.

15. The compound is tert-butyl (2-(2-(2-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetamido)ethoxy)ethoxy)ethyl)carbamate (Example 63); N-(2-(2-(2-aminoethoxy)ethoxy)ethyl)-2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetamide·TFA (Example 64); N-(2-(2-(2-(2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetamido)ethoxy)ethoxy)ethyl)-3’,6’-dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9’-xanthene]-5-carboxamide (Example 65); 2,2-dimethyl-4-oxo-3,8,11-trioxa-5-azatridecan-13-yl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate (Example 66); 2-(2-(2-Aminoethoxy)ethoxy)ethyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate·TFA (Example 67); or 2-(2-(2-(3’,6’-Dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9’-xanthene]-5-carboxamide)ethoxy)ethoxy)ethyl 2-((2R,5S)-4-(6-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)-2,5-dimethylpiperazin-1-yl)-2-(4-fluorophenyl)acetate (Example 68) The compound according to claim 14 or a salt thereof, which is as described above.

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