Triazolo-pyrimidine analogues for treating diseases associated with inhibition of Werner syndrome RecQ helicase (WRN)

Abstract

The present invention relates to a compound of formula (I) or a pharma- ceutically acceptable salt thereof: [Formula 1] TIFF2024522345000952.tif39170 (In the formula, R1, R2, R3, R4, R5, R 26 , R 27 , y, R, M, W, L, V, T, Y, J, K and A are as defined in the specification), therapeutic uses of the compounds, use of the compounds as research chemicals, pharmaceutical compositions and combinations comprising the compounds, and methods of making the compounds of the invention.

Description

[Technical Field]

[0001] The present invention provides bicyclic compounds, such as 7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl) compounds and analogs and derivatives thereof, their use to inhibit Werner syndrome RecQ DNA helicase (WRN), and methods of using such compounds to treat disease, particularly in the treatment of cancer, particularly cancers characterized by microsatellite instability-high (MSI-H) or mismatch repair deficiency (dMMR), including colorectal, gastric, and endometrial cancers. The invention also provides uses of such compounds as research chemicals, intermediate compounds, combinations, processes, and formulations. [Background technology]

[0002] Loss of DNA mismatch repair function is a common initiating event in cancer development, occurring in 10-30% of colorectal, endometrial, ovarian, and gastric cancers (Aaltonen, LA et al. Clues to the pathogenesis of familial colorectal cancer, Science 260, 812-816 (1993); Bonneville R et al., Landscape of Microsatellite Instability Across 39 Cancer Types. JCO Precis Oncol. 1:PO.17.00073 (2017). Cancers that have lost their mismatch repair (MMR) capacity have a high mutational burden and frequent deletion and insertion events in repetitive DNA tracts, a phenotype known as microsatellite instability (MSI). Advances in the treatment of microsatellite instability-high (MSI-H) cancers have been made, with pembrolizumab (anti-PD1) recently approved as a first-line treatment for MSI-H-dMMR metastatic colorectal cancer after demonstrating significantly longer progression-free survival compared with chemotherapy. However, significant unmet medical needs remain in CRC and other MSI-H indications (Andre T., et al. Pembrolizumab in Microsatellite-Instability-High Advanced Colorectal Cancer. N Engl J Med;383(23):2207-2218(2020)).Several large-scale functional genomics screens across a large panel of cell lines, including Novartis, using 398 cell lines from the Cancer Cell Line Encyclopedia (CCLE) (McDonald ER et al., Project DRIVE: A Compendium of Cancer Dependencies and Synthetic Lethal Relationships Uncovered by Large-Scale, Deep RNAi Screening. Cell 170(3):577-592 (2017)), identified the Werner syndrome RecQ helicase (WRN) as being selectively required for the survival of cell lines with defective mismatch repair function that became MSI-H (Behan, FM et al., Prioritization of cancer therapeutic targets using CRISPR-Cas9 screens. Nature 568, 511-516 (2019); Chan, EM et al., WRN helicase is a synthetic lethal target in microsatellite unstable cancers. Nature 568, 551-556 (2019), Kategaya, L., Perumal, SK, Hager, JH & Belmont, LD. Werner syndrome helicase is required for the survival of cancer cells with microsatellite instability. iScience 13, 488-497 (2019), Lieb, S. et al. Werner syndrome helicase is a selective vulnerability of microsatellite instability-high tumor cells. eLife8, e43333 (2019). WRN is synthetically lethal in MSI cancers. Depletion of WRN has antiproliferative effects, resulting in activation of multiple DNA damage signaling markers, induction of cell cycle arrest, and apoptosis in MMR cancer models but not in cancer cells with an intact MMR pathway.These findings indicate that WRN provides DNA repair and maintenance functions essential for cell survival in MSI cancers. Recently, the mechanism of WRN dependence has been elucidated. Dinucleotide TA repeats have been shown to be selectively unstable and undergo large-scale expansions in MSI cells. These expanded TA repeats form secondary DNA structures that require WRN helicase for unwinding (van Wietmarschen, N. et al. Repeat expansions confer WRN dependence in microsatellite-unstable cancers. Nature 586, 292-298, 2020). In the absence of WRN (or when WRN helicase is inhibited), expanded TA repeats in MSI cells undergo nuclease cleavage and chromosome breakage. Therefore, inhibiting WRN helicase is an attractive strategy for treating mismatch repair-deficient cancers. Summary of the Invention

[0003] There remains a need for new therapies and treatments for cancer, particularly cancers characterized by microsatellite instability-high (MSI-H) or deficient mismatch repair (dMMR), including colorectal, gastric, or endometrial cancer. The present invention provides compounds, pharmaceutically acceptable salts thereof, pharmaceutical compositions thereof, and combinations thereof, which are inhibitors of Werner syndrome RecQ DNA helicase (WRN). The present invention further provides methods for treating, preventing, or ameliorating a disease or condition, comprising administering to a subject in need thereof an effective amount of a WRN inhibitor. The present invention also provides compounds, pharmaceutically acceptable salts thereof, pharmaceutical compositions thereof, and combinations thereof, which are useful in treating cancer, particularly cancers characterized by microsatellite instability-high (MSI-H) or deficient mismatch repair (dMMR). Also provided are compounds that bind to and / or inhibit WRN and are therefore useful as research chemicals, for example, as chemical probes and tool compounds. Various embodiments of the present invention are described herein. In certain aspects, provided herein are compounds of formula (I) or a pharmaceutically acceptable salt thereof: [ka] [In the formula, R, M, W, L, V and T are independently selected from C, CH and N; Subformulas 1a, 1b, 1c, 1d, 1e and 1f: [ka] Forming; A is -C(O)-, -S(O)-, -S(O)2- and [ka] is a linker selected from: Y is N, C or CH; y is 0, 1, 2, 3 or 4; [ka] means that when Y is CH, it is connected to the adjacent carbon atom via a single bond, or when Y is C, it is connected to the adjacent atom via a double bond, [ka] is a single bond, then Y is unsubstituted or substituted by OH or F; If Y is N, [ka] is a single bond; [ka] means that K is connected to the adjacent carbon atom via a single or double bond; where: [ka] If is a double bond, [ka] is a single bond, K is CH, J is C, A is -C(O)-, -S(O)-, -S(O)2- and [ka] is a linker selected from: or [ka] When is a single bond, K is -CH2-, -CH2CH2-, -NH- and (5-membered ring: [ka] and J is N, A is selected from -C(O)-, -S(O)-, -S(O)2- and [ka] is a linker selected from: or [ka] is a single bond, K is -CH2-, J is CH, A is -S(O)-, -S(O)2- and [ka] is a linker selected from: R5 is independent, -(C 1~ C4) alkyl, -(C 3~ C5) cycloalkyl, where two R5 substituents on the same ring carbon atom, together with the carbon atom to which they are attached, form (C 3~ C4) a cycloalkylspiro ring or a 3- or 4-membered heterocyclylspiro ring may be formed, wherein the heterocyclylspiro ring contains ring carbon atoms and one ring heteroatom selected from O, N, and S; · [ka] is a carbon-nitrogen single bond, K and the R5 substituents on the adjacent carbon atom are joined to form ring C: [ka] wherein ring C is a fused (C3-C6) cycloalkyl ring, a fused (C3-C6) heterocyclyl ring or a fused phenyl ring, wherein the fused (C3-C6) heterocyclyl ring contains ring carbon atoms and one ring heteroatom selected from O, N and S; · [ka] is a carbon-carbon single bond, Y is N, [ka] is a single bond, A is -S(O)-, -S(O)2- and [ka] When K and the R5 substituent on the adjacent carbon atom are taken together to form ring C: [ka] may form where K is -CH2- and J is N, two R5 substituents may be linked to form a (C1-C3) alkylene or heteroalkylene bridge, where the heteroalkylene bridge is one heteroatom selected from N and O, or is -CH2-O-CH2-. Selected from Here, the ring: [ka] One or more H atoms on the above may be replaced by deuterium: R1 is cycloalkenyl, wherein said cycloalkenyl is a partially unsaturated monocyclic ring containing 5 or 6 ring carbon atoms, said cycloalkenyl being unsubstituted or having 1, 2, 3 or 4, preferably 1 or 2, R 33 where R 33 is halo, and said cycloalkenyl or halo-substituted cycloalkenyl has 0, 1 or 2 R 15 Is substituted by a substituent, Alternatively, R is heterocyclyl, wherein the heterocyclyl is a fully saturated or partially unsaturated 5- or 6-membered group comprising ring carbon atoms and 1 or 2 ring heteroatoms independently selected from N, O and S, wherein the heterocyclyl is unbridged or bridged, the bridge being 1 or 2 carbon atoms, wherein the heterocyclyl is unsubstituted or has 1, 2, 3 or 4, preferably 1 or 2, R 33 where R 33 is halo, and said heterocyclyl or halo-substituted heterocyclyl is independently R 15 , R 16 , R 17 , R 18 , R 19 , R 20 , R 22 and R 23 or substituted by 0, 1 or 2 substituents selected from Alternatively, said heterocyclyl or halo-substituted heterocyclyl is fused to a cyclopropyl ring, wherein said cyclopropyl ring is unsubstituted or substituted with 1, 2 or 3 F; Alternatively, said heterocyclyl or halo-substituted heterocyclyl has two substituents at the same ring carbon atom which are joined to form a cyclopropyl spiro ring; Alternatively, the heterocyclyl or halo-substituted heterocyclyl is fused to a (C3-C5) heterocycloalkyl ring, wherein the (C3-C5) heterocycloalkyl ring contains ring carbon atoms and one ring O atom; or Alternatively, R is heteroaryl, wherein said heteroaryl is a 5- or 6-membered fully unsaturated monocyclic group containing ring carbon atoms and 1, 2, 3, or 4 ring heteroatoms independently selected from N, O, and S, preferably 1 or 2 ring heteroatoms, wherein the total number of ring S atoms does not exceed 1 and the total number of ring O atoms does not exceed 1, and wherein said heteroaryl is unsubstituted or R 21 and R 30 and is substituted by 1, 2, or 3 substituents independently selected from 21 and R 30 is independently selected from halo and (C1-C4) alkyl, wherein said (C1-C4) alkyl is unsubstituted or substituted with 1, 2 or 3 halo; Alternatively, R1 is phenyl, wherein said phenyl is unsubstituted or has 1, 2, 3 or 4, preferably 1 or 2, R 33 where R 33 is halo, and said phenyl or halo-substituted phenyl is selected from 0, 1 or 2 R 15 Is substituted with a substituent, Alternatively, R1 is (C2-C4)alkynyl or (C2-C4)alkenyl, wherein said (C2-C4)alkynyl and (C2-C4)alkenyl are unsubstituted or substituted by (C1-C4)alkyl-OC(O)- or morpholinyl; Each R 15 , R 16 , R 17 , R 18 , R19 , R 20 , R 22 and R 23 is, independently, Hello, unsubstituted or substituted with 1, 2 or 3 halo (C 1~ C4) alkyl-O-, (C1-C4)alkyl unsubstituted or substituted by OH, —O—(C1-C2)alkyl or 1, 2 or 3 halo, HOC(O)-(CH2) n -, H3C-C(O)(CH2) n -, (C1-C4) alkyl-OC(O)(CH2) n , =O, azetidinyl or pyrrolidinyl, wherein the azetidinyl and pyrrolidinyl are linked to the rest of the molecule via an N atom and are each unsubstituted or substituted with one or two F, ·R 25 (R 24 )N-(where R 24 is H or (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo, and R 25 is H or (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo OH is selected from where n is 0, 1 or 2; R 26 is CH3, H or deuterium; R 27 is CH3, H or deuterium; Or, R 26 and R 27 together with the carbon atoms to which they are attached form a cyclopropyl ring; R2 is part: [ka] and; R6 is H, Hello, (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo, (C3-C5)cycloalkyl unsubstituted or substituted by 1, 2 or 3 halo; -O-(C1-C4)alkyl, unsubstituted or substituted with 1, 2 or 3 halo; ·OH, and ·CN Selected from; R8 is selected from H, halo, and (C1-C4) alkyl unsubstituted or substituted with 1, 2, or 3 halo; R9 is selected from H, O-CH3, OH, CN, CH3, and halo; R 28 teeth, SF5, H, ·-C(O)H, Hello, (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo, ·(C1-C4)alkynyl; (C1-C4) alkenyl, (C3-C5)cycloalkyl unsubstituted or substituted with 1, 2 or 3 halo, and OCF3 Selected from; X is selected from C-R7 and N, where R7 is H or halo, or R 28 or R6 together with the atoms to which they are attached form a fused (C4-C6)cycloalkyl ring, wherein said fused (C4-C6)cycloalkyl ring is unsubstituted or substituted with 1, 2 or 3 halo; or R2 is [ka] is selected from where: R 31is selected from H, halo, and CH3; R 32 is selected from H, halo, and CH3; R3 is Cyclopropyl, ·O-CH3, N(CH3)2, ·S-CH3, (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 substituents independently selected from halo and OH; R4 is [ka] Selected from where: R 10 , R 11 , R 12 , R 13 and R 14 is, independently, H, Hello, (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents, -O-(C1-C2)alkyl or (C1-C2)alkyl substituted by OH, -S-(C1-C3) alkyl, -O-(C1-C4)alkyl, unsubstituted or substituted with 1, 2 or 3 halo substituents; ·OH, (C3-C5)cycloalkyl (wherein said (C3-C5)cycloalkyl is unsubstituted or substituted with 1 or 2 halo), -O-(C3-C5)cycloalkyl, -NR 34 R 35 (where R 34 and R 35 is independent ohhh, o(C1-C4)alkyl, wherein said (C1-C4)alkyl is unsubstituted or substituted with OH or —O(C1-C2)alkyl; o and where R 34 and R 35 can be taken together with the carbon atoms to which they are attached to form azetidine, pyrrolidinyl, and piperidine rings, wherein the azetidine, pyrrolidinyl, and piperidine are unsubstituted or substituted with CH3. selected from ·CN, -(C2-C4)alkenyl, -(C2-C4)alkynyl, -C(O)H, and -C(O)(C1-C4) alkyl Selected from; and * indicates point of attachment].

[0004] In another aspect, the present invention provides pharmaceutical compositions comprising a compound of the present invention and one or more pharmaceutically acceptable carriers.

[0005] In another aspect, the present invention provides combinations, in particular pharmaceutical combinations, comprising a compound of formula (I) according to the present invention and one or more therapeutically active agents.

[0006] In another aspect, the present invention provides compounds of formula (I) according to the invention for use as a pharmaceutical, particularly for the treatment of disorders or diseases that can be treated by WRN inhibition.

[0007] In another aspect, the present invention provides a compound of formula (I) of the present invention for use in the treatment of cancer, particularly wherein the cancer is characterised by microsatellite instability high (MSI-H) or deficient mismatch repair (dMMR).

[0008] In another aspect, the present invention provides a method for treating a disorder or disease in a subject that can be treated by WRN inhibition, comprising administering to the subject a therapeutically effective amount of a compound of formula (I) of the present invention.

[0009] In another aspect, the present invention provides a method of treating cancer in a subject, more particularly wherein the cancer is characterized by microsatellite instability-high (MSI-H) or deficient mismatch repair (dMMR), comprising administering to the subject a therapeutically effective amount of a compound of formula (I) of the present invention.

[0010] In another aspect, there is provided the use of a compound of formula (I) of the present invention in the manufacture of a medicament for the treatment of a disorder or disease that can be treated by WRN inhibition.

[0011] In another aspect, the present invention provides a compound of formula (I) of the invention for use as a research chemical, for example as a chemical probe or as a tool compound.

[0012] In another aspect, the invention provides a solid form, process, or intermediate described herein. [Brief explanation of the drawings]

[0013] [Figure 1] 1 shows the powder X-ray diffractogram of Example 42. [Figure 2] 1 shows the powder X-ray diffractogram of Example 86. [Figure 3] 1 shows the powder X-ray diffractogram of Example 47. [Figure 4] 1 shows the powder X-ray diffractogram of Example 57. [Figure 5] 1 shows the powder X-ray diffractogram of Example 96. [Figure 6] Showing efficacy and tolerability after once daily (qd) administration of Example 58 in female nude mice bearing SW48 xenografts [Figure 7] 1 shows the powder X-ray diffractogram of Example 42. [Figure 8] 1 shows colony formation assay data for the compound of Example 42 using MSI and MSS cells. [Figure 9] Shows efficacy of Example 42 after administration in female nude mice bearing SW48 xenografts [Figure 10] Shows efficacy of Example 96 after administration in female nude mice bearing SW48 xenografts [Figure 11] Shows efficacy of Example 57 after administration in female nude mice bearing SW48 xenografts DETAILED DESCRIPTION OF THE INVENTION

[0014] The present disclosure provides compounds of formula (I): [ka] (In the formula, R1, R2, R3, R4, R5, R 26 , R 27 , y, R, M, W, L, V, T, Y, J, K, and A are as described in the Summary of the Invention above.

[0015] Unless otherwise specified, the term "compounds of the disclosure" or "compounds of Formula (I)" refers to compounds of Formula (I), subformulas thereof, exemplified compounds, and salts thereof, as well as all zwitterions, stereoisomers (including diastereoisomers and enantiomers), rotamers, tautomers, and isotopically labeled compounds (including deuterium substitution), and inherently formed moieties, and combinations or mixtures of those embodiments described above.

[0016] Various (enumerated) embodiments of the present invention are described herein, and it will be understood that the features specified in each embodiment may be combined with other specified features to provide further embodiments of the present invention.

[0017] Embodiment 1. A compound of formula (I) or a pharmaceutically acceptable salt thereof, as described above.

[0018] Embodiment 2: When R1 is a ring, each R ring atom adjacent to the R ring atom at which the R ring is attached to the remainder of the molecule is independently unsubstituted or substituted only with halo, in particular independently unsubstituted or substituted with one F substituent, preferably The R1 ring is connected to the rest of the molecule via an R1 ring nitrogen atom or an R1 ring carbon atom that is double-bonded to an adjacent ring atom A compound of formula (I) as defined in embodiment 1 or a pharmaceutically acceptable salt thereof.

[0019] Embodiment 3.R1 is cycloalkenyl, wherein said cycloalkenyl is a partially unsaturated monocyclic ring containing 5 or 6 ring carbon atoms, said cycloalkenyl is unsubstituted or has 1, 2, 3 or 4, preferably 1 or 2, R 33 where R 33 is halo, and said cycloalkenyl or halo-substituted cycloalkenyl has 0, 1 or 2 R 15 Is substituted by a substituent, Alternatively, R is heterocyclyl, wherein the heterocyclyl is a fully saturated or partially unsaturated 5- or 6-membered group comprising ring carbon atoms and 1 or 2 ring heteroatoms independently selected from N, O and S, the heterocyclyl is unbridged or bridged, the bridge being 1 or 2 carbon atoms, the heterocyclyl is unsubstituted or has 1, 2, 3 or 4, preferably 1 or 2, R 33 is substituted with R 33 is halo, and said heterocyclyl or halo-substituted heterocyclyl is independently R 15 , R 16 , R 17 , R 18 , R 19 , R 20 , R 22 and R 23 or substituted by 0, 1 or 2 substituents selected from Alternatively, R is heteroaryl, wherein said heteroaryl is a 5- or 6-membered fully unsaturated monocyclic group containing ring carbon atoms and 1, 2, 3, or 4 ring heteroatoms independently selected from N, O, and S, preferably 1 or 2 ring heteroatoms, wherein the total number of ring S atoms does not exceed 1 and the total number of ring O atoms does not exceed 1, and wherein said heteroaryl is unsubstituted or R 21 and R 30 and wherein R is substituted by 1, 2, or 3 substituents independently selected from 21 and R 30 is independently selected from halo and (C1-C4) alkyl, wherein said (C1-C4) alkyl is unsubstituted or substituted with 1, 2 or 3 halo; And each R 15 , R 16 , R 17 , R 18 , R 19 , R 20 , R 22 and R 23 is, independently, Hello, unsubstituted or substituted with 1, 2 or 3 halo (C 1~ C4) alkyl-O-, (C1-C4)alkyl unsubstituted or substituted by OH, —O—(C1-C2)alkyl or 1, 2 or 3 halo, HOC(O)-(CH2) n -, H3C-C(O)(CH2) n -, (C1-C4) alkyl-OC(O)(CH2) n , =O azetidinyl or pyrrolidinyl, wherein said azetidinyl and pyrrolidinyl are linked to the rest of the molecule via an N atom and are each unsubstituted or substituted with one or two F, ·R 25 (R 24 )N-(where R 24 is H or (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo, and R 25is H or (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo OH is selected from where n is 0, 1 or 2 A compound of formula (I) or a pharmaceutically acceptable salt thereof as defined in embodiment 1 or 2.

[0020] Embodiment 4.R1 is cycloalkenyl, wherein the cycloalkenyl is a partially unsaturated monocyclic ring containing 5 or 6 ring carbon atoms, and the cycloalkenyl is unsubstituted or contains one or two R 33 where R 33 is halo, preferably F, and said cycloalkenyl or halo-substituted cycloalkenyl has zero or one R 15 is substituted by a substituent, preferably one substituent, and R 15 teeth, a) unsubstituted or substituted with 1, 2 or 3 halo (C 1~ C2) alkyl-O-, b) (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo; c) HOC(O)-(CH2) n -, d) H3C-C(O)(CH2) n -, e) H3C-OC(O)(CH2) n , f) O, and g)R 25 (R 24 )N-, H(where R 24 is H or (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo, and R 25 is H or (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo is selected from n is 0 or 1, where: R of the cycloalkenyl or halo-substituted cycloalkenyl 15Substituents a) through g) are not present on the ring atom adjacent to the ring atom at which the cycloalkenyl or halo-substituted cycloalkenyl is attached to the remainder of the molecule, and preferably said cycloalkenyl or halo-substituted cycloalkenyl has one R in the ring para position relative to the remainder of the molecule. 15 is a 6-membered ring having a substituent, the cycloalkenyl or halo-substituted cycloalkenyl is linked to the remainder of the compound via an R1 ring carbon atom that is double-bonded to an adjacent R1 ring carbon atom; Alternatively, R is heterocyclyl, wherein the heterocyclyl is a fully saturated or partially unsaturated 5- or 6-membered group containing ring carbon atoms and one or two ring heteroatoms independently selected from N, NH, O, and S, and wherein the heterocyclyl is unbridged or bridged, the bridge being one or two carbon atoms, and wherein the heterocyclyl is unsubstituted or contains one or two R 33 where R 33 is halo, preferably F, and said heterocyclyl or halo-substituted heterocyclyl is independently R 15 , R 16 , R 17 , R 18 , R 19 , R 20 , R 22 and R 23 and wherein R is substituted by 0 or 1 substituent selected from 15 , R 16 , R 17 , R 18 , R 19 , R 20 , R 22 and R 23 is, independently, a) unsubstituted or substituted with 1, 2 or 3 halo (C 1~ C4) alkyl-O-, b) (C1-C4) alkyl unsubstituted or substituted with OH, —O—(C1-C2) alkyl or 1, 2 or 3 halo; c) HOC(O)-(CH2) n -, d) H3C-C(O)(CH2) n -, e) H3C-OC(O)(CH2) n , f)=O, g)R 25 (R 24 )N-(where R 24 is H, (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo, and R 25 is H, (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo h)OH is selected from n is 0 or 1, where: the substituents a) to h) of said heterocyclyl or halo-substituted heterocyclyl are not present on the ring atom at which the heterocyclyl or halo-substituted heterocyclyl is attached to the remainder of the molecule, preferably said heterocyclyl or halo-substituted heterocyclyl is a 6-membered ring which has 0 or 1 substituent selected from a) to h) in the meta or para position relative to the remainder of the molecule, preferably in the para position; the heterocyclyl is linked to the remainder of the compound via an R1 ring nitrogen atom or an R1 ring carbon atom that is double bonded to an adjacent ring atom; Alternatively, R is heteroaryl, wherein said heteroaryl is a 5- or 6-membered fully unsaturated monocyclic group containing ring carbon atoms and 1 or 2 ring heteroatoms independently selected from N, O and S, preferably N, wherein the total number of ring S atoms does not exceed 1 and the total number of ring O atoms does not exceed 1, and wherein said heteroaryl is unsubstituted or R 21 and R 30 and is substituted by one or two substituents independently selected from 21 and R 30are independently selected from (C1-C2) alkyl, wherein said (C1-C2) alkyl is unsubstituted or substituted with 1, 2 or 3 halo, and wherein preferably said alkyl or haloalkyl substituent is not on the R1 ring atom adjacent to the R1 ring atom at which the heteroaryl is attached to the remainder of the molecule, and more preferably, when the heteroaryl is a 6-membered ring, said alkyl or haloalkyl substituent is in the ring para position relative to the remainder of the molecule. A compound of formula (I) according to any one of embodiments 1, 2 or 3, or a pharmaceutically acceptable salt thereof.

[0021] Embodiment 5.R1 is [ka] Selected from; R 33 is F; R 15 is halo, azetidinyl, or pyrrolidinyl, wherein said azetidinyl and pyrrolidinyl are linked to the remainder of the molecule via an N atom and are unsubstituted or substituted with one or two F; R 16 is R 25 (R 24 )N- (where R 24 is H or (C1-C2) alkyl, and R 25 is H or (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo, especially F; R 17 is a halo; R 18 is a halo; R 19 is a halo; R 20 is a halo; R 21 is (C1-C2) alkyl; R 22 and R 23 are each independently (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo, HOC(O)-(CH2) n -, H3C-C(O)(CH2) n -, (H3C)3C-OC(O)(CH2) n - Selected from; where n is 0, 1 or 2; and R 30 is CH3 A compound of formula (I) according to any one of embodiments 1 to 3, or a pharmaceutically acceptable salt thereof.

[0022] Embodiment 6.R1 is [ka] Selected from; R 15 is F; R 16 is R 25 (R 24 )N- and; R 17 is F; R 18 is F; R 19 is F; R 20 is F; R 21 is CH3; R 22 are CF3, CHF2CH2, HOC(O)-CH2-, H3C-C(O)-, (H3C)3C-OC(O)-; R 23 are CF3, CHF2CH2-, (H3C)3C-OC(O)-; R 24 is CH3; and R 25 is CHF2CH2- A compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of embodiments 1 to 5.

[0023] Embodiment 7.R2 comprises the moiety: [ka] and; R6 is H, Hello, (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo, (C3-C5)cycloalkyl unsubstituted or substituted by 1, 2 or 3 halo; -O-(C1-C4)alkyl, unsubstituted or substituted with 1, 2 or 3 halo; ·OH, and ·CN Selected from; R8 is selected from H, halo, and (C1-C4) alkyl unsubstituted or substituted with 1, 2, or 3 halo; R9 is selected from H, O-CH3, OH, CN, CH3, and halo; R 28 teeth, SF5, H, ·-C(O)H, Hello, (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo, ·(C1-C4)alkynyl; (C1-C4) alkenyl, (C3-C5)cycloalkyl unsubstituted or substituted with 1, 2 or 3 halo, and Selected from OCF3; and X is selected from C-R7 and N (wherein R7 is H or halo). A compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of embodiments 1 to 6.

[0024] Embodiment 8.R2 comprises the moiety: [ka] and; where: R6 is selected from H, halo, (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo; R8 is selected from H, halo, (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo; R9 is selected from H, O-CH3, OH, CN, CH3, and halo; R 28 is selected from SF5, halo, (C1-C4)alkyl unsubstituted or substituted with 1, 2 or 3 halo, and -C(O)H; X is selected from C-R7 and N; R7 is selected from H and halo A compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of embodiments 1 to 7.

[0025] Embodiment 9.R2 comprises the moiety: [ka] and; R6 is selected from H, Cl, CH3, F, and Br; R8 is selected from H, Cl, F, and CF3; R9 is selected from H, CH3, and Cl; R 28 is selected from CF3, CF2H, -CH2CH3, Cl, SF5, Br, and -C(O)H; X is selected from C-R7 and N; R7 is selected from H and F A compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of embodiments 1 to 8.

[0026] Embodiment 10.R 26 is H and R 27 10. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of embodiments 1 to 9, wherein

[0027] Embodiment 11. A compound of Formula (I) according to any one of embodiments 1 to 10, or a pharmaceutically acceptable salt thereof, wherein R3 is (C1-C4)alkyl unsubstituted or substituted with 1, 2 or 3 substituents each independently selected from halo and OH.

[0028] Embodiment 12. A compound of formula (I) according to any of embodiments 1 to 11, or a pharmaceutically acceptable salt thereof, wherein R3 is (C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 substituents independently selected from halo and OH, preferably -CH2CH3 or CH3, more preferably -CH2CH3.

[0029] Embodiment 13. Y is N; [ka] 13. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any of embodiments 1 to 12, wherein is Y linked by a single bond.

[0030] Embodiment 14. [ka] is K connected by a single bond, and K is selected from -CH2-, -CH2CH2-, -NH-, and (5-membered ring: [ka] and J is N, and A is selected from -C(O)-, -S(O)-, -S(O) 2- , and [ka] 14. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of embodiments 1 to 13, wherein the linker is selected from:

[0031] Embodiment 15. [ka] are Ks linked by a single bond, where K is -CH2-, J is N, and A is -C(O)-, -S(O)-, -S(O)2-, and [ka] 15. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of embodiments 1 to 14, wherein the linker is selected from:

[0032] Embodiment 16. A compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of embodiments 1 to 15, wherein A is a linker selected from -C(O)- and -S(O)2-, preferably -C(O)-.

[0033] Embodiment 17. R5 independently comprises: -(C 1~ C4) alkyl, preferably methyl; where two R5 substituents on the same ring carbon atom, together with the carbon atom to which they are attached, form (C 3~ C4) may form a cycloalkyl spiro ring or a 3- or 4-membered heterocyclyl spiro ring, wherein the heterocyclyl spiro ring contains ring carbon atoms and one ring heteroatom selected from O, N, and S; · [ka] is a carbon-nitrogen single bond, K and the R5 substituents on the adjacent carbon atom are joined to form ring C: [ka] wherein ring C is a fused (C3-C6) cycloalkyl ring, in particular a fused cyclobutyl ring, a fused (C3-C6) heterocyclyl ring or a fused phenyl ring, said fused (C3-C6) heterocyclyl ring containing ring carbon atoms and one ring heteroatom selected from O, N and S, and when K is -CH2- and J is N, the two R5 substituents may be linked to form a (C1-C3) alkylene or heteroalkylene bridge, where the heteroalkylene bridge is one heteroatom selected from N and O, or is -CH2-O-CH2-. 17. The compound of formula (I) according to any one of embodiments 1 to 16, or a pharmaceutically acceptable salt thereof, selected from:

[0034] Embodiment 18. R5 independently represents: -(C 1~ C4) alkyl, preferably methyl; · [ka] is a carbon-nitrogen single bond, K and the R5 substituents on the adjacent carbon atom are joined to form ring C: [ka] wherein ring C is a fused (C3-C6) cycloalkyl ring, in particular a fused cyclobutyl ring, a fused (C3-C6) heterocyclyl ring, said fused (C3-C6) heterocyclyl ring containing ring carbon atoms and one ring heteroatom selected from O, N and S, and when K is -CH2- and J is N, the two R5 substituents may be linked to form a (C1-C3) alkylene or heteroalkylene bridge, where the heteroalkylene bridge is one heteroatom selected from N and O, or is -CH2-O-CH2-. 18. The compound of formula (I) according to any one of embodiments 1 to 17, or a pharmaceutically acceptable salt thereof, selected from:

[0035] Embodiment 19. R5 independently comprises: -(C 1~ C2) alkyl, preferably methyl, and · [ka] is a carbon-nitrogen single bond, K and the R5 substituents on the adjacent carbon atom are joined to form ring C: [ka] wherein ring C is a fused (C3-C4) cycloalkyl ring, particularly a fused cyclobutyl ring. 19. The compound of formula (I) according to any one of embodiments 1 to 18, or a pharmaceutically acceptable salt thereof, selected from:

[0036] Embodiment 20.R5 independently comprises: CH3 and y is 1 or 2, and · [ka] is a carbon-nitrogen single bond, K and the R5 substituents on the adjacent carbon atom are joined to form ring C: [ka] wherein ring C is a fused cyclobutyl ring. 20. The compound of formula (I) according to any one of embodiments 1 to 19, or a pharmaceutically acceptable salt thereof, selected from:

[0037] Embodiment 21. A compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of embodiments 1 to 20, wherein y is 0, 1, 2 or 3, preferably 0, 1 or 2.

[0038] Embodiment 22.R4 is [ka] (In the formula, R 10 is selected from H, halo, (C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents, —O—(C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; R 11 is selected from H, halo, (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; R 12 is selected from H, halo, (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; R 13 is selected from H, —S—CH, halo, (C-C) alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; and R 14 is selected from H, halo, (C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents, —O—(C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents, and cyclopropyl 22. The compound of formula (I) according to any one of embodiments 1 to 21, or a pharmaceutically acceptable salt thereof, selected from:

[0039] Embodiment 23.R4 is [ka] (In the formula, R 10 is selected from H, F, Cl, CH3, and OCF3; R 11 is selected from H, Cl, F, and CH3; R 12 is selected from H, Cl, and CH3; R 13 is selected from H, —S—CH3, and CH3; and R 14is selected from H, CH, —CHCH, cyclopropyl, —OCHF, OCF, and Cl. 23. The compound of formula (I) according to any one of embodiments 1 to 22, or a pharmaceutically acceptable salt thereof, selected from:

[0040] Embodiment 24. Formula (I) is a compound of formula 1a: [ka] is (Preferably, formula (I) is formula 1a), A compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of embodiments 1 to 23.

[0041] Embodiment 25. Formula (I) is a compound of formula 1b: [ka] or a pharmaceutically acceptable salt thereof,

[0042] Embodiment 26. Formula (I) is a compound of formula 1c: [ka] or a pharmaceutically acceptable salt thereof,

[0043] Embodiment 27. Formula (I) is a compound of formula 1d: [ka] or a pharmaceutically acceptable salt thereof,

[0044] Embodiment 28. Formula (I) is represented by formula 1e [ka] or a pharmaceutically acceptable salt thereof,

[0045] Embodiment 29. Formula (I) is a compound of formula 1f: [ka] or a pharmaceutically acceptable salt thereof,

[0046] Embodiment 30. Formula (I) is a compound of formula 1g: [ka] or a pharmaceutically acceptable salt thereof, More preferably, formula (I) is formula 1g.

[0047] Embodiment 31. Formula (I) is a compound of formula 1h: [ka] or a pharmaceutically acceptable salt thereof. Most preferably, formula (I) is formula 1h.

[0048] Embodiment 32. A compound of formula (Ig) or a pharmaceutically acceptable salt thereof as defined in any one of embodiments 1, 24, or 30. [ka] (Wherein R1 is [ka] Selected from; R 15 is H or F; R 16is H or R 25 (R 24 )N- and; R 17 is H or F; R 18 is H or F; R 19 is H or F; R 20 is H or F; R 21 is H or CH3; R 22 are H, CF3, CHF2CH2, HOC(O)-CH2-, H3C-C(O)-, (H3C)3C-OC(O)-; R 23 are H, CF3, CHF2CH2-, (H3C)3C-OC(O)-; R 24 is CH3; R 25 is CHF2CH2-; R 26 is CH3, H or deuterium; R 27 is H or deuterium; R2 is part: [ka] and R6 is selected from H, Cl, CH3, F, and Br; R8 is selected from H, Cl, F, and CF3; R9 is selected from H, CH3, and Cl; R 28 is selected from CF3, CF2H, -CH2CH3, Cl, SF5, Br, and -C(O)H; X is selected from C-R7 and N; R7 is selected from H and F; R3 is selected from CH3, CH2CH3, cyclopropyl, hydroxyethyl; R4 is [ka] Selected from; During the ceremony, R 10 is selected from H, F, Cl, CH3, and OCF3; R 11 is selected from H, Cl, F, and CH3; R 12 is selected from H, Cl, and CH3; R 13 is selected from H, —S—CH3, and CH3; R 14 is selected from H, CH3, -CH2CH3, cyclopropyl, OCHF2, OCF3, and Cl; Y is N or CH, preferably N; y is 0, 1 or 2; R5 is CH3, or two R5 groups on adjacent carbon atoms, together with the carbon atoms to which they are attached, form a fused cyclobutyl ring: [ka] Forming and optionally a ring: [ka] wherein one or more H is replaced by deuterium; * indicates the point of attachment).

[0049] Embodiment 33. A compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of embodiments 1, 24, or 30. [ka] (In the formula, R1 is [ka] Selected from; R 15is H or F; R 16 is H or R 25 (R 24 )N- and; R 17 is H or F; R 18 is H or F; R 19 is H or F; R 20 is H or F; R 21 is H or CH3; R 22 are CF3, CHF2CH2, HOC(O)-CH2-, H3C-C(O)-, (H3C)3C-OC(O)-; R 23 are CF3, CHF2CH2-, (H3C)3C-OC(O)-; R 24 is CH3; R 25 is CHF2CH2-; R 26 is CH3, H or deuterium; R 27 is H or deuterium; R2 is part: [ka] and During the ceremony, R6 is selected from H, Cl, CH3, F, and Br; R8 is selected from H, Cl, F, and CF3; R9 is selected from H, CH3, and Cl; R 28 is selected from CF3, CF2H, -CH2CH3, Cl, SF5, Br, and -C(O)H; X is selected from C-R7 and N; R7 is selected from H and F; R3 is selected from CH3, CH2CH3, cyclopropyl, and hydroxyethyl; R4 is [ka] Selected from; During the ceremony, R 10 is selected from H, F, Cl, CH3, and OCF3; R 11 is selected from H, Cl, F, and CH3; R 12 is selected from H, Cl, and CH3; R 13 is selected from H, and CH3; R 14 is selected from H, CH3, -CH2CH3, cyclopropyl, -OCHF2, OCF3, and Cl; y is 0, 1 or 2; R5 is CH3, or two R5 groups on adjacent carbon atoms, together with the carbon atoms to which they are attached, form a fused cyclobutyl ring: [ka] and * indicates the point of attachment).

[0050] Embodiment 34.R1 is [ka] 34. The compound of formula (I) according to any one of embodiments 1 to 33, or a pharmaceutically acceptable salt thereof, selected from:

[0051] Embodiment 35.R1 is [ka] or a pharmaceutically acceptable salt thereof.

[0052] Embodiment 36.R1 is [ka] or a pharmaceutically acceptable salt thereof.

[0053] Embodiment 37.R1 is [ka] or a pharmaceutically acceptable salt thereof.

[0054] Embodiment 38.R 28 38. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any of embodiments 1 to 37, wherein is selected from CF3, SF5 and Br.

[0055] Embodiment 39.R 28 or a pharmaceutically acceptable salt thereof.

[0056] Embodiment 40. A compound of Formula (I) or a pharmaceutically acceptable salt thereof according to any one of embodiments 1 to 39, wherein X is CR7.

[0057] Embodiment 41. A compound of Formula (I) or a pharmaceutically acceptable salt thereof according to any one of embodiments 1 to 40, wherein R7 is H.

[0058] Embodiment 42. A compound of Formula (I) or a pharmaceutically acceptable salt thereof according to any one of Embodiments 1 to 41, wherein R6 is Cl or CH3.

[0059] Embodiment 43. A compound of Formula (I) or a pharmaceutically acceptable salt thereof as described in embodiment 42, wherein R6 is Cl.

[0060] Embodiment 44. A compound of Formula (I) according to any one of Embodiments 1 to 43, or a pharmaceutically acceptable salt thereof, wherein R8 is F or H.

[0061] Embodiment 45. A compound of Formula (I) according to embodiment 44, or a pharmaceutically acceptable salt thereof, wherein R8 is H.

[0062] Embodiment 46. A compound of Formula (I) or a pharmaceutically acceptable salt thereof according to any one of embodiments 1 to 45, wherein R9 is H.

[0063] Embodiment 47.R 26 47. The compound of formula (I) according to any one of embodiments 1 to 46, or a pharmaceutically acceptable salt thereof, wherein:

[0064] Embodiment 48 R 27 48. The compound of formula (I) according to any one of embodiments 1 to 47, or a pharmaceutically acceptable salt thereof, wherein:

[0065] Embodiment 49.R 26 and R 27 and R are deuterium.

[0066] Embodiment 50. A compound of Formula (I) or a pharmaceutically acceptable salt thereof according to any one of Embodiments 1 to 49, wherein R3 is -CH2-CH3.

[0067] Embodiment 51. A compound of Formula (I) or a pharmaceutically acceptable salt thereof according to any one of embodiments 1 to 50, wherein y is 0.

[0068] Embodiment 52. Part: [ka] teeth, [ka] 52. The compound of formula (I) according to any one of embodiments 1 to 51, or a pharmaceutically acceptable salt thereof, selected from:

[0069] Embodiment 53. Part: [ka] teeth, [ka] especially [ka] Selected from or or [ka] or a pharmaceutically acceptable salt thereof.

[0070] Embodiment 54. Part: [ka] teeth, [ka] 52. The compound of formula (I) according to any one of embodiments 1 to 51, or a pharmaceutically acceptable salt thereof, selected from:

[0071] Embodiment 55. Part: [ka] teeth, [ka] 55. The compound of formula (I) according to any one of embodiments 1 to 54, or a pharmaceutically acceptable salt thereof, selected from:

[0072] Embodiment 56. Part: [ka] teeth, [ka] or a pharmaceutically acceptable salt thereof.

[0073] Embodiment 57. Part: [ka] teeth, [ka] or a pharmaceutically acceptable salt thereof.

[0074] Embodiment 58. Part: [ka] teeth, [ka] or a pharmaceutically acceptable salt thereof.

[0075] Embodiment 59.R 10 The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any of embodiments 1-58, wherein is H, F or Cl.

[0076] Embodiment 60.R 11 59. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of embodiments 1 to 59, wherein

[0077] Embodiment 61.R 1261. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of embodiments 1 to 60, wherein

[0078] Embodiment 62.R 13 62. The compound of formula (I) according to any one of embodiments 1 to 61, or a pharmaceutically acceptable salt thereof, wherein

[0079] Embodiment 63.R 14 is CH3 or H. The compound of Formula (I) or a pharmaceutically acceptable salt thereof according to any of embodiments 1-62.

[0080] Embodiment 64.R 14 64. The compound of Formula (I) or a pharmaceutically acceptable salt thereof according to any of embodiments 1-63, wherein:

[0081] Embodiment 65.R4 is [ka] 65. The compound of formula (I) according to any one of embodiments 1 to 64, or a pharmaceutically acceptable salt thereof, selected from:

[0082] Embodiment 66.R4 is [ka] 66. The compound of formula (I) according to any of embodiments 65, selected from:

[0083] Embodiment 67.R4 is [ka] 67. The compound of formula (I) according to any of embodiments 66, selected from:

[0084] Embodiment 68.R1 is [ka] Selected from; R 15 is H or F; R 16 is H or R 25 (R 24 )N- and; R 17 is H or F; R 18 is H or F; R 19 is H or F; R 20 is H or F; R 21 is H or CH3; R 22 are H, CF3, CHF2CH2, HOC(O)-CH2-, H3C-C(O)-, (H3C)3C-OC(O)-; R 23 are H, CF3, CHF2CH2-, (H3C)3C-OC(O)-; R 24 is CH3; R 25 is CHF2CH2-; and R4 is [ka] Selected from: where: R 10 is selected from H, F, Cl, CH3, and OCF3; R 11 is selected from H, Cl, F, and CH3; R 12 is selected from H, Cl, and CH3; R 13 is selected from H, and CH3; R 14 is selected from H, CH3, -CH2CH3, cyclopropyl, -OCHF2, OCF3, and Cl A compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of embodiments 1 and 24 to 31. or a pharmaceutically acceptable salt thereof.

[0085] Embodiment 69.R1 teeth, [ka] Selected from; portion: [ka] teeth, [ka] (Especially here [ka] teeth, [ka] [ka] is) Selected from; portion [ka] teeth, [ka] Selected from; and R4 is [ka] 32. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of embodiments 1 and 24 to 31, selected from

[0086] Embodiment 70.R1 is [ka] Selected from; portion: [ka] teeth, [ka] especially [ka] and [ka] Selected from; portion: [ka] teeth, [ka] especially, [ka] Selected from; And R4 is [ka] especially, [ka] Selected from: A compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of embodiments 1 and 24 to 31.

[0087] Embodiment 71. The compound is [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] 32. The compound of formula (I) according to any of embodiments 1, 24, 30 and 31, or a pharmaceutically acceptable salt thereof, selected from:

[0088] Embodiment 72. The compound is

[0089] [ka] [ka] [ka] 40. The compound of formula (I) according to any one of embodiments 1 to 39, or a pharmaceutically acceptable salt thereof, selected from:

[0090] Embodiment 73. The compound is [ka] 41. The compound of formula (I) according to embodiment 1, 39 or 40, selected from:

[0091] Embodiment 74. A compound of Formula (I) or a pharmaceutically acceptable salt thereof, according to any one of embodiments 1 to 73, wherein the compound is in non-zwitterionic form.

[0092] Embodiment 75. A compound of Formula (I) or a pharmaceutically acceptable salt thereof according to any one of Embodiments 1 to 73, wherein the compound is in zwitterionic form.

[0093] Embodiment 76. A compound of Formula (I) or a pharmaceutically acceptable salt thereof, according to any of embodiments 1 to 73, wherein the compound is a mixture of zwitterionic and non-zwitterionic forms.

[0094] Embodiment 77. A compound of Formula (I) or a pharmaceutically acceptable salt thereof according to any of Embodiments 1 to 73, wherein the R4 group exists in zwitterionic form (d) or (e). [ka]

[0095] Embodiment 78.R4 is [ka] 74. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any of embodiments 1-73, which exists in a zwitterionic form selected from:

[0096] Embodiment 79. A compound of formula (I) or a pharmaceutically acceptable salt thereof as described in any of embodiments 1 to 73, wherein R4 exists as a mixture of zwitterionic forms (a) and (b) as described in embodiment 78.

[0097] Embodiment 80.R4 is Non-zwitterionic form (e) and zwitterionic form (a) or (b) [ka] or The non-zwitterionic form (e) and the zwitterionic forms (a) and (b) [ka] 74. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any of embodiments 1-73, wherein

[0098] Embodiment 81. R4 is in the zwitterionic form (c): [ka] 74. The compound of formula (I) according to any one of embodiments 1 to 73, wherein:

[0099] Embodiment 82. R4 can be obtained in a zwitterionic form (c) and a non-zwitterionic form (d): [ka] 74. The compound of formula (I) according to any of embodiments 1-73, or a pharmaceutically acceptable salt thereof, wherein

[0100] Embodiment 83. The compound N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide is Non-zwitterionic forms: [ka] or zwitterionic form: [ka] or zwitterionic form: [ka] 32. The compound of formula (I) according to any of embodiments 1, 24, 30 or 31, wherein the compound is a mixture of any two or three of said forms.

[0101] Embodiment 84. The compound (R)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide is Non-zwitterionic forms: [ka] or zwitterionic form: [ka] or zwitterionic form: [ka] 32. The compound of formula (I) according to any of embodiments 1, 24, 30 or 31, wherein the compound is a mixture of any two or three of said forms.

[0102] Embodiment 85. The compound N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide is Non-zwitterionic forms: [ka] or zwitterionic form: [ka] or zwitterionic form: [ka] 32. The compound of formula (I) according to any of embodiments 1, 24, 30 or 31, wherein the compound is a mixture of any two or three of said forms.

[0103] Embodiment 86. The compound 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide is Non-zwitterionic forms: [ka] or zwitterionic form: [ka] or zwitterionic form: [ka] 32. The compound of formula (I) according to any of embodiments 1, 24, 30 or 31, wherein the compound is a mixture of any two or three of said forms.

[0104] Embodiment 87: The compound N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide is Non-zwitterionic forms: [ka] or zwitterionic form: [ka] 32. The compound of formula (I) according to any of embodiments 1, 24, 30 or 31, or a mixture of said forms.

[0105] Embodiment 88. A compound of Formula (I) or a pharmaceutically acceptable salt thereof, according to any one of embodiments 1 to 73, wherein the compound is the sodium salt.

[0106] Embodiment 89. A compound of formula (I) or a pharmaceutically acceptable salt thereof, as described in any of embodiments 1 to 88, wherein the compound is in amorphous form. For example, the compound is a sodium salt in amorphous form.

[0107] Embodiment 90. A compound of Formula (I) or a pharmaceutically acceptable salt thereof, as described in any of embodiments 1 to 88, wherein the compound is in a crystalline form.

[0108] Embodiment 91. The compound is in crystalline form [ka] 32. The compound of formula (I) according to any of embodiments 1, 24, 30 and 31, wherein

[0109] Embodiment 92. The compound is in crystalline form [ka] 32. The compound of formula (I) according to any of embodiments 1, 24, 30 and 31, wherein

[0110] Embodiment 93. The compound is in crystalline form. [ka] 32. The compound of formula (I) according to any of embodiments 1, 24, 30 and 31, wherein

[0111] Embodiment 94. A compound of Formula I according to any of embodiments 1 to 31. The compound is in crystalline form [ka] 32. The compound of formula (I) according to any of embodiments 1, 24, 30 and 31, wherein

[0112] Embodiment 95. The compound is in crystalline form. [ka] 32. The compound of formula (I) according to any of embodiments 1, 24, 30 and 31, wherein

[0113] Embodiment 96. A compound of formula (I) as described in embodiments 91-95, wherein the compound is in substantially pure form.

[0114] Embodiment 97. The crystalline form of embodiment 91 comprises four or more 2θ values ​​selected from the group consisting of 11.85±0.2, 13.71±0.2, 14.46±0.2, 15.33±0.2, 17.03±0.2, 18.33±0.2, 19.98±0.2, 22.42±0.2, 22.95±0.2, and 27.20±0.2 at a temperature of about 22°C; or 6.78±0.2, 8.97±0.2, 11.88±0.2, 13.55±0.2, 13.74±0.2, 14.48±0.2 at a temperature of about 22°C. 2. The compound of formula (I) according to embodiment 1, characterized by an X-ray powder diffraction pattern comprising four or more 2θ values ​​selected from the group consisting of 0.2, 15.34±0.2, 16.83±0.2, 17.03±0.2, 18.30±0.2, 19.49±0.2, 19.94±0.2, 21.28±0.2, 21.51±0.2, 22.38±0.2, 22.91±0.2, 23.27±0.2, 25.41±0.2, 27.26±0.2, 29.03±0.2, 29.78±0.2 and 29.96±0.2.

[0115] Embodiment 98. The crystalline form of embodiment 91 comprises five or more 2θ values ​​selected from the group consisting of 11.85±0.2, 13.71±0.2, 14.46±0.2, 15.33±0.2, 17.03±0.2, 18.33±0.2, 19.98±0.2, 22.42±0.2, 22.95±0.2, and 27.20±0.2 at a temperature of about 22°C; or 6.78±0.2, 8.97±0.2, 11.88±0.2, 13.55±0.2, 13.74±0.2, 14.48±0.2 at a temperature of about 22°C. 0.2, 15.34±0.2, 16.83±0.2, 17.03±0.2, 18.30±0.2, 19.49±0.2, 19.94±0.2, 21.28±0.2, 21.51±0.2, 22.38±0.2, 22.91±0.2, 23.27±0.2, 25.41±0.2, 27.26±0.2, 29.03±0.2, 29.78±0.2 and 29.96±0.2.

[0116] Embodiment 99. The compound of formula (I) as described in embodiment 1, wherein the crystalline form as described in embodiment 91 is characterized by an X-ray powder diffraction pattern substantially the same as the X-ray powder diffraction pattern shown in FIG. 1 or FIG. 7.

[0117] Embodiment 100. The crystalline form described in embodiment 92 is a compound of formula (I) according to embodiment 1, characterized by a powder X-ray diffraction pattern at a temperature of about 22°C comprising four or more 2θ values ​​selected from the group consisting of 10.29±0.2, 13.31±0.2, 14.01±0.2, 15.26±0.2, and 17.34±0.2.

[0118] Embodiment 101. The compound of formula (I) as defined in embodiment 1, wherein the crystalline form as defined in embodiment 92 is further characterized by a powder X-ray diffraction pattern at a temperature of about 22°C comprising five 2θ values ​​selected from the group consisting of 10.29±0.2, 13.31±0.2, 14.01±0.2, 15.26±0.2, and 17.34±0.2.

[0119] Embodiment 102. The compound of formula (I) as defined in embodiment 1, wherein the crystalline form as defined in embodiment 92 is characterized by an X-ray diffraction spectrum substantially identical to the powder X-ray diffraction pattern shown in FIG. 2.

[0120] Embodiment 103. The crystalline form of embodiment 93 is the compound of formula (I) of embodiment 1, characterized by an X-ray powder diffraction pattern at a temperature of about 22°C comprising four or more 2θ values ​​selected from the group consisting of 9.45±0.2, 12.75±0.2, 13.28±0.2, 21.69±0.2, 25.25±0.2, and 26.85±0.2.

[0121] Embodiment 104. The compound of formula (I) according to embodiment 1, wherein the crystalline form according to embodiment 93 is further characterized by an X-ray powder diffraction pattern at a temperature of about 22°C comprising five or more 2θ values ​​selected from the group consisting of 9.45±0.2, 12.75±0.2, 13.28±0.2, 21.69±0.2, 25.25±0.2, and 26.85±0.2.

[0122] Embodiment 105. The compound of formula (I) as defined in embodiment 1, wherein the crystalline form as defined in embodiment 93 is characterized by an X-ray powder diffraction pattern substantially the same as the X-ray powder diffraction pattern shown in FIG. 3.

[0123] Embodiment 106. The crystalline form of embodiment 94 is the compound of formula (I) of embodiment 1, characterized by a powder X-ray diffraction pattern at a temperature of about 22°C comprising four or more 2θ values ​​selected from the group consisting of 11.81±0.2, 13.75±0.2, 14.45±0.2, 15.32±0.2, 17.04±0.2, 17.40±0.2, 18.27±0.2, 19.95±0.2, 22.92±0.2, and 27.13±0.2.

[0124] Embodiment 107. The crystalline form of embodiment 94 is the compound of formula (I) of embodiment 1, further characterized by a powder X-ray diffraction pattern at a temperature of about 22°C comprising five or more 2θ values ​​selected from the group consisting of 11.81±0.2, 13.75±0.2, 14.45±0.2, 15.32±0.2, 17.04±0.2, 17.40±0.2, 18.27±0.2, 19.95±0.2, 22.92±0.2, and 27.13±0.2.

[0125] Embodiment 108. The compound of formula (I) as defined in embodiment 1, wherein the crystalline form as defined in embodiment 94 is characterized by an X-ray diffraction spectrum substantially identical to the powder X-ray diffraction pattern shown in FIG. 4.

[0126] Embodiment 109. The crystalline form of embodiment 95 is the compound of formula (I) of embodiment 1, characterized by an X-ray powder diffraction pattern at a temperature of about 22°C comprising four or more 2θ values ​​selected from the group consisting of 13.20±0.2, 14.78±0.2, 15.97±0.2, 16.91±0.2, 19.95±0.2, 20.85±0.2, 24.43±0.2, 25.47±0.2, and 31.06±0.2.

[0127] Embodiment 110. The compound of formula (I) of embodiment 1, wherein the crystalline form of embodiment 95 is further characterized by an X-ray powder diffraction pattern at a temperature of about 22°C comprising five or more 2θ values ​​selected from the group consisting of 13.20±0.2, 14.78±0.2, 15.97±0.2, 16.91±0.2, 19.95±0.2, 20.85±0.2, 24.43±0.2, 25.47±0.2, and 31.06±0.2.

[0128] Embodiment 111. The compound of formula (I) as defined in embodiment 1, wherein the crystalline form as defined in embodiment 95 is characterized by an X-ray diffraction spectrum substantially identical to the powder X-ray diffraction pattern shown in FIG.

[0129] Embodiment 112. Formula (I) is a compound of formula 1a: [ka] [Wherein R1 is [ka] Selected from; R 33 is F; R 15 is halo, azetidinyl, or pyrrolidinyl, wherein said azetidinyl and pyrrolidinyl are linked to the remainder of the molecule via an N atom and are unsubstituted or substituted with one or two F; R 16 is R 25 (R 24 )N- (where R 24 is H or (C1-C2) alkyl, and R 25 is H or (C1-C2) alkyl; R 17 is a halo; R 18 is a halo; R 19 is a halo; R 20 is a halo; R 21 is (C1-C2) alkyl; R 22 and R 23 are each independently (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo, HOC(O)-(CH2) n -, H3C-C(O)(CH2) n -, (H3C)3C-OC(O)(CH2) n -; where n is 0, 1, or 2 Selected from; and R 30 is CH3; R2 is the part [ka] and where: R6 is selected from H, halo, (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo; R8 is selected from H, halo, and (C1-C4) alkyl unsubstituted or substituted with 1, 2, or 3 halo; R9 is selected from H, halo, (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo; R 28 is selected from SF5, halo, (C1-C4)alkyl unsubstituted or substituted with 1, 2 or 3 halo, and -C(O)H; X is selected from C-R7 and N; R7 is selected from H and halo; R 26 is H and R 27 is H; R3 is -CH2CH3 or CH3; A is a linker selected from -C(O)- and -S(O)-, preferably -C(O)-; and R4 is [ka] is selected from where: R 10 is selected from H, halo, (C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents, —O—(C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; R 11 is selected from H, halo, (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; R 12 is selected from H, halo, (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; R 13is selected from H, —S—CH halo, (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; and R 14 is selected from H, halo, (C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents, —O—(C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents, and cyclopropyl or a pharmaceutically acceptable salt thereof. Preferably, formula (I) or 1a is 1g: [ka] and more preferably 1h: [ka] or a pharmaceutically acceptable salt thereof.

[0130] Embodiment 113. Formula (I) is a compound of formula 1b: [ka] [Wherein R1 is [ka] Selected from; R 33 is F; R 15 is halo, azetidinyl, or pyrrolidinyl, wherein said azetidinyl and pyrrolidinyl are linked to the remainder of the molecule via an N atom and are unsubstituted or substituted with one or two F; R 16 is R 25 (R 24 )N- (where R 24 is H or (C1-C2) alkyl, and R 25is H or (C1-C2) alkyl; R 17 is a halo; R 18 is a halo; R 19 is a halo; R 20 is a halo; R 21 is (C1-C2) alkyl; R 22 and R 23 are each independently (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo, HOC(O)-(CH2) n -, H3C-C(O)(CH2) n -, (H3C)3C-OC(O)(CH2) n -; where n is 0, 1, or 2 Selected from; and R 30 is CH3; R2 is part: [ka] (In the formula, R6 is selected from H, halo, (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo; R8 is selected from H, halo, and (C1-C4) alkyl unsubstituted or substituted with 1, 2, or 3 halo; R9 is selected from H, halo, (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo; R 28 is selected from SF5, halo, (C1-C4)alkyl unsubstituted or substituted with 1, 2 or 3 halo, and -C(O)H; X is selected from C-R7 and N; and R7 is selected from H and halo and; R 26 is H and R 27 is H; R3 is -CH2CH3 or CH3; A is a linker selected from -C(O)- and -S(O)-, preferably -C(O)-; and R4 is [ka] (In the formula, R 10 is selected from H, halo, (C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents, —O—(C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; R 11 is selected from H, halo, (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; R 12 is selected from H, halo, (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; R 13 is selected from H, —S—CH halo, (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; and R 14 is selected from H, halo, (C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents, —O—(C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents, and cyclopropyl Select from or a pharmaceutically acceptable salt thereof. Formula 1b is preferably 1b1: [ka] and In particular, where Y is N.

[0131] Embodiment 114. Formula (I) is a compound of formula 1c: [ka] [Wherein R1 is [ka] Selected from; R 33 is F; R 15 is halo, azetidinyl, or pyrrolidinyl, wherein said azetidinyl and pyrrolidinyl are linked to the remainder of the molecule via an N atom and are unsubstituted or substituted with one or two F; R 16 is R 25 (R 24 )N- (where R 24 is H or (C1-C2) alkyl, and R 25 is H or (C1-C2) alkyl; R 17 is a halo; R 18 is a halo; R 19 is a halo; R 20 is a halo; R 21 is (C1-C2)] alkyl; R 22 and R 23 are each independently (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo, HOC(O)-(CH2) n -, H3C-C(O)(CH2) n -, (H3C)3C-OC(O)(CH2) n -; where n is 0, 1, or 2 Selected from; and R 30 is CH3; R2 is part: [ka] (In the formula, R6 is selected from H, halo, (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo; R8 is selected from H, halo, and (C1-C4) alkyl unsubstituted or substituted with 1, 2, or 3 halo; R9 is selected from H, halo, (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo; R 28 is selected from SF5, halo, (C1-C4)alkyl unsubstituted or substituted with 1, 2 or 3 halo, and -C(O)H; X is selected from C-R7 and N; and R7 is selected from H and halo and; R 26 is H and R 27 is H; R3 is -CH2CH3 or CH3; A is a linker selected from -C(O)- and -S(O)-, preferably -C(O)-; and R4 is [ka] (In the formula, R 10 is selected from H, halo, (C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents, —O—(C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; R 11 is selected from H, halo, (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; R 12is selected from H, halo, (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; R 13 is selected from H, —S—CH halo, (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; and R 14 is selected from H, halo, (C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents, —O—(C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents, and cyclopropyl (selected from or a pharmaceutically acceptable salt thereof. Formula 1c is preferably 1c1: [ka] and In particular, where Y is N.

[0132] Embodiment 115. Formula (I) is a compound of formula 1d: [ka] [Wherein R1 is [ka] Selected from; R 33 is F; R 15 is halo, azetidinyl, or pyrrolidinyl, wherein said azetidinyl and pyrrolidinyl are linked to the remainder of the molecule via an N atom and are unsubstituted or substituted with one or two F; R 16 is R 25 (R 24 )N- (where R 24 is H or (C1-C2) alkyl, and R 25is H or (C1-C2) alkyl; R 17 is a halo; R 18 is a halo; R 19 is a halo; R 20 is a halo; R 21 is (C1-C2) alkyl; R 22 and R 23 teeth, Each independently, (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo, HOC(O)-(CH2) n -, H3C-C(O)(CH2) n -, (H3C)3C-OC(O)(CH2) n -; where n is 0, 1, or 2 Selected from; and R 30 is CH3; R2 is part: [ka] (In the formula, R6 is selected from H, halo, (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo; R8 is selected from H, halo, and unsubstituted or substituted with 1, 2 or 3 halo (C1-C4); R9 is selected from H, halo, unsubstituted or substituted with 1, 2 or 3 halo (C1-C4); R 28 is selected from SF5, halo, unsubstituted or substituted with 1, 2 or 3 halo (C1-C4), and —C(O)H; X is selected from C-R7 and N; and R7 is selected from H and halo and; R 26 is H and R 27 is H; R3 is -CH2CH3 or CH3; A is a linker selected from -C(O)- and -S(O)-, preferably -C(O)-; and R4 is [ka] (In the formula, R 10 is selected from H, halo, (C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents, —O—(C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; R 11 is selected from H, halo, (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; R 12 is selected from H, halo, (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; R 13 is selected from H, —S—CH halo, (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; and R 14 is selected from H, halo, (C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents, —O—(C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents, and cyclopropyl Select from or a pharmaceutically acceptable salt thereof. Formula 1d is preferably 1d1: [ka] and In particular, where Y is N.

[0133] Embodiment 116. Formula (I) is represented by formula 1e: [ka] [Wherein R1 is [ka] Selected from; R 33 is F; R 15 is halo, azetidinyl, or pyrrolidinyl, wherein said azetidinyl and pyrrolidinyl are linked to the remainder of the molecule via an N atom and are unsubstituted or substituted with one or two F; R 16 is R 25 (R 24 )N- (where R 24 is H or (C1-C2) alkyl, and R 25 is H or (C1-C2) alkyl; R 17 is a halo; R 18 is a halo; R 19 is a halo; R 20 is a halo; R 21 is (C1-C2) alkyl; R 22 and R 23 are each independently (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo, HOC(O)-(CH2) n -, H3C-C(O)(CH2) n -, (H3C)3C-OC(O)(CH2) n -; where n is 0, 1, or 2 Selected from; and R 30 is CH3; R2 is part: [ka] (In the formula, R6 is selected from H, halo, (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo; R8 is selected from H, halo, unsubstituted or substituted with 1, 2 or 3 halo (C1-C4); R9 is selected from H, halo, unsubstituted or substituted with 1, 2 or 3 halo (C1-C4); R 28 is selected from SF5, halo, unsubstituted or substituted with 1, 2 or 3 halo (C1-C4), and —C(O)H; X is selected from C-R7 and N; and R7 is selected from H and halo and; R 26 is H and R 27 is H; R3 is -CH2CH3 or CH3; A is a linker selected from -C(O)- and -S(O)-, preferably -C(O)-; and R4 is [ka] (In the formula, R 10 is selected from H, halo, (C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents, —O—(C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; R 11 is selected from H, halo, (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; R 12is selected from H, halo, (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; R 13 is selected from H, —S—CH halo, (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; and R 14 is selected from H, halo, (C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents, —O—(C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents, cyclopropyl Select from or a pharmaceutically acceptable salt thereof. Formula 1e is preferably 1e1: [ka] and In particular, where Y is N.

[0134] Embodiment 117. Formula (I) is a compound of formula 1f: [ka] [Wherein R1 is [ka] Selected from; R 33 is F; R 15 is halo, azetidinyl, or pyrrolidinyl, wherein said azetidinyl and pyrrolidinyl are linked to the remainder of the molecule via an N atom and are unsubstituted or substituted with one or two F; R 16 is R 25 (R 24 )N- (where R 24 is H or (C1-C2) alkyl, and R 25is H or (C1-C2) alkyl; R 17 is a halo; R 18 is a halo; R 19 is a halo; R 20 is a halo; R 21 is (C1-C2) alkyl; R 22 and R 23 are each independently (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo, HOC(O)-(CH2) n -, H3C-C(O)(CH2) n -, (H3C)3C-OC(O)(CH2) n -; where n is 0, 1, or 2 Selected from; and R 30 is CH3; R2 is part: [ka] (In the formula, R6 is selected from H, halo, (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo; R8 is selected from H, halo, (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo; R9 is selected from H, halo, (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo; R 28 is selected from SF5, halo, (C1-C4)alkyl unsubstituted or substituted with 1, 2 or 3 halo, and -C(O)H; X is selected from C-R7 and N; R7 is selected from H and halo and; R 26 is H and R 27 is H; R3 is -CH2CH3 or CH3; A is a linker selected from -C(O)- and -S(O)-, preferably -C(O)-; and R4 is [ka] (In the formula, R 10 is selected from H, halo, (C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents, —O—(C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; R 11 is selected from H, halo, (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; R 12 is selected from H, halo, (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; R 13 is selected from H, —S—CH halo, (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; and R 14 is selected from H, halo, (C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents, —O—(C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents, cyclopropyl Select from or a pharmaceutically acceptable salt thereof. Formula 1f is preferably 1f1: [ka] In particular, where Y is N.

[0135] Embodiment 1.1. A compound of formula (I) according to embodiment 1, or a pharmaceutically acceptable salt thereof, wherein: [ka] (I) is 1 g, and R1 is [ka] Selected from; R 15 is H or F; R 16 is H or R 25 (R 24 )N- and; R 17 is H or F; R 18 is H or F; R 19 is H or F; R 20 is H or F; R 21 is H or CH3; R 22 are CF3, CHF2CH2, HOC(O)-CH2-, H3C-C(O)-, (H3C)3C-OC(O)-; R 23 are CF3, CHF2CH2-, (H3C)3C-OC(O)-; R 24 is CH3; R 25 is CHF2CH2-; R 26 is CH3, H or deuterium; R 27 is H or deuterium; R2 is part: [ka] wherein R6 is selected from H, Cl, CH3, F, and Br; R8 is selected from H, Cl, F, and CF3; R9 is selected from H, CH3, and Cl; R 28 is selected from CF3, CF2H, -CH2CH3, Cl, SF5, Br, and -C(O)H; X is selected from C-R7 and N; R7 is selected from H and F and; R3 is selected from CH3, CH2CH3, cyclopropyl, and hydroxyethyl; R4 is [ka] (In the formula, R 10 is selected from H, F, Cl, CH3, and OCF3; R 11 is selected from H, Cl, F, and CH3; R 12 is selected from H, Cl, and CH3; R 13 is selected from H, and CH3; R 14 is selected from H, CH, —CHCH, cyclopropyl, —OCHF, OCF, and Cl. Selected from; y is 0, 1 or 2; R5 is CH3, or two R5 groups on adjacent carbon atoms, together with the carbon atoms to which they are attached, form a fused cyclobutyl ring: [ka] Forming; and * indicates the attachment point A compound of formula (I) or a pharmaceutically acceptable salt thereof as described in embodiment 1. or a pharmaceutically acceptable salt thereof.

[0136] Embodiment 2.1.R1 is [ka] or a pharmaceutically acceptable salt thereof.

[0137] Embodiment 3.1.R1 is [ka] or a pharmaceutically acceptable salt thereof.

[0138] Embodiment 4.1.R1 is [ka] or a pharmaceutically acceptable salt thereof.

[0139] Embodiment 5.1.R1 is [ka] or a pharmaceutically acceptable salt thereof,

[0140] Embodiment 6.1.R 28 is selected from CF3, SF5 and Br; or a pharmaceutically acceptable salt thereof;

[0141] Embodiment 7.1.R 28 The compound of formula (I) or a pharmaceutically acceptable salt thereof according to embodiments 1-6, wherein is selected from CF3.

[0142] Embodiment 8.1.X is a compound of Formula (I) as described in any one of embodiments 1 to 7, or a pharmaceutically acceptable salt thereof, which is CR7.

[0143] Embodiment 9.1. A compound of Formula (I) or a pharmaceutically acceptable salt thereof as described in Embodiments 1-8 wherein R7 is H.

[0144] Embodiment 10.1. A compound of Formula (I) or a pharmaceutically acceptable salt thereof according to any of embodiments 1 to 9, wherein R6 is Cl or CH3.

[0145] Embodiment 11.1. A compound of Formula (I) or a pharmaceutically acceptable salt thereof according to any one of Embodiments 1 to 10, wherein R6 is Cl.

[0146] Embodiment 12.1. A compound of Formula (I) according to any one of Embodiments 1 to 11, or a pharmaceutically acceptable salt thereof, wherein R8 is F or H.

[0147] Embodiment 13.1. A compound of Formula (I) or a pharmaceutically acceptable salt thereof as defined in any one of Embodiments 1 to 12, wherein R8 is H.

[0148] Embodiment 14.1. A compound of Formula (I) or a pharmaceutically acceptable salt thereof as described in any one of Embodiments 1 to 13, wherein R9 is H.

[0149] Embodiment 15.1.R 26 15. The compound of formula (I) or a pharmaceutically acceptable salt thereof as defined in embodiments 1-14, wherein

[0150] Embodiment 16.1.R 27 is H; or a pharmaceutically acceptable salt thereof.

[0151] Embodiment 17.1.R 26 and R 27 and R are each deuterium. 15. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of embodiments 1 to 14.

[0152] Embodiment 18.1. A compound of Formula (I) or a pharmaceutically acceptable salt thereof as described in any one of Embodiments 1 to 18, wherein R3 is -CH2-CH3.

[0153] Embodiment 19.1. A compound of Formula (I) or a pharmaceutically acceptable salt thereof according to any of embodiments 1 to 18, wherein y is 0.

[0154] Embodiment 20.1. Part: [ka] teeth, [ka] 19. The compound of formula (I) according to embodiments 1-18, selected from:

[0155] Embodiment 21.1 Part: [ka] teeth, [ka] 21. The compound of formula (I) according to embodiments 1 to 18 and 20, selected from: or a pharmaceutically acceptable salt thereof.

[0156] Embodiment 22.1 Part: [ka] teeth, [ka] 21. The compound of formula (I) according to embodiments 1 to 18 and 20, selected from: or a pharmaceutically acceptable salt thereof.

[0157] Embodiment 23.1 Part: [ka] teeth, [ka] 23. The compound of formula (I) according to embodiments 1-22, selected from: or a pharmaceutically acceptable salt thereof.

[0158] Embodiment 24.1 Part: [ka] teeth, [ka] 24. The compound of formula (I) according to embodiments 1-23, selected from: or a pharmaceutically acceptable salt thereof.

[0159] Embodiment 25.1 Part: [ka] teeth, [ka] 25. The compound of formula (I) according to embodiments 1-24, selected from: or a pharmaceutically acceptable salt thereof.

[0160] Embodiment 26.1 Part: [ka] teeth, [ka] 26. The compound of formula (I) according to embodiments 1 to 25, selected from: or a pharmaceutically acceptable salt thereof.

[0161] Embodiment 27.1 R 10 27. Compounds of formula (I) or pharmaceutically acceptable salts thereof according to embodiments 1-26, wherein is H, F or Cl.

[0162] Embodiment 28.1 R 11 is H or CH3; or a pharmaceutically acceptable salt thereof.

[0163] Embodiment 29.1 R 12 is H; or a pharmaceutically acceptable salt thereof.

[0164] Embodiment 30.1 R 13 is H; or a pharmaceutically acceptable salt thereof.

[0165] Embodiment 31.1 R 14 is CH3 or H. The compound of Formula (I) or a pharmaceutically acceptable salt thereof according to embodiments 1-30.

[0166] Embodiment 32.1 R 14 32. The compound of Formula (I) or a pharmaceutically acceptable salt thereof according to embodiments 1-31, wherein is CH3. Embodiment 33.1 R4 is [ka] 33. The compound of formula (I) according to embodiments 1-32, selected from: or a pharmaceutically acceptable salt thereof.

[0167] Embodiment 34.1 R4 is [ka] 34. The compound of formula (I) according to embodiments 1-33, selected from: or a pharmaceutically acceptable salt thereof.

[0168] Embodiment 35.1 R4 is [ka] 35. The compound of formula (I) according to embodiments 1-34, selected from: or a pharmaceutically acceptable salt thereof.

[0169] Embodiment 36.1 R1 is [ka] Selected from; R 15 is H or F; R 16 is H or R 25 (R 24 )N- and; R 17 is H or F; R 18 is H or F; R 19 is H or F; R 20 is H or F; R 21 is H or CH3; R 22 are H, CF3, CHF2CH2, HOC(O)-CH2-, H3C-C(O)-, (H3C)3C-OC(O)-; R 23 are H, CF3, CHF2CH2-, (H3C)3C-OC(O)-; R 24 is CH3; R 25 is CHF2CH2-; and R4 is [ka] (In the formula, R 10 is selected from H, F, Cl, CH3, and OCF3; R 11 is selected from H, Cl, F, and CH3; R 12 is selected from H, Cl, and CH3; R 13 is selected from H, and CH3; R 14 is selected from H, CH, —CHCH, cyclopropyl, —OCHF, OCF, and Cl. Selected from A compound of formula (I) or a pharmaceutically acceptable salt thereof according to embodiments 1 and 6 to 26, or a pharmaceutically acceptable salt thereof.

[0170] Embodiment 37.1 R1 is [ka] Selected from; And part: [ka] teeth, [ka] Selected from: [ka] teeth, [ka] Selected from; and R4 is [ka] Selected from: A compound of formula (I) as defined in embodiment 1 or a pharmaceutically acceptable salt thereof.

[0171] Embodiment 38.1 R1 is [ka] Selected from: [ka] teeth, [ka] Selected from: [ka] teeth, [ka] especially, [ka] Selected from; And R4 is [ka] especially, [ka] Selected from: A compound of formula (I) or a pharmaceutically acceptable salt thereof as defined in embodiment 37.

[0172] In a further aspect, the invention is as claimed herein.

[0173] In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof: [ka] [In the formula, R1 is cycloalkenyl, wherein said cycloalkenyl is a partially unsaturated monocyclic ring containing 5 or 6 ring carbon atoms, said cycloalkenyl being unsubstituted or having 1, 2, 3 or 4, preferably 1 or 2, R 33 where R 33 is halo, and said cycloalkenyl or halo-substituted cycloalkenyl has 0, 1 or 2 R 15 Is substituted by a substituent, Alternatively, R is heterocyclyl, wherein the heterocyclyl is a fully saturated or partially unsaturated 5- or 6-membered group comprising ring carbon atoms and 1 or 2 ring heteroatoms independently selected from N, O and S, wherein the heterocyclyl is unbridged or bridged, the bridge being 1 or 2 carbon atoms, wherein the heterocyclyl is unsubstituted or has 1, 2, 3 or 4, preferably 1 or 2, R 33 where R 33 is halo, and said heterocyclyl or halo-substituted heterocyclyl is independently R 15 , R 16 , R 17 , R 18 , R 19 , R 20 , R 22 and R 23 or substituted by 0, 1 or 2 substituents selected from Alternatively, R is heteroaryl, wherein said heteroaryl is a 5- or 6-membered fully unsaturated monocyclic group containing ring carbon atoms and 1, 2, 3, or 4 ring heteroatoms independently selected from N, O, and S, preferably 1 or 2 ring heteroatoms, wherein the total number of ring S atoms does not exceed 1 and the total number of ring O atoms does not exceed 1, and wherein said heteroaryl is unsubstituted or R 21 and R 30 and is substituted by 1, 2, or 3 substituents independently selected from 21 and R 30 is independently selected from halo and (C1-C4) alkyl, wherein said (C1-C4) alkyl is unsubstituted or substituted with 1, 2 or 3 halo; And each R 15 , R 16 , R 17 , R 18 , R 19 , R 20 , R 22 and R 23 is, independently, Halo unsubstituted or substituted with 1, 2 or 3 halo (C 1~ C4) alkyl-O-, (C1-C4)alkyl unsubstituted or substituted by OH, —O—(C1-C2)alkyl or 1, 2 or 3 halo, HOC(O)-(CH2) n -, H3C-C(O)(CH2) n -, (C1-C4) alkyl-OC(O)(CH2) n , =O azetidinyl or pyrrolidinyl, wherein the azetidinyl and pyrrolidinyl are linked to the rest of the molecule via an N atom and are each unsubstituted or substituted with one or two F, ·R 25 (R 24 )N-(where R 24 is H or (C-C) alkyl unsubstituted or substituted with 1, 2 or 3 halo, and R 25 is H or (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo. OH Selected from where n is 0, 1 or 2; Especially R1, cycloalkenyl, wherein the cycloalkenyl is a partially unsaturated monocyclic ring containing 5 or 6 ring carbon atoms, and the cycloalkenyl is unsubstituted or contains one or two R 33 where R 33 is halo, preferably F, and said cycloalkenyl or halo-substituted cycloalkenyl has zero or one R 15is substituted by a substituent, preferably one substituent, and R 15 teeth, h) unsubstituted or substituted with 1, 2 or 3 halo (C 1~ C2) alkyl-O-, i) (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo; j) HOC(O)-(CH2) n -, k) H3C-C(O)(CH2) n -, l) H3C-OC(O)(CH2) n , m)=O, and n)R 25 (R 24 )N-, H(where R 24 is H or (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo, and R 25 is H or (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo. is selected from n is 0 or 1; where: R of the cycloalkenyl or halo-substituted cycloalkenyl 15 Substituents a) through g) are not present on the ring atom adjacent to the ring atom at which the cycloalkenyl or halo-substituted cycloalkenyl is attached to the remainder of the molecule, and preferably said cycloalkenyl or halo-substituted cycloalkenyl has one R in the ring para position relative to the remainder of the molecule. 15 is a 6-membered ring having a substituent, the cycloalkenyl or halo-substituted cycloalkenyl is linked to the remainder of the compound via an R1 ring carbon atom that is double-bonded to an adjacent R1 ring carbon atom; Alternatively, R is heterocyclyl, wherein the heterocyclyl is a fully saturated or partially unsaturated 5- or 6-membered group containing ring carbon atoms and one or two ring heteroatoms independently selected from N, NH, O, and S, and wherein the heterocyclyl is unbridged or bridged, the bridge being one or two carbon atoms, and wherein the heterocyclyl is unsubstituted or contains one or two R 33 where R 33 is halo, preferably F, and said heterocyclyl or halo-substituted heterocyclyl is independently R 15 , R 16 , R 17 , R 18 , R 19 , R 20 , R 22 and R 23 and wherein R is substituted by 0 or 1 substituent selected from 15 , R 16 , R 17 , R 18 , R 19 , R 20 , R 22 and R 23 is, independently, i) unsubstituted or substituted with 1, 2 or 3 halo (C 1~ C4) alkyl-O-, j) (C1-C4) alkyl unsubstituted or substituted with OH, —O—(C1-C2) alkyl or 1, 2 or 3 halo; k) HOC(O)-(CH2) n -, l) H3C-C(O)(CH2) n -, m) H3C-OC(O)(CH2) n , n)=O o)R 25 (R 24 )N-(where R 24 is H, (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo, and R 25 is H, (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo p)OH is selected from where n is 0 or 1; and the substituents a) to h) of said heterocyclyl or halo-substituted heterocyclyl are not present on the ring atom at which the heterocyclyl or halo-substituted heterocyclyl is attached to the remainder of the molecule, preferably said heterocyclyl or halo-substituted heterocyclyl is a 6-membered ring which has 0 or 1 substituent selected from a) to h) in the meta or para position relative to the remainder of the molecule, preferably in the para position; and the heterocyclyl is linked to the remainder of the compound via an R1 ring nitrogen atom or an R1 ring carbon atom that is double bonded to an adjacent ring atom; Alternatively, R is heteroaryl, wherein said heteroaryl is a 5- or 6-membered fully unsaturated monocyclic group containing ring carbon atoms and 1 or 2 ring heteroatoms independently selected from N, O and S, preferably N, wherein the total number of ring S atoms does not exceed 1 and the total number of ring O atoms does not exceed 1, and wherein said heteroaryl is unsubstituted or R 21 and R 30 and is substituted by one or two substituents independently selected from 21 and R 30 are independently selected from (C1-C2)alkyl, said (C1-C2)alkyl being unsubstituted or substituted with 1, 2 or 3 halo, wherein preferably said alkyl or haloalkyl substituent is not on the R1 ring atom adjacent to the R1 ring atom at which the heteroaryl is attached to the remainder of the molecule, and more preferably, when the heteroaryl is a 6-membered ring, said alkyl or haloalkyl substituent is in the ring para position relative to the remainder of the molecule. is provided.

[0174] More particularly, R1 is [ka] Selected from; R 33 is F; R 15is halo, azetidinyl, or pyrrolidinyl, wherein said azetidinyl and pyrrolidinyl are linked to the remainder of the molecule via an N atom and are unsubstituted or substituted with one or two F; R 16 is R 25 (R 24 )N- (where R 24 is H or (C1-C2) alkyl, and R 25 is H or (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo, especially F; R 17 is a halo; R 18 is a halo; R 19 is a halo; R 20 is a halo; R 21 is (C1-C2) alkyl; R 22 and R 23 are each independently (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo, HOC(O)-(CH2) n -, H3C-C(O)(CH2) n -, (H3C)3C-OC(O)(CH2) n -; where n is 0, 1, or 2 Selected from; and R 30 is CH3. More particularly, R1 is [ka] Selected from; R 15 is F; R 16 is R 25 (R 24 )N- and; R 17 is F; R 18 is F; R 19 is F; R 20 is F; R 21 is CH3; R 22 are CF3, CHF2CH2, HOC(O)-CH2-, H3C-C(O)-, (H3C)3C-OC(O)-; R 23 are CF3, CHF2CH2-, (H3C)3C-OC(O)-; R 24 is CH3; and R 25 is CHF2CH2-.

[0175] Preferably, R1 is [ka] is selected from.

[0176] In another embodiment, R2 is the moiety: [ka] and; R6 is H, Hello, (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo, (C3-C5)cycloalkyl unsubstituted or substituted by 1, 2 or 3 halo; -O-(C1-C4)alkyl, unsubstituted or substituted with 1, 2 or 3 halo; ·OH, and ·CN Selected from; R8 is selected from H, halo, and unsubstituted or substituted with 1, 2 or 3 halo (C1-C4); R9 is selected from H, O-CH3, OH, CN, CH3, and halo; R 28 teeth, SF5, H, ·-C(O)H, ·Halo; (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo, ·(C1-C4)alkynyl; (C1-C4) alkenyl, (C3-C5)cycloalkyl unsubstituted or substituted with 1, 2 or 3 halo, and OCF3 Selected from; and X is selected from C-R7 and N, where R7 is H or halo.

[0177] In particular, R2 is the part: [ka] (In the formula, R6 is selected from H, halo, (C1-C4) alkyl unsubstituted or substituted with 1, 2 or 3 halo; R8 is selected from H, halo, and unsubstituted or substituted with 1, 2 or 3 halo (C1-C4); R9 is selected from H, O-CH3, OH, CN, CH3, and halo; R 28 is selected from SF5, halo, unsubstituted or substituted with 1, 2 or 3 halo (C1-C4), and —C(O)H; X is selected from C-R7 and N; R7 is selected from H and halo is.

[0178] More particularly, R2 is the moiety: [ka] and; R6 is selected from H, Cl, CH3, F, and Br; R8 is selected from H, Cl, F, and CF3; R9 is selected from H, CH3, and Cl; R 28 is selected from CF3, CF2H, -CH2CH3, Cl, SF5, Br, and -C(O)H; X is selected from C-R7 and N; R7 is selected from H and F. More particularly, R2 is [ka] , preferably [ka] is a moiety selected from

[0179] In another embodiment, R 26 is H and R 27 is H.

[0180] In another embodiment, R3 is (C1-C4)alkyl unsubstituted or substituted with 1, 2 or 3 substituents independently selected from halo and OH, particularly (C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 substituents independently selected from halo and OH, preferably -CH2CH3 or CH3, more preferably -CH2CH3.

[0181] In another embodiment, Y is N, and [ka] is a Y linked by a single bond.

[0182] In another embodiment, [ka] is K connected by a single bond, and K is -CH2-, -CH2CH2-, -NH-, and (5-membered ring: [ka] (forming a bond), J is N, and A is selected from -C(O)-, -S(O)-, -S(O)2-, and [ka] is a linker selected from

[0183] especially, [ka] are Ks linked by a single bond, where K is -CH2-, J is N, and A is -C(O)-, -S(O)-, -S(O)2-, and [ka] is a linker selected from

[0184] In another embodiment, A is a linker selected from -C(O)- and -S(O)2-, preferably -C(O)-.

[0185] In another embodiment, R5 is independently -(C 1~ C4) alkyl, preferably methyl; where two R5 substituents on the same ring carbon atom, together with the carbon atom to which they are attached, form (C 3~ C4) may form a cycloalkylspiro ring or a 3- or 4-membered heterocyclylspiro ring, wherein the heterocyclylspiro ring contains ring carbon atoms and one ring heteroatom selected from O, N, and S; · [ka] is a carbon-nitrogen single bond, K and the R5 substituents on the adjacent carbon atom are joined to form ring C: [ka] wherein ring C is a fused (C3-C6) cycloalkyl ring, in particular a fused cyclobutyl ring, a fused (C3-C6) heterocyclyl ring or a fused phenyl ring, said fused (C3-C6) heterocyclyl ring containing ring carbon atoms and one ring heteroatom selected from O, N and S, and when K is -CH2- and J is N, the two R5 substituents may be linked to form a (C1-C3) alkylene or heteroalkylene bridge, where the heteroalkylene bridge is one heteroatom selected from N and O, or is -CH2-O-CH2-. is selected from.

[0186] In particular, R5 can independently -(C 1~ C4) alkyl, preferably methyl; · [ka] is a carbon-nitrogen single bond, K and the R5 substituents on the adjacent carbon atom are joined to form ring C: [ka] wherein ring C is a fused (C3-C6) cycloalkyl ring, in particular a fused cyclobutyl ring, or a fused (C3-C6) heterocyclyl ring, said fused (C3-C6) heterocyclyl ring containing ring carbon atoms and one ring heteroatom selected from O, N and S, and when K is -CH2- and J is N, the two R5 substituents may be linked to form a (C1-C3) alkylene bridge or a heteroalkylene bridge, wherein the heteroalkylene bridge is one heteroatom selected from N and O, or is -CH2-O-CH2-. is selected from.

[0187] More particularly, R5 is independently -(C 1~ C2) alkyl, preferably methyl, and · [ka] is a carbon-nitrogen single bond, K and the R5 substituents on the adjacent carbon atom are joined to form ring C: [ka] wherein ring C is a fused (C3-C4) cycloalkyl ring, particularly a fused cyclobutyl ring. is selected from.

[0188] More particularly, R5 is independently CH3 and y is 1 or 2, and · [ka] is a carbon-nitrogen single bond, K and the R5 substituents on the adjacent carbon atom are joined to form ring C: [ka] wherein ring C is a fused cyclobutyl ring. is selected from.

[0189] In another embodiment, y is 0, 1, 2 or 3, preferably 0, 1 or 2.

[0190] In another embodiment, R4 is [ka] (In the formula, R 10 is selected from H, halo, (C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents, —O—(C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; R 11 is selected from H, halo, (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; R 12 is selected from H, halo, (C1-C2) alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; R 13 is selected from H, —S—CH, halo, (C-C) alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents; and R 14 is selected from H, halo, (C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents, —O—(C1-C2)alkyl unsubstituted or substituted with 1, 2 or 3 halo substituents, and cyclopropyl is selected from.

[0191] In particular, R4 is [ka] (In the formula, R 10 is selected from H, F, Cl, CH3, and OCF3; R 11 is selected from H, Cl, F, and CH3; R 12 is selected from H, Cl, and CH3; R 13 is selected from H, —S—CH3, and CH3; and R14 is selected from H, CH, —CHCH, cyclopropyl, —OCHF, OCF, and Cl. is selected from.

[0192] More particularly, R4 is [ka] is selected from.

[0193] More particularly, R4 is [ka] Preferably [ka] is selected from.

[0194] In another embodiment, Formula (I) is represented by Formula 1a: [ka] is.

[0195] In another embodiment, the moiety: [ka] teeth, [ka] especially, [ka] is selected from.

[0196] synthesis Also provided is a method for preparing a compound of formula (I) as described herein, particularly 1a, 1b, 1c, 1d, 1e, 1f, 1h or 1g, preferably 1a, more preferably 1g or 1h, or a pharmaceutically acceptable salt thereof as described herein.

[0197] Additionally provided are intermediate compounds used in the chemical synthesis of compounds of formula (I) described herein, e.g., formula 1a, preferably 1g or 1h, or a pharmaceutically acceptable salt thereof described herein.

[0198] In another aspect, there is provided an intermediate compound, or a process including an intermediate compound, as described below.

[0199] In particular, compounds or formulae such as: [ka] (Sodium salt of the compound of Example 42)

[0200] A compound of formula A or a salt thereof: [ka] (In the formula, R1, R2, R3, R 26 , R 27 , R5, y and Y are as defined herein). Compound: [ka]

[0201] Formula B: [ka] (In the formula, R1, R2, R3, R 26 , R 27, R5, y, and Y are as defined herein, and PG1 is a protecting group, or a salt thereof. Suitable protecting groups are well known to those skilled in the art and include BOC.

[0202] [ka] A compound or a salt thereof,

[0203] Formula C: [ka] (wherein R1, R3, R5, y, and Y are as defined herein, and PG2 is a protecting group) or a salt thereof. Suitable protecting groups are well known to those skilled in the art. Such protecting groups PG2 include BOC.

[0204] [ka] A compound or a salt thereof,

[0205] Formula D: [ka] or a salt thereof, wherein R3, R5, y, and Y are as defined herein, and PG3 is a protecting group. Suitable protecting groups are well known to those skilled in the art.

[0206] [ka] A compound or a salt thereof,

[0207] [ka] A compound or a salt thereof,

[0208] Also, compounds: [ka] 1. A method for making a compound of formula: [ka] The compound formula: [ka] and a compound of The present invention provides a method for the preparation of a hydroxybenzoate comprising reacting with coupling reagents and conditions known to those skilled in the art, particularly HOAt, EDCI, and DIPEA. Such reagents have the advantage of being suitable for scale-up given their availability and low cost, while enabling a robust process with good yields. Protected or unprotected forms can be used, as known to those skilled in the art.

[0209] Also, compounds: [ka] 1. A method for making a compound of formula: [ka] and / or [ka] and / or [ka] The present invention provides a method for preparing the compound of formula (I) via an intermediate of formula (I).

[0210] formulation In another aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. In a further embodiment, the composition comprises at least two pharmaceutically acceptable carriers, such as those described herein. The pharmaceutical composition may be formulated for a specific route of administration, such as oral administration, parenteral administration (e.g., by injection, infusion, transdermal, or topical administration), and rectal administration. Topical administration may also involve inhalation or intranasal application. The pharmaceutical composition of the present invention may be in the form of a solid formulation (including, but not limited to, capsules, tablets, pills, granules, powders, or suppositories), or a liquid formulation (including, but not limited to, solutions, suspensions, or emulsions). Tablets may be film-coated or enteric-coated according to methods known in the art. Typically, the pharmaceutical composition is a tablet or gelatin capsule containing the active ingredient in combination with one or more of the following: a) diluents, such as lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, and / or glycine; b) lubricants, such as silica, talc, stearic acid, its magnesium or calcium salts, and / or polyethylene glycol; in the case of tablets, c) binders, such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, and / or polyvinylpyrrolidone; optionally d) disintegrants, such as starch, agar, alginic acid or its sodium salt, or effervescent mixtures; and e) Absorbents, colorants, flavorings, and sweeteners.

[0211] Compounds intended for parenteral or oral administration can be solubilized using a variety of methods, including nanosuspensions, solid dispersions, and liposomes (van Hoogevest P., Xiangli L., and Alfred F. "Drug delivery strategies for poorly water-soluble drugs: The industrial perspective," Expert Opinion on Drug Delivery 2011, 8(11), 1481-1500).

[0212] Solid dispersion technology has been used to improve the dissolution properties and bioavailability of orally administered drugs (Dhirendra K et al: 'Solid dispersions: A review', Pakistan Journal of Pharmaceutical Sciences, Faculty of Pharmacy, University of Karachi, Pakistan, vol. 22, no. 2, 30 April 2000, pages 234-246).

[0213] Common methods for solubilizing compounds for parenteral administration are pH optimization or the use of cosolvents (e.g., PEG300, PEG400, propylene glycol, or ethanol). If these approaches are not feasible for some reason, the use of surfactants can be considered (e.g., Tween® 80 or Cremophor EL®). Cyclodextrins have been established as safe solubilizing agents. Compounds with high solubility in natural oils (e.g., propofol) can be solubilized in parenteral fat emulsions.

[0214] Also provided are pharmaceutical compositions comprising a compound of formula (I) described herein, e.g., formula 1a, preferably 1g or 1h, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.

[0215] In another embodiment, there is provided a pharmaceutical composition that is an amorphous solid dispersion comprising the compound N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide, or a pharmaceutically acceptable salt thereof.

[0216] In a further embodiment, the pharmaceutical composition comprises the compound N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide, or a pharmaceutically acceptable salt thereof, and an amino methacrylate copolymer, in particular Eudragit® E PO.

[0217] use The compounds of formula (I) of the present invention, in free form or in pharmaceutically acceptable salt form, exhibit valuable pharmacological properties, e.g., WRN inhibitory properties, as shown, for example, in the in vitro and in vivo tests provided in the following sections, and are therefore of use in therapy or as research chemicals, e.g., chemical probes and tool compounds.

[0218] Also provided are compounds of formula (I) as described herein, particularly 1a, 1b, 1c, 1d, 1e, 1f, 1h, or 1g. The compounds can be used as research chemicals, such as tool compounds or chemical probes, particularly for the study of WRN. In another embodiment, there is provided the use of compounds of formula (I) as described herein, particularly 1a, 1b, 1c, 1d, 1e, 1f, 1h, or 1g, as research chemicals, such as tool compounds or chemical probes, particularly for the study of WRN.

[0219] Also provided are compounds of formula (I) described herein, particularly compounds 1a, 1b, 1c, 1d, 1e, 1f, 1h, or 1g, preferably compounds 1a, more preferably compounds 1g or 1h, or pharmaceutically acceptable salts thereof, for use in treating cancer. Cancers that can be treated by WRN inhibition include cancers characterized by microsatellite instability-high (MSI-H) or deficient mismatch repair (dMMR). In particular, compounds of formula (I) described herein, such as compounds of formula 1a, preferably compounds 1g or 1h, or pharmaceutically acceptable salts thereof, can be useful in treating cancers characterized by microsatellite instability-high (MSI-H) or deficient mismatch repair (dMMR).

[0220] There is also provided a compound of formula (I) as described herein, in particular compound 1a, 1b, 1c, 1d, 1e, 1f, 1h or 1g, preferably compound 1a, more preferably 1g or 1h, or a pharmaceutically acceptable salt thereof, for use as a pharmaceutical. In particular, said use comprises For diseases treated by WRN inhibition, For the treatment of cancer, For the treatment of cancers characterized by microsatellite instability-high (MSI-H) or deficient mismatch repair (dMMR), For the treatment of cancers characterized by microsatellite instability-high (MSI-H) or mismatch repair deficient (dMMR), such as colorectal, gastric, prostate, endometrial, adrenocortical, uterine, cervical, esophageal, breast, renal, and ovarian cancers, for the treatment of cancers characterized by microsatellite instability-high (MSI-H) or mismatch repair deficient (dMMR), selected from colorectal cancer, gastric cancer, prostate cancer, and endometrial cancer; or for the treatment of cancer, wherein the cancer characterized by microsatellite instability-high (MSI-H) or deficient mismatch repair (dMMR) is selected from endometrial carcinoma of the uterine corpus, colon adenocarcinoma, gastric adenocarcinoma, rectal adenocarcinoma, adrenocortical carcinoma, uterine carcinosarcoma, cervical squamous cell carcinoma, cervical adenocarcinoma, esophageal carcinoma, breast cancer, renal clear cell carcinoma, prostate cancer, and ovarian serous cystadenocarcinoma. is.

[0221] The following methods are also provided: a method for modulating WRN activity in a subject, the method comprising administering to the subject a therapeutically effective amount of a compound of formula (I) as described herein, in particular compound 1a, 1b, 1c, 1d, 1f, 1h or 1g, preferably compound 1a, more preferably 1g or 1h, or a pharmaceutically acceptable salt thereof, a method of inhibiting WRN in a subject, the method comprising administering to the subject a therapeutically effective amount of a compound of formula (I) as described herein, in particular compound 1a, 1b, 1c, 1d, 1e, 1f, 1h or 1g, preferably compound 1a, more preferably 1g or 1h, or a pharmaceutically acceptable salt thereof; a method for treating a disorder or disease in a subject that can be treated by WRN inhibition, the method comprising administering to the subject a therapeutically effective amount of a compound of formula (I) as described herein, in particular compound 1a, 1b, 1c, 1d, 1e, 1f, 1h or 1g, preferably compound 1a, more preferably compound 1g or 1h, or a pharmaceutically acceptable salt thereof; a method of treating cancer in a subject, the method comprising administering to the subject a therapeutically effective amount of a compound of formula (I) as described herein, in particular 1a, 1b, 1c, 1d, 1e, 1f, 1h or 1g, preferably 1a, more preferably 1g or 1h, or a pharmaceutically acceptable salt thereof; A method for treating cancer in a subject, comprising administering a therapeutically effective amount of a compound of Formula (I) as described herein, particularly 1a, 1b, 1c, 1d, 1e, 1f, 1h, or 1g, preferably 1a, more preferably 1g or 1h, or a pharmaceutically acceptable salt thereof, wherein the cancer is characterized by microsatellite instability-high (MSI-H) or deficient mismatch repair (dMMR). In particular, the cancer characterized by microsatellite instability-high (MSI-H) or deficient mismatch repair (dMMR) is selected from colorectal cancer, gastric cancer, prostate cancer, endometrial cancer, adrenocortical cancer, uterine cancer, cervical cancer, esophageal cancer, breast cancer, kidney cancer, and ovarian cancer. More particularly, the cancer characterized by microsatellite instability-high (MSI-H) or deficient mismatch repair (dMMR) is selected from colorectal cancer, gastric cancer, prostate cancer, and endometrial cancer. Examples include endometrial carcinoma of the uterine corpus, colon adenocarcinoma, gastric adenocarcinoma, rectal adenocarcinoma, adrenocortical carcinoma, uterine carcinosarcoma, cervical squamous cell carcinoma, cervical adenocarcinoma, esophageal carcinoma, breast carcinoma, renal clear cell carcinoma, prostate carcinoma, and ovarian serous cystadenocarcinoma.

[0222] of a compound of formula (I) as described herein, in particular 1a, 1b, 1c, 1d, 1e, 1f, 1h or 1g, preferably 1a, more preferably 1g or 1h, or a pharmaceutically acceptable salt thereof, In treatment, In pharmaceutical manufacturing, in the manufacture of a medicament for the treatment of cancer, in particular cancers characterized by microsatellite instability-high (MSI-H) or deficient mismatch repair (dMMR), In the manufacture of a medicament for the treatment of a disease that can be treated by inhibition of WRN, Use, In particular, the cancer is characterized by microsatellite instability-high (MSI-H) or deficient mismatch repair (dMMR), such as colorectal cancer, gastric cancer, prostate cancer, endometrial cancer, adrenocortical carcinoma, uterine cancer, cervical cancer, esophageal cancer, breast cancer, kidney cancer, and ovarian cancer, in particular colorectal cancer, gastric cancer, prostate cancer, or endometrial cancer, or endometrial cancer of the uterine corpus, colon adenocarcinoma, rectal adenocarcinoma, adrenocortical carcinoma, uterine carcinosarcoma, cervical adenocarcinoma, esophageal cancer, breast cancer, renal clear cell carcinoma, and ovarian serous cystadenocarcinoma. Uses are also provided.

[0223] In some embodiments, the subject has or has been diagnosed as having a microsatellite instability (MSI-H) cancer, e.g., compared to a control, e.g., a normal subject. In one embodiment, the subject has an MSI-H advanced solid tumor, colorectal cancer (CRC), endometrial cancer, uterine cancer, gastric cancer, or another MSI-H cancer. In some embodiments, the subject has colorectal cancer (CRC), endometrial cancer, or gastric cancer, which has or has been diagnosed as having microsatellite instability (MSI-H), e.g., compared to a control, e.g., a normal subject. Such diagnostic techniques are known in the art.

[0224] form Depending on the selection of starting materials and procedures, compounds may exist in one of the possible stereoisomers or as a mixture thereof, for example, as pure optical isomers or as stereoisomeric mixtures, such as racemic and diastereomeric mixtures, depending on the number of asymmetric carbon atoms. The present disclosure is intended to include all such possible stereoisomers, including racemic mixtures, diastereomeric mixtures, and optically pure forms. Optically active (R)- and (S)-stereoisomers may be prepared using chiral synthons or chiral reagents or separated using conventional techniques. When compounds contain double bonds, the substituents may be in the E- or Z-configuration. When compounds contain disubstituted cycloalkyls, the cycloalkyl substituents may have cis- or trans-configuration. All tautomeric forms are also intended to be included.

[0225] As used herein, the term "salt" or "salts" refers to acid addition or base addition salts of the compounds of the present disclosure. "Salt" specifically includes "pharmaceutically acceptable salts." The term "pharmaceutically acceptable salts" refers to salts that retain the biological effectiveness and properties of the compounds of this invention and that are not typically biologically or otherwise undesirable. In many cases, the compounds of the present invention are capable of forming acid and / or base salts by virtue of the presence of amino and / or carboxyl groups or groups similar thereto.

[0226] Pharmaceutically acceptable acid addition salts can be formed with inorganic and organic acids.

[0227] Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like.

[0228] Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, sulfosalicylic acid, and the like.

[0229] Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases.

[0230] Inorganic bases from which salts can be derived include, for example, ammonium salts and metals from columns I to XII of the periodic table. In particular embodiments, salts are derived from sodium, potassium, ammonium, calcium, magnesium, iron, silver, zinc, and copper; particularly suitable salts include ammonium, potassium, sodium, calcium, and magnesium salts.

[0231] Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, etc. Particular organic amines include isopropylamine, benzathine, cholinate, diethanolamine, diethylamine, lysine, meglumine, piperazine, and tromethamine.

[0232] In another aspect, the present invention provides an anti-inflammatory agent, comprising an anti-inflammatory agent, selected from the group consisting of acetate, ascorbate, adipate, aspartate, benzoate, besylate, bromide / hydrobromide, bicarbonate / carbonate, bisulfate / sulfate, camphorsulfonate, caprate, chloride / hydrochloride, chlortheophyllonate, citrate, ethanedisulfonate, fumarate, gluceptate, gluconate, glucuronate, glutamate, glutarate, glycolate, hippurate, hydroiodide / iodide, isethionate, lactate, lactobiolate, and the like. and the like. In some embodiments, the compounds of the present invention are provided in the form of a salt of the following: phosphate, lauryl sulfate, malate, maleate, malonate, mandelate, mesylate, methylsulfate, mucate, naphthoate, napsylate, nicotinate, nitrate, octadecanoate, oleate, oxalate, palmitate, pamoate, phosphate / hydrogenphosphate / dihydrogenphosphate, polygalacturonate, propionate, sebacate, stearate, succinate, sulfosalicylate, sulfate, tartrate, tosylate, triphenylacetate, trifluoroacetate, or xinafoate.

[0233] Any formula given herein is intended to represent unlabeled forms of the compounds as well as isotopically labeled forms, in addition to the deuteration specifically required by formula (I). Isotopically labeled compounds have structures depicted by the formulas given herein, except that one or more atoms are replaced with an atom having a selected atomic mass or mass number. Isotopes that can be incorporated into the compounds of the present invention include, for example, isotopes of hydrogen.

[0234] Additionally, certain isotopes, particularly deuterium (i.e.2 Incorporation of H or D) may result in certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life, or reduced dosage requirements, or improved therapeutic index or tolerability. It is understood that in this context deuterium is considered a substituent of the compounds of the present invention. The concentration of deuterium can be defined by the isotopic enrichment factor. As used herein, the term "isotopic enrichment factor" refers to the ratio between the isotopic abundance and the natural abundance of a particular isotope. When a substituent in a compound of the invention is designated as deuterium, such compound has, for each designated deuterium atom, an isotopic enrichment factor of at least 3500 (52.5% deuterium incorporation at each designated deuterium atom), at least 4000 (60% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium incorporation), or at least 6633.3 (99.5% deuterium incorporation). It is understood that the term "isotopic enrichment factor" can be applied to any isotope in a manner similar to that described for deuterium.

[0235] Examples of isotopes that may be incorporated into compounds of the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, such as 3 H, 11 C. 13 C. 14 C. 15 N, 18 F, 31 P, 32 P, 35 S, 36 Cl, 123 I, 124 I, 125 Therefore, the present invention includes, for example, 3 H and 14 Radioactive isotopes such as C, or 2 H and13 It should be understood that this includes compounds incorporating one or more of any of the foregoing isotopes, including those in which non-radioactive isotopes such as C are present. Such isotopically labeled compounds may be used in metabolic studies ( 14 C), reaction rate tests (e.g., 2 H or 3 H), detection or imaging techniques such as positron emission tomography (PET) or single photon emission computed tomography (SPECT), including drug or substrate tissue distribution assays, or in radiation treatment of patients. 18 F or labeled compounds may be particularly desirable for PET or SPECT studies. Isotopically labeled compounds of the present invention can generally be prepared by conventional techniques known to those skilled in the art, or by methods analogous to those described in the accompanying Examples and Preparations, substituting the appropriate isotopically labeled reagent for the previously used non-labeled reagent.

[0236] Deuterated compounds of formula (I) include the deuterated form of Example 42, Example 43: [ka] and [ka] and, Example 86: [ka] Example 50: [ka] Example 57: [ka] and Example 47: [ka] Deuterated forms of the compounds are included.

[0237] definition A "compound of the invention" or a "compound of Formula (I)" or a "compound of Formula 1a" or 1g or h, etc. includes a zwitterion thereof, a non-zwitterion thereof (uncharged form), or a pharmaceutically acceptable salt of said zwitterion or non-zwitterionic form thereof.

[0238] "Zwitterion" or "zwitterionic form" refers to a compound that contains both positively charged and negatively charged functional groups.

[0239] For example, compounds of formula (I) described herein can include the following forms, where R4 is in a zwitterionic form (c) or a non-zwitterionic form (d): [ka] or a mixture thereof.

[0240] The compounds of formula (I) described herein may also include the following forms, wherein R4 is in a zwitterionic form (a) or (b) or in a non-zwitterionic form (e), [ka] Or a mixture of two of them, or a mixture of all three of them. Halo means fluoro, chloro or bromo, especially fluoro or chloro. The alkyl and alkoxy groups containing the required number of carbon atoms can be unbranched or branched. Examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, sec-butyl, and t-butyl. Examples of alkoxy include, but are not limited to, methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy, sec-butoxy, and t-butoxy.

[0241] "=O" means an oxo substituent.

[0242] When R1 is substituted or unsubstituted cycloalkenyl, said cycloalkenyl includes, but is not limited to, groups such as cyclohexenyl, particularly cyclohex-1-en-1-yl.

[0243] When R1 is a substituted or unsubstituted heterocyclyl, the heterocyclyl includes, but is not limited to, groups such as morpholinyl, piperidinyl, pyrrolidinyl, 6-oxa-3-azabicyclo[3.1.1]heptan-3-yl, 5,6-dihydro-1,4-dioxin-2-yl, dihydropyranyl, particularly 3,4-dihydro-2H-pyran-6-yl, 5,6-dihydro-2H-pyran-3-yl and 3,6-dihydro-2H-pyran-4-yl, piperazinyl, tetrahydropyridinyl, such as 1,4,5,6-tetrahydropyridin-3-yl and 1,2,3,6-tetrahydropyridin-4-yl, and dihydropyridinyl, such as 3,6-dihydropyridinyl.

[0244] When R1 is substituted or unsubstituted heteroaryl, said heteroaryl includes, but is not limited to, groups such as pyridinyl, particularly pyridin-3-yl.

[0245] The term "cancer" refers to a disease characterized by the rapid and uncontrolled growth of abnormal cells. Cancer cells can spread locally or through the bloodstream and lymphatic system to other parts of the body. Examples of various cancers are described herein, including, but not limited to, colon cancer, gastric cancer, endometrial cancer, prostate cancer, adrenocortical cancer, uterine cancer, cervical cancer, esophageal cancer, breast cancer, kidney cancer, ovarian cancer, etc.

[0246] The terms "tumor" and "cancer" are used interchangeably herein, e.g., both terms encompass solid and liquid, e.g., diffuse or circulating, tumors. As used herein, the term "cancer" or "tumor" includes pre-cancerous as well as malignant cancers and tumors.

[0247] As used herein, a "WRN inhibitor" or "WRN helicase inhibitor" refers to a compound that inhibits Werner syndrome RecQ DNA helicase (WRN). As used herein, the term "WRN" refers to the Werner syndrome RecQ DNA helicase protein. The term "WRN" includes mutants, fragments, variants, isoforms, and homologs of full-length wild-type WRN. In one embodiment, the protein is encoded by the WRN gene (Entrez gene ID 7486; Ensembl ID ENSG00000165392). An exemplary WRN sequence is available from the Uniprot database under accession number Q14191.

[0248] A "WRN-mediated disease or condition" includes diseases or conditions, such as cancer, that are treated by WRN inhibition. In particular, this can include cancers characterized by microsatellite instability-high (MSI-H) or deficient mismatch repair (dMMR).

[0249] As used herein, the terms "microsatellite unstable cancer", "microsatellite unstable high cancer", "microsatellite high cancer", and "MSI high cancer", "MSI hi " and "MSI-H" are used interchangeably to describe cancers that have multiple alterations within the length of simple repeat genomic sequences within microsatellites.

[0250] Determining a patient's MSI-H or dMMR tumor status can be performed, for example, using a polymerase chain reaction (PCR) test for MSI-H status or an immunohistochemistry (IHC) test for dMMR. Methods for identifying MSI-H or dMMR tumor status are described, for example, in Ryan et al. Crit Rev Oncol Hematol. 2017;116:38-57; Dietmaier and Hofstadter. Lab Invest 2001,81:1453-1456; and Kawakami et al. Curr Treat Options Oncol. 2015:16(7):30).

[0251] Microsatellite instability can also be found in colorectal, gastric, and endometrial cancers, particularly adrenocortical carcinoma, uterine cancer, cervical cancer, esophageal cancer, breast cancer, kidney cancer, prostate cancer, and ovarian cancer. Examples of frequent microsatellite cancers include endometrial cancer of the uterine corpus, colon adenocarcinoma, gastric adenocarcinoma, rectal adenocarcinoma, adrenocortical carcinoma, uterine carcinosarcoma, cervical squamous cell carcinoma, cervical adenocarcinoma, esophageal cancer, breast cancer, renal clear cell carcinoma, prostate cancer, and ovarian serous cystadenocarcinoma.

[0252] Cancers with "defective mismatch repair" (dMMR) or "dMMR signature" include cancers associated with reported mutations or epigenetic silencing of MLH1, PMS2, MSH2, MSH3, MSH6, MLH3, and PMS1, microsatellite instability, or other gene inactivation mechanisms, including, but not limited to, lung cancer, breast cancer, kidney cancer, colorectal cancer, ovarian cancer, prostate cancer, upper aerodigestive tract cancer, gastric cancer, endometrial cancer, liver cancer, pancreatic cancer, hematopoietic and lymphoid tissue cancer, skin cancer, thyroid cancer, pleural cancer, autonomic ganglion cancer, central nervous system cancer, soft tissue cancer, pediatric rhabdoid sarcoma, melanoma, and other cancers. Cells or cancers with "defective" mismatch repair have a significant reduction in mismatch repair (e.g., at least about 25%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% reduction). In some cases, cells or cancers with defective mismatch repair function do not perform mismatch repair.

[0253] As used herein, the term "pharmaceutical composition" refers to a compound of the present invention or a pharmaceutically acceptable salt thereof, combined with at least one pharmaceutically acceptable carrier in a form suitable for oral or parenteral administration.

[0254] As used herein, the term "pharmaceutically acceptable carrier" refers to a substance useful in the preparation or use of a pharmaceutical composition, and includes, for example, suitable diluents, solvents, dispersion media, surfactants, antioxidants, preservatives, isotonic agents, buffers, emulsifiers, absorption delaying agents, salts, drug stabilizers, binders, excipients, disintegrants, lubricants, wetting agents, sweeteners, flavoring agents, dyes, and combinations thereof, as known to those skilled in the art (see, e.g., Remington The Science and Practice of Pharmacy, 22nd Ed. Pharmaceutical Press, 2013, pp. 1049-1070).

[0255] The terms "synthetic lethal" and "synthetic lethal" are used to refer to a reduction in cell viability and / or a reduction in cell proliferation rate caused by a combination of mutations or approaches that result in loss of function in two or more genes (e.g., RNA interference or protein function inhibition), but not by loss of function of only one of those genes.

[0256] The term "therapeutically effective amount" of a compound of the invention refers to that amount of a compound of the invention that elicits a biological or medical response in a subject, such as a reduction or inhibition of enzyme or protein activity, or that ameliorates symptoms, alleviates pathology, slows or retards the progression of a disease, or prevents a disease.

[0257] In one embodiment, the term "therapeutically effective amount" refers to an amount of a compound of the present invention that, when administered to a subject, is effective to (1) at least partially alleviate, prevent, and / or ameliorate a condition, or a disorder or disease (i) mediated by WRN, or (ii) associated with WRN activity, or (iii) characterized by WRN activity (normal or abnormal), or (2) reduce or inhibit the activity of WRN.

[0258] In another embodiment, the term "therapeutically effective amount" refers to an amount of a compound of the invention that, when administered to a cell, or tissue, or non-cellular biological material, or culture medium, is effective to at least partially reduce or inhibit the activity of WRN or reduce WRN protein levels.

[0259] As used herein, the term "subject" refers to primates (e.g., humans, male or female), dogs, rabbits, guinea pigs, pigs, rats, and mice. In certain embodiments, the subject is a primate, rat, or mouse. In yet other embodiments, the subject is a human.

[0260] As used herein, the terms "inhibit," "inhibition," or "inhibiting" refer to the alleviation or suppression of a given condition, symptom, or disorder or disease, or a significant decrease in the baseline activity of a biological activity or process.

[0261] As used herein, the terms "treat," "treating," or "treatment" of any disease or disorder refers to alleviating or ameliorating the disease or disorder (i.e., slowing or halting the progression of the disease or at least one of its clinical symptoms); or alleviating or ameliorating at least one physical parameter or biomarker associated with the disease or disorder (including those that may not be discernible by the patient).

[0262] As used herein, the terms "prevent," "preventing," or "prevention" of any disease or disorder refers to the prophylactic treatment of the disease or disorder; or the delay in the onset or progression of the disease or disorder.

[0263] As used herein, a subject is "in need of" a treatment if such subject would benefit biologically, medically, or in quality of life from such treatment.

[0264] As used herein, the terms "a," "an," "the," and similar terms used in connection with the present invention (particularly in connection with the claims) are to be construed to encompass both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context.

[0265] "Can be linked" means to link or not to link.

[0266] "Can be substituted by deuterium" means either substituted by deuterium or not substituted by deuterium.

[0267] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. Furthermore, the materials, methods, and examples are illustrative only and not limiting. Unless otherwise indicated herein or clearly contradicted by context, all methods described herein can be performed in any suitable order. The use of any and all examples, or exemplary language (e.g., "etc.") provided herein is intended merely to further clarify the invention and does not limit the scope of the invention as otherwise claimed.

[0268] Isomers Any asymmetric atom (e.g., carbon, etc.) of the compounds of the present invention can be present in racemic or enantiomerically enriched form, for example, in the (R)-, (S)-, or (R,S)-configuration. In certain embodiments, each asymmetric atom has an enantiomeric excess of at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% in the (R)- or (S)-configuration. Substituents at atoms having unsaturated double bonds may, where possible, be present in cis-(Z)- or trans-(E)-form.

[0269] Thus, as used herein, the compounds of the invention may be in the form of one of the possible stereoisomers, rotamers, atropisomers, tautomers or mixtures thereof, for example, as substantially pure geometric (cis or trans) stereoisomers, diastereomers, optical isomers (enantiomers), racemates, or mixtures thereof.

[0270] Any resulting mixture of stereoisomers can be separated into pure or substantially pure geometric or optical isomers, diastereomers, racemates, for example, by chromatography and / or fractional crystallization on the basis of the physical chemical differences of the constituent components.

[0271] Any resulting racemic forms of the compounds or intermediates of the present invention can be resolved into their optical antipodes by known methods, for example, by separating their diastereomeric salts obtained with optically active acids or bases, and liberating the optically active acidic or basic compounds. In particular, compounds of the present invention can be resolved into their optical antipodes by, for example, fractional crystallization of salts formed with optically active acids, such as tartaric acid, dibenzoyltartaric acid, diacetyltartaric acid, di-O,O'-p-toluoyltartaric acid, mandelic acid, malic acid, or camphor-10-sulfonic acid, using basic moieties in this way. Racemic compounds or racemic intermediates of the present invention can also be resolved by chiral chromatography, for example, high-pressure liquid chromatography (HPLC) using a chiral adsorbent.

[0272] The compounds of the present invention, i.e., compounds of formula (I) containing groups capable of acting as donors and / or acceptors for hydrogen bonds, can form co-crystals with suitable co-crystal formers. These co-crystals can be prepared from compounds of formula (I) by known co-crystal formation procedures. Such procedures include grinding, heating, co-sublimation, co-melting, or contacting a compound of formula (I) in solution with a co-crystal former under crystallization conditions and isolating the co-crystal formed thereby. Suitable co-crystal formers include those described in WO 2004 / 078163. Accordingly, the present invention further provides co-crystals comprising compounds of formula (I).

[0273] Furthermore, the compounds of the invention, including their salts, can also be obtained in the form of their hydrates, or include other solvents used for their crystallization.

[0274] The compounds of the present invention may inherently or intentionally form solvates with pharmaceutically acceptable solvents (including water), and therefore the present invention encompasses both solvated and non-solvated forms. The term "solvate" refers to a molecular complex of the compounds of the present invention (including pharmaceutically acceptable salts thereof) with one or more solvent molecules. Such solvent molecules are commonly used in the pharmaceutical field and are known to be harmless to recipients, such as water, ethanol, etc. The term "hydrate" refers to a complex in which the solvent molecule is water.

[0275] Dosage form The pharmaceutical composition or combination of the present invention may be administered in a unit dose of, for example, about 1 to 1000 mg of the active ingredient for a subject weighing about 50 to 70 kg.

[0276] combination "Combination" refers to a fixed combination in one unit dosage form, or to combined administration in which a compound of Formula (I) or a pharmaceutically acceptable salt thereof and a combination partner (e.g., another drug, as described below, also referred to as a "therapeutic agent" or "adjunct") are administered simultaneously and independently, or may be administered separately within a time interval (particularly when these time intervals allow the combination partners to exhibit a cooperative, e.g., synergistic, effect). The single components may be packaged as a kit or packaged individually. One or both of the components (e.g., powder or liquid) may be reconstituted or diluted to the desired dosage before administration. The terms "co-administration" or "administration in combination," or the like, as used herein, are meant to encompass the administration of selected combination partners to a single subject (e.g., patient) in need thereof, and are intended to include therapeutic regimens in which the agents need not necessarily be administered by the same route of administration or at the same time. The term "pharmaceutical combination," as used herein, refers to a product resulting from the mixing or combining of multiple therapeutic agents, and includes both fixed and non-fixed combinations of therapeutic agents. The term "fixed combination" means that both therapeutic agents, e.g., a compound of the invention and a combination partner, are administered to a patient simultaneously in the form of a single entity or dosage. The term "non-fixed combination" means that both therapeutic agents, e.g., a compound of the invention and a combination partner, are administered to a patient as separate entities simultaneously, in parallel, or sequentially with no specific time limit, such administration resulting in therapeutically effective levels of the two compounds in the patient's body. The latter also applies to cocktail therapy, e.g., the administration of three or more therapeutic agents.

[0277] The combinations described herein may include compositions of the invention co-formulated and / or co-administered with a compound of Formula (I) and one or more additional therapeutic agents, such as one or more anti-cancer agents, cytotoxic or cytostatic agents, hormonal therapy, vaccines, and / or other immunotherapy. In other embodiments, the combination is further administered or used in conjunction with other therapeutic modalities, including surgery, radiation, cryosurgery, and / or hyperthermia. Such combination therapies may advantageously utilize lower dosages of the administered therapeutic agents, thereby avoiding potential toxicities or complications associated with the treatments.

[0278] Also provided are combinations comprising a compound of formula (I) as described herein, in particular compounds 1a, 1b, 1c, 1d, 1e, 1f, 1h or 1g as described herein, preferably 1a, more preferably 1g or 1h, or a pharmaceutically acceptable salt thereof, and one or more additional therapeutically active agents. The additional therapeutic agent is, for example, a chemical compound, peptide, antibody, antibody fragment, or nucleic acid that is therapeutically active or enhances therapeutic activity when administered to a patient in combination with a compound of the present disclosure. In particular, the additional therapeutically active agent is Anticancer drugs, chemotherapy drugs, Anastrozole (Arimidex®), bicalutamide (Casodex®), bleomycin sulfate (Blenoxane®), busulfan (Myleran®), busulfan injection (Busulfex®), capecitabine (Xeloda®), N4-pentoxycarbonyl-5-deoxy-5-fluorocytidine, carboplatin (Paraplatin®), carmustine (BiCNU®), chlorambucil (Leukeran®), cisplatin cytarabine (Platinol®), cladribine (Leustatin®), cyclophosphamide (Cytoxan® or Neosar®), cytarabine, cytosine arabinoside (Cytosar-U®), cytarabine liposome injection (DepoCyt®), dacarbazine (DTIC-Dome®), dactinomycin (actinomycin D, Cosmegan), daunorubicin hydrochloride (Cerubidine®), daunorubicin citrate liposome injection (Daun oXome®), dexamethasone, docetaxel (Taxotere®), doxorubicin hydrochloride (Adriamycin®, Rubex®), etoposide (Vepesid®), fludarabine phosphate (Fludara®), 5-fluorouracil (Adrucil®, Efudex®), flutamide (Eulexin®), tezacitibine, gemcitabine (difluorodeoxycytidine), hydroxyurea (Hydrea®), idarubicin (I damycin (registered trademark), ifosfamide (IFEX (registered trademark), irinotecan (Camptosar (registered trademark), L-asparaginase (ELSPAR (registered trademark), leucovorin calcium, melphalan (Alkeran (registered trademark), 6-mercaptopurine (Purinethol (registered trademark), methotrexate (Folex (registered trademark), mitoxantrone (Novantrone (registered trademark), Mylotarg, paclitaxel (Taxol (registered trademark), Phoenix (yttrium 90 / MX-DTPA),chemotherapeutic agents selected from pentostatin, polifeprosan 20 (Gliadel®) including carmustine implant, tamoxifen citrate (Nolvadex®), teniposide (Vumon®), 6-thioguanine, thiotepa, tirapazamine (Tirazone®), topotecan hydrochloride for injection (Hycamptin®), vinblastine (Velban®), vincristine (Oncovin®) and vinorelbine (Navelbine®), in particular fluorouracil (5-FU) and irinotecan (Camptosar®), PD-1 inhibitors, anti-PD-1 antibody molecules, or PDR001 (Novartis), nivolumab (Bristol-Myers Squibb), pembrolizumab (Merck & Co), pidilizumab (CureTech), MEDI0680 (Medimmune), cemiplimab (REGN2810, Regeneron), dostallimab (TSR-042, Tesaro), PF-06801591 (Pfizer), tislelizumab (BGB-A317, Beigene), BGB-108 (Beigene), INCSHR1210 (Incyte), balstilimab (AGEN2035, Agenus), sintilimab (InnoVent), toripalimab (Shanghai Junshi Bioscience), camrelizumab (Jiangsu Hengrui Medicine Co.), and PD-1 inhibitors selected from AMP-224 (Amplimmune), penprimab (Akeso Biopharma Inc), dimvelelimab (Arcus Biosciences Inc) and prorugolimab (Biocad Ltd), in particular PDR001, more in particular tislelizumab (BGB-A317, Beigene). is.

[0279] In a further embodiment, the additional therapeutically active agent is the chemotherapy irinotecan (Camptosar®).

[0280] In another embodiment, the additional therapeutically active agent is an inhibitor of PD-1, e.g., human PD-1. In another embodiment, the immunomodulator is an inhibitor of PD-L1, e.g., human PD-L1. In one embodiment, the PD-1 or PD-L1 inhibitor is an antibody molecule directed against PD-1 or PD-L1. In another embodiment, the additional therapeutically active agent is an anti-PD-1 antibody molecule. In a further embodiment, the PD-1 inhibitor is an anti-PD-1 antibody molecule described in U.S. Patent Application Publication No. 2015 / 0210769, entitled "Antibody Molecules to PD-1 and Uses Thereof," published July 30, 2015, which is incorporated by reference in its entirety.

[0281] In another embodiment, a combination of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, a chemotherapeutic agent, and a PD-1 inhibitor is provided. In particular, the chemotherapy and PD-1 inhibitor are selected from those described above. More particularly, the chemotherapeutic agent is irinotecan (Camptosar®) and the PD-1 inhibitor is PDR001 or tislelizumab. The tislelizumab may have a heavy chain of SEQ ID NO: 3 and a light chain of SEQ ID NO: 4. In some embodiments, the anti-PD-1 antibody is administered at 100 mg / week. In some embodiments, tislelizumab is administered IV at 300 mg on day 1 of each 28-day cycle. In some embodiments, tislelizumab may be administered at 500 mg once every four weeks.

[0282] In another embodiment, the anti-PD-1 antibody molecule, e.g., tislelizumab, comprises the following heavy and / or light chain: VH, VL, HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3.

[0283] [Table 1]

[0284] In some embodiments, the PD-1 inhibitor comprises the HCDR and LCDR of tislelizumab set forth in SEQ ID NOs: 7-12.

[0285] In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) is administered at a flat rate of about 100 mg to about 600 mg. In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) is administered at a dose of about 100 mg to about 500 mg. In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) is administered at a dose of about 100 mg to about 400 mg. In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) is administered at a dose of about 100 mg to about 300 mg. In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) is administered at a dose of about 100 mg to about 200 mg. In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) is administered at a dose of about 200 mg to about 600 mg. In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) is administered at a dose of about 200 mg to about 500 mg. In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) is administered at a dose of about 200 mg to about 400 mg. In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) is administered at a dose of about 200 mg to about 300 mg. In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) is administered at a dose of about 300 mg to about 600 mg. In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) is administered at a dose of about 300 mg to about 500 mg. In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) is administered at a dose of about 300 mg to about 400 mg. In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) is administered at a dose of about 400 mg to about 600 mg. In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) is administered at a dose of about 400 mg to about 500 mg. In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) is administered at a dose of about 500 mg to about 600 mg. In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) is administered at a dose of about 600 mg to about 700 mg. In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) is administered at a dose of about 700 mg to about 800 mg.In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) is administered at a dose of about 800 mg to about 900 mg. In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) is administered at a dose of about 900 mg to about 1000 mg.

[0286] The PD-1 inhibitor (e.g., tislelizumab) is administered at a flat dose of about 100 mg. The PD-1 inhibitor (e.g., tislelizumab) is administered at a dose of about 200 mg. The PD-1 inhibitor (e.g., tislelizumab) is administered at a dose of about 300 mg. The PD-1 inhibitor (e.g., tislelizumab) is administered at a dose of about 400 mg. The PD-1 inhibitor (e.g., tislelizumab) is administered at a dose of about 500 mg. The PD-1 inhibitor (e.g., tislelizumab) is administered at a dose of about 600 mg. The PD-1 inhibitor (e.g., tislelizumab) is administered at a dose of about 700 mg. The PD-1 inhibitor (e.g., tislelizumab) is administered at a dose of about 800 mg. The PD-1 inhibitor (e.g., tislelizumab) is administered at a dose of about 900 mg. PD-1 inhibitors (e.g., tislelizumab) are administered at doses of approximately 1000 mg.

[0287] In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) is administered once every 10 weeks. In some embodiments, the PD-1 inhibitor is administered once every 9 weeks. In some embodiments, the PD-1 inhibitor is administered once every 8 weeks. In some embodiments, the PD-1 inhibitor is administered once every 7 weeks. In some embodiments, the PD-1 inhibitor is administered once every 6 weeks. In some embodiments, the PD-1 inhibitor is administered once every 5 weeks. In some embodiments, the PD-1 inhibitor is administered once every 4 weeks. In some embodiments, the PD-1 inhibitor is administered once every 3 weeks. In some embodiments, the PD-1 inhibitor is administered once every 2 weeks. In some embodiments, the PD-1 inhibitor is administered once every week.

[0288] In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) is administered intravenously.

[0289] In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) is administered over about 20 to 40 minutes (e.g., about 30 minutes). In some embodiments, the PD-1 inhibitor is administered over about 30 minutes. In some embodiments, the PD-1 inhibitor is administered over about 1 hour. In some embodiments, the PD-1 inhibitor is administered over about 2 hours. In some embodiments, the PD-1 inhibitor is administered over about 3 hours. In some embodiments, the PD-1 inhibitor is administered over about 4 hours. In some embodiments, the PD-1 inhibitor is administered over about 5 hours. In some embodiments, the PD-1 inhibitor is administered over about 6 hours.

[0290] In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) is administered intravenously at a dose of about 300 mg to about 500 mg (e.g., about 400 mg) once every four weeks. In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) is administered intravenously at a dose of about 200 mg to about 400 mg (e.g., about 300 mg) once every three weeks. In some embodiments, tislelizumab is administered at a dose of 400 mg once every four weeks. In some embodiments, tislelizumab is administered at a dose of 300 mg once every three weeks.

[0291] In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) is administered intravenously over about 20 to about 40 minutes (e.g., about 30 minutes) at a dose of about 300 mg to about 500 mg (e.g., about 400 mg) once every two weeks. In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) is administered intravenously over about 20 to about 40 minutes (e.g., about 30 minutes) at a dose of about 200 mg to about 400 mg (e.g., about 300 mg) once every three weeks.

[0292] In some embodiments, the PD-1 inhibitor (e.g., tislelizumab) is administered at a dose of about 100 mg / week. For example, if a patient is administered a 10-week dose, the PD-1 inhibitor (e.g., tislelizumab) can be administered at 1000 mg. If a 9-week dose is administered, the PD-1 inhibitor (e.g., tislelizumab) can be administered at 900 mg. If an 8-week dose is administered, the PD-1 inhibitor (e.g., tislelizumab) can be administered at 800 mg. If a 7-week dose is administered, the PD-1 inhibitor (e.g., tislelizumab) can be administered at 700 mg. If a 6-week dose is administered, the PD-1 inhibitor (e.g., tislelizumab) can be administered at 600 mg. If a 5-week dose is administered, the PD-1 inhibitor (e.g., tislelizumab) can be administered at 500 mg. If a 4-week dose is administered, the PD-1 inhibitor (e.g., tislelizumab) can be administered at 400 mg. If a 3-weekly dose is administered, the PD-1 inhibitor (e.g., tislelizumab) can be administered at 300 mg. If a 2-weekly dose is administered, the PD-1 inhibitor (e.g., tislelizumab) can be administered at 200 mg. If a 1-weekly dose is administered, the PD-1 inhibitor (e.g., tislelizumab) can be administered at 100 mg.

[0293] For example, when using an anti-PD-1 antibody such as tislelizumab, it can be administered at a dose of 200 mg as an intravenous infusion once every three weeks. Alternatively, it can be administered at a dose of 300 mg as an intravenous infusion once every four weeks. When using an anti-PD-1 antibody such as tislelizumab, it can be administered at a dose of 300 mg as an intravenous infusion once every three weeks. Alternatively, it can be administered at a dose of 400 mg as an intravenous infusion once every four weeks.

[0294] The structures of the active compounds, identified by code numbers, generic names or trade names, can be obtained from the current edition of the standard compendium "The Merck Index" or from databases such as Patents International (e.g., IMS World Publications). The above-mentioned compounds that can be used in combination with the compounds of the present disclosure can be prepared and administered as described in the art, for example, in the documents cited above.

[0295] In one embodiment, the present invention provides a product comprising a compound of the present invention and at least one other therapeutic agent as a combined preparation for simultaneous, separate or sequential use in therapy. In one embodiment, the therapy is treatment of a disease or condition mediated by WRN. Products provided as combined preparations include compositions comprising a compound of formula (I) and the other therapeutic agent together in the same pharmaceutical composition, or in separate forms, e.g., in the form of a kit.

[0296] In one embodiment, the present invention provides a kit comprising two or more separate pharmaceutical compositions, at least one of which contains a compound of the present invention. In one embodiment, the kit comprises a means for retaining the compositions separately, such as a container, a divided bottle, or a divided foil packet. An example of such a kit is a blister pack, such as those commonly used for packaging tablets, capsules, and the like.

[0297] The kits of the present invention can be used to administer different dosage forms, e.g., oral and parenteral, to administer the separate compositions at different dosage intervals, or to titrate the separate compositions relative to each other. To aid in compliance, the kits of the present disclosure typically include directions for administration.

[0298] In the combination therapy of the present invention, the compound of the present invention and the other therapeutic agent may be manufactured and / or formulated by the same or different manufacturers. Furthermore, the compound of the present invention and the other therapeutic agent may be combined into a combination therapy (i) prior to delivery of the combination product to the physician (e.g., in the case of a kit containing the compound of the present invention and the other therapeutic agent); (ii) by the physician (or under the physician's guidance) immediately prior to administration; or (iii) in the patient himself, e.g., during sequential administration of the compound of the present invention and the other therapeutic agent.

[0299] Thus, the present invention provides the use of a compound of the present invention for treating a disease or condition mediated by WRN, wherein the medicament is prepared for administration in combination with another therapeutic agent. The present invention also provides the use of a compound of the present invention for treating a disease or condition mediated by WRN, wherein the medicament is administered in combination with a compound of the present invention.

[0300] The present invention also provides a compound of the present invention for use in the treatment of a disease or condition mediated by WRN, wherein the compound of the present invention is prepared for administration with another therapeutic agent. The present invention also provides a second therapeutic agent for use in the treatment of a disease or condition mediated by WRN, wherein the second therapeutic agent is prepared for administration with a compound of the present invention. The present invention also provides a compound of the present invention for use in the treatment of a disease or condition mediated by WRN, wherein the compound of the present invention is administered for administration with another therapeutic agent. The present invention also provides a second therapeutic agent for use in the treatment of a disease or condition mediated by WRN, wherein the second therapeutic agent is administered with a compound of the present invention.

[0301] The invention also provides the use of a compound of the invention for treating a disease or condition mediated by WRN, wherein the patient has been previously treated (e.g., within 24 hours) with another therapeutic agent. The invention also provides the use of a compound of the invention for treating a disease or condition mediated by WRN, wherein the patient has been previously treated (e.g., within 24 hours) with another therapeutic agent. [Example]

[0302] Formulation example The compounds of formula (I) may be formulated as amorphous solid dispersion tablets as described below.

[0303] A. Amorphous Solid Dispersions:

[0304] [Table 2]

[0305] *Eudragit® E PO is described in dx.doi.org / 10.1021 / mp4000635 I Mol.Pharmaceutics 2013,10,2630-2641 and has the registered CAS registry number 24938-16-7.

[0306] Dichloromethane was added to a container, followed by the compound of Example 42 in free form (not the sodium salt), basic polymethacrylate, and copovidone to obtain a slurry. Dichloromethane was added again to the container, and the mixture was stirred until a clear solution was obtained. The solution was spray-dried, and the resulting powder was then dried in a stirred-bed vacuum dryer and then sieved.

[0307] B. Film-coated tablets:

[0308] [Table 3]

[0309] Mannitol, croscarmellose sodium, colloidal silicon dioxide, and sodium stearyl fumarate were added to the Example 42 compound powder obtained in Part A above, and the mixture was blended and then compressed into tablets. The coating agent was dispersed in water to obtain a uniform suspension, which was used to coat the tablets.

[0310] Biological Assays and Data The activity of the compounds according to the invention can be assessed by the following in vitro and in vivo methods.

[0311] Materials and Methods Molecular biology and virus production. DNA encoding human Werner helicase (UniProt Q14191, WRN, amino acids S2-S1432) was engineered as four codon-optimized DNA strings for expression in E. coli. Strings were ordered from GeneArt (LifeTechnologies, Regensburg, Germany) or prepared by subcloning overlapping oligonucleotides.

[0312] Baculovirus from expression plasmid pLAF1202 (SEQ ID NO: 1) encoding His-ZZ-3C-WRN (aa N517-P1238, encoded by nucleotides 578-2743 in the sequence) was generated for transfection using the FlashBac Ultra system (Oxford Expression Technologies 100302) with 540 ng of plasmid DNA, 5.4 μg of FlashBac Ultra DNA, and 5.4 microliters of Lipofectin (Life Technologies 18292-011) according to the manufacturer's instructions. After 5 hours of incubation, the solution was diluted with 500 microliters of TC100 medium (Life Technologies 13055-025) and incubated at 27°C for 7 days.

[0313] Cells were harvested by centrifugation at 800 × g for 10 minutes, and the virus-containing supernatant was transferred to a new sterile tube. For initial viral amplification, 500 microliters of virus was added to 25 mL of SF9 cells at 1 million cells / mL and incubated at 27°C (200 rpm) for 5 days. Cell viability, density, and diameter were measured, and virus showing signs of infection was harvested by centrifugation at 3000 rpm for 15 minutes.

[0314] Baculovirus-infected insect cells (BIIC) were generated as described in Wasilko et al, 2009, DOI:10.1016 / j.pep.2009.01.002. Briefly, 100 million SF9 cells (1 million cells / mL) in 100 mL of ESF921 medium (Expression System-96-001-01, supplemented with 0.5x streptomycin / penicillin) were infected with 300 million baculovirus particles of each construct (estimated MOI = 3) in an Erlenmeyer flask and incubated at 27°C and 130 rpm for 24 h. Infected cells were transferred to a 50 mL tube and harvested by centrifugation at 100 x g for 10 min at RT.

[0315] Cells were resuspended at 10 million / mL in ESF921 (0.5x streptomycin / penicillin) medium containing BSA (final 10 mg / mL) and 10% DMSO. 500 μL aliquots of cells were transferred to 1.8 mL cryotubes and frozen overnight at -80°C in a Nunc Cryo 1°C freezing container.

[0316] SEQ ID NO: 1 [ka] [ka] [ka]

[0317] Protein expression and purification A BIIC aliquot for the Werner helicase protein His-ZZ-3C-WRN (aa N517-P1238, pLAF1202) was diluted 1 / 100 in ESF921 medium and further diluted 1 / 100 into an expression / production flask containing Sf21 cells (1 million cells / mL) in 1 L of ESF921 medium and incubated for 96 hours (27°C, 130 rpm) for protein expression.

[0318] WRN protein was purified using the following protocol. Cell pellets were thawed and resuspended in 80 mL of buffer A (50 mM Tris, 300 mM NaCl, 20 mM imidazole, 1 mM TCEP, 10% glycerol, pH 7.8) supplemented with Turbonuclease (final concentration 40 units / mL, Merck) and cOmplete protease inhibitor tablets (1 tablet / 50 mL, Roche). Cells were lysed by passing three times through a homogenizer (Avestin, Emulsiflex C3) at 800-1000 bar. The lysed sample was centrifuged at 48,000 × g for 40 min (Sorvall RC5B, SS-34 rotor), and the supernatant was passed through a 0.45 μm filter.

[0319] The lysate was loaded onto a 5 mL HisTrap crude FF column (GE Healthcare) attached to an AeKTA Pure 25 chromatography system (GE Healthcare). Contaminating proteins were washed off with buffer A, and bound proteins were eluted with a linear gradient over 10 column volumes to 100% buffer B (50 mM Tris, 300 mM NaCl, 300 mM imidazole, 1 mM TCEP, 10% glycerol, pH 7.8). 1% (w / w) HRV 3C protease (His-MBP tagged, produced in-house) was added to the eluted protein. The N-terminal purification tag was cleaved by the protease during overnight dialysis against 2 L of buffer (50 mM Tris pH 7.0, 150 mM NaCl, 1 mM TCEP, 10% glycerol, 0.02% CHAPS) at 5 °C. The protein solution was then carefully diluted by adding two volumes of 20 mM Tris pH 7.0, 10% glycerol, and 0.02% CHAPS. The slightly turbid protein solution was passed through a 0.45 μm filter. The cleaved protein was loaded onto a Resource S 6 mL column (GE Healthcare) pre-equilibrated with 20 mM Tris, 20 mM NaCl, 1 mM TCEP, and 10% glycerol, pH 7.0. The cleaved tag and contaminating proteins were washed away with equilibration buffer. The bound target protein was eluted with a linear gradient of the same buffer containing 1 M sodium chloride over 20 column volumes and then injected onto a HiLoad 16 / 600 Superdex 75 pg column (GE Healthcare) pre-equilibrated with 50 mM Tris pH 7.4, 300 mM NaCl, and 10% glycerol. Fractions containing pure protein were identified by SDS-PAGE and pooled. The purified protein was finally divided into aliquots and frozen on dry ice. Protein purity, quantity, and identity were determined by RP-HPLC and LC-MS.

[0320] In vitro enzyme activity assay for WRN helicase An ATPase assay was set up to measure the DNA-dependent ATP hydrolysis activity of WRN helicase, and this assay was also used to evaluate the inhibitory properties of the compounds of the present invention against DNA-dependent WRN ATPase activity.

[0321] The core helicase motif of the WRN protein (aa N517-P1238) was generated for this assay (protein was generated as described above). The 45-nucleotide oligonucleotide sequence (TTTTTTTTTTTTTTTTTTTTTTCCAAGTAAAACGACGGCCAGTGC; SEQ ID NO: 2) designated "FLAP26," described in Brosh et al., 2009, DOI: 10.1074 / jbc.M111446200, was purchased from IDT (Integrated DNA Technologies, Leuven, Belgium) and used as the signal-strand DNA substrate. The ADP-Glo ​​assay kit (Promega, Madison, WI), which allows for quantification of ADP generated in the ATP hydrolysis reaction, was used to set up this assay.

[0322] A time course experiment was first performed to determine the best enzyme assay conditions (including buffer conditions, reaction time, and concentrations of protein, ATP, and DNA substrate). A typical reaction consisted of 10 nM WRN protein, 0.2 nM FLAP26, and 300 micromolar ATP in the following assay buffer: 30 mM Tris pH 7.5, 2 mM MgCl, 0.02% BSA, 50 mM NaCl, 0.1% pluronic F127 (prepared in DNAse-free water).

[0323] To evaluate the inhibitory properties of compounds of the present invention, serial dilutions were prepared in DMSO (10 half-log dilutions from a 10 mM DMSO solution). Fifty nanoliters of each concentration was preincubated for 3 hours in assay buffer containing 600 micromolar ATP in a 384-well small-volume assay plate (Greiner #784075) containing 2.5 microliters of 20 nM WRN helicase protein. Control wells included a "high control" (no inhibition) containing DMSO without test compound and a "low control" (maximum inhibition) containing buffer without protein. The reaction was initiated by the addition of 2.5 microliters of FLAP26 at 0.4 nM and incubated for 30 minutes at room temperature. The reaction was stopped by the addition of 5 microliters of the first ADP-Glo ​​reagent and incubated for 1 hour to remove excess ATP. Subsequently, 10 microliters of ATP detection reagent was added and incubated for an additional hour before reading. Luminescence output was recorded using a Tecan 1000 reader with a 5-minute delay before reading. Each concentration of compound was tested in duplicate on the assay plate.

[0324] Data analysis was performed using in-house developed software (Novartis Helios software application, Novartis Institutes for BioMedical Research, unpublished) using the method described by Formenko et al., 2006, DOI: 10.1016 / j.cmpb.2006.01.008. After normalizing well activity values ​​to % inhibition (% inhibition = [(high control - sample) / (high control - low control)] x 100), IC values ​​were calculated from duplicate measurements on each plate according to [4]. 50 Data analysis was also performed using a four-parameter fit (e.g., GraphPad Prism, XLfit) to determine the IC 50 This can be done using commercially available software designed to derive the reported IC values. 50 Values ​​are the geometric means of at least two independent replicates.

[0325] Methods for detecting effects on cell proliferation The colon cancer cell lines SW48 (RRID: CVCL_1724), HCT116 (RRID: CVCL_0291), and SNU-407 (RRID: CVCL_5058) were obtained from ATCC. The WRN knockdown-insensitive colon cancer cell line DLD-1 (RRID: CVCL_0248) was obtained from the Korean Cell Line Bank (KCLB) and used to generate derivatives in which the endogenous WRN gene copy was knocked out by CRISPR-mediated editing using standard CRISPR methods. The resulting cell line, DLD1-WRN-KO, was used to evaluate potential off-target compound effects.

[0326] SW48, SNU-407, and DLD1-WRN-KO cells were cultured in growth medium consisting of RPMI-1640 (Amimed catalog no. 1-41F22-I), 2 mM L-glutamine (Amimed catalog no. 5-10K50), 10 mM HEPES (Gibco catalog no. 15630-056), 1 mM sodium pyruvate (Amimed catalog no. 5-60F00-H), 1× penicillin-streptomycin (Amimed catalog no. 4-01F00-H), and 10% fetal bovine serum (Amimed catalog no. 2-01F30-G, lot no. LB11566P). HCT 116 cells were cultured in growth medium consisting of McCoys 5A (Amimed catalog number 1-18F01-I), 2 mM L-glutamine (Amimed catalog number 5-10K50), 1x penicillin-streptomycin (Amimed catalog number 4-01F00-H), and 10% fetal bovine serum (Amimed catalog number 2-01F30-G, lot number LB11566P). All cells were maintained at 37°C in a humidified 5% CO2 incubator.

[0327] After filtration through a Steriflip-NY 20 μm filter (Millipore catalog no. SCNY00020), trypsinized cells were seeded into white, clear-bottom 96-well plates (Costar catalog no. 3903) at 2,000 (SW48) or 1,500 (SNU-407, DLD1-WRN-KO, HCT 116) cells / well in 100 microliters of growth medium. Three replicate plates were prepared for each compound treatment condition. Additionally, one plate (designated "day 0") was prepared for quantitating viable cell numbers at the time of compound addition. After overnight incubation at 37°C in a humidified 5% CO2 atmosphere, eight three-fold serial dilutions of a given compound stock (obtained at a concentration of 10 mM in DMSO and stored at 4°C) were dispensed directly into each of the triplicate assay plates using an HP 300D non-contact digital dispenser (TECAN). The final concentration of DMSO was normalized to 0.1% in all wells. Ninety-six hours after compound addition, cellular ATP levels, as a surrogate for cell viability, were assessed after the addition of 50 microliters of CellTiterGlo (Promega catalog number G7573) reagent, and luminescence was quantified on an MPLEX multimode plate reader (TECAN) after a 10-minute incubation at room temperature. The number of viable cells in the "day 0" plate was similarly quantified on the day of compound addition.

[0328] For data analysis, the assay background signal determined in wells containing medium but no cells was subtracted from all data points before further calculations. The degree of growth inhibition and potential cell death were evaluated by comparing the ATP levels (measured using CellTiterGlo, Promega) in compound-treated cells to those present at the time of compound addition. For this purpose, the following conditional concepts were programmatically applied in HELIOS. HELIOS is in-house software that applies a multi-stage decision tree to reach the optimal concentration-response curve fit (Gubler et al, SLAS DOI: 10.1177 / 2472555217752140), and calculates the growth % (G%) of each well treated with a compound: G% = ((T - V0) / V0)) * 100 (when T < V0), G% = ((T - V0) / (V - V0)))*100 (when T ≥ V0) (where V0 is the survival level at the time of compound addition, and V and T are the vehicle control survival level and compound-treated survival level, respectively, at the end of compound incubation). 100%, 0%, and -100% indicate, respectively, the absence of growth inhibition, growth arrest, and complete cell death. The compound concentration (GI50) that results in 50% growth inhibition and the residual cell survival rate (data (cmax), expressed as a percentage) at the highest concentration of the test compound were routinely calculated. Data analysis can also be performed using commercially available software designed to derive IC50 values using a 4-parameter fit (e.g., GraphPad Prism, XL Fit). The reported GI 50 values are the geometric mean of at least two independent replicates.

[0329] In Vivo Demonstration of the Efficacy of the Compounds of the Invention Experiments were performed on female Crl:NU(NCr)-Foxn1NU-homozygous nude mice (Charles River). Animals were housed in Allentown XJ cages (IVC, maximum 6 mice per cage) under optimized hygienic conditions, with food and water ad libitum and a 12-h light:dark cycle. Animals were allowed to acclimate for at least 1 week before enrollment in the experimental design. The studies described herein were conducted in accordance with License 2275 approved by the Basel Cantonal Veterinary Office.

[0330] SW48 human colon cancer cells were obtained from ATCC. Cells were cultured in RPMI-1640 medium (BioConcept, Amimed, #1-41F01-I) supplemented with 10% FCS (BioConcept, #2-01F30-I), 2 mM L-glutamine (BioConcept Ltd., Amimed, #5-10K50-H), 1 mM sodium pyruvate (BioConcept #5-60F00-H), and 10 mM HEPES (BioConcept #5-31F00-H) at 37°C in a 5% CO atmosphere. To establish SW48 xenografts, cells were harvested and resuspended in HBSS (Gibco, #14175). 100 μL of 5 million cells were injected subcutaneously into the right flank of isoflurane-anesthetized animals.

[0331] Tumor growth was monitored periodically after cell inoculation, and when tumor volume reached an appropriate volume, animals were randomized into treatment groups (n=7). Tumor volume was measured approximately twice a week during the treatment period. Tumor size (mm 3 ) was calculated from ((L × W × π / 6), where W = tumor width and L = tumor length.

[0332] Depending on the target concentration, 50–200 mg of amorphous sodium salt of the test compound (corrected for salinity) was dissolved in 8 mL of a 10% w / v aqueous solution of 2-hydroxypropyl-beta-cyclodextrin (HPBCD). The pH was adjusted to pH 7.4 with 0.1 M HCl (approximately 1 equivalent), and the resulting solution was filled to a total volume of 10 mL with 10% aqueous HPBCD. If cloudy, the solution was filtered. The resulting solution formulation was used for in vivo studies.

[0333] Tumor-bearing animals were screened until their tumors reached 186 mm 3 When tumors reached an adequate size to form groups with a mean tumor volume of 100 mg / kg, they were enrolled into treatment groups (n=7). Animals were then treated with vehicle (10% hydroxypropyl-beta-cyclodextrin) or a compound of the invention at 240 mg / kg QD by oral gavage at 20 mL / kg. Animals were weighed twice weekly and frequently examined for overt signs of adverse effects.

[0334] Tumor and body weight change data were statistically analyzed using GraphPad Prism 7.00 (GraphPad Software). If the data variance was normally distributed, one-way ANOVA with Dunnett's post-hoc test was used to analyze data for comparisons between treatment and control groups. Where applicable, results are presented as mean ± SEM.

[0335] As a measure of efficacy, %T / C% values ​​are calculated at the end of the experiment according to the following: (Δtumor volume 処置 / Δtumor volume 対照 )*100 Tumor regression was calculated as follows: -(Δtumor volume 処置 / Δtumor volume 処置開始時 )*100. Here, Δtumor volume indicates (mean tumor volume on the day of evaluation)−(mean tumor volume at the start of the experiment).

[0336] Treatment was performed, for example, on tumors with a mean tumor volume of 186 mm3 In the case of 10% hydroxypropyl-beta-cyclodextrin in water, QD oral administration was initiated at 240 mg / kg. The treatment period with Example 58 was 18 days, after which overall efficacy and tolerability were assessed based on tumor volume and body weight changes observed during the treatment period (Figure 6). Orally administered Example 58 at 240 mg / kg qd induced an antitumor response against SW48 xenografts in nude mice (Figure 6). The T / C% value on day 18 was -9 (p=<0.05 compared to vehicle control, one-way ANOVA with Dunnett's post-hoc test). The mean tumor volume on day 18 in 4 / 7 surviving animals showed a regression of -26%. Based on body weight, QD administration of Example 58 at 240 mg / kg was well tolerated (Figure 6). Data for compounds of Examples 42, 96 and 57 are shown in Figures 9, 10 and 11, respectively.

[0337] This application contains a Sequence Listing that has been submitted electronically in ASCII format and is incorporated herein by reference in its entirety. The ASCII copy, created on May 21, 2021, is named PAT059096_SL.txt and is 13184 bytes in size.

[0338] The following table shows the IC in the WRN ATPase assay for compounds of the present invention. 50 Data and GI in proliferation assays using SW48 and DLD1-WRN-KO cell lines 50 For example, Example 1 shows a proliferation GI of 50 nM in the SW48 cell line and >10 micromolar in the DLD1 WRN-KO cell line. 50 , a 50 nM WRN ATPase inhibitor.

[0339] [Table 4]

[0340] [Table 5]

[0341] [Table 6]

[0342] Data are geometric means from at least duplicate determinations.

[0343] In another aspect, the present invention provides compounds of formula (I) for use in the treatment of cancer or as research chemicals, such as chemical probes, where some of the WRN inhibitory activity is via a mechanism not assessed by the biochemical assays described above.

[0344] Methods for detecting clonogenic effects on cell proliferation (CFA = colony formation assay) Cell lines were obtained from ATCC, and media and culture conditions were used as recommended by ATCC. All cells were maintained at 37°C in a humidified 5% CO2 incubator. Cells were seeded in 12-well plates at 250–2,000 cells per well in 1 ml of media. The WRN inhibitor compound from Example 42 was added at a starting concentration of 10 μM. After overnight incubation at 37°C in a humidified 5% CO2 atmosphere, 10 three-fold serial dilutions of a given compound stock (obtained at a concentration of 10 mM in DMSO and stored at 4°C) were dispensed directly into each assay plate using an HP 300D non-contact digital dispenser (TECAN). The final DMSO concentration was normalized to 0.1% in all wells. Cells were left in the incubator for 8–20 days, with media changes every 3–4 days. After this, 100 μl of 37% formaldehyde was added directly to each test well and incubated for 15 minutes at room temperature. After rinsing twice with 5 ml of water, 0.5 ml of 0.05% methylene blue was added for 20 minutes at room temperature. The wells were rinsed three times with water, and 1 ml of 3% HCl was added to the plate and shaken until the color was completely dissolved. 200 μl of this solution was transferred to a 96-well plate, and absorbance was measured at 650 nM using a microtiter plate reader (Synergy HT). The compound concentration resulting in 50% growth inhibition (GI50) was calculated using XLfit with Dose Response One Site Model 201: fit = (A + ((BA) / (1 + ((x / C)^D)))). For non-adherent cell lines, cellular ATP levels, as a surrogate for cell viability, were assessed after removing 600 μl and adding 200 μl of CellTiterGlo (Promega Cat. No. G7573) reagent to the cultures. Luminescence quantification was performed on a Synergy HT plate reader after a 15-minute incubation at room temperature. Data were analyzed similarly to methylene blue staining. Results are shown in Figure 8. Further information on MSI-H and MSS is available from the following references: ·Chan et Al.,WRN helicase is a synthetic lethal target in microsatellite unstable cancers.Nature.2019 Apr;568(7753):551-556.doi:10.1038 / s41586-019-1102-x.Epub 2019 Apr 10.PMID:30971823;PMCID:PMC6580861. ·McDonald ERet al.,Project DRIVE:A Compendium of Cancer Dependencies and Synthetic Lethal Relationships Uncovered by Large-Scale,Deep RNAi Screening.Cell 170(3):577-592(2017)).

[0345] Preparation of compounds The compounds of the present disclosure can be prepared as described in the Examples below. The following examples are intended to illustrate the present invention and should not be construed as limiting the present invention.

[0346] device Microwave: All microwave reactions were carried out in a Biotage Initiator or Anton Paar monowave 450 irradiating from a 2.45 GHz magnetron with a Robot Eight / Robot Sixty / Robot Twentyfour processing capacity at 0-400 W unless otherwise specified.

[0347] UPLC-MS Method: A Waters Acquity UPLC equipped with a Waters SQ detector is used unless otherwise specified.

[0348] UPLC-MS 1: Column CORTECS (trademark) C18+2.7μm, Column dimensions: 2.1 x 50 mm Column temperature: 80°C Eluent A: Water + 4.76% isopropanol + 0.05% FA + 3.75mM AA B: Isopropanol + 0.05% FA Flow rate 1.0mL / min Gradient: 1 to 50% B in 1.4 min, 50 to 98% B in 0.3 min

[0349] UPLC-MS 2: Column: ACQUITY UPLC® BEH C18 1.7 μm Column dimensions: 2.1 x 100 mm Column temperature: 80°C Eluent A: Water + 4.76% isopropanol + 0.05% FA + 3.75mM AA B: Isopropanol + 0.05% FA Flow rate 0.4mL / min Gradient: 1 to 60% B in 8.4 min, 60 to 98% B in 1.0 min

[0350] UPLC-MS 3: Column: ACQUITY UPLC® BEH C18 1.7 μm Column dimensions: 2.1 x 50 mm Column temperature: 80°C Eluent A: Water + 4.76% isopropanol + 0.05% FA + 3.75mM AA B: Isopropanol + 0.05% FA Flow rate 0.6mL / min Gradient: 1 to 98% B in 1.7 min

[0351] UPLC-MS 4: Column: ACQUITY UPLC® BEH C18 1.7 μm Column dimensions: 2.1 x 50 mm Column temperature: 80°C Eluent A: Water+0.05% FA+3.75mM AA B: Isopropanol + 0.05% FA Flow rate 0.6 / 0.7mL / min Gradient: 5 to 98% B in 1.7 min

[0352] UPLC-MS 5: Column: XBridge® BEH™ C18 2.5 μm Column dimensions: 2.1 x 50 mm Column temperature: 80°C Eluent A: Water + 5mM NH4OH B: Acetonitrile + 5mM NH4OH Flow rate 1.0mL / min Gradient: 2 to 98% B in 1.4 min

[0353] UPLC-MS 6: Column: Ascentis® Express C18 2.7 μm Column dimensions: 2.1 x 50 mm Column temperature: 80°C Eluent A: Water + 4.76% isopropanol + 0.05% FA + 3.75mM AA B: Isopropanol + 0.05% FA Flow rate 1.0mL / min Gradient: 1 to 50% B in 1.4 min, 50 to 98% B in 0.3 min

[0354] UPLC-MS 7: Column: Acquity UPLC® HSS T3 1.8 μm Column dimensions: 2.1 x 50 mm Column temperature: 60°C Eluent A: Water + 0.05% formic acid + 3.75 mM ammonium acetate B: Acetonitrile + 0.04% FA Flow rate 1.0mL / min Gradient: 2 to 98% B in 1.4 min

[0355] UPLC-MS 8: Column: Acquity UPLC® HSS T3 1.8 μm Column dimensions: 2.1 x 50 mm Column temperature: 60°C Eluent A: Water + 0.05% formic acid + 3.75 mM ammonium acetate B: Acetonitrile + 0.04% FA Flow rate 1.0mL / min Gradient: 5 to 98% B in 1.4 min

[0356] UPLC-MS 9: Column: XBridge® BEH™ C18 2.5 μm Column dimensions: 2.1 x 50 mm Column temperature: 80°C Eluent A: Water + 5mM NH4OH B: Acetonitrile + 5mM NH4OH Flow rate 1.0mL / min Gradient: 2 to 98% B in 9.4 min

[0357] UPLC-MS 10: Column CORTECS (trademark) C18+2.7μm, Column dimensions: 2.1 x 50 mm Column temperature: 80°C Eluent A: Water+0.05% FA+3.75mM AA B: Isopropanol + 0.05% FA Flow rate 1.0mL / min 1.4 minutes concave 1%~98%B

[0358] UPLC-MS 11: Equipment: Shimadzu NEXERA UPLC PDA with Shimadzu LCMS 2020 as MSD Column: Mercury MS Synergi C12 2.5 μm Column dimensions: 20 x 4.0 mm Column temperature: 40°C Eluent A: Water + 0.1% FA B: Acetonitrile Flow rate 2.0mL / min Gradient Time / %B: 0.01 / 5, 0.5 / 5, 1.0 / 95, 1.5 / 95, 2.0 / 5, 3.0 / 5

[0359] UPLC-MS 12: Instrumentation: Agilent 1200 HPLC PDA with AB Sciex API2000 TQ as MSD Column: Mercury MS Synergi C12 2.5μm Column dimensions: 20 x 4.0 mm Column temperature: 30°C Eluent A: Water + 0.1% FA B: Acetonitrile Flow rate 2.0mL / min Gradient Time / %B: 0.01 / 30, 0.5 / 30, 1.0 / 95, 2.4 / 95, 2.5 / 30, 3.0 / 30

[0360] UPLC-MS 13: Instrumentation: Agilent 1200 HPLC PDA with AB Sciex API3200 QTRAP as MSD Column: Kinetex EVO C18 2.6 μm Column dimensions: 50 x 4.6 mm Column temperature: 30°C Eluent A: Water + 0.1% FA B: Acetonitrile + 0.1% FA Flow rate 1.5mL / min Gradient Time / B: 0 / 20, 0.2 / 50, 1 / 95, 2.7 / 95, 2.8 / 20, 4 / 20

[0361] UPLC-MS 14: Column: Acquity UPLC® HSS T3 1.8 μm Column dimensions: 2.1 x 100 mm Column temperature: 60°C Eluent A: Water + 0.05% formic acid + 3.75 mM ammonium acetate B: Acetonitrile + 0.04% FA Flow rate 1.0mL / min Gradient: 5 to 98% B in 9.4 min

[0362] HPLC method: HPLC 1: Instruments: Agilent 1100 series with PDA detector Column: Kinetex C-18, 5 μm Column dimensions: 150 x 4.6 mm Column temperature: 40°C Eluent A: Water + 0.01% TFA B: Acetonitrile Flow rate 1.0mL / min Gradient Time / B: 0 / 30, 2 / 40, 5 / 90, 8 / 100, 10 / 100, 11 / 30, 12 / 30

[0363] HPLC 2: Instrumentation: Acquity Arc Waters UHPLC with PDA detector (2998PDA) Column: Kinetex EO, 2.6 μm Column dimensions: 100 x 4.6 mm Column temperature: 40°C Eluent A: Water + 0.01% TFA B: Acetonitrile Flow rate 1.0mL / min Gradient Time / B: 0 / 5, 2 / 5, 6 / 70, 10 / 100, 13 / 100, 13.5 / 5, 15 / 5

[0364] HPLC 3: Instrument Agilent 1260 HPLC Column: Agilent Poroshell 120 C18, 2.7 μm Column dimensions: 4.6 x 50 mm Column temperature: 40°C Eluent A: Water + 0.01% TFA B: Acetonitrile + 0.01% TFA Flow rate 1.2mL / min Gradient: 0%B to 50%B in 5 minutes, hold for 2 minutes

[0365] HPLC 4: Instrument Agilent 1260 Column: Agilent Poroshell 120 EC-C18, 2.7 μm Column dimensions: 4.6 x 50 mm Column temperature: 40°C Eluent A: Water + 0.1% TFA B: Acetonitrile + 0.1% TFA Flow rate 1.2mL / min Gradient: 5%B to 95%B in 5 minutes, hold for 2 minutes

[0366] HPLC 5: Instrument Agilent 1260 HPLC Column: InertSustain C18, 5 μm Column dimensions: 4.6 x 150 mm Column temperature: 30°C Eluent A: Water + 5mmol(NH4)2CO3 B: Acetonitrile Flow rate 1.0mL / min Gradient: 10%B to 90%B in 8 minutes, hold for 2 minutes

[0367] HPLC 6: Instrument: Agilent 1260 infinity series HPLC system with DAD / ELSD Column: Atlantis dC18, 5 μm Column dimensions: 4.6 x 250 mm Column temperature: 25°C Eluent A: Water + 0.1% TFA B: Acetonitrile Flow rate 1.0mL / min Gradient: 10%B to 100%B in 15 minutes, hold for 5 minutes

[0368] Chiral analytical HPLC method C-HPLC 1: Instrument: Agilent 1260 Infinity II Series with PDA detector Injection: 3μL Mobile phase: A: Hexane B: 0.1% HCOOH (in EtOH) Flow rate: 20mL / min Column: CELLULOSE 4 (150 x 4.6 mm, 5 μm) Detection UV: 210nm Gradient: Uniform concentration: 50:50

[0369] C-HPLC 2: Instrument: Agilent 1260 Infinity II Series with PDA detector Injection: 10μL Mobile phase: A: Hexane B: 0.1% HCOOH (in EtOH) Flow rate: 15mL / min Column: CELLULOSE 4 (150 x 4.6 mm, 5 μm) Detection UV: 210 nm. Gradient: uniform concentration 50:50

[0370] C-HPLC 3: Instrument: Agilent 1260 Infinity II Series with PDA detector Injection volume: 8μL Mobile phase: A: Hexane B: 0.1% HCOOH (in EtOH) Flow rate: 1mL / min Column: CELLULOSE 4 (150 x 4.6 mm, 5 μm) Detection UV: 210 or 254 nm Gradient: Uniform concentration: 50:50

[0371] C-HPLC 4: Instrument: Agilent 1260 Infinity II Series with PDA detector Injection volume: 3μL Mobile phase: A: Hexane B: 0.1% HCOOH (in EtOH) Flow rate: 1mL / min Column: CELLULOSE 4 (150 x 4.6 mm, 5 μm) Detection UV: 210 or 254 nm Gradient: Uniform concentration: 50:50

[0372] C-HPLC 5: Instrument: Agilent 1260 Infinity II Series with PDA detector Injection volume: 2μL Mobile phase: A: Hexane B: 0.1% TFA in EtOH Flow rate: 1mL / min Column: CELLULOSE 4 (150 x 4.6 mm, 5 μm) Detection: 210 or 254 nm Gradient: Uniform concentration: 50:50

[0373] C-HPLC 6: Instrument: Agilent 1260 Infinity II Series with PDA detector Injection volume: 3μL Mobile phase: A: Hexane B: 0.1% TFA in EtOH Flow rate: 1mL / min Column: CELLULOSE 4 (150 x 4.6 mm, 5 μm) Detection UV: 210 or 254 nm Gradient: uniform concentration 50:50

[0374] C-HPLC 7: Equipment:Waters UPC 2 MS Injection volume: 5μL Mobile phase: A: 25% (MeOH+0.1% NH3); B: 75% scCO2 uniform concentration Flow rate: 3mL / min Column: Chiralpak IB-N 5μm, 100×4.6mm Detection UV: 190~400nm Oven temperature: 40°C

[0375] C-HPLC 8: Equipment:Waters UPC 2 MS Injection volume: 5μL Mobile phase: A: 35% (MeOH + 0.1% NH3); B: 65% scCO2 uniform concentration Flow rate: 3mL / min Column: Chiralpak IB-N 5μm, 100×4.6mm Detection: 190~400nm Oven temperature: 40°C

[0376] C-HPLC 9: Instrument: Waters UPC 2 Injection volume: 1 μL Mobile phase: A: CO2; B: MeOH + 0.05% DEA Flow rate: 2.4 mL / min Column: Chiralpak IC, 3 μm, 100 × 4.6 mm Detection: 220 nm Column temperature: 35 °C Back pressure: 100 bar

[0377] C-HPLC 10: Instrument: Waters UPC 2 -MS Injection volume: 5 μL. Mobile phase: Uniform concentration A = 35% (IPA + 0.1% NH3) B = 65% scCO2 Flow rate: 3 mL / min Column: Chiralpak IB, 5 μm, 100 × 4.6 mm Detection: DAD 210~400 nm Back pressure: 1800 psi

[0378] C-HPLC 11: Instrument: Waters Acquity UPC Injection volume: 5 μL Mobile phase: A: 35% (MeOH + 0.05% NH3); B: 65% scCO2 Flow rate: 3 mL / min Column: Chiralpak IB-N 5 μm, 100 × 4.6 mm Detection UV: 240 nm Oven temperature: 40 °C

[0379] C-HPLC 12: Instrument: Waters Acquity UPC Injection volume: 5 μL Mobile phase: A: 35% (MeOH + 0.05% NH3); B: 65% scCO2 Flow rate: 3 mL / min Column: Chiralpak IB-N 5μm, 100×4.6mm Detection UV: 240nm Oven temperature: 40°C

[0380] C-HPLC 13: Equipment:Waters UPC 2 -MS Injection volume: 5μL Mobile phase: uniform concentration A=25%(MeOH+0.05% NH3) B=75% scCO2 Flow rate: 3mL / min Column: Chiralpak IB, 5μm, 100×4.6mm Detection: DAD 210~400nm: Back pressure: 1800 psi

[0381] Adjustment method: Column chromatography: Column chromatography was carried out on silica gel using prepacked columns as detailed below, or using glass columns according to standard flash chromatography techniques, unless otherwise stated. System 1 TeledyneISCO, CombiFlash Rf, CombiFlash Rf+ System 2 Biotage Isolera Column: Pre-packed RediSep Rf cartridge or SNAP cartridge Adsorption of sample onto Isolute, onto silica gel, or as a solution

[0382] Supercritical Fluid Chromatography (SFC) Purification was performed on a Waters preparative SFC-100-MS system with ABSYS updates, equipped with a Waters 2998 photodiode array detector and a Waters MS single quadrupole detector.

[0383] SFC 1: Equipment: WATERS SFC 100 with ABSYS update Mobile phase: A: CO2, B: MeOH: Flow rate: 150mL / min MeOH + 30mL / min CO2, 180mL / min constant flow Column: 250 x 30 Reprospher PEI 100A 5μm Temperature 50℃ Back pressure: 100bar Detection UV: 210~400nm). Gradient: 18% B to 26% B in 6.86 minutes

[0384] SFC 2: Equipment: WATERS SFC 100 with ABSYS update Mobile phase: A: CO2; B: MeOH Flow rate: 150mL / min MeOH + 30mL / min CO2, 180mL / min constant flow Column: 100 x 30 Reprosil NH2100A 3 μm Temperature 50℃ Back pressure: 100bar Detection UV: 210~400nm Gradient: 34%B to 42%B in 2.8 minutes

[0385] SFC 3: Equipment: WATERS SFC 100 with ABSYS update Mobile phase: A: CO2, B: MeOH Flow rate: 150mL / min MeOH + 30mL / min CO2, 180mL / min constant flow Column: 100 x 30 Reprosil NH2100A 3 μm Temperature: 50℃ Back pressure: 100bar Detection UV: 210~400nm Gradient: (First run) 21% B to 29% B in 2.8 minutes (Second run to improve detection) 25% B to 33% B in 2.8 minutes

[0386] SFC 4: Equipment: WATERS SFC 100 Mobile phase: A.CO2, B:MeOH: Flow rate: 100mL / min Column: 250 x 30 Reprospher PEI 100A 5μm Temperature 32℃ Back pressure: 120bar Detection UV: 210~400nm Gradient: 40% B to 55% B in 10.2 min

[0387] SFC 5: Equipment: WATERS SFC 100 Mobile phase: A: CO2, B: MeOH Flow rate: 100mL / min MeOH Column: 250 x 30 Reprospher PEI 100A 5μm Temperature 32℃ Back pressure: 120bar Detection UV: 210~400nm Gradient: 35% B to 45% B in 10.3 min

[0388] SFC 6: Equipment: WATERS SFC 100 with ABSYS update Mobile phase: A: CO2, B: MeOH: Flow rate: 150mL / min MeOH+30mL / min CO2, constant flow of 180mL / min Column: 100 x 30 Reprosil NH2100A 3 μm Temperature: 50℃ Back pressure: 100bar Detection UV: 210~400nm Gradient: 31% B to 39% B in 2.8 min

[0389] SFC 7: Equipment: WATERS SFC 100 with ABSYS update Mobile phase: A: CO2, B: MeOH Flow rate: 150mL / min MeOH+30mL / min CO2, constant flow of 180mL / min Column: 250 x 30 waters Torus2-PIC 130A 5μm Temperature: 50℃ Back pressure: 100bar Detection UV: 210~400nm Gradient: 26% B to 34% B in 5.48 minutes

[0390] Reverse phase HPLC: RP-HPLC Basic 1: System Gilson Column: Waters X-Bridge Prep C18 OBD (100 mm x 30 mm), 5 μm Eluent A: Water + 7.3 mM NH4OH, B: Acetonitrile Flow rate 40mL / min

[0391] RP-HPLC Basic 2: System: Agilent 1200 Series with single quadrupole mass spectrometer Column: XBRIDGE (150 mm x 20 mm), 5.0 μm Eluent A: 0.02% ammonia (in water), B: acetonitrile Flow rate 20mL / min

[0392] RP-HPLC acidic 1: System Gilson Column: Waters SunFire Prep C18 OBD (100 mm x 30 mm), 5 μm Eluent A: Water + 0.1% TFA, B: Acetonitrile Flow rate 40mL / min

[0393] RP-HPLC acidic 2: System: Agilent 1200 Series with single quadrupole mass spectrometer Column: LUNA C18 (250 mm x 19 mm), 4.0 μm Eluent A: 0.1% HCOOH (in water), B: acetonitrile Flow rate 20mL / min

[0394] RP-HPLC acidic 3: System: Agilent 1200 Series with single quadrupole mass spectrometer Column: LUNA (250 mm x 21.2 mm), 5.0 μm Eluent A: 0.1% HCOOH (in water), B: acetonitrile Flow rate 18mL / min

[0395] RP-HPLC acidic 4: System: Agilent 1200 Series with single quadrupole mass spectrometer Column: LUNA (250 mm x 21.2 mm), 5.0 μm Eluent A: 0.1% HCOOH (in water), B: acetonitrile Flow rate 20mL / min

[0396] RP-HPLC acidic 5: System: Agilent 1200 Series with single quadrupole mass spectrometer Column: LUNA Phenomenex (250 mm x 21.2 mm), 5.0 μm Eluent A: 0.1% HCOOH (in water), B: acetonitrile Flow rate 18mL / min

[0397] RP-HPLC acid 6: System: Agilent 1200 Series with single quadrupole mass spectrometer Column: Atlantis (250 mm x 19 mm), 5.0 μm Eluent A: 0.1% HCOOH (in water), B: acetonitrile:MeOH Flow rate 18mL / min

[0398] RP-HPLC acidic 7: System: Agilent 1200 Series with single quadrupole mass spectrometer Column: LUNA C18 (250 mm x 21.2 mm), 5.0 μm Eluent A: 0.1% HCOOH (in water), B: acetonitrile Flow rate 20mL / min

[0399] RP-HPLC acidic 8: System: Agilent 1200 Series with single quadrupole mass spectrometer Column: Atlantis (250 mm x 19 mm), 5.0 μm Eluent A: 0.1% HCOOH (in water), B: acetonitrile Flow rate 18mL / min

[0400] RP-HPLC acidic 9: System: Agilent 1200 Series with single quadrupole mass spectrometer Column: Atlantis (250 mm x 21.2 mm), 5.0 μm Eluent A: 0.1% HCOOH (in water), B: acetonitrile Flow rate 20mL / min

[0401] RP-HPLC acidic 10: System: Agilent 1200 Series with single quadrupole mass spectrometer Column: Atlantis (250 mm x 21.2 mm), 5.0 μm Eluent A: 0.1% HCOOH (in water), B: acetonitrile Flow rate 18mL / min

[0402] RP-HPLC acidic 11: System: Agilent 1200 Series with single quadrupole mass spectrometer Column: LUNA OMEGA (250 mm x 21.2 mm), 5.0 μm Eluent A: 0.1% HCOOH (in water), B: acetonitrile:MeOH (1:1) Flow rate 20mL / min

[0403] RP-HPLC acidic 12: System: Agilent 1200 Series with single quadrupole mass spectrometer Column: LUNA C18 (250 mm x 21.2 mm), 5.0 μm Eluent A: 0.1% HCOOH (in water), B: acetonitrile Flow rate 18mL / min

[0404] RP-HPLC acidic 13: System Gilson Column: Nucleodur C18 (21mm x 250mm) Eluent A: Water + (0.1% COOH), B: Acetonitrile + 0.1% COOH Flow rate 40mL / min

[0405] RP-HPLC acidic 14: Teledyne / Isco AccqPrep HP150 prep system Column: 50 x 100 Xbridge C18 (50 mm x 100 mm), 5 μm Eluent A: Water + 0.1% TFA, B: Acetonitrile Flow rate 100mL / min

[0406] RP-HPLC neutral 1: System: Agilent 1200 Series with single quadrupole mass spectrometer Column: KINETEX (150 mm x 21.2 mm), 5 μm Eluent A: water, B: acetonitrile Flow rate 20mL / min

[0407] RP-HPLC Neutral 2: System: Agilent 1200 Series with single quadrupole mass spectrometer Column: Atlantis (250mm x 21.2mm), 5μm Eluent A: water, B: acetonitrile Flow rate 17mL / min

[0408] RP-HPLC Neutral 3: System: Agilent 1200 Series with single quadrupole mass spectrometer Column: LUNA C18 (250 mm x 21.2 mm), 5 μm Eluent A: water, B: acetonitrile Flow rate 20mL / min

[0409] RP-HPLC Neutral 4: System: Agilent 1200 Series with single quadrupole mass spectrometer Column: LUNA Phenomenex (250 mm x 21.2 mm), 5 μm Eluent A: water, B: acetonitrile Flow rate 18mL / min

[0410] RP-HPLC neutral 5: System: Agilent 1200 Series with single quadrupole mass spectrometer LUNA column (250mm x 21.2mm) Eluent A: water, B: acetonitrile Flow rate 20mL / min

[0411] Preparation of compounds The following examples are intended to illustrate the present invention and should not be construed as limiting it. Temperatures are given in degrees Celsius. Unless otherwise indicated, all evaporations are carried out under reduced pressure, typically between about 15 mmHg and 100 mmHg (= 20 to 133 mbar). Abbreviations used are those commonly used in the art.

[0412] All starting materials, building blocks, reagents, acids, bases, dehydrating agents, solvents, and catalysts utilized to synthesize the compounds of the present invention are either commercially available or can be prepared by organic synthesis methods known to those skilled in the art. Additionally, the compounds of the present invention can be prepared by organic synthesis methods known to those skilled in the art, as shown in the examples below.

[0413] The structures of final products, intermediates, and starting materials are confirmed by standard analytical spectroscopic characteristics, such as MS, IR, or NMR. The absolute stereochemistry of specific isomers has been determined by analysis of the X-ray crystal structure of a complex of the respective compound bound to WRN or by the small molecule X-ray crystal structure of a precursor to the final compound. Amines synthesized by acidic deprotection of the Boc precursor were often obtained as HCl or TFA salts. The corresponding free bases can be isolated by partitioning between DCM and saturated aqueous NaHCO3, as described for intermediate Y.

[0414] General conditions: Mass spectra were acquired on LC-MS systems using electrospray, chemical, or electron impact ionization from various instruments configured as follows: a Waters Acquity UPLC equipped with a Waters SQ detector, a Shimadzu NEXERA UPLC PDA equipped with a Shimadzu LCMS 2020 as the MSD, an Agilent 1200 HPLC PDA equipped with an AB Sciex API 2000 TQ as the MSD, and an Agilent 1200 HPLC PDA equipped with an AB Sciex API 3200 QTRAP as the MSD. [M+H] + refers to the protonated molecular ion of a chemical species. NMR spectra were performed using Bruker Ultrashield™ 400 (400 MHz), Bruker Ultrashield™ 400 Plus (400 MHz), Bruker Ultrashield™ 600 (600 MHz), and Bruker Ascend™ 400 (400 MHz) spectrometers, all with and without tetramethylsilane as an internal standard. Chemical shifts (d values) are reported in ppm downfield from tetramethylsilane, and spectral splitting patterns are designated as singlets (s), doublets (d), triplets (t), multiplets, unresolved or further overlapping signals (m), or broad signals (br). Solvents are indicated in parentheses.

[0415] Celite:Celite R (Celite Corporation) = Filter aid based on diatomaceous earth Phase separator: Biotage Isolute Phase Separator (Part Number: 120-1906-D (for 15 mL), Part Number: 120-1908-F (for 70 mL) and Part number: 120-1909-J (for 150mL) SiliaMetS® Thiol: SiliCYCLE Thiol Metal Scavenger (Part Number: R51030B, Loading: 1.31 mmol / g, Particle Size: 40-63 μm) ISOLUTE® Si-Thiol: Biotage thiol metal scavenger (Part Number: 9180-0100, Loading: 1.3 mmol / g) PL-BnSH MP Resin: Agilent thiol metal scavenger (Part Number: PL3582-6689, 2.2 mmol / g 100A, 150-1 kg) ISOLUTE® Si-TMT: Biotage Thiol Metal Scavenger (Part Number: 9538) Smopex®-301: Alfa Aesar thiol metal scavenger (Part Number: 45902) PL-HCO3MP SPE Cartridge (500mg / 6mL) - (Part Number: PL3540-C603) PL-HCO3MP SPE Cartridge (100mg / 6mL) - (Part Number: PL3540-A603)

[0416] Selected compounds were crystallized and further characterized. Experimental procedures are outlined in the examples below, and descriptions of equipment and methods are outlined below.

[0417] [Table 7]

[0418] [Table 8]

[0419] Sodium salt formation: The compound was suspended in tert-butanol. 0.1M NaOH (1 equivalent) was added. The mixture was stirred / sonicated overnight at room temperature. If the suspension turned into a clear solution, it was freeze-dried. If the suspension was still cloudy, water was added and the resulting solution was freeze-dried. If no change occurred, 0.1M NaOH was added up to a total of 2 equivalents until a clear solution was observed, which was then freeze-dried. If the NMR of the resulting solid still contained tert-butanol, the solid was dissolved in a small amount of water and freeze-dried again. The final sodium salt was obtained as a colorless powder. Its amorphous state was confirmed by XRPD.

[0420] Synthesis scheme In general, compounds of formula (I) can be prepared according to the schemes provided below. The examples outlining specific synthetic routes, and the general schemes below, are intended to provide guidance to synthetic chemists of ordinary skill in the art, who will readily understand that variations in solvents, concentrations, reagents, protecting groups, order of synthetic steps, times, temperatures, and the like may be made as necessary. The schemes provided below are intended to represent single diastereomers / enantiomers as well as isomeric mixtures thereof. Resolution of diastereomers / enantiomers may be carried out according to the techniques described herein.

[0421] The present invention includes the novel processes described herein, and further includes any variations of the processes, in which intermediates obtained at any stage thereof are used as starting materials to carry out the remaining steps, or in which starting materials are formed in situ under the reaction conditions, or in which reaction components are used in the form of their salts or optically pure substances. The compounds and intermediates of the present disclosure can also be converted into each other according to methods commonly known to those skilled in the art. In another aspect, the present invention provides novel intermediate compounds described herein.

[0422] [Table 9]

[0423] [Table 10]

[0424] [Table 11]

[0425] [Table 12]

[0426] [Table 13]

[0427] Synthesis of the final compound Scheme 1: Preparation of final compounds [ka]

[0428] Example 1: 2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-5-ethyl-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide [ka] N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (Intermediate B) (630 mg, 1.11 mmol) was suspended in DMF (6 mL). 4-Chloro-3-hydroxypicolinic acid (384 mg, 2.21 mmol), DIPEA (967 μL, 5.54 mmol), HOBt (299 mg, 2.21 mmol), and EDC·HCl (425 mg, 2.21 mmol) were added to the RM and stirred at room temperature for 12 h. Water was added to the RM, and the suspension was filtered. The resulting solid was purified by reverse-phase preparative HPLC (RP-HPLC acidic 5: 20-30% B in 2 min, 30-60% B in 8 min) to give the title compound. LC-MS: Rt = 1.09 min; MS m / z [M+H] + 724.6 / 726.6, m / z [MH] - 722.3 / 724.3; UPLC-MS 1 LC-MS: Rt=5.33 min; MS m / z [M+H] + 724.2 / 726.2, m / z [MH] - 722.3 / 724.2; UPLC-MS 2 1 H NMR(400MHz,DMSO-d6)δ 10.83(s,br,1H),10.34(s,1H),8.05(m,2H),7.96(d,J=2.1Hz,1H),7.72(dd,J=2.1Hz,8.8Hz,1H),7.55(d,J=5.1Hz,1H),5.21(s, 2H),4.53(m,1H),3.66(m,4H),3.54(m,3H),3.38(m,4H),3.23(m,1H),2.96(m,3H),2.78(m,1H),2.60(m,1H),1.16(t,J=7.3Hz,3H)

[0429] Example 2: N-(2-chloro-4-(trifluoromethyl)phenyl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide [ka] N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (Intermediate B) (200 mg, 352 μmol) was suspended in DMF (5 mL). 3-Hydroxypicolinate perfluorophenyl (Intermediate CT) (215 mg, 703 μmol) and EtN (97.0 μL, 703 μmol) were added, and the RM was stirred at 70 °C for 3 h. The RM was concentrated under reduced pressure. The crude product was first purified by column chromatography (silica gel column: 12 g silica, eluent: DCM:MeOH 100:0 to 90:10). A second purification by reverse phase preparative HPLC (RP-HPLC acidic 9: 40-50% B in 2 min, 50-55% B in 10 min) then afforded the title compound. LC-MS: Rt=0.98 min; MS m / z [M+H] + 690.6 / 692.6, m / z [MH] - 688.4 / 690.3; UPLC-MS 1 LC-MS: Rt=4.84 min; MS m / z [M+H] + 690.2 / 692.2 m / z [MH] - 688.3 / 690.3; UPLC-MS 2 1 H NMR(400MHz,DMSO-d6)δ 10.37(s,br,1H),10.34(s,br,1H),8.05(m,2H),7.96(d,J=2.1Hz,1H),7.72(dd,J=2.1Hz,8.7Hz,1H),7.28(m,2H),5.21(s,2H) ,4.53(m,1H),3.66(m,4H),3.46(m,3H),3.38(m,4H),3.20(m,1H),2.92(m,3H),2.76(m,1H),2.58(m,1H),1.16(t,J=7.5Hz,3H)

[0430] Example 3: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(4-fluoro-3-hydroxypicolinoyl)piperazin-1-yl)-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide [ka]

[0431] Step 1: 2-(6-(4-(3-(benzyloxy)-4-fluoropicolinoyl)piperazin-1-yl)-5-ethyl-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide To N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide.TFA (Intermediate B) (620 mg, 908 μmol) in DMF (3 mL) was added EtN (503 μL, 3.63 mmol), followed by 3-(benzyloxy)-4-fluoropicolinic acid (Intermediate CU) (224 mg, 908 μmol), and then HATU (380 mg, 999 μmol). The RM was stirred at room temperature for 30 min. The RM was diluted with water (5 mL), and the resulting suspension was stirred at room temperature for 90 min. The suspension was filtered. The cake was washed with water (20 mL) and dried under vacuum to give the title compound as an off-white solid. LC-MS: Rt=1.21 min; MS m / z [M+H] + 798.5 / 800.5, m / z [MH] - 796.5 / 798.5;UPLC-MS 1.

[0432] Step 2: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(4-fluoro-3-hydroxypicolinoyl)piperazin-1-yl)-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 2-(6-(4-(3-(benzyloxy)-4-fluoropicolinoyl)piperazin-1-yl)-5-ethyl-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide (617 mg, 773 μmol) was dissolved in HBr (48% aqueous solution) (1.00 mL, 8.84 mmol) and allowed to stand at room temperature for 3 h. It was then stored in a stoppered flask in the refrigerator over the weekend. The RM was allowed to warm to room temperature and then heated at 35° C. with stirring for 140 min. The RM was neutralized to pH 6 by the addition of 1 M aqueous NaOH and extracted with DCM (2×30 mL). The combined organic phase was dried over NaSO, filtered, and concentrated under reduced pressure. The crude product was purified in two batches by reverse-phase preparative HPLC (RP-HPLC acidic 1: 30-50% B in 20 min, 50% plateau for 1 min, and RP-HPLC acidic 1: 20-47% B in 20 min, 20% plateau for 1 min). The product-containing fractions were combined and partitioned between DCM (30 mL) and saturated aqueous NaHCO3 (5 mL). The organic layer was separated by filtration through a phase separator and concentrated under reduced pressure. The residue was recrystallized from MeOH / water to give the title compound as colorless crystals. LC-MS: Rt=0.98 min; MS m / z [M+H] + 708.5 / 710.5, m / z [MH] - 706.4 / 708.4; UPLC-MS 1 LC-MS: Rt=4.89 min; MS m / z [M+H] + 708.2 / 710.2, m / z [MH] - 706.2 / 708.2; UPLC-MS 2 1H NMR (400 MHz, DMSO-d6) δ 10.72(s,br,1H),10.33(s,1H),8.08(dd,J=5.3Hz,6.9Hz,1H),8.04(d,J=8.7 Hz,1H),7.96(d,J=2.1Hz,1H),7.72(dd,J=2.2Hz,8.8Hz,1H),7.35(dd,J=5.3 Hz,11.9Hz,1H),5.21(s,2H),4.52(m,1H),3.66(m,4H),3.45(m,3H),3.38(m, 4H),3.21(m,1H),2.94(m,3H),2.77(m,1H),2.59(m,1H),1.16(t,J=7.4Hz,3H)

[0433] Example 4: N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide [ka] N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (Intermediate C) (400 mg, 686 μmol) was suspended in DMF (5 mL). 3-Hydroxypicolinic acid (191 mg, 1.37 mmol), DIPEA (599 μL, 3.43 mmol), HOBt (185 mg, 1.37 mmol), and EDC·HCl (263 mg, 1.37 mmol) were added to the RM and stirred at room temperature for 12 h. Water was added to the RM and filtered. The solid was purified by reverse-phase preparative HPLC (RP-HPLC Neutral 5: 10-20% B in 2 min, 20-45% B in 10 min) to give the title compound. LC-MS: Rt = 1.00 min; MS m / z [M+H] + 704.1 / 706.2, m / z [MH] - 702.2 / 704.2; UPLC-MS 1 LC-MS: Rt = 5.01 min; MS m / z [M+H] + 704.2 / 706.1, m / z [MH] - 702.3 / 704.3; UPLC-MS 2 1 H NMR(400MHz,DMSO-d6)δ 10.37(s,br,1H),10.07(s,1H),8.06(m,1H),7.93(s,1H),7.75(s,1H),7.28(m,2H),5.16(s,2H),4.53(m,1H),3.65(m ,4H),3.46(m,3H),3.39(m,4H),3.20(m,1H),2.94(m,3H),2.75(m,1H),2.58(m,1H),2.34(s,3H),1.16(t,J=7.1Hz,3H)

[0434] Example 5: 2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide [ka] 2-(5-Ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide.HCl (Intermediate D) (230 mg, 381 μmol) and DIPEA (333 μL, 1.91 mmol) were dissolved in DCM (10 mL), and then 3-hydroxypicolinoyl chloride (Intermediate CV) (90.0 mg, 572 μmol) was added at 0° C. and stirred for 1 hour. 3-Hydroxypicolinoyl chloride (Intermediate CV) (90.0 mg, 572 μmol) was added again at 0° C. and stirred for 1 hour. The RM was diluted with DCM, washed with water, aqueous NaHCO (2 × 20 mL), washed again with water and brine, dried over NaSO, filtered, and concentrated. The crude product was purified by reverse-phase preparative HPLC (RP-HPLC acidic 7: 30–40% B in 2 min, 40–50% B in 9 min) to give the title compound as a light brown solid. LC-MS: Rt=0.96 min; MS m / z [M+H] + 688.2, m / z [MH] - 686.3;UPLC-MS1 LC-MS: Rt=4.81 min; MS m / z [M+H] + 688.2, m / z [MH] - 686.3;UPLC-MS2 1 H NMR(400MHz,DMSO-d6)δ 10.35(s,boad,1H),10.06(s,br,1H),8.05(m,1H),7.77(d,J=13.0Hz,1H),7.66(d,J=8.2Hz,1H),7.28(m,2H),5.18(s,2H),4.53(m ,1H),3.66(m,4H),3.46(m,3H),3.38(m,4H),3.20(m,1H),2.92(m,3H),2.76(m,1H),2.58(m,1H),2.33(s,3H),1.15(t,J=7.2Hz,3H)

[0435] Example 6: 2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-5-ethyl-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide [ka] 2-(5-Ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (Intermediate D) (220 mg, 388 μmol) was suspended in DMF (5 mL). 4-Chloro-3-hydroxypicolinic acid (101 mg, 582 μmol), DIPEA (339 μL, 1.94 mmol), HOBt (105 mg, 777 μmol), and EDC·HCl (149 mg, 777 μmol) were added to the RM and stirred at room temperature for 12 h. Water was added to the RM, and the precipitate was filtered off. The resulting solid was purified by reverse-phase preparative HPLC (RP-HPLC acidic 5: 20-30% B in 2 min, 30-60% B in 8 min) to give the title compound. LC-MS: Rt=1.07 min; MS m / z [M+H] + 722.3 / 724.3, m / z [MH] - 720.3 / 722.3; UPLC-MS 1 LC-MS: Rt=5.32 min; MS m / z [M+H] + 722.2 / 724.2 m / z[MH] - 720.2 / 722.2; UPLC-MS 2 1H NMR(400MHz,DMSO-d6)δ 10.82(s,br,1H),10.05(s,1H),8.06(d,J=5.1Hz,1H),7.77(d,J=12.8Hz,1H),7.67(d,J=8.1Hz,1H),7.55(d,J=4.8Hz,1H),5.18(s,2H),4 .53(m,1H),3.65(m,4H),3.53(m,3H),3.38(m,4H),3.24(m,1H),2.93(m,3H),2.78(m,1H),2.60(m,1H),2.33(s,3H),1.16(t,J=7.1Hz,3H)

[0436] Example 7: N-(2-chloro-5-fluoro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide [ka] N-(2-chloro-5-fluoro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide HCl (Intermediate E) (300 mg, 481 μmol) and DIPEA (420 μL, 2.41 mmol) were dissolved in DCM (15 mL), and then 3-hydroxypicolinoyl chloride (Intermediate CV) (152 mg, 962 μmol) was added at 0° C. and stirred for 2 h. 3-hydroxypicolinoyl chloride (Intermediate CV) (152 mg, 962 μmol) was added again, and stirring was continued for 14 h. The RM was diluted with DCM, washed with water, aqueous NaHCO (2 × 20 mL), washed again with water and brine, dried over NaSO, filtered, and concentrated. The crude product was purified by column chromatography (silica gel column: 12 g silica, eluent DCM:MeOH 100:0 to 99:1). The product-containing fractions were concentrated and then further purified by reverse-phase preparative HPLC (RP-HPLC acidic 7: 40–50% B in 2 min, 50–60% B in 8 min) to afford the title compound as an off-white solid. LC-MS: Rt = 1.01 min; MS m / z [M+H] + 708.4 / 710.4, m / z [MH] - 706.4 / 708.4; UPLC-MS 1 LC-MS: Rt = 5.11 min; MS m / z [M+H] + 708.2 / 710.1 m / z[MH] - 706.2 / 708.2; UPLC-MS 2 1 H NMR(400MHz,DMSO-d6)δ 10.44(s,1H),10.37(s,1H),8.07(m,2H),8.01(d,J=7.3Hz,1H),7.28(m,2H),5.25(s,2H),4.53(m,1H),3.65 (m,4H),3.46(m,3H),3.37(m,4H),3.22(m,1H),2.93(m,3H),2.76(m,1H),2.58(m,1H),1.15(t,J=7.4Hz,3H)

[0437] Example 8: 2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide [ka] To a stirred solution of 3-hydroxypicolinic acid (143 mg, 1.03 mmol) in DMF (2.5 mL) was added EDC·HCl (197 mg, 1.03 mmol) and HOBt (13.0 mg, 1.03 mmol) at room temperature. In a separate flask, 2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide·HCl (Intermediate F) (300 mg, 513 μmol) in DMF (2.5 mL) was mixed with DIPEA (537 μL, 3.08 mmol) at room temperature. This solution was added to the first RM at room temperature and stirred for 12 h. This RM was concentrated, water was added, and the mixture was extracted with EtOAc. The organic layer was washed with aqueous NaHCO (twice), dried over anhydrous NaSO, and concentrated under reduced pressure. The crude product was purified by reverse-phase preparative HPLC (RP-HPLC Neutral 4: 30-40% B in 2 min, 40-50% B in 8 min) to give the title compound. LC-MS: Rt=0.92 min; MS m / z [M+H] + 670.4, m / z [MH] - 668.3;UPLC-MS 1 LC-MS: Rt=4.64 min; MS m / z [M+H] + 670.3, m / z [MH] - 668.3;UPLC-MS 2 1H NMR(400MHz,DMSO-d6)δ 10.40(s,1H,br),9.98(s,1H),8.05(m,1H),7.70(d,J=8.4Hz,1H),7.62(s,br,1H),7.53(d,J=8.3Hz,1H),7.28(m,2H),5.14(s,2H),4.5 3(m,1H),3.66(m,4H),3.46(m,3H),3.39(m,4H),3.20(m,1H),2.95(m,3H),2.76(m,1H),2.58(m,1H),2.33(s,3H),1.16(t,J=7.1Hz,3H)

[0438] Example 9: 2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-5-ethyl-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide [ka] 4-Chloro-3-hydroxypicolinic acid (120 mg, 692 μmol) was dissolved in DMF (3 mL). 2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide·HCl (Intermediate F) (270 mg, 462 μmol), EDC·HCl (177 mg, 923 μmol), DIPEA (403 μL, 2.31 mmol), and HOBt (125 mg, 923 μmol) were added at 0 °C and stirred at room temperature for 14 h. The RM was diluted with water, extracted with 5% MeOH in DCM, and washed with saturated aqueous NaHCO3 and brine. The combined organic layers were dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by reverse-phase preparative HPLC (RP-HPLC acidic 7: 30-40% B in 2 min, 40-60% B in 8 min) to give the title compound. LC-MS: Rt=1.02 min; MS m / z [M+H] + 704.2 / 706.2, m / z [MH] -702.2 / 704.2; UPLC-MS 1 LC-MS: Rt = 5.05 min; MS m / z [M+H] + 704.2 / 706.2, m / z [MH] - 702.3 / 704.2; UPLC-MS 2 1 H NMR(400MHz,DMSO-d6)δ 10.85(s,br,1H),9.99(s,1H),8.03(d,J=5.1Hz,1H),7.70(d,J=8.4Hz,1H),7.62(s,br,1H),7.53(m,2H),5.14(s,2H),4.53(m,1 H),3.66(m,4H),3.53(m,3H),3.39(m,4H),3.24(m,1H),2.95(m,3H),2.77(m,1H),2.60(m,1H),2.34(s,3H),1.17(t,J=7.1Hz,3H)

[0439] Example 10: N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide [ka] N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (Intermediate G) (900 mg, 1.58 mmol) was suspended in DMF (5 mL). 3-Hydroxypicolinate perfluorophenyl (Intermediate CT) (964 mg, 3.16 mmol) and EtN (438 μL, 3.16 mmol) were added, and the RM was stirred at 70 °C for 3 h. The RM was concentrated under reduced pressure. The crude product was purified by column chromatography (silica gel column: 24 g silica, eluent DCM:MeOH 100:0 to 99:1) and recrystallized from isopropanol to give the title compound. LC-MS: Rt=0.85 min; MS m / z [M+H] + 691.4 / 693.4, m / z [MH] - 689.5 / 691.5; UPLC-MS 1 1 H NMR(400MHz,DMSO-d6)δ 10.52(s,1H),10.36(s,1H),8.54(d,J=8.3Hz,1H),8.06(m,1H),7.96(d,J=8.5Hz,1H),7.28(m,2H),5.25(s,2H), 4.53(m,1H),3.65(m,4H),3.46(m,3H),3.37(m,4H),3.20(m,1H),2.94(m,3H),2.76(m,1H),2.58(m,1H),1.15(t,J =7.1Hz,3H)

[0440] Example 11: 2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamide [ka] To a stirred solution of 3-hydroxypicolinic acid (101 mg, 724 μmol), EDC·HCl (139 mg, 724 μmol), and HOBt (98.0 mg, 724 μmol) in DMF (3 mL) was added 2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamide (Intermediate H) (200 mg, 362 μmol) followed by DIPEA (379 μL, 2.17 mmol) at room temperature. The RM was stirred at room temperature for 16 h. The reaction was concentrated under reduced pressure, and water was added. The resulting brown solid was filtered and dried under vacuum. The crude product was purified by reverse-phase preparative HPLC (RP-HPLC neutral 1: 25-35% B in 2 min, 35-50% B in 9 min) to give the title compound. LC-MS: Rt=0.95 min; MS m / z [M+H] + 674.6, m / z [MH] - 672.4; UPLC-MS 1 LC-MS: Rt = 4.68 min; MS m / z [M+H] + 674.2, m / z [MH] - 672.3;UPLC-MS 2 1 H NMR(400MHz,DMSO-d6)δ 10.58(s,br,2H),8.21(m,1H),8.04(m,1H),7.79(d,J=10.9Hz,1H),7.57(d,J=8.7Hz,1H),7.27(m,2H),5.19(s,2H),4.53 (m,1H),3.65(m,4H),3.46(m,3H),3.37(m,4H),3.20(m,1H),2.92(m,3H),2.76(m,1H),2.57(m,1H),1.14(t,J=7.2Hz,3H)

[0441] Example 12: N-(4-chloro-2-methyl-5-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-morpholino-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide [ka] 3-Hydroxypicolinic acid (84.0 mg, 605 μmol) was dissolved in DMF (8 mL). N-(4-chloro-2-methyl-5-(trifluoromethyl)phenyl)-2-(5-ethyl-2-morpholino-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide·HCl (Intermediate I) (250 mg, 404 μmol), EDC·HCl (116 mg, 605 μmol), DIPEA (423 μL, 2.42 mmol), and HOBt (82.0 mg, 605 μmol) were added at 0 °C and stirred at room temperature for 16 h. A portion of the RM was concentrated, diluted with water, extracted with EtOAc, washed with saturated aqueous NaHCO and brine, and the combined organic layers were dried over NaSO, filtered, and concentrated under reduced pressure. The crude product was sonicated in ACN and MeOH (1:1) (5 mL) and then filtered. The resulting solid was washed with EtOAc and pentane and dried to give the title compound as an off-white solid. LC-MS: Rt = 1.00 min; MS m / z [M+H] + 704.6 / 706.5, m / z [MH] - 702.4 / 704.4; UPLC-MS 1 LC-MS: Rt=4.94 min; MS m / z [M+H] + 704.2 / 706.2, m / z [MH] - 702.3 / 704.3; UPLC-MS 2 1 H NMR(400MHz,DMSO-d6)δ 10.41(s,br,1H),10.07(s,br,1H),8.04(m,1H),7.94(s,1H),7.66(s,1H),7.27(m,2H),5.12(s,2H),4.53(m,1H),3.65( m,4H),3.46(m,3H),3.39(m,4H),3.20(m,1H),2.94(m,3H),2.75(m,1H),2.58(m,1H),2.33(s,3H),1.16(t,J=7.4Hz,3H)

[0442] Example 13: rac-N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide [ka] Rac-N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide·HCl (Intermediate K) (360 mg, 566 μmol), EDC·HCl (163 mg, 850 μmol), 3-hydroxypicolinic acid (118 mg, 850 μmol), and HOBt (115 mg, 850 μmol) were dissolved in DMF (5 mL). DIPEA (594 μL, 3.40 mmol) was added at 0° C. and the mixture was stirred at room temperature for 16 hours. The RM was diluted with water, extracted with EtOAc, washed with saturated aqueous NaHCO3 and brine, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was first purified by column chromatography (2 x silica gel columns: 12 g silica, eluent DCM:MeOH 100:0 to 95:5) to give the title compound as an off-white solid. LC-MS: Rt=1.08 min; MS m / z [M+H] + 720.1 / 722.0 m / z[MH] - 718.3 / 720.2; UPLC-MS 1 LC-MS: Rt=5.42 min; MS m / z [M+H] + 720.1 / 722.1, m / z [MH] - 718.3 / 720.3; UPLC-MS 2 1H NMR(400MHz,DMSO-d6)δ 10.38(s,1H),10.08(s,1H),8.06(m,1H),7.92(s,1H),7.76(s,1H),7.2 9(m,2H),5.15(s,2H),4.75(d,br,J,47.5Hz,1H),4.53(m,1H),3.73(m, 1H),3.58(m,2H),3.44(m,3H),3.23(m,1H),2.93(m,3H),2.76(m,1H),2 .58(m,1H),2.35(s,3H),1.81(m,4H),1.52(m,1H),1.16(t,J=7.2Hz,3H)

[0443] Example 14: rac-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide [ka] Rac-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide·HCl (Intermediate L) (300 mg, 499 μmol) was dissolved in DMF (4 mL). 3-Hydroxypicolinic acid (174 mg, 1.25 mmol), EDC·HCl (239 mg, 1.25 mmol), HOBt (169 mg, 1.25 mmol), and DIPEA (436 μL, 2.50 mmol) were added at 0 °C, and the RM was stirred at room temperature for 24 h. The RM was diluted with water, extracted with 5% MeOH in DCM, washed with saturated aqueous NaHCO3, brine, and the organic layer was dried and concentrated under reduced pressure. The crude product was purified by reverse-phase preparative HPLC (RP-HPLC acidic 6: 40-50% B in 2 min, 50-70% B in 8 min) to give the title compound. LC-MS: Rt = 1.01 min; MS m / z [M+H] +686.2, m / z [MH] - 684.3; UPLC-MS 1 LC-MS: Rt = 5.05 min; MS m / z [M+H] + 686.2, m / z [MH] - 684.3; UPLC-MS 2 1 H NMR(400MHz,DMSO-d6)δ 10.32(s,br,1H),9.99(s,1H),8.06(m,1H),7.69(d,J=8.8Hz,1H),7.62(s,br,1H ),7.53(d,J=8.2Hz,1H),7.28(m,2H),5.13(s,2H),4.75(d,br,J=48.2Hz,1H),4. 53(m,1H),3.72(m,1H),3.58(m,2H),3.43(m,3H),3.21(m,2H),2.93(m,3H),2.75 (m,1H),2.58(m,1H),2.34(s,3H),1.82(m,3H),1.53(m,1H),1.16(t,J=7.4Hz,3H)

[0444] Example 14a: (R)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide and Example 14b: (S)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide [ka] Chiral separation of rac-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide: Preparative chiral HPLC (Instrument: Agilent 1200 Series, equipped with a single quadrupole mass spectrometer: LUX CELLULOSE-4, 250 mm × 21.2 mm; Eluent: A = hexane, B = 0.1% HCOOH (in MeOH:EtOH 1:1); Flow rate: 15.0 mL / min; Detection: 210 nm; Injection volume: 0.9 mL; Gradient: isocratic 70(A):30(B)).

[0445] Example 14a: The first eluting isomer was stirred in Et2O (20 mL), filtered, and the resulting solid was dried under vacuum to give the title compound. Chiral HPLC (C-HPLC 3): Rt = 6.17 min LC-MS: Rt = 1.01 min; MS m / z [M+H] + 686.2, m / z [MH] - 684.3; UPLC-MS 1 LC-MS: Rt = 5.05 min; MS m / z [M+H] + 686.2, m / z [MH] - 684.3; UPLC-MS 2 1 H NMR(400MHz,DMSO-d6)δ 10.36(s,br,1H),10.00(s,1H),8.05(m,1H),7.69(d,J=8.5Hz,1H),7.62(s,br,1H),7.53(d,J=8.4Hz,1H),7.28(m,2H),5.13(s,2H),4.85 - 4.65(d,br,J=47.5Hz,1H),4.53(m,1H),3.72(m,1H),3.65-3.35(m,6H),3.20(m,1H),2.94(m ,3H),2.75(m,1H),2.57(m,1H),2.34(s,3H),1.81(m,3H),1.52(m,1H),1.16(t,J=7.3Hz,3H)

[0446] Example 14b: The second eluting isomer was purified by reverse-phase preparative HPLC (RP-HPLC acidic 5: 20-30% B in 2 min, 30-60% B in 8 min) to give the title compound. Chiral HPLC (C-HPLC 4): Rt = 8.20 min LC-MS: Rt=1.02 min; MS m / z [M+H] + 686.4, m / z [MH] - 684.4;UPLC-MS1 LC-MS: Rt=5.12 min; MS m / z [M+H] + 686.3, m / z [MH] - 684.3; UPLC-MS 2 1 H NMR(400MHz,DMSO-d6)δ 10.43(s,br,1H),10.00(s,1H),8.05(m,1H),7.69(d,J=8.5Hz,1H),7.62(s,br,1H),7.53(d,J=8.5Hz,1H),7.28(m,2H),5.13(s,2H),4.85 - 4.65(d,br,J=47.7Hz,1H),4.53(m,1H),3.74(m,1H),3.65-3.35(m,6H),3.21(m,1H),2.94(m ,3H),2.75(m,1H),2.58(m,1H),2.34(s,3H),1.78(m,3H),1.54(m,1H),1.16(t,J=7.4Hz,3H)

[0447] Example 15: rac-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide [ka] Rac-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (Intermediate M) (160 mg, 274 μmol) was suspended in DMF (2 mL), and 3-hydroxypicolinic acid (95.0 mg, 684 μmol), DIPEA (239 μL, 1.37 mmol), HOBt (92.0 mg, 684 μmol), and EDC·HCl (131 mg, 684 μmol) were added to RM and stirred at room temperature for 12 h. Water was added to the RM, the precipitate was filtered off, and the resulting crude product was purified by reverse-phase preparative HPLC (RP-HPLC neutral 2: 30-40% B in 2 min, 40-75% B in 9 min) to give the title compound. LC-MS: Rt=1.06 min; MS m / z [M+H] + 706.3 / 708.2, m / z [MH] - 704.3 / 706.4; UPLC-MS 1 LC-MS: Rt=5.34 min; MS m / z [M+H] + 706.2 / 708.1, m / z [MH] - 704.3 / 706.3; UPLC-MS 2 1 H NMR(400MHz,DMSO-d6)δ 10.36(2s,2H),8.04(m,2H),7.96(s,br,1H),7.72(d,J=8.7Hz,1H),7.28(m,2H),5.20(s,2H),4.85 - 4.5(d,br,J=48.6Hz,1H),4.53(m,1H),3.72(m,1H),3.65 - 3.35(m,5H),3.23(m,2H),2.94(m,3H),2.75(m,1H),2.57(m,1H),1.77(m,3H),1.52(m,1H),1.15(t,J=7.2Hz,3H)

[0448] Example 15a: ((R)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide) or ((S)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide) and and Example 15b: ((R)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide), or ((S)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide). [ka] Chiral separation of rac-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide: Preparative chiral HPLC (Instrument: SEPIATEC SFC100; Column: OVEN3 Chiralpak IB-N 250×30mm 5μm; Eluent: A: 28% [MeOH+0.1%NH3] B: 72% scCO2; Flow rate: 90.0 mL / min; Detection: 236 nm; Injection volume: 0.30 mL; Gradient: Isodic concentration A: 28%, B: 72% scCO2

[0449] Example 15a: The first eluting stereoisomer was concentrated under reduced pressure at 35° C. to give a white solid. Chiral HPLC (C-HPLC 7): Rt = 3.33 min, 99% ee LC-MS: Rt=1.05 min; MS m / z [M+H] + 706.3 / 708.3, m / z [MH] - 704.3 / 706.3; UPLC-MS 3 1 H NMR(600MHz,DMSO-d6)δ 10.35(2s,2H),8.05(m,1H),8.03(d,J=8.8Hz,1H),7.96(d,J=2.1Hz,1H),7.71(dd,J=2.2Hz,8.7H z,1H),7.28(m,2H),5.20(s,2H),4.80-4.65(d,br,J=46.9Hz,1H),4.53(m,1H),3.72(m,1H),3.65 - 3.35(m,5H),3.29(m,1H),3.19(m,1H),2.93(m,3H),2.75(m,1H),2.58(m,1H),1.95 - 1.7(m,3H),1.52(m,1H),1.15(t,J=7.3Hz,3H)

[0450] Example 15b: The second eluting isomer was concentrated under reduced pressure at 35° C. to give a beige solid. Chiral HPLC (C-HPLC7): Rt = 3.89 min, 94% ee LC-MS: Rt=1.05 min; MS m / z [M+H] + 706.3 / 708.3, m / z [MH] - 704.3 / 706.2; UPLC-MS 3 1H NMR(600MHz,DMSO-d6)δ 10.35(2s,2H),8.05(m,2H),7.96(d,J=2.1Hz,1H),7.71(dd,J=2.1Hz,8.8Hz,1H),7.2 8(m,2H),5.20(s,2H),4.80-4.70(d,br,J=47.8Hz,1H),4.53(m,1H),3.73(m,1H),3.65 - 3.35(m,5H),3.28(m,1H),3.20(m,1H),2.93(m,3H),2.75(m,1H),2.58(m,1H),1.95 - 1.7(m,3H),1.52(m,1H),1.15(t,J=7.3Hz,3H)

[0451] Example 16: rac-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide [ka] Rac-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide·HCl (Intermediate N) (100 mg, 161 μmol) and DIPEA (140 μL, 803 μmol) were dissolved in DCM (5 mL), and then 3-hydroxypicolinoyl chloride (Intermediate CV) (50.6 mg, 321 μmol) was added at 0 °C and stirred for 2 h. The RM was diluted with DCM and washed with water and saturated aqueous NaHCO (2 × 20 mL), followed by water and brine. The organic layer was dried over NaSO, filtered, and concentrated under reduced pressure. The crude product was purified by reverse-phase preparative HPLC (RP-HPLC acidic 7: 30-40% B in 2 min, 40-50% B in 8 min) to give the title compound as a light brown solid. LC-MS: Rt=0.94 min; MS m / z [M+H]+ 707.6 / 709.6, m / z [MH] - 705.4 / 707.4; UPLC-MS 1 LC-MS: Rt = 4.59 min; MS m / z [M+H] + 707.2 / 709.2, m / z [MH] - 705.3 / 707.2; UPLC-MS 2 1 H NMR(400MHz,DMSO-d6)δ 10.54(s,1H),10.37(s,1H),8.54(d,J=8.3Hz,1H),8.06(m,1H),7.96(d,J=8.0Hz,1H),7.28(m,2H),5.25(s,2H),4.85 - 4.65(d,br,J=47.2Hz,1H),4.53(m,1H),3.73(m,1H),3.60(m,1H),3.46(m,4H),3.23(m,2 H),2.94(m,3H),2.75(m,1H),2.57(m,1H),1.79(m,3H),1.52(m,1H),1.15(t,J=7.3Hz,3H)

[0452] Example 16a: (R)—N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide and Example 16b: (S)—N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide [ka] Chiral separation of rac-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-2-(3-fluoropiperidin-1-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide: Preparative chiral HPLC (Instrument: Agilent 1200 Series equipped with a single quadrupole mass spectrometer; Column: CELLULOSE-4, 250 mm × 21.2 mm; Eluent: A = hexane, B = 0.1% HCOOH (in MeOH:EtOH 1:1); Flow rate: 18.0 mL / min; Detection: 210 nm; Injection volume: 0.9 mL; Gradient: isocratic 70(A):30(B)). The separated and concentrated chiral isomers were removed, washed with n-hexane, decanted, dried, and analyzed.

[0453] Example 16a: First eluting stereoisomer, off-white solid. Chiral HPLC (C-HPLC5): Rt = 6.189 min LC-MS: Rt=0.93 min; MS m / z [M+H] + 707.1 / 709.1, m / z [MH] - 705.3 / 705.2; UPLC-MS 1 LC-MS: Rt = 4.60 min; MS m / z [M+H] + 707.1 / 709.0, m / z [MH] - 705.3 / 705.2; UPLC-MS 2 1 H NMR(400MHz,DMSO-d6)δ 10.63,(s,br,2H),8.53(d,J=7.6Hz,1H),8.05(m,1H),7.89(d,J=7.6Hz,1H),7.28(m,2H),5.19(s,2H),4.85 -4.65(d,br,J=48.0Hz,1H),4.53(m,1H),3.71(m,1H),3.65 - 3.15(m,7H),2.93(m,3H),2.75(m,1H),2.58(m,1H),1.81(m,3H),1.52(m,1H),1.15(t,J=7.3Hz,3H)

[0454] Example 16b: Second eluting stereoisomer, off-white solid. Chiral HPLC (C-HPLC6): Rt = 7.575 min LC-MS: Rt=0.93 min; MS m / z [M+H] + 707.1 / 709.0, m / z [MH] - 705.3 / 705.2; UPLC-MS 1 LC-MS: Rt = 4.60 min; MS m / z [M+H] + 707.1 / 709.0, m / z [MH] - 705.3 / 705.2; UPLC-MS 2 1 H NMR(400MHz,DMSO-d6)δ 10.43,(s,broad,2H),8.54(d,J=8.2Hz,1H),8.05(m,1H),7.94(d,J=8.2Hz,1H),7.28(m,2H),5.23(s,2H),4.85 -4.65(d,br,J=48.3Hz,1H),4.53(m,1H),3.71(m,1H),3.60(m,1H),3.46(m,4H),3.21(m,2 H),2.93(m,3H),2.75(m,1H),2.58(m,1H),1.81(m,3H),1.52(m,1H),1.15(t,J=7.3Hz,3H)

[0455] Example 17: 2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-2-(pyrrolidin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide [ka] 3-Hydroxypicolinic acid (183 mg, 1.32 mmol) was dissolved in DMF (10 mL). Then, 2-(5-ethyl-7-oxo-6-(piperazin-1-yl)-2-(pyrrolidin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide·HCl (Intermediate P) (500 mg, 879 μmol), EDC·HCl (337 mg, 1.76 mmol), DIPEA (767 μL, 4.39 mmol), and HOBt (237 mg, 1.76 mmol) were added at 0° C. and stirred at room temperature for 16 h. The RM was diluted with water, extracted with 5% MeOH in DCM, washed with saturated aqueous NaHCO and brine, and the combined organic layers were dried over NaSO, filtered, and concentrated under reduced pressure. The crude product was purified twice by column chromatography (2× silica gel columns: 12 g silica, eluent DCM:MeOH 100:0 to 98:2). The resulting solid was stirred with 5% ACN and MeOH in EtO for 30 min, then sonicated for 10 min, filtered off, washed with n-pentane, and dried to give the title compound as an off-white solid. LC-MS: Rt=1.04 min; MS m / z [M+H] + 654.6, m / z [MH] - 652.4; UPLC-MS 1 LC-MS: Rt = 5.08 min; MS m / z [M+H] + 654.3, m / z [MH] - 652.3;UPLC-MS 2 1 H NMR(400MHz,DMSO-d6)δ 10.37(s,1H),9.99(s,1H),8.06(m,1H),7.71(d,J=8.4Hz,1H),7.62(m,1H),7.53(d,J=8.4Hz,1H),7.28(m,2H),5.14(s,2H),4.54(m ,1H),3.49(m,3H),3.37(m,4H),3.20(m,1H),2.93(m,3H),2.75(m,1H),2.58(m,1H),2.35(s,3H),1.90(m,4H),1.16(t,J=7.1Hz,3H)

[0456] Example 18: N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-2-(pyrrolidin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide [ka] To a stirred solution of 3-hydroxypicolinic acid (166 mg, 1.19 mmol), EDC·HCl (228 mg, 1.19 mmol), and HOBt (161 mg, 1.19 mmol) in DMF (3 mL) was added N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-7-oxo-6-(piperazin-1-yl)-2-(pyrrolidin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (Intermediate Q) (330 mg, 596 μmol) and DIPEA (624 μL, 3.57 mmol), and the RM was stirred at room temperature for 16 h. The RM was concentrated under reduced pressure, and water was added. The resulting brown solid was filtered off and dried under vacuum. The crude product was purified by reverse-phase preparative HPLC (RP-HPLC acidic 4: 35-40% B in 2 min, 40-45% B in 10 min) to give the title compound. LC-MS: Rt=0.94 min; MS m / z [M+H] + 675.3 / 677.3, m / z [MH] - 673.3 / 675.3; UPLC-MS 1 LC-MS: Rt = 4.68 min; MS m / z [M+H] + 675.2 / 677.2, m / z [MH] - 673.2 / 675.2; UPLC-MS 2 1H NMR(400MHz,DMSO-d6)δ 10.54(s,br,1H),10.38(s,br,1H),8.55(d,J=8.4Hz,1H),8.06(m,1H),7.95(d,J=8.4Hz,1H),7.28(m,2H),5.25(s,2H) ),4.53(m,1H),3.46(m,3H),3.35(m,4H),3.19(m,1H),2.91(m,3H),2.75(m,1H),2.57(m,1H),1.89(m,4H),1.15(t,3H)

[0457] Example 19: N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-2-(pyrrolidin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide [ka] To a stirred solution of N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)-2-(5-ethyl-7-oxo-6-(piperazin-1-yl)-2-(pyrrolidin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide·HCl (Intermediate R) (150 mg, 249 μmol) in DCM (2 mL) was added 3-hydroxypicolinoyl chloride (Intermediate CV) (43.0 mg, 273 μmol) at 0 °C, followed by the dropwise addition of DIPEA (217 μL, 1.24 mmol). The RM was stirred at room temperature for 45 min. 3-Hydroxypicolinoyl chloride (Intermediate CV) (43.0 mg, 273 μmol) was added, and the RM was stirred at room temperature for 12 h. 3-Hydroxypicolinoyl chloride (Intermediate CV) (43.0 mg, 273 μmol) was added, and the RM was stirred at RT for 16 h. The RM was concentrated under reduced pressure. The crude product was purified by reverse-phase preparative HPLC (RP-HPLC Neutral 3: 25-35% B in 2 min, 35-60% B in 8 min) to give the title compound. LC-MS: Rt = 1.11 min; MS m / z [M+H] +688.7 / 690.6, m / z [MH] - 686.4 / 688.3; UPLC-MS 1 LC-MS: Rt=5.52 min; MS m / z [M+H] + 688.2 / 690.2, m / z [MH] - 686.3 / 688.3; UPLC-MS 2 1 H NMR(400MHz,DMSO-d6)δ 10.37(s,br,1H),10.08(s,1H),8.06(m,1H),7.95(s,1H),7.75(s,1H),7.28(m,2H),5.17(s,2H),4.53(m,1H),3.46(m ,3H),3.36(m,4H),3.20(m,1H),2.93(m,3H),2.75(m,1H),2.58(m,1H),2.35(s,3H),1.90(m,4H),1.15(t,J=7.3Hz,3H)

[0458] Example 20: N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(4-((2,2-difluoroethyl)(methyl)amino)piperidin-1-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide [ka] To N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(4-((2,2-difluoroethyl)(methyl)amino)piperidin-1-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (Intermediate S) (310 mg, 309 μmol) in DCM (3 mL) was added 3-hydroxypicolinoyl chloride (Intermediate CV) (61.0 mg, 387 μmol) at 5° C. and the RM was stirred for 3 min. DIPEA (162 μL, 928 μmol) was added and the RM was warmed to room temperature and stirred for 1 h. 3-Hydroxypicolinoyl chloride (Intermediate CV) (61.0 mg, 387 μmol) and DIPEA (162 μL, 928 μmol) were added, and the RM was further stirred at room temperature for 1 h 40 min. The RM was partitioned between DCM (20 mL) and 5% aqueous NaHCO3 (20 mL). The organic layer was separated by filtration through a phase separator. The aqueous layer was extracted with DCM (20 mL). The organic layers were combined and evaporated in vacuo to give a brown gum. The crude product was purified by reverse-phase preparative HPLC (RP-HPLC acidic 1: 25 mL / min, 5–35% over 20 min, 35% plateau for 1 min). The product-containing fractions were combined and basified with 5% aqueous NaHCO3. Extraction with DCM (5 × 30 mL) was performed. The combined organic layers were filtered through a phase separator and evaporated in vacuo to give an off-white foam. Further purification by column chromatography (RediSep column: silica 4 g, eluent DCM:MeOH 100:0 to 94:6) gave a colorless solid which was recrystallized from MeOH / water to give the title compound as a colorless powder. LC-MS: Rt=0.73 min; MS m / z [M+H] + 782.4 / 784.3, m / z [MH] - 780.5 / 782.5; UPLC-MS 1 1H NMR (400 MHz, DMSO-d6) δ 10.52(s,1H),10.37(s,1H),8.54(d,J=8.2Hz,1H),8.07(m,1H),7.96(d,J= 8.6Hz,1H),7.29(m,2H),6.15-5.85(m,br,1H),5.25(s,2H),4.54(m,1H),4 .10(m,2H),3.45(m,3H),3.22(m,1H),2.94(m,3H),2.78(m,4H),2.59(m,1H) ),2.27(s,3H),2.01(m,1H),1.71(m,2H),1.38(m,3H),1.16(t,J=7.4Hz,3H)

[0459] Example 21: 2-(2-(6-oxa-3-azabicyclo[3.1.1]heptan-3-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide [ka] 3-Hydroxypicolinic acid (141 mg, 992 μmol) was dissolved in DCM (5.5 mL) at room temperature under argon. 1-Chloro-N,N,2-trimethylprop-1-en-1-amine (149 mg, 1.09 mmol) was added, and the RM was stirred at room temperature for 1.25 h. 2-(2-(6-oxa-3-azabicyclo[3.1.1]heptan-3-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (Intermediate U) (278 mg, 496 μmol) in DCM (2.5 mL) and DIPEA (260 μL, 1.49 mmol) was added to the brown suspension. The resulting brown solution was stirred at room temperature for 3.5 h. The RM was quenched with water (5 mL) and saturated aqueous NaHCO3 (5 mL). It was extracted four times with DCM (4 x 40 mL). The combined organic layers were washed twice with water, dried through a phase separator, and concentrated under reduced pressure. The residue was adsorbed onto Isolute and purified by column chromatography (RediSep column: 24 g silica, eluent DCM:MeOH 100:0 to 90:10). The product-containing fractions were combined and concentrated. The solid was subjected to SFC (SFC 5). The product-containing fractions were combined and concentrated to give the title compound as an off-beige solid. A portion of the solid was crystallized from MeOH (1.5 mL) and DCM (2 mL). The resulting solid was dried under high vacuum to give the title compound. LC-MS: Rt=0.97 min; MS m / z [M+H] + 682.4, m / z [MH] - 680.3;UPLC-MS 3 1H NMR(600MHz,DMSO-d6)δ 10.40(s,1H),10.01(s,1H),8.07(m,1H),7.72(d,J=8.1Hz,1H),7.63(s,br,1H),7.5 4(d,J=8.3Hz,1H),7.29(m,2H),5.17(s,2H),4.66(m,2H),4.54(m,1H),3.67(d,J=12H) z,2H),3.58(d,J=11.8Hz,2H),3.49(m,2H),3.40(m,1H),3.21(m,1H),3.13(m,1H),2 .96(m,3H),2.77(m,1H),2.59(m,1H),2.35(s,3H),1.89(m,1H),1.15(t,J=7.6Hz,3H)

[0460] Example 22: 2-(2-(6-oxa-3-azabicyclo[3.1.1]heptan-3-yl)-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-5-methyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamide [ka] 1-Chloro-N,N,2-trimethylprop-1-en-1-amine (141 mg, 1.06 mmol) was added to a solution of 3-hydroxypicolinic acid (134 mg, 961 μmol) in DCM (5 mL) under argon, and the RM was stirred at room temperature for 2 h. Then, 2-(2-(6-oxa-3-azabicyclo[3.1.1]heptan-3-yl)-5-methyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamide (Intermediate V) (441 mg, 481 μmol) dissolved in DCM (2.8 mL) was added, followed by DIPEA (420 μL, 2.40 mmol). The RM was stirred at room temperature for 2.2 h. The activated 0.5 equivalents of 3-hydroxypicolinic acid solution from above was added to the RM again, followed by DIPEA (77.0 μL, 441 μmol). The RM was stirred at room temperature for 2 h. The RM was quenched with water (6 mL), and saturated aqueous NaHCO3 (6 mL) was added. The mixture was extracted with DCM (4 × 40 mL). The organic layer was washed with saturated aqueous NaHCO3 and water, dried through a phase separator, and concentrated under reduced pressure. The crude product was adsorbed onto Isolute and purified by column chromatography (RediSep column: 24 g silica, eluent DCM:MeOH 100:0 to 90:10). The product-containing fractions were combined and concentrated under reduced pressure. The solid was further purified by reverse-phase preparative HPLC (RP-HPLC acidic 1: 15–85% B in 20 min, 1 min at 85% plateau). The product-containing fractions were combined and basified with a small amount of saturated aqueous NaHCO3. The ACN was removed under reduced pressure and the residue was extracted with DCM (3 x 40 mL). The combined organic layers were washed with water (10 mL), then dried over a phase separator and concentrated under reduced pressure to give the title compound as a beige solid. LC-MS: Rt=0.93 min; MS m / z [M+H] + 672.4, m / z [MH] - 670.4;UPLC-MS 3 1H NMR(600MHz,DMSO-d6)δ 10.69(s,br,1H),10.39(s,br,1H),8.22(t,J=8.1Hz,1H),8.06(t,J=3.1Hz,1H),7.8 0(dd,J=2.1Hz,10.9Hz,1H),7.56(dd,J=1.7Hz,8.4Hz,1H),7.29(m,2H),5.24(s,2H) ,4.65(m,2H),4.52(m,1H),3.66(m,2H),3.57(m,2H),3.46(m,2H),3.38(m,1H),3.22 (m,1H),3.12(m,1H),2.96(m,1H),2.75(m,1H),2.57(m,1H),2.47(s,3H),1.89(m,1H)

[0461] Example 23: 2-(2-((3R,4S)-3,4-difluoropyrrolidin-1-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide [ka] 3-Hydroxypicolinic acid (152 mg, 1.07 mmol) was dissolved in DCM (6 mL) at room temperature under argon. 1-Chloro-N,N,2-trimethylprop-1-en-1-amine (161 mg, 1.18 mmol) was added, and the RM was stirred at room temperature for 1.2 h. 2-(2-((3R,4S)-3,4-difluoropyrrolidin-1-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (Intermediate W) (430 mg, 537 μmol) in DCM (3.5 mL) and DIPEA (281 μL, 1.61 mmol) was added to the brown suspension. The resulting brown solution was stirred at room temperature for 1 h. The RM was quenched with water (10 mL), saturated aqueous NaHCO3 (5 mL), and extracted with DCM (4 x 40 mL). The combined organic layers were washed twice with water, dried through a phase separator, and concentrated under reduced pressure. The residue was adsorbed onto Isolute and purified by column chromatography (RediSep column: 40 g silica, eluent DCM:MeOH 100:0 to 90:10). The product-containing fractions were combined and concentrated under reduced pressure to give the title compound. A portion of the solid was crystallized from MeOH (1.5 mL) and DCM (2 mL). The resulting gray solid was suspended in Et2O, filtered, and then dried under high vacuum to give the title compound. LC-MS: Rt = 1.01 min; MS m / z [M+H] + 690.3, m / z [MH] - 688.3; UPLC-MS 3 1H NMR (600 MHz, DMSO-d6) δ 10.39(s,br,1H),10.01(s,1H),8.06(m,1H),7.72(d,J=8.3Hz,1H),7.64(s,b r,1H),7.54(d,J=8.3Hz,1H),7.29(m,2H),5.45(m,1H),5.36(m,1H),5.16(s,2 H),4.54(m,1H),3.81(m,2H),3.57(m,2H),3.47(m,2H),3.39(m,1H),3.21(m, 1H),2.95(m,3H),2.76(m,1H),2.58(m,1H),2.36(s,3H),1.16(t,J=7.5Hz,3H)

[0462] Example 24a: ((R)-2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)acetamine (S)-2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)acetamide) and and Example 24b: ((R)-2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)acetate amide) or ((S)-2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)acetamide) [ka] To a stirred solution of N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (Intermediate Y) (300 mg, 504 μmol), 4-chloro-3-hydroxypicolinic acid (140 mg, 807 μmol), HOBt (136 mg, 1.01 mmol), and EDC·HCl (193 mg, 1.01 mmol) in DCM (20 mL) was added pyridine (122 μL, 1.51 mmol) at 0 °C. The RM was stirred at room temperature for 16 h. The RM was quenched with NaHCO and extracted with DCM. The organic layer was dried over Na2SO4 and concentrated under reduced pressure, and the crude product was purified by column chromatography (silica gel column: silica 4 g, eluent DCM:MeOH 100:0 to 98:2). The residue was separated by preparative chiral HPLC (instrument: Agilent 1200 Series equipped with a single quadrupole mass spectrometer; column: LUX CELLULOSE-4, 250 mm × 21.1 mm, 5.0 μm; eluent: A = hexane, B = 0.1% HCOOH (in EtOH); flow rate: 15 mL / min, detection: 210 nm; injection volume: 0.9 mL; gradient: isocratic 50(A):50(B)).

[0463] Example 24a: The product-containing fractions were concentrated at 40° C., washed with n-pentane (5×10 mL), decanted, and dried to give the off-white title compound—first eluting stereoisomer. Chiral HPLC (C-HPLC 2): Rt = 10.764 min LC-MS: Rt=1.08 min; MS m / z [M+H] + 750.5 / 752.5, m / z [MH] - 748.4 / 750.4; UPLC-MS 1 LC-MS: Rt=5.29 min; MS m / z [M+H] + 750.2 / 752.2, m / z [MH]- 748.2 / 750.2; UPLC-MS 2 1 H NMR(400MHz,DMSO-d6)δ 10.68(s,br,2H),8.56(d,J=8.1Hz,1H),7.98(d,J=5.6Hz,1H),7.94(d,J=8 .1Hz,1H),7.50(d,J=5.1Hz,1H),6.72(m,1H),5.34(s,2H),4.53(m,1H),3. 52(m,4H),3.28(m,4H),2.98(m,3H),2.80(m,1H),2.63(m,1H),2.55(m,1H) ,2.46(m,1H),2.16(m,2H),1.95(m,1H),1.68(m,1H),1.17(t,J=7.3Hz,3H)

[0464] Example 24b: The product-containing fractions were concentrated at 40° C., washed with n-pentane (5×10 mL), decanted, and dried to give the off-white title compound—second eluting stereoisomer. Chiral HPLC (C-HPLC 2): Rt = 18,800 min LC-MS: Rt=1.08 min; MS m / z [M+H] + 750.1 / 752.1, m / z [MH] - 748.2 / 750.2; UPLC-MS 1 LC-MS: Rt = 5.30 min; MS m / z [M+H] + 750.1 / 752.1, m / z [MH] - 748.2 / 750.2; UPLC-MS 2 1H NMR (400 MHz, DMSO-d6) δ 10.83(s,br,1H),10.55(s,br,1H),8.56(d,J=8.2Hz,1H),8.06(d,J=5.3Hz,1H ),7.92(d,J=8.2Hz,1H),7.55(d,J=5.3Hz,1H),6.72(m,1H),5.35(s,2H),4.54( m,1H),3.54(m,4H),3.28(m,3H),3.25(m,1H),2.99(m,3H),2.81(m,1H),2.62(m ,1H),2.41(m,2H),2.16(m,2H),1.96(m,1H),1.66(m,1H),1.18(t,J=7.3Hz,3H)

[0465] Example 25a: ((R)—N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide) or ((S)—N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide) and and Example 25b: ((R)—N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide) or ((S)—N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide). [ka] N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide.HCl Intermediate Y) (120 mg, 190 μmol) and DIPEA (166 μL, 950 μmol) were dissolved in DCM (5 mL), and then 3-hydroxypicolinoyl chloride Intermediate CV) (59.9 mg, 380 μmol) was added at 0° C. and stirred for 2 hours. 3-Hydroxypicolinoyl chloride (Intermediate CV) (59.9 mg, 380 μmol) was added again, and the reaction was continued with stirring for 12 h. The RM was diluted with DCM, washed with water and NaHCO (2 × 20 mL), washed with water and brine, dried over NaSO, filtered, and concentrated. This was combined with the product from another experiment and purified by column chromatography (silica gel column: 4 g silica, eluent DCM:MeOH 100:0 to 99:1), and then further purified by reverse-phase preparative HPLC (RP-HPLC acidic 10: 40–50% B in 2 min, 50–60% B in 8 min) to give the title compound as an off-white solid. The racemate was purified by preparative chiral HPLC (instrument: Agilent 1200 Series equipped with a single quadrupole mass spectrometer; column: CELLULOSE-4, 250 mm × 21.2 mm; eluent: A = hexane, B = 0.1% HCOOH (MeOH:EtOH 1:1; flow rate: 20 mL / min; gradient: 210 nm; injection volume: 0.9 mL; gradient: isocratic 60(A):40(B)).

[0466] Example 25a: First eluting stereoisomer, off-white solid. Chiral HPLC (C-HPLC 1): Rt = 10.070 min LC-MS: Rt=0.98 min; MS m / z [M+H] + 716.5 / 718.6, m / z [MH] - 714.3 / 716.3; UPLC-MS 1 LC-MS: Rt=4.76 minutes; MS m / z[M+H] + 716.2 / 718.2,m / z[MH] - 714.2 / 716.2;UPLC-MS 2 1 H NMR(400MHz,DMSO-d6)δ 10.46(s,br,2H),8.56(d,J=8.5Hz,1H),8.05(m,1H),7.90(d,J=8.4Hz,1H),7.28(m,2H),6.72(m,1H),5.30(s,2H),4.54(m,1H),3.47(m,4H) ,3.27(s,3H),3.21(m,1H),2.96(m,3H),2.79(m,1H),2.59(m,3H),2.4 3(m,1H),2.14(m,1H),1.95(m,1H),1.67(m,1H),1.17(t,J=7.2Hz,3H)

[0467] Example 25b: The second dissolution stereoisotropic body, オフホワイト-colored solid. KIRAL HPLC (C-HPLC 1): Rt=16.023min LC-MS: Rt=0.96 min; MS m / z[M+H] + 716.3 / 718.3,m / z[MH] - 714.3 / 716.3;UPLC-MS 1 LC-MS: Rt=4.77 min; MS m / z[M+H] + 716.2 / 718.2,m / z[MH] - 714.2 / 716.2;UPLC-MS 2 1 H NMR(400MHz,DMSO-d6)δ 10.39(s,br,2H),8.56(d,J=8.0Hz,1H),8.06(m,1H),7.93(d,J=8.1Hz,1H),7.28(m,2H),6.72(m,1H),5.32(s,2H),4.54(m,1H),3.46(m,4H) ,3.27(s,3H),3.20(m,1H),2.96(m,3H),2.79(m,1H),2.59(m,3H),2.4 1(m,1H),2.14(m,1H),1.95(m,1H),1.68(m,1H),1.17(t,J=7.1Hz,3H)

[0468] Example 26: rac-2-(5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide [ka] 4-Chloro-3-hydroxypicolinic acid (118 mg, 851 μmol) was dissolved in DMF (5 mL), and then rac-2-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide·HCl (Intermediate Z) (260 mg, 425 μmol), EDC·HCl (163 mg, 851 μmol), DIPEA (372 μL, 2.13 mmol), and HOBt (115 mg, 851 μmol) were added at 0 °C, and the RM was stirred at room temperature for 14 h. The RM was diluted with water, extracted with 5% MeOH in DCM, washed with saturated aqueous NaHCO3 and brine, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by reverse-phase preparative HPLC (RP-HPLC acidic 5: 30-40% B in 2 min, 40-50% B in 8 min) to give the title compound. LC-MS: Rt = 0.90 min; MS m / z [M+H] + 696.3, m / z [MH] - 694.3; UPLC-MS 1 LC-MS: Rt=4.38 min; MS m / z [M+H] + 696.3, m / z [MH] - 694.4; UPLC-MS 2 1H NMR(400MHz,DMSO-d6)δ 10.26(m,2H),8.19(d,J=8.3Hz,1H),8.06(m,1H),7.72(d,J=8.2Hz,1H),7.28(m,2H),6.72(m,1H),5.27(s,2H),4.54(m,1H), 3.47(m,4H),3.28(m,4H),2.98(m,3H),2.80(m,1H),2.57(m,6H),2.15(m,1H),1.96(m,1H),1.68(m,1H),1.19(t,J=7.5Hz,3H)

[0469] Example 27: N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(5,6-dihydro-1,4-dioxin-2-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide [ka] N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(5,6-dihydro-1,4-dioxin-2-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (Intermediate AA) (492 mg, 692 μmol) was dissolved in DCM (7 mL) under argon at 0° C. 3-Hydroxypicolinoyl chloride (Intermediate CV) (163 mg, 1.04 mmol) was added to the suspension, followed by the slow addition of DIPEA (483 μL, 2.77 mmol), and the solution was stirred at room temperature for 1.5 hours. 3-Hydroxypicolinoyl chloride (Intermediate CV) (25.0 mg, 159 μmol) and DIPEA (320 μL, 1.84 mmol) were added again, and the RM was continued to stir at room temperature for 2.3 h. The reaction was quenched by adding water (5 mL) and saturated aqueous NaHCO3 (5 mL). It was then extracted four times with DCM (4 × 40 mL). The organic layer was washed with water (5 mL), saturated aqueous NaHCO3 (5 mL), and again with water (10 mL). The organic layer was dried over a phase separator and concentrated under reduced pressure. The residue was adsorbed onto Isolute and purified by column chromatography (RediSep column: silica 40 g Gold, eluent DCM:MeOH 100:0 to 90:10). The product-containing fractions were combined and concentrated under reduced pressure. The resulting solid was purified by reverse-phase preparative HPLC (RP-HPLC acidic 1: 15-85% B in 20 min, 85% plateau for 1 min). The product-containing fractions were combined and basified with saturated aqueous NaHCO3 (5 mL). The ACN was removed under reduced pressure. The aqueous layer was washed four times with DCM (4 × 35 mL). The combined organic layers were washed with water (10 mL), dried over a phase separator, and concentrated under reduced pressure to give the title compound as a white solid. LC-MS: Rt=0.87 min; MS m / z [M+H] + 690.3 / 692.3, m / z [MH] - 688.1 / 690.1; UPLC-MS 1

[0470] Example 28: N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide [ka]

[0471] Step 1: N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-6-(4-(5-methoxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide To a stirred solution of 5-methoxy-6-methylpyrimidine-4-carboxylic acid (Intermediate CW) (133 mg, 794 μmol) in DCM (9 mL) at 0 °C, EDC·HCl (203 mg, 1.06 mmol), pyridine (128 μL, 1.59 mmol), and HOBt (143 mg, 1.06 mmol) were added. The RM was stirred at 0 °C for 10 min, then N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (Intermediate AC) (300 mg, 529 μmol) was added and the RM was stirred at RT for 16 h. The RM was diluted with DCM, washed with saturated NaHCO3, aqueous solution, and washed with water, and the combined organic layers were dried over Na2SO4, concentrated, and dried. The crude product was purified by column chromatography (silica gel column: 12 g silica, eluent DCM:MeOH 100:0 to 99:1) to give the title compound. LC-MS: Rt=1.51 min; MS m / z [M+H] + 717.2 / 719.2; UPLC-MS 11

[0472] Step 2: N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide To a stirred solution of N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-6-(4-(5-methoxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (250 mg, 310 μmol) in DMF (3 mL) was added LiCl (132 mg, 3.10 mmol) and the RM was heated at 150° C. for 3 h. LiCl (132 mg, 3.10 mmol) was added again and the RM was stirred at 150° C. for 4 h. The RM was quenched with water, extracted with 10% MeOH in DCM (3 × 50 mL), dried over NaSO, filtered, and concentrated to dryness. The crude product was purified by column chromatography (silica gel column: 12 g silica, eluent DCM:MeOH 100:0 to 98:2). The residue was purified by reverse-phase preparative HPLC (RP-HPLC acidic 4: 15–25% B in 2 min, 25–55% B in 7 min), and the product-containing fractions were concentrated below 40 °C and dried to give the title compound as an off-white solid. LC-MS: Rt=0.97 min; MS m / z [M+H] + 703.2 / 705.2, m / z [MH] - 701.3 / 703.2; UPLC-MS 1 LC-MS: Rt=4.73 min; MS m / z [M+H] + 703.2 / 705.1, m / z [MH] - 701.3 / 703.2; UPLC-MS 2 1H NMR(400MHz,DMSO-d6)δ 10.52(s,br,1H),10.25(s,br,1H),8.57(m,2H),7.95(d,J=8.3Hz,1H),5.86(m,1H),5.35(s,2H),4.52(m,1H),4.10(m,2H),3 .49(m,3H),3.25(m,1H),2.99(m,3H),2.81(m,1H),2.64(m,1H),2.44(s,3H),2.17(m,2H),1.84(m,2H),1.18(t,J=7.6Hz,3H)

[0473] Example 29: N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide [ka] N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (Intermediate AC) (700 mg, 1.24 mmol) was suspended in DMF (12 mL), and perfluorophenyl 3-hydroxypicolinate (Intermediate CT) (754 mg, 2.47 mmol) and EtN (342 μL, 2.47 mmol) were added at room temperature, and the RM was stirred at 80° C. for 16 h. The RM was extracted three times with DCM. The combined organic layers were washed with brine, dried over NaSO, and concentrated under reduced pressure. The crude product was purified by column chromatography (silica gel column: 12 g silica, eluent hexane: EtOAc 100:0 to 60:40). The product-containing fractions were concentrated and dried under high vacuum to give the title compound. LC-MS: Rt=0.94 min; MS m / z [M+H] + 688.5 / 690.5, m / z [MH] -686.2 / 688.2; UPLC-MS 1 LC-MS: Rt=4.58 min; MS m / z [M+H] + 688.2 / 690.2, m / z [MH] - 686.2 / 688.2; UPLC-MS 2 1 H NMR(400MHz,DMSO-d6)δ 10.54(s,1H),10.38(s,1H),8.57(d,J=8.5Hz,1H),8.06(m,1H),7.95(d,J=8.5Hz,1H),7.28(m,2H),5.85(m,1H),5.35(s,2H),4.54( m,1H),4.10(m,2H),3.42(m,3H),3.21(m,1H),2.96(m,3H),2.79(m,1H),2.61(m,1H),2.16(m,2H),1.84(m,2H),1.17(t,J=7.3Hz,3H)

[0474] Example 30: 2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)acetamide [ka] To a stirred solution of 4-chloro-3-hydroxypicolinic acid (282 mg, 1.62 mmol) in DMF (5 mL) was added DIPEA (354 μL, 2.03 mmol) and PyAOP (635 mg, 1.22 mmol) at 0° C. After 10 min, N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (Intermediate AC) (460 mg, 811 μmol) was added to the RM at 0° C. The RM was stirred at room temperature for 16 h. Water was added, and the mixture was extracted with EtOAc. The organic layer was dried over NaSO and concentrated under reduced pressure. The crude product was purified by reverse-phase preparative HPLC (RP-HPLC acidic 5: 20-30% B in 2 min, 30-60% B in 8 min) to give the title compound. LC-MS: Rt=1.04 min; MS m / z [M+H] + 722.5 / 724.5 / 726.5, m / z [MH] - 720.3 / 722.2 / 724.2; UPLC-MS 1 LC-MS: Rt = 5.07 min; MS m / z [M+H] + 722.2 / 724.1 / 726.2, m / z [MH] - 720.2 / 722.2 / 724.2; UPLC-MS 2 1 H NMR(400MHz,DMSO-d6)δ 10.83(s,1H),10.55(s,1H),8.58(d,J=8.4Hz,1H),8.07(d,J=5.0Hz,1H),7.96(d,J=8.4Hz,1H),7.56(d,J=5.0Hz,1H),5.86(m,1H),5.36(s,2 H),4.54(m,1H),4.10(m,2H),3.54(m,3H),3.24(m,1H),2.98(m,3H),2. 81(m,1H),2.65(m,1H),2.16(m,2H),1.84(m,2H),1.18(t,J=7.1Hz,3H)

[0475] Example 31: 2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide [ka]

[0476] Step 1: 2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-6-(4-(5-methoxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide To a stirred solution of 2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide (Intermediate AD) (600 mg, 1.10 mmol), 5-methoxy-6-methylpyrimidine-4-carboxylic acid (Intermediate CW) (222 mg, 1.32 mmol), and HATU (626 mg, 1.65 mmol) in DMF (15 mL) was added DIPEA (288 μL, 1.65 mmol) at 0° C. The RM was stirred at room temperature for 16 h. The RM was concentrated under reduced pressure to provide the title compound. LC-MS: Rt=1.48 min; MS m / z [M+H] + 697.3; UPLC-MS 11

[0477] Step 2: 2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide To a stirred solution of 2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-6-(4-(5-methoxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide (250 mg, 255 μmol) in DMF (2 mL) was added LiCl (108 mg, 2.55 mmol) and the RM was stirred in a MW at 200° C. for 1 h. The RM was quenched with water and extracted with 10% MeOH in DCM (3×50 mL), dried over NaSO, filtered, and concentrated to dryness. The crude product was purified by column chromatography (silica gel column: 12 g silica, eluent DCM:MeOH 100:0 to 98:2). The residue after column chromatography was purified by reverse-phase preparative HPLC (RP-HPLC acidic 10: 15-25% B in 2 min, 15-60% B in 10 min). The product-containing fractions were concentrated to give 45 mg of a solid, which was combined with another batch, washed with 30% EtO in n-hexane, and dried to give the title compound as an off-white solid. LC-MS: Rt = 0.90 min; MS m / z [M+H] + 683.6, m / z [MH] - 681.4; UPLC-MS 1 LC-MS: Rt=4.34 min; MS m / z [M+H] + 683.3, m / z [MH] - 681.3;UPLC-MS 2 1H NMR(400MHz,DMSO-d6)δ 10.22(s,br,2H),8.57(s,1H),8.20(d,J=8.0Hz,1H),7.73(d,J=8.2Hz,1H),5.86(m,1H),5.27(s,2H),4.52(m,1H),4.10(m,2H),3.50 (m,3H),3.27(m,1H),2.99(m,3H),2.81(m,1H),2.65(m,1H),2.58(s,3H),2.44(s,3H),2.18(m,2H),1.85(m,2H),1.19(t,J=7.2Hz,3H)

[0478] Example 32: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide [ka] N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (Intermediate AE) (590 mg (80% pure), 833 μmol) was dissolved in DCM (10 mL) and 3-hydroxypicolinoyl chloride (Intermediate CV) (197 mg, 1.25 mmol) was added, followed by DIPEA (437 μL, 2.50 mmol). The RM was stirred at room temperature for 1.5 h. 3-Hydroxypicolinoyl chloride (Intermediate CV) (36.0 mg, 228 μmol) was added. The RM was stirred at room temperature for 1 h. DIPEA (1.00 mL, 5.73 mmol) was added. The RM was stirred at room temperature for 2 h. 3-Hydroxypicolinoyl chloride (Intermediate CV) (116 mg, 736 μmol) was added. The RM was stirred at room temperature for 1 h. Water (10 mL), saturated aqueous NaHCO3 (10 mL), and DCM (10 mL) were added. The aqueous layer was washed twice with DCM (2 × 10 mL). The combined organic layers were dried over a phase separator and concentrated under reduced pressure. The crude product was purified in five portions by reverse-phase preparative HPLC (5 × RP-HPLC acidic 1:5 to 100% B). All product-containing fractions with purity >95% were combined, basified with saturated aqueous NaHCO3, extracted twice with DCM (2 × 15 mL), dried over a phase separator, and concentrated under reduced pressure. The concentrated fractions were suspended in MeOH and sonicated for 1 min. It was then filtered, and the cake was washed with MeOH (500 μL) and dried under high vacuum to give the title compound. At Rt = 1.05 min, all impure product-containing fractions were combined, basified with saturated aqueous NaHCO3, extracted twice with DCM (2 × 15 mL), dried on a phase separator, and concentrated under reduced pressure. At Rt = 1.13 min, all impure product-containing fractions were combined, basified with saturated aqueous NaHCO3, extracted twice with DCM (2 × 15 mL), dried on a phase separator, and concentrated under reduced pressure. Both impure fractions were combined and suspended in MeOH (10 mL). It was then sonicated for 30 min and filtered. The cake was washed with a small amount of MeOH (10 mL) and dried under high vacuum to give the product in 90% purity.The cake was suspended in MeOH (10 mL) and ACN (10 mL) and stirred for 2 h at 40° C. It was filtered, the cake washed with MeOH (1 mL) and dried under HV to give the title compound. Both pure fractions were dissolved in DCM (10 mL) and EtOH (10 mL) and left at room temperature for 5 days. The solid was filtered off and washed with a small amount of EtO. The cake was dried under HV to give the title compound. LC-MS: Rt=1.08 min; MS m / z [M+H] + 687.2 / 689.2, m / z [MH] - 685.4 / 687.5; UPLC-MS 1 1 H NMR(400MHz,DMSO-d6)δ 10.38(s,1H),10.33(s,1H),8.07(m,2H),7.96(s,1H),7.71(d,J=8.6Hz,1H),7.29(m,2H),5.87(m,1H),5.31(s,2H),4.55(m,1H) ),4.10(m,2H),3.43(m,3H),3.23(m,1H),2.97(m,3H),2.80(m,1H),2.62(m,1H),2.17(m,2H),1.84(m,2H),1.18(t,J=7.6Hz,3H)

[0479] Example 33: N-(4-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide [ka] N-(4-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (Intermediate AG) (340 mg, 600 μmol) was suspended in DMF (10 mL), and perfluorophenyl 3-hydroxypicolinate (Intermediate CT) (366 mg, 1.20 mmol) and EtN (166 μL, 1.20 mmol) were added. The RM was stirred at 70 °C for 3 h. The crude product was purified by reverse-phase preparative HPLC (RP-HPLC acidic 3: 10–20% B in 2 min, 20–60% B in 10 min) to give the title compound. LC-MS: Rt=0.84 min; MS m / z [M+H] + 688.3 / 690.3, m / z [MH] - 686.3 / 688.3; UPLC-MS 1 LC-MS: Rt = 4.15 min; MS m / z [M+H] + 688.2 / 690.2, m / z [MH] - 686.2 / 688.2; UPLC-MS 2 1 H NMR(600MHz,DMSO-d6)δ 10.65(s,1H),10.36(s,1H),9.06(s,1H),8.22(s,1H),8.02(m,1H),7.25(m,2H),6.72(m,1H),5.29(s,2H),4.50(m,1H),4.21(m,2 H),3.76(m,2H),3.42(m,2H),3.35(m,1H),3.18(m,1H),2.93(m,3H),2.76(m,1H),2.58(m,1H),2.52(m,2H),1.15(t,J=7.6Hz,3H)

[0480] Example 34: 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide [ka] 2-(2-(3,6-Dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide (Intermediate AH) (450 mg, 823 μmol) was suspended in DMF (5 mL). Perfluorophenyl 3-hydroxypicolinate (Intermediate CT) (503 mg, 1.65 mmol) and EtN (228 μL, 1.65 mmol) were added, and the RM was stirred at 70° C. for 3 h. The RM was concentrated under reduced pressure. The crude product was purified by reverse-phase preparative HPLC (RP-HPLC acidic 11: 30-40% B in 2 min, 40-70% B in 10 min) to give the title compound. LC-MS: Rt=0.77 min; MS m / z [M+H] + 668.3, m / z [MH] - 666.3;UPLC-MS 1 LC-MS: Rt=3.79 min; MS m / z [M+H] + 668.3, m / z [MH] - 666.3;UPLC-MS 2 1 H NMR(400MHz,DMSO-d6)δ 10.36(m,br,2H),8.78(s,1H),8.05(m,1H),7.85(s,1H),7.28(m,2H),6.83(m,1H),5.25(s,2H),4.55(m,1H),4.26(m,2H),3 .81(m,2H),3.45(m,3H),3.22(m,1H),3.00(m,3H),2.80(m,1H),2.62(m,1H),2.50(m,2H),2.38(s,3H),1.20(t,J=7.3Hz,3H)

[0481] Example 35: 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide [ka] To a solution of 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide·TFA (Intermediate AI) (400 mg, 606 μmol) in DMF (4 mL) was added EtN (252 μL, 1.82 mmol), followed by the dropwise addition of perfluorophenyl 3-hydroxypicolinate (Intermediate CT) (185 mg, 606 μmol) at 0° C. The RM was then warmed to room temperature and stirred at 80° C. for 14 h. The crude product was concentrated under reduced pressure, extracted with water, and purified by column chromatography (silica gel column: silica 12 g, eluent hexane: EtOAc 100:0 to 20:80). The product was recrystallized with ACN to give the title compound. LC-MS: Rt=0.81 min; MS m / z [M+H] + 668.6, m / z [MH] - 666.4;UPLC-MS 1 LC-MS: Rt=3.91 min; MS m / z [M+H] + 668.3, m / z [MH] - 666.3;UPLC-MS 2 1H NMR(400MHz,DMSO-d6)δ 10.38(s,1H),10.23(s,1H),8.19(d,J=8.4Hz,1H),8.06(m,1H),7.73(d,J=8.4Hz,1H),7.28(m,2H),6.82(m,1H),5.28(s,2H),4.55(m,1H), 4.26(m,2H),3.80(m,2H),3.44(m,3H),3.22(m,1H),2.97(m,3H),2.8 0(m,1H),2.62(m,1H),2.57(s,3H),2.52(m,2H),1.19(t,J=7.2Hz,3H)

[0482] Example 36: 2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide [ka] To 4-chloro-3-hydroxypicolinic acid (423 mg, 2.44 mmol) in DCM (10 mL) was added DIPEA (500 μL, 2.86 mmol) and PyAOP (829 mg, 1.59 mmol). After the color changed to dark brown, the RM was stirred for 10 min, followed by the dropwise addition of 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-6-(trifluoromethyl)pyridin-3-yl)acetamide·TFA (Intermediate AI) (700 mg, 1.06 mmol) in DCM (10 mL) and DIPEA (500 μL, 2.86 mmol). The RM was stirred at room temperature for 18 h. The RM was poured into saturated aqueous NaHCO3 and extracted several times with EtOAc. The combined organic layers were dried over a phase separator and concentrated. The crude product was purified by column chromatography (silica gel column: 12 g silica, eluent hexane:EtOAc 100:0 to 0:100). The product-containing fractions were combined and concentrated under reduced pressure. Then, it was washed with Et2O. The solid was purified again by column chromatography (silica gel column: 12 g silica, eluent hexane:EtOAc 100:0 to 5:95) to give the title compound. LC-MS: Rt = 0.91 min; MS m / z [M+H] + 702.3 / 704.3, m / z [MH] - 700.3 / 702.3; UPLC-MS 1 LC-MS: Rt=4.49 min; MS m / z [M+H] + 702.2 / 704.2, m / z [MH] - 700.3 / 702.3; UPLC-MS 2 1H NMR (400 MHz, DMSO-d6) δ 10.83(s,br,1H),10.22(s,br,1H),8.19(d,J=8.8Hz,1H),8.06(d,J=5.0Hz, 1H),7.73(d,J=8.4Hz,1H),7.55(d,J=5.0Hz,1H),6.82(m,1H),5.28(s,2H), 4.54(m,1H),4.26(m,2H),3.80(m,2H),3.50(m,3H),3.15(m,1H),3.00(m,3H) ),2.81(m,1H),2.65(m,1H),2.57(s,3H),2.48(m,2H),1.19(t,J=7.0Hz,3H)

[0483] Example 37: 2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide [ka] 4-Chloro-3-hydroxypicolinic acid (233 mg, 1.34 mmol) was suspended in DCM (7 mL) and 1-chloro-N,N,2-trimethylprop-1-en-1-amine (190 μL, 1.44 mmol) was added. After 5 min, most of the solid dissolved. The light suspension was stirred at room temperature for 2.5 h. The RM was cooled to 0 °C and 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (Intermediate AJ) (431 mg, 765 μmol) was added, followed by DIPEA (601 μL, 3.44 mmol). The yellow solution turned to a black solution. The RM was stirred at room temperature for 20 min. Water (10 mL), saturated aqueous NaHCO3 (10 mL), and DCM (10 mL) were added. The aqueous layer was washed twice with DCM (2 × 10 mL). The combined organic layers were dried over a phase separator and concentrated under reduced pressure. The crude product was purified by column chromatography (RediSep column: 40 g silica, eluent: DCM:DCM / MeOH (8 / 2) 100:0 to 50:50). The product-containing fractions were combined, concentrated under vacuum, and dried under high vacuum to give a brown solid. This was then purified in three portions by reverse-phase preparative HPLC (RP-HPLC acidic 1: 20 min, 20–75% B, 1 min at 75% plateau; RP-HPLC acidic 1: 20 min, 25–80% B, 1 min at 80% plateau; and RP-HPLC acidic 1: 20 min, 40–80% B, 1 min at 80% plateau). The product-containing fractions were combined, basified with saturated aqueous NaHCO3, extracted twice with DCM (2 × 15 mL), dried on a phase separator, and concentrated under reduced pressure. All impure fractions were combined, basified with saturated aqueous NaHCO3, extracted twice with DCM, dried on a phase separator, and concentrated under reduced pressure. This was then suspended in ACN (2 mL), sonicated for 2 minutes, filtered, and the pure fractions were combined and concentrated under reduced pressure to give the title compound. The solid was dissolved in EtOH (10 mL) and DCM (15 mL), filtered, and left to crystallize at room temperature for 3 days. This was then filtered and washed with Et2O.The cake was dried under HV to give the title compound. The sodium salt was prepared similarly to the general procedure. LC-MS: Rt = 1.11 min; MS m / z [M+H] + 719.4 / 721.4, m / z [MH] - 717.5 / 719.5; UPLC-MS 1 1 H NMR(400MHz,DMSO-d6)δ 10.83(s,1H),10.20(s,1H),8.06(d,J=5.1Hz,1H),7.57(m,3H),6.82(m,1H),5.26(s,2H),4.54(m,1H),4.26(m,2H),3.81 (m,2H),3.55(m,3H),3.25(m,1H),3.00(m,3H),2.81(m,1H),2.65(m,1H),2.50(m,2H),2.24(s,3H),1.19(t,J=7.4Hz,3H)

[0484] Example 38: 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide [ka] 2-(2-(3,6-Dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (Intermediate AJ) (504 mg, 894 μmol) and 3-hydroxypicolinoyl chloride (Intermediate CV) (254 mg, 1.61 mmol) were mixed in DCM (7 mL), and DIPEA (312 μL, 1.79 mmol) was added. The dark solution was stirred at room temperature for 1 h. Water (10 mL), saturated aqueous NaHCO3 (10 mL), and DCM (10 mL) were added. The aqueous layer was washed twice with DCM (2 × 10 mL). The combined organic layers were dried over a phase separator and concentrated under reduced pressure. The crude product was purified by column chromatography (RediSep column: 24 g silica, eluent: DCM: DCM / MeOH (8 / 2) 100:0 to 35:65). The product-containing fractions were combined, concentrated under vacuum, and dried under high vacuum to give a white solid. The solid was dissolved in EtOH (5 mL) and DCM (15 mL) and left to crystallize to give the title compound. LC-MS: Rt=1.02 min; MS m / z [M+H] + 685.3, m / z [MH] - 683.4; UPLC-MS 1 1 H NMR(400MHz,DMSO-d6)δ 10.38(s,1H),10.20(s,1H),8.06(m,1H),7.58(m,2H),7.29(m,2H),6.82(m,1H),5.25(s,2H),4.55(m,1H),4.26(m,2H),3. 81(m,2H),3.44(m,3H),3.22(m,1H),2.98(m,3H),2.80(m,1H),2.62(m,1H),2.52(m,2H),2.24(s,3H),1.19(t,J=7.3Hz,3H)

[0485] Example 39: 2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide [ka] The reaction was carried out in four batches obtained as follows: 4-Chloro-3-hydroxypicolinic acid (400 mg, 2.31 mmol) was dissolved in DCM (36 mL) at room temperature under argon. 1-Chloro-N,N,2-trimethylprop-1-en-1-amine (360 mg, 2.66 mmol) was added, and the RM was stirred at room temperature for 2.5 hours. At 0°C, N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (Intermediate AK) (1.10 g, 1.58 mmol) and DIPEA (1.23 mL, 7.08 mmol) were added. The resulting brown solution was stirred at room temperature for 1.3 h. The combined RM from four batches was quenched with water (40 mL) and saturated aqueous NaHCO3 (40 mL). This was extracted four times with DCM (4 × 200 mL). The combined organic layers were washed with water (50 mL) and brine (50 mL), dried through a phase separator, and concentrated under reduced pressure. The residue was adsorbed onto Isolute and purified in two portions by column chromatography (RediSep column: Silica 120 g Gold, eluent DCM:DCM / MeOH (1 / 1) 100:0 to 60:40) and (RediSep column: Silica 120 g Gold, eluent DCM:DCM / MeOH (1 / 1) 100:0 to 90:10). The product-containing fractions were combined and concentrated under reduced pressure to give the title compound as a beige solid. LC-MS: Rt=1.13 min; MS m / z [M+H] +721.4 / 723.4 / 725.4, m / z [MH] - 719.5 / 721.5 / 723.5; UPLC-MS 1 1 H NMR(400MHz,DMSO-d6)δ 10.83(s,1H),10.35(s,1H),8.06(m,2H),7.96(s,1H),7.71(dd,J=2.1Hz,8.7Hz,1H),7.55(d,J=5.1Hz,1H),6.83(m,1H),5.32(s,2H),4. 54(m,1H),4.25(m,2H),3.80(m,2H),3.53(m,3H),3.26(m,1H),2.99(m,3H),2.82(m,1H),2.65(m,1H),2.52(m,2H),1.19(t,J=7.1Hz,3H)

[0486] Example 40: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(4-fluoro-3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide [ka]

[0487] Step 1: 2-(6-(4-(3-(benzyloxy)-4-fluoropicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide To a stirred solution of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide·TFA (Intermediate AK) (300 mg, 441 μmol), 3-(benzyloxy)-4-fluoropicolinic acid (Intermediate CU) (115 mg, 463 μmol), and HATU (201 mg, 529 μmol) in DMF (10 mL) was added DIPEA (385 μL, 2.21 mmol) at room temperature, and the RM was stirred at room temperature for 5 min. The RM was diluted with EtOAc / water and extracted twice with EtOAc. The combined organic extracts were dried over Na2SO4 and concentrated. The crude product was purified by column chromatography (RediSep column: 24 g silica, eluent DCM:DCM / MeOH (8 / 2) 100:0 to 50:50). The product-containing fractions were combined and concentrated to give a white foam. LC-MS: Rt=1.23 min; MS m / z [M+H] + 795.3 / 797.3, m / z [MH] - 793.4 / 795.4; UPLC-MS 1

[0488] Step 2: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(4-fluoro-3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide To a stirred solution of 2-(6-(4-(3-(benzyloxy)-4-fluoropicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide (289 mg, 363 μmol) in DCM (10 mL) was added boron trichloride methyl sulfide complex (363 μL, 727 μmol) at room temperature, and the RM was stirred at room temperature for 20 h. The RM was quenched with MeOH. It was diluted with DCM / NaHCO and extracted twice with DCM, and the combined organic extracts were washed with water and brine, dried over NaSO, and concentrated. The crude product was purified by column chromatography (RediSep column: silica 24 g, eluent DCM:DCM / MeOH (8 / 2) 100:0 to 60:40). The product-containing fractions were combined and concentrated to give an off-white solid. This solid was dissolved in EtOH (6 mL) at 50 °C, and the RM was left at room temperature for 4 h. The white solid was filtered to give the title compound. The sodium salt was prepared similarly to the general procedure. LC-MS: Rt=1.03 min; MS m / z [M+H] + 705.4 / 707.4, m / z [MH] - 703.5 / 705.5; UPLC-MS 1 1 H NMR(400MHz,DMSO-d6)δ 10.80(s,br,1H),10.36(s,1H),8.06(m,2H),7.96(m,1H),7.71(dd,J=2.1Hz,8.8Hz,1H),7.34(dd,J=5.3Hz,10.9Hz,1H),6.83(m,1H),5.31(s, 2H),4.54(m,1H),4.25(m,2H),3.80(m,2H),3.46(m,3H),3.23(m,1H),2 .98(m,3H),2.80(m,1H),2.63(m,1H),2.51(m,2H),1.18(t,J=7.5Hz,3H)

[0489] Example 41: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(4-fluoro-3-hydroxy-6-methylpicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide [ka]

[0490] Step 1: 2-(6-(4-(3-(benzyloxy)-4-fluoro-6-methylpicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (Intermediate AK) (227 mg, 334 μmol), 3-(benzyloxy)-4-fluoro-6-methylpicolinic acid (Intermediate CX) (110 mg, 371 μmol), and HATU (140 mg, 367 μmol) were suspended in DCM (5 mL) and cooled to 0 °C. DIPEA (204 μL, 1.17 mmol) was then added, and the RM was stirred at room temperature for 2.5 h. Water (10 mL), saturated aqueous NaHCO (10 mL), and DCM (10 mL) were added. The aqueous layer was washed twice with DCM (2 × 10 mL). The combined organic layers were dried through a phase separator and concentrated under reduced pressure. The crude product was purified by reverse-phase preparative HPLC (RP-HPLC basic 1: 5–95% B in 20 min, 95% plateau for 1 min). The product-containing fractions were combined, basified with saturated aqueous NaHCO3, extracted twice with DCM, dried through a phase separator, and concentrated under reduced pressure to give the title compound. LC-MS: Rt=1.27 min; MS m / z [M+H] +809.5 / 811.5, m / z [MH] - 807.2 / 809.2; UPLC-MS 1

[0491] Step 2: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(4-fluoro-3-hydroxy-6-methylpicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 2-(6-(4-(3-(benzyloxy)-4-fluoro-6-methylpicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide (242 mg, 292 μmol) was dissolved in DCM (5 mL), TFA (5.00 mL, 64.9 mmol) was added, and the RM was stirred overnight at 60° C. After overnight, approximately 50% conversion. The RM was continued stirring at 60° C. overnight. The RM was concentrated under reduced pressure. The crude product was purified by reverse-phase preparative ISCO (RediSep column: C18 50 g Gold, eluent water + 0.1% TFA:ACN 100:0 to 0:100). The product-containing fractions were combined, basified with saturated aqueous NaHCO3, extracted twice with DCM, dried over a phase separator, and concentrated under reduced pressure to give the title compound. LC-MS: Rt=1.06 min; MS m / z [M+H] + 719.5 / 721.6, m / z [MH] - 717.4 / 719.4; UPLC-MS 1 LC-MS: Rt=5.31 min; MS m / z [M+H] + 719.5 / 721.5, m / z [MH] - 717.5 / 719.3; UPLC-MS 2 1H NMR(400MHz,DMSO-d6)δ 10.35(s,2H),8.05(d,J=8.3Hz,1H),7.96(d,J=2.1Hz,1H),7.71(dd,J=2 .1Hz,8.7Hz,1H),7.22(d,J=11.8Hz,1H),6.83(m,1H),5.31(s,2H),4.52( m,1H),4.25(m,2H),3.80(m,2H),3.47(m,3H),3.22(m,1H),2.98(m,3H), 2.80(m,1H),2.64(m,1H),2.51(m,2H),2.39(s,3H),1.18(t,J=7.1Hz,3H)

[0492] Example 42: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyridin-4(7H)-yl)acetamide [ka]

[0493] Step 1: 2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyridin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide To a stirred solution of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyridin-4(7H)-yl)acetamide (Intermediate AK) (300 mg, 429 μmol), 5-(benzyloxy)-6-methylpyrimidine-4-carboxylic acid (Intermediate CY) (115 mg, 472 μmol), and HATU (245 mg, 644 μmol) in DMF (3 mL) was added DIPEA (375 μL, 2.15 mmol) at room temperature, and the RM was stirred at room temperature for 15 min. The RM was diluted with EtOAc / water and extracted twice with EtOAc, and the combined organic layers were dried over NaSO and concentrated. The crude product was purified by column chromatography (RediSep column: 24 g silica, eluent DCM:MeOH 100:0 to 90:10). The product-containing fractions were combined and concentrated to give the title compound as a beige foam. The product was dissolved in EtOH and stirred at 60° C. for 18 hours, then cooled to 0° C., filtered off, and washed with EtOH to give the title compound as a white solid. LC-MS: Rt = 1.20 min; MS m / z [M+H] + 792.4 / 794.4, m / z [MH] - 790.6 / 792.6; UPLC-MS 1

[0494] Step 2: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyridin-4(7H)-yl)acetamide To a stirred solution of 2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyridin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide (266 mg, 312 μmol) in DCM (6 mL) was added boron trichloride methyl sulfide complex (312 μL, 625 μmol) at room temperature, and the RM was stirred at room temperature for 14 h. The RM was quenched with MeOH. It was then diluted with DCM / water and extracted twice with DCM, and the combined organic extracts were washed with water and brine, dried over Na2SO4, and concentrated. The crude product was purified by column chromatography (RediSep column: 24 g silica, eluent DCM:MeOH 100:0 to 93:07). Product-containing fractions were combined and concentrated to give the title compound as an off-white solid. The product was dissolved in EtOH and stirred at 60° C. for 18 hours, then cooled to 0° C., filtered, and washed with EtOH to give the title compound as a white solid characterized by the XRPD diffractogram in FIG. 1. Ex 42a in the table below indicates the most prominent peak (2 theta°) in the XRPD diffractogram in FIG. 1. XRPD was repeated on a sample of the title compound prepared by the procedure described in Example 42 above, and then purified by column chromatography (RediSep 150 g, eluent DCM:MeOH 100:0 to 90:10), followed by trituration in EtOH. The sample was characterized by the XPRD diffractogram shown in Figure 7. Ex42b in the table below indicates the most prominent peak (2 theta°) in the XRPD diffractogram of Figure 7. LC-MS: Rt=1.05 min; MS m / z [M+H] + 702.4 / 704.4, m / z [MH] - 700.5 / 702.5; UPLC-MS 1 11H NMR (600 MHz, DMSO-d6) δ 10.40 (s, 1H), 10.20 (br s, 1H), 8.57 (s, 1H), 8.06 (d, J = 8.5 Hz, 1H), 7.97 (d, J = 2.0 Hz, 1H), 7.72 (dd, J = 8.7 Hz, J = 2.1 Hz, 1H), 6.83 (m, 1H), 5.32 (m, 2H), 4.52 (m, 1H), 4.25 (m, 2H), 3.80 (t, J = 5.5 Hz, 2H), 3.50 (m, 3H), 3.25 (m, 1H), 2.99 (m, 3H), 2.81 (m, 1H), 2.63 (m, 1H), 2.51 (m, 2H), 2.44 (s, 3H), 1.18 (t, J = 7.5 Hz, 3H).

[0495] The sodium salt was prepared in the same manner as the general procedure. 1 1H NMR (400 MHz, DMSO-d6) δ 10.36 (s, br, 2H), 8.55 (s, 1H), 8.05 (d, J = 8.5 Hz, 1H), 7.96 (m, 1H), 7.71 (dd, J = 2.1 Hz, 8.8 Hz, 1H), 6.83 (m, 1H), 5.32 (s, 2H), 4.52 (m, 1H), 4.25 (m, 2H), 3.80 (m, 2H), 3.48 (m, 3H), 3.25 (m, 1H), 2.99 (m, 3H), 2.81 (m, 1H), 2.64 (m, 1H), 2.52 (m, 2H), 2.43 (s, 3H), 1.18 (t, J = 7.4 Hz, 3H).

[0496] [Table 14]

[0497] [Table 15]

[0498] Example 43: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyridin-4(7H)-yl)acetamide-2,2-d2 [ka] A beige suspension of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyridin-4(7H)-yl)acetamide (Example 42) (70.0 mg, 100 μmol) and KCO (13.8 mg, 100 μmol) in THF (997 μL) / DMSO-d (282 μL, 3.99 mmol) / DO (180 μL, 9.97 mmol) was heated to 65° C. and stirred for 45 min. A small amount of water was added to the RM, followed by extraction three times with EtOAc. The combined organic layers were washed once with brine, dried over Na2SO4, concentrated, and dried under reduced pressure to give a beige solid. The aqueous layer still contained the product. The aqueous layer was concentrated to remove THF and then extracted twice with DCM. The aqueous layer was diluted with saturated aqueous NaHCO3 and extracted three times with DCM. The combined organic layers were dried on a phase separator, concentrated, and dried under vacuum to give the title compound. LC-MS: Rt=1.04 min; MS m / z [M+H] + 704.5 / 706.5, m / z [MH] - 702.2 / 704.2; UPLC-MS 1 1H NMR(400MHz,DMSO-d6)δ 10.36(s,2H),8.56(s,1H),8.06(d,J=8.4Hz,1H),7.96(d,J=2.1Hz,1H),7.71(dd,J=2.1Hz,8.8Hz,1H),6.83(m,1H),4.52(m,1H),4.25 (m,2H),3.80(m,2H),3.49(m,3H),3.25(m,1H),2.98(m,3H),2.81(m,1H),2.64(m,1H),2.52(m,2H),2.44(s,3H),1.19(t,J=7.2Hz,3H)

[0499] Example 44: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxypyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyridin-4(7H)-yl)acetamide [ka]

[0500] Step 1: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-methoxypyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyridin-4(7H)-yl)acetamide N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyridin-4(7H)-yl)acetamide (Intermediate AK) (508 mg, 727 μmol), 5-methoxypyrimidine-4-carboxylic acid (134 mg, 872 μmol), and HATU (427 mg, 1.09 mmol) were mixed in DMF (10 mL) under argon at room temperature. DIPEA (635 μL, 3.64 mmol) was added, and the RM was stirred at room temperature for 2 h. The reaction was quenched with water (10 mL). Saturated aqueous NaHCO3 (15 mL) was added, and the mixture was extracted with EtOAc (3 × 70 mL). The organic layer was washed with water (20 mL) and brine (20 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was adsorbed onto Isolute and purified by column chromatography (RediSep column: 40 g silica, eluent DCM:DCM / MeOH (8 / 2) 100:0 to 50:50). The product-containing fractions were combined, concentrated, mixed with Et2O, sonicated, filtered, and dried under high vacuum to give the title compound as a beige solid. LC-MS: Rt = 0.99 min; MS m / z [M+H] + 702.2 / 704.2, m / z [MH] - 700.4 / 702.4; UPLC-MS 1

[0501] Step 2: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxypyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyridin-4(7H)-yl)acetamide N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-methoxypyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyridin-4(7H)-yl)acetamide (178 mg, 254 μmol) was mixed with DMF (2 mL), LiCl (43.0 mg, 1.01 mmol) was added, and the RM was stirred at 150 °C for 18.3 h. The mixture was then allowed to cool to room temperature. The reaction was diluted with water and saturated aqueous NaHCO (5 mL). It was extracted with EtOAc (3 × 20 mL). The organic layer was washed with water (5 mL) and brine (2 × 5 mL). The organic layer was washed again with EtOAc (3 × 20 mL). The combined organic layers were dried over MgSO4, filtered through a phase separator, and concentrated under reduced pressure. The residue was adsorbed onto Isolute and purified by column chromatography (RediSep column: 40 g silica, eluent DCM:DCM / MeOH (8 / 2) 100:0 to 35:65). The product-containing fractions were combined and concentrated under reduced pressure. The solid was purified by reverse-phase preparative HPLC (RP-HPLC basic 1: 15 to 65% B in 20 min, 65% plateau for 1 min). The product-containing fractions were combined, the ACN removed under reduced pressure, and the residue was lyophilized to give the title compound as a beige solid. LC-MS: Rt=0.98 min; MS m / z [M+H] + 688.3 / 690.3, m / z [MH] - 686.3 / 688.3; UPLC-MS 1 1H NMR(400MHz,DMSO-d6)δ 10.75(s,br,1H),10.35(s,br,1H),8.70(s,1H),8.44(s,1H),8.05(d,J=8. 5Hz,1H),7.96(d,J=2.2Hz,1H),7.71(dd,J=2.1Hz,8.9Hz,1H),6.83(m,1H), 5.31(s,2H),4.51(m,1H),4.25(m,2H),3.80(m,2H),3.47(m,3H),3.23(m,1H) ),2.97(m,3H),2.81(m,1H),2.63(m,1H),2.50(m,2H),1.18(t,J=7.4Hz,3H)

[0502] Example 45: N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyridin-4(7H)-yl)acetamide [ka] N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyridin-4(7H)-yl)acetamide (Intermediate AL) (212 mg, 337 μmol) and 3-hydroxypicolinoyl chloride (Intermediate CV) (89.0 mg, 565 μmol) were dissolved in DCM (5 mL) and DIPEA (294 μL, 1.68 mmol) was added. The RM was stirred at room temperature for 30 min. 3-Hydroxypicolinoyl chloride (Intermediate CV) (89.0 mg, 565 μmol) was added again and the RM was continued to stir at room temperature for 1 h. Water (10 mL), saturated aqueous NaHCO3 (10 mL), and DCM (10 mL) were added. The aqueous layer was washed twice with DCM (2 × 10 mL). The combined organic layers were dried on a phase separator and concentrated under reduced pressure. The crude product was purified by column chromatography (RediSep column: 40 g silica, eluent DCM:DCM / MeOH (9 / 1) 100:0 to 60:40). The product-containing fractions were combined, concentrated under vacuum, and dried under high vacuum to give a beige solid. The crude product was purified by reverse-phase preparative HPLC (RP-HPLC acidic 1: 20–80% in 20 min, 80% plateau for 1 min). The product-containing fractions were combined, basified with saturated aqueous NaHCO3, extracted twice with DCM (2 × 15 mL), dried on a phase separator, and concentrated under reduced pressure. The product was dissolved in DCM (5 mL) and MeOH (2 mL) and left at room temperature overnight. The resulting crystals were filtered off and washed with EtO (4 × 3 mL). The cake was dried under high vacuum to give the title compound as white crystals. LC-MS: Rt=0.92 min; MS m / z [M+H] + 688.4 / 690.4, m / z [MH] - 686.4 / 688.3; UPLC-MS 3 1H NMR(400MHz,DMSO-d6)δ 10.56(s,1H),10.38(s,1H),8.56(d,J=8.0Hz,1H),8.07(m,1H),7.96(d,J=8.0Hz,1H),7.29(m,2H),6.82(m,1H),5.36(s,2H),4.55( m,1H),4.25(m,2H),3.80(m,2H),3.46(m,3H),3.23(m,1H),2.98(m,3H),2.80(m,1H),2.62(m,1H),2.51(m,2H),1.18(t,J=7.3Hz,3H)

[0503] Example 46: 2-(6-(4-(4-chloro-3-hydroxypicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyridin-4(7H)-yl)-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)acetamide [ka] 4-Chloro-3-hydroxypicolinic acid (1.48 g, 8.54 mmol) was suspended in DCM (140 mL) and 1-chloro-N,N,2-trimethylprop-1-en-1-amine (1.29 mL, 9.73 mmol) was added. After 1 h, the reddish-brown solution transformed into a yellow solution. The RM was stirred at room temperature for 2.5 h. N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyridin-4(7H)-yl)acetamide (Intermediate AL) (5.31 g, 8.43 mmol) and DIPEA (3.90 mL, 22.3 mmol) were dissolved in DCM (80 mL), and the prepared acid chloride solution was added dropwise over 1 h. 5% aqueous NaHCO3 and DCM were added. The aqueous layer was washed twice with DCM (2 × 100 mL). The combined organic layers were dried over a phase separator and concentrated under reduced pressure. The crude product was purified by column chromatography (RediSep column: 330 g silica, eluent DCM:MeOH 100:0 to 90:10). The product-containing fractions were combined, concentrated, and dried under high vacuum. The residue was purified by reverse-phase preparative ISCO (RediSep column: C18 240 g, eluent water:ACN 100:0 to 0:100). The product-containing fractions were combined, concentrated, and dried under reduced pressure. The resulting solid was suspended in EtOH (25 mL) at 55 °C, stirred at 55 °C overnight, cooled to room temperature, stirred at 0 °C for 1 h, filtered, and dried under high vacuum to give the title compound. LC-MS: Rt = 0.99 min; MS m / z [M+H] + 722.3 / 724.3 / 726.3, m / z [MH] - 720.2 / 722.2 / 724.2; UPLC-MS 1 1H NMR(400MHz,DMSO-d6)δ 10.79(s,br,1H),10.59(s,br,1H),8.57(d,J=8.1Hz,1H),8.05(d,J=4.9 Hz,1H),7.95(d,J=8.5Hz,1H),7.54(d,J=4.9Hz,1H),6.82(m,1H),5.36( s,2H),4.54(m,1H),4.25(m,2H),3.80(m,2H),3.53(m,3H),3.23(m,1H), 2.99(m,3H),2.81(m,1H),2.66(m,1H),2.51(m,2H),1.18(t,J=7.4Hz,3H)

[0504] Example 47: N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyridin-4(7H)-yl)acetamide [ka]

[0505] Step 1: N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-methoxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyridin-4(7H)-yl)acetamide To a stirred solution of 5-methoxy-6-methylpyrimidine-4-carboxylic acid (Intermediate CW) (311 mg, 1.85 mmol) in DMF (10 mL) was added EDC·HCl (355 mg, 1.85 mmol), HOBt (250 mg, 1.85 mmol), and pyridine (300 μL, 3.71 mmol) at room temperature. N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyridin-4(7H)-yl)acetamide (Intermediate AL) (700 mg, 1.24 mmol) was then added, and the mixture was stirred at room temperature for 14 h. Water (20 mL) was added to the RM, which was then extracted with 10% MeOH in DCM (2 × 20 mL). The organic phase was washed with brine (20 mL), dried over NaSO, filtered, and concentrated. The crude product was purified by column chromatography (silica gel column: 12 g silica, eluent DCM:MeOH 100:0 to 95:5) to give the title compound. The product was heated in EtOH:DCM (1:2) until completely dissolved. The solution was evaporated to dryness at room temperature and then dried under vacuum to give the title compound as a beige solid. LC-MS: Rt=1.48 min; MS m / z [M+H] + 717.1 / 719.1; UPLC-MS 11

[0506] Step 2: N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyridin-4(7H)-yl)acetamide To a stirred solution of N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-methoxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyridin-4(7H)-yl)acetamide (700 mg, 820 μmol) in DMF (10 mL) was added LiCl (348 mg, 8.20 mmol) at room temperature, and the RM was stirred at 140 °C for 14 h. Water (20 mL) was added to the RM, which was extracted twice with DCM (2 × 20 mL). The organic phase was washed with brine (20 mL), dried over NaSO, filtered, and concentrated. The crude product was purified by reverse-phase preparative HPLC (RP-HPLC acidic 4: 30-40% B in 2 min, 40-55% B in 8 min) to give the title compound. The product was heated in EtOH:DCM (1:2) until completely dissolved. The solution was evaporated to dryness at room temperature and then dried under vacuum to give the title compound as a beige solid, characterized by the XRPD diffractogram in Figure 3. The following table shows the most prominent peaks (2 theta°) in the XRPD diffractogram in Figure 3. The sodium salt was prepared similarly to the general procedure. LC-MS: Rt=0.92 min; MS m / z [M+H] + 703.4 / 705.4, m / z [MH] - 701.5 / 703.5; UPLC-MS 1 1 H NMR(400MHz,DMSO-d6)δ 10.46(s,br,2H),8.56(m,2H),7.95(d,J=8.4Hz,1H),6.82(m,1H),5.35(s,2H),4.52(m,1H),4.25(m,2H),3.80(m,2) H),3.48(m,3H),3.25(m,1H),2.99(m,3H),2.81(m,1H),2.65(m,1H),2.51(m,2H),2.44(s,3H),1.18(t,J=7.7Hz,3H)

[0507] [Table 16]

[0508] Example 48: N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(4-fluoro-3-hydroxypicolinoyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyridin-4(7H)-yl)acetamide [ka]

[0509] Step 1: 2-(6-(4-(3-(benzyloxy)-4-fluoropicolinoyl)piperazin-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyridin-4(7H)-yl)-N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)acetamide To a stirred solution of N-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyridin-4(7H)-yl)acetamide (Intermediate AL) (445 mg, 785 μmol), 3-(benzyloxy)-4-fluoropicolinic acid (Intermediate CU) (204 mg, 824 μmol), and HATU (358 mg, 942 μmol) in DMF (10 mL) was added DIPEA (685 μL, 3.92 mmol) at room temperature, and the RM was stirred at room temperature for 5 min. The RM was diluted with EtOAc / water and extracted twice with EtOAc. The combined organic extracts were washed with water and brine, dried over anhydrous NaSO, and concentrated. The crude product was purified by column chromatography (RediSep column: 24 g silica, eluent DCM:DCM / MeOH (8 / 2) 100:0 to 50:50). The product-containing fractions were combined and concentrated to give the title compound as a yellow foam. LC-MS: Rt=1.14 min; MS m / z [M+H] +796.5 / 798.5, m / z [MH] - 794.5 / 796.5; UPLC-MS 1

[0510] Step 2: N-(2-...

Claims

【Request 1】 【Chemical 72-1】 【change】 or a pharmaceutically acceptable salt thereof, selected from:

2. 10. The compound of claim 1, or a pharmaceutically acceptable salt thereof, in non-zwitterionic form.

3. 10. The compound of claim 1, or a pharmaceutically acceptable salt thereof, in zwitterionic form.

4. 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is a sodium salt.

5. 10. The compound of claim 1, or a pharmaceutically acceptable salt thereof, in amorphous form.

6. 10. The compound of claim 1, or a pharmaceutically acceptable salt thereof, in crystalline form.

7. 10. A combination comprising a compound of claim 1 or a pharmaceutically acceptable salt thereof and one or more additional therapeutically active agents.

8. 10. A pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers.

9. 1. A pharmaceutical composition for use in a method for modulating WRN activity in a subject, comprising: The pharmaceutical composition comprises the compound of claim 1 or a pharmaceutically acceptable salt thereof, 10. A pharmaceutical composition, wherein the method comprises administering to the subject a therapeutically effective amount of the compound of claim 1 or a pharmaceutically acceptable salt thereof.

10. 1. A pharmaceutical composition for use in a method of inhibiting WRN in a subject, comprising: The pharmaceutical composition comprises the compound of claim 1 or a pharmaceutically acceptable salt thereof, 10. A pharmaceutical composition, wherein the method comprises administering to the subject a therapeutically effective amount of the compound of claim 1 or a pharmaceutically acceptable salt thereof.

11. 1. A pharmaceutical composition for use in a method of treating cancer in a subject, comprising: The pharmaceutical composition comprises the compound of claim 1 or a pharmaceutically acceptable salt thereof, 10. A pharmaceutical composition, wherein the method comprises administering to the subject a therapeutically effective amount of the compound of claim 1 or a pharmaceutically acceptable salt thereof.

12. The pharmaceutical composition of claim 11, wherein the cancer is characterized by microsatellite instability-high (MSI-H) or deficient mismatch repair (dMMR).

13. The following compound or a pharmaceutically acceptable salt thereof: 【Chemistry 101】

14. 14. The compound of claim 13, or a pharmaceutically acceptable salt thereof, in non-zwitterionic form.

15. 14. The compound of claim 13, or a pharmaceutically acceptable salt thereof, in zwitterionic form.

16. 14. The compound of claim 13, or a pharmaceutically acceptable salt thereof, wherein the compound is a sodium salt.

17. 14. The compound of claim 13, or a pharmaceutically acceptable salt thereof, in amorphous form.

18. 14. The compound of claim 13, or a pharmaceutically acceptable salt thereof, in crystalline form.

19. 14. A combination comprising a compound of claim 13 or a pharmaceutically acceptable salt thereof and one or more additional therapeutically active agents.

20. 14. A pharmaceutical composition comprising a compound of claim 13 or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers.

21. The following compound or a pharmaceutically acceptable salt thereof: 【Chemistry 102】

22. 22. The compound of claim 21, or a pharmaceutically acceptable salt thereof, in non-zwitterionic form.

23. 22. The compound of claim 21, or a pharmaceutically acceptable salt thereof, in zwitterionic form.

24. 22. The compound of claim 21, or a pharmaceutically acceptable salt thereof, wherein the compound is a sodium salt.

25. 22. The compound of claim 21, or a pharmaceutically acceptable salt thereof, in amorphous form.

26. 22. The compound of claim 21, or a pharmaceutically acceptable salt thereof, in a crystalline form.

27. 22. A combination comprising a compound of claim 21 or a pharmaceutically acceptable salt thereof and one or more additional therapeutically active agents.

28. 22. A pharmaceutical composition comprising a compound of claim 21, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.

29. (R)—N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide; and N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide; or a pharmaceutically acceptable salt thereof.

30. 1. A pharmaceutical composition for use in a method for modulating WRN activity in a subject, comprising: The pharmaceutical composition comprises the compound of claim 21 or a pharmaceutically acceptable salt thereof; 22. A pharmaceutical composition, wherein the method comprises administering to the subject a therapeutically effective amount of the compound of claim 21 or a pharmaceutically acceptable salt thereof.

31. 1. A pharmaceutical composition for use in a method of inhibiting WRN in a subject, comprising: The pharmaceutical composition comprises the compound of claim 21 or a pharmaceutically acceptable salt thereof; 22. A pharmaceutical composition, wherein the method comprises administering to the subject a therapeutically effective amount of the compound of claim 21 or a pharmaceutically acceptable salt thereof.

32. 1. A pharmaceutical composition for use in a method of treating a disorder or disease that can be treated by inhibiting WRN in a subject, comprising: The pharmaceutical composition comprises the compound of claim 21 or a pharmaceutically acceptable salt thereof; 22. A pharmaceutical composition, wherein the method comprises administering to the subject a therapeutically effective amount of the compound of claim 21 or a pharmaceutically acceptable salt thereof.

33. 1. A pharmaceutical composition for use in a method of treating cancer in a subject, comprising: The pharmaceutical composition comprises the compound of claim 21 or a pharmaceutically acceptable salt thereof; 22. A pharmaceutical composition, wherein the method comprises administering to the subject a therapeutically effective amount of the compound of claim 21 or a pharmaceutically acceptable salt thereof.

34. 34. The pharmaceutical composition of claim 33, wherein the cancer is characterized by microsatellite instability-high (MSI-H) or deficient mismatch repair (dMMR).

35. 35. The pharmaceutical composition of claim 34, wherein the cancer is selected from colon cancer, gastric cancer, prostate cancer, endometrial cancer, adrenocortical cancer, uterine cancer, cervical cancer, esophageal cancer, breast cancer, renal cancer, and ovarian cancer.

36. 35. The pharmaceutical composition of claim 34, wherein the cancer is colon cancer.

37. The following compounds: 【Chemistry 103】

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