Piperidinyl pyridinyl formonitrile derivatives as inhibitors of glutamyl peptide cyclotransferases and glutamyl peptide cyclotransferase-like proteins

Abstract

The present invention provides certain piperidinyl pyridyl formonitrile derivatives and pharmaceutically acceptable salts thereof, which are inhibitors of glutamyl peptide cyclotransferase (QPCT) and glutamyl peptide cyclotransferase-like protein (QPCTL), and are thus useful in the treatment of diseases that can be treated by inhibition of QPCT / L. The invention also provides pharmaceutical compositions containing them and processes for preparing the compounds.

Description

TECHNICAL FIELD

[0001] The present invention provides certain piperidinylpyridyl cyanide derivatives and pharmaceutically acceptable salts thereof, which are inhibitors of glutaminyl peptide cyclotransferase (QPCT) and glutaminyl peptide cyclotransferase-like protein (QPCTL) and thus are useful in the treatment of diseases that can be treated by inhibition of QPCT / L. The present invention also provides pharmaceutical compositions containing them and processes for preparing the compounds. BACKGROUND

[0002] Glutaminyl peptide cyclotransferase (QPCT) and glutaminyl peptide cyclotransferase-like protein (QPCTL) catalyze the intramolecular cyclization of N-terminal glutamine (Q) residues into pyroglutamic acid (pE) and release of ammonia [Stephan Schilling et al., “Identification of Human Glutaminyl Cyclase as a Metalloenzyme POTENT INHIBITION BY IMIDAZOLE DERIVATIVES AND HETEROCYCLIC CHELATORS,” Journal of Biological Chemistry 278, no. 50 (2003): 49773-79, https: / / doi.org / 10.1074 / jbc.m309077200; Holger Cynis et al., “Isolation of an Isoenzyme of Human Glutaminyl Cyclase: Retention in the Golgi Complex Suggests Involvement in the Protein Maturation Machinery,” Journal of Molecular Biology 379, no. 5 (2008): 966-80, https: / / doi.org / 10.1016 / j.jmb.2008.03.078; Anett Stephan et al., “Mammalian Glutaminyl Cyclases and Their Isoenzymes Have Identical Enzymatic Characteristics,” FEBS Journal276, no. 22 (2009): 6522-36, https: / / doi.org / 10.1111 / j.1742-4658.2009.07337.x.] Although QPCT is a secreted protein, QPCTL remains within the Golgi complex. Both enzymes have high homology in the active site and similar catalytic specificity. Due to the high homology of the active site, inhibition of the active site blocks the enzymatic activity of both enzymes, QPCT and QPCTL. Therefore the term “QPCT / L” describes both enzymes. Due to their different cellular localization, differences in their relevance for the modification of biological substrates have been reported. Known substrates of intracellular QPCTL and / or extracellular QPCT are CD47 [Meike E. W. Logtenberg et al., “Glutaminyl Cyclase Is an Enzymatic Modifier of the CD47-SIRPa Axis and a Target for Cancer Immunotherapy,” Nature Medicine 25, no. 4 (2019): 612-19,https: / / doi.org / 10.1038 / s41591-019-0356-z.] different chemokines (e.g. CCL2 and 7 or CX3CL1) [Rosa Barreira da Silva et al., “Loss of the Intracellular Enzyme QPCTL Limits Chemokine Function and Reshapes Myeloid Infiltration to Augment Tumor Immunity,” Nature Immunology 23, no. 4 (2022): 568-80, https: / / doi.org / 10.1038 / s41590-022-01153-x; Astrid Kehlen et al., “N-Terminal Pyroglutamate Formation in CX3CL1 Is Essential for Its Full Biologic Activity,” Bioscience Reports37, no.4 (2017): BSR20170712, https: / / doi.org / 10.1042 / bsr20170712.], amyloid-β peptide [Cynis et al., “Isolation of an Isoenzyme of Human Glutaminyl Cyclase: Retention in the Golgi Complex Suggests Involvement in the Protein Maturation Machinery.”] or hormones like TRH [Andreas Becker et al., “IsoQC (QPCTL) Knock-out Mice Suggest Differential Substrate Conversion by Glutaminyl Cyclase Isoenzymes,”] Biological Chemistry 397, no. 1 (2016): 45-55, https: / / doi.org / 10.1515 / hsz-2015-0192.]. N-terminal glutamine modification to pyroglutamate on substrates has functional effects on proteins and can influence different pathomechanisms of several diseases. CD47 is expressed on the surface of almost all body cells, including apoptotic cells, senescent cells, or cancer cells. [Meike EW Logtenberg, Ferenc A. Scheeren, and Ton N. Schumacher, “The CD47-SIRPα Immune Checkpoint,” Immunity 52, no. 5 (2020): 742-52, https: / / doi.org / 10.1016 / j.immuni.2020.04.011]. The major ligand of CD47 is signal regulatory protein α (SIRPα), an inhibitory transmembrane receptor found on bone marrow-like cells (e.g., macrophages, monocytes, neutrophils, dendritic cells, and other cells). QPCTL-mediated N-terminal pyroglutamate modification on CD47 requires SIRPα binding [Deborah Hatherley et al., “Paired Receptor Specificity Explained by Structures of Signal Regulatory Proteins Alone and Complexed with CD47,”]. Molecular Cell31, no. 2 (2008): 266-77, https: / / doi.org / 10.1016 / j.molcel.2008.05.026; Meike EW Logtenberg et al., “Glutaminyl Cyclase Is an Enzymatic Modifier of the CD47- SIRPα Axis and a Target for Cancer Immunotherapy,” Nature Medicine 25, no. 4 (2019): 612-19, https: / / doi.org / 10.1038 / s41591-019-0356-z.] This signaling axis induces a "Don't Eat Me Signal," thereby preventing macrophages from phagocytosing CD47-expressing cells. Therefore, high CD47 expression is associated with the pathogenesis of diseases such as cancer [Logtenberg et al., "Glutaminyl Cyclase Is an Enzymatic Modifier of the CD47-SIRPα Axis and a Target for Cancer Immunotherapy," 2019; Meike EWLogtenberg, Ferenc A. Scheeren, and Ton N. Schumacher, "The CD47-SIRPα Immune Checkpoint," Immunity 52, no. 5 (2020): 742-52, https: / / doi.org / 10.1016 / j.immuni.2020.04.011.], COVID-19 [Katie-May McLaughlin et al., “A Potential Role of the CD47 / SIRPalpha Axis in COVID-19 Pathogenesis,” Current Issues in Molecular Biology 43, no. 3 (2021): 1212-25, https: / / doi.org / 10.3390 / cimb43030086.], pulmonary fibrosis [Gerlinde Wernig et al., “Unifying Mechanism for Different Fibrotic Diseases,” Proceedings of the National Academy of Sciences114, no. 18(2017): 4757-62, https: / / doi.org / 10.1073 / pnas.1621375114; Lu Cui et al., "Activation of JUN in Fibroblasts Promotes Pro-Fibrotic Program andModulates Protective Immunity," Nature Communications 11, no. 1 (2020): 2795, https: / / doi.org / 10.1038 / s41467-020-16466-4.], Systemic sclerosis [Wernig et al., “Unifying Mechanism for Different Fibrotic Diseases”; Tristan Lerbs et al., “CD47 Prevents the Elimination of Diseased Fibroblasts in Scleroderma,” JCI Insight 5, no. 16 (2020): e140458, https: / / doi.org / 10.1172 / jci.insight.140458.] and liver fibrosis [Taesik Gwag et al., “Anti-CD47 Antibody Treatment Attenuates Liver Inflammation and Fibrosis in Experimental Non-alcoholic Steatohepatitis Models,” Liver International 42, no. 4 (2022): 829-41, https: / / doi.org / 10.1111 / liv.15182.]. Because enhanced CD47 expression blocks the clearance of apoptotic cells, there is an accumulation of apoptotic lung epithelial cells, leading to profibrotic stimulation and accelerating lung inflammation and scarring [Alexandra L. McCubrey and Jeffrey L. Curtis, “Efferocytosis and Lung Disease,” Chest143, no.6 (2013): 1750-57, https: / / doi.org / 10.1378 / chest.12-2413; Brennan D. Gerlach et al., “Efferocytosis Induces Macrophage Proliferation to Help Resolve TissueInjury,” Cell Metabolism [2021, https: / / doi.org / 10.1016 / j.cmet.2021.10.015.]. Because the half-life and function of CD47 are primarily dependent on QPCTL enzyme activity, inhibition of QPCT and QPCTL may be a suitable mechanism for the treatment of pulmonary fibrosis such as IPF or SSC-ILD [Lerbs et al., "CD47 Prevents the Elimination of Diseased Fibroblasts in Scleroderma."], and it can be used alone or in combination with current standard treatments for pulmonary fibrosis (such as nintedanib) [Luca Richeldi et al., "Efficacy and Safety of Nintedanib in Idiopathic Pulmonary Fibrosis,"]. The New England Journal of Medicine 370, no. 22 (2014): 2071-82, https: / / doi.org / 10.1056 / nejmoa1402584; Kevin RFlaherty et al., "Nintedanib in Progressive Fibrosing Interstitial LungDiseases," New England Journal of Medicine 381, no. 18 (2019): 1718-27, https: / / doi.org / 10.1056 / nejmoa1908681.] or future treatments such as PDE4 inhibitors [Luca Richeldi et al., “Trial of a Preferential Phosphodiesterase 4B Inhibitor for Idiopathic Pulmonary Fibrosis,” New England Journal of Medicine 386, no. 23 (2022): 2178-87, https: / / doi.org / 10.1056 / nejmoa2201737] for common use.

[0003] By expressing CD47, cancer cells can evade destruction by the immune system or immune surveillance, for example, by evading phagocytosis by immune cells [Stephen B. Willingham et al., "The CD47-Signal Regulatory Protein Alpha (SIRPa) Interaction Is a Therapeutic Target for Human Solid Tumors,"]. Pr oceedingsoftheNationalAcademyofSciences 109, no. 17 (2012): 6662-67, https: / / doi.org / 10.1073 / pnas.1121623109].

[0004] In addition to CD47, chemokines (such as CCL2 and CX3CL1) have been identified as substrates for QPCTL and / or QPCT [Holger Cynis et al., “The Isoenzyme of Glutaminyl Cyclase Is an Important Regulator of Monocyte Infiltration under Inflammatory Conditions,”] EMBOMolecularMedicine 3, no. 9 (2011): 545-58, https: / / doi.org / 10.1002 / emmm.201100158]. N-terminal pGlu formation shows increased in vivo activity by both conferring resistance to aminopeptidases and by increasing its ability to induce chemokine receptor signaling. Due to the intracellular mechanism of N-terminal cyclization mediated by the Golgi-related enzyme QPCTL, the two major monocyte chemoattractants CCL2 and CCL7 are insensitive to DPP4 inactivation in vivo. QPCTLs have been confirmed as key regulators of monocyte migration to solid tumors [Kaspar Bresser et al., “QPCTL Regulates Macrophage and Monocyte Abundance and Inflammatory Signatures in the Tumor Microenvironment,”]. Oncoimmunology11, no. 1 (2022):2049486, https: / / doi.org / 10.1080 / 2162402x.2022.2049486; Rosa Barreira da Silva et al., "Loss of the Intracellular Enzyme QPCTL Limits Chemokine Function andReshapes Myeloid Infiltration to Augment Tumor Immunity," Nature Immunology [2022, 1-13, https: / / doi.org / 10.1038 / s41590-022-01153-x]. Targeting chemokines has long been promoted as a potential strategy for regulating cell transport in various disease conditions.

[0005] Therefore, there is a desire to provide potent QPCT / L inhibitors.

[0006] Jimenez-Sanchez et al., Nature Chemical Biology, 2015, 11, 347-357, (hereinafter “JS, NCB 2015”) disclosed human glutamine acyl cyclase (hQC) inhibitors SEN177 and SEN180:

[0007] The supplementary information discloses the IC for the SEN177 to separate hQC. 50 IC with a capacity of 53 nM and for discrete QPCTL 50 The value is 13 nM. (Supplementary information) discloses the IC for SEN180 and hQC. 50 IC with 170 nM and QPCTL 50 It is 58 nM.

[0008] Pozzi, C et al., Journal of Biological Inorganic Chemistry, 2018, 23,(8), 1219-1226, (hereinafter “P, JBIC 2018”) further disclosed SEN177 and its binding mode in the hQC cavity. Among them, the K-type binding mode of SEN177 to isolated hQC was disclosed. i It is 20 nM.

[0009] WO 2018 / 178384 discloses QPCTL inhibitors of general formula ABDE, which include Examples 1094 and 1095 (Formula (XIIa) on page 123 and Table on page 125):

[0010] WO 2018 / 178384 does not disclose any biological data for Examples 1094 and 1095.

[0011] WO 2022 / 086920 discloses QPCTL inhibitors with the following general formula.

[0012] It includes compounds 3 and 6: .

[0013] The chemical name of compound 3 disclosed in WO 2022 / 086920 is "1-(1-(6'-chloro-[3,3'-bipyridin]-2-yl)piperidin-4-yl)-1H-1,2,3-triazol-4-amine". This name does not correspond to the chemical structure disclosed therein, but it corresponds to the substitution structure in which the fluorine atom is replaced by a chlorine atom:

[0014] WO 2022 / 086920 discloses, in which

[00343] , the inhibitory activity of compound 3 (including substituted compound 3) and 6 pairs of isolated QPCTLs is IC50. 50 <1µM.

[0015] CN 114874186 discloses a glutamine acyl cyclase isoenzyme inhibitor with the following general formula.

[0016] It includes Examples 21 and 23 (tables on page 17):

[0017] The ICs given in CN 114874186 for embodiments 21 and 23 50 The values ​​are 29.22 nM and 11.26 nM, respectively. Detailed Implementation

[0018] The present invention discloses novel piperidinylpyridinyl carboxylate derivatives of formula (I).

[0019] It is an inhibitor of glutamine cyclotransferase (QPCT) and glutamine cyclotransferase-like protein (QPCTL), and has appropriate pharmacological and pharmacokinetic properties that enable it to be used as a drug for treating conditions and / or diseases that can be treated by inhibiting QPCT / L.

[0020] The compounds of the present invention can provide several advantages, such as enhanced potency, cellular efficacy, high metabolic and / or chemical stability, high selectivity, safety and tolerability, enhanced solubility, enhanced permeability, desired plasma protein binding, enhanced bioavailability, suitable pharmacokinetic characteristics, and the possibility of forming stable salts.

[0021] Compounds of the present invention The present invention provides novel piperidinylpyridinyl carboxylate derivatives that are surprisingly potent inhibitors of QPCT and QPCTL (Assay A), as well as potent inhibitors of QPCT / L in cells associated with (but not limited to) lung disease or cancer (Assay B).

[0022] Furthermore, the novel piperidinylpyridinyl carboxylate derivatives of the present invention have suitable membrane permeability and low in vitro efflux (determination C).

[0023] Therefore, the compounds of the present invention are more suitable for human use.

[0024] The compounds of this invention differ structurally from SEN177 and SEN180 in JS, NCB 2015 in that the nitrile substituent is attached ortho-to the piperidinyl ring of the pyridinyl ring. Furthermore, the pyridinyl ring (including the piperidinyl ring connecting the pyridinyl ring) is generally trisubstituted or tetrasubstituted. Further, R 1 Not limited to hydrogen, and A represents a substituted heterocyclic system other than pyridyl.

[0025] The compounds of this invention differ structurally from Examples 1094 and 1095 in WO 2018 / 178384 in that the nitrile substituent is attached at an ortho position to the piperidinyl ring of the pyridinyl ring. Furthermore, the pyridinyl ring (including the piperidinyl ring connecting the pyridinyl ring) is generally trisubstituted or tetrasubstituted. Further, R 1 Not limited to hydrogen, and A represents a heterocyclic system other than pyridinyl. Furthermore, the 5-membered heterocycle attached to the piperidinyl ring at the 4-position relative to the piperidinyl nitrogen is an aminothiazolyl ring in Example 1094 and an aminothiadiazoleyl ring in Example 1095; however, in the compounds of the present invention, it is a 3-substituted 4-methyl-4H-1,2,4-triazolyl ring.

[0026] The compounds of this invention differ structurally from compounds 3 (including substituted compound 3) and 6 in WO 2022 / 086920 in that the nitrile substituent is attached ortho-to the piperidinyl ring of the pyridinyl ring. Furthermore, the pyridinyl ring (including the piperidinyl ring connecting the pyridinyl ring) is generally trisubstituted or tetrasubstituted. Further, R 1 Not limited to hydrogen, and A represents a heterocyclic system other than pyridyl. Furthermore, the 5-membered heterocycle "M" in the general formula WO 2022 / 086920 is a regioisomer of the 3-substituted 4-methyl-4H-1,2,4-triazolyl ring in compound 3, and the 5-membered heterocycle "M" in the general formula WO 2022 / 086920 is a 3-substituted 4-methyl-4H-1,2,4-triazolyl ring in compound 4, as in the compound of the present invention, but with an amino group.

[0027] The compounds of the present invention differ structurally from compounds 21 and 23 in CN114874186 in that the central sulfonamide portion connecting the piperidinyl ring and the benzene ring is directly replaced by a bond, and the benzene ring is replaced by a pyridinyl ring. Furthermore, a nitrile substituent is attached ortho-to the piperidinyl ring linking position of the pyridinyl ring. Additionally, the pyridinyl ring (including the piperidinyl ring linking the pyridinyl ring) is generally trisubstituted or tetrasubstituted. Further, the compounds of the present invention do not contain an amino linker between the benzene ring and the other cyclic ring.

[0028] These structural differences between the compounds of the present invention and those of the prior art unexpectedly result in an advantageous combination of the following: (i) potent inhibition of QPCT and QPCTL, (ii) potent inhibition of QPCT / L in cells relating to (but not limited to) lung disease or cancer, and (iii) appropriate membrane permeability and low efflux.

[0029] Therefore, the compounds of the present invention are superior to those disclosed in the prior art in terms of the combination of the following parameters: • Strong inhibition of QPCT and QPCTL (Assay A) • Strong inhibition of QPCT / L in cells associated with (but not limited to) lung disease or cancer (assay B) • Appropriate membrane permeability and low efflux (measured by C) This invention provides novel compounds of formula (I).

[0030] in A is A1a, which is a 5- or 6-membered monoheteroaryl ring containing one or two heteroatom members selected from nitrogen and oxygen; At least one of the heteroatom members is nitrogen; Or A is A1b, which is a 9- or 10-membered fused bicyclic heteroaryl ring containing one to four heteroatom members selected from nitrogen and oxygen; At least one of the heteroatom members is nitrogen; And A can be independently selected via one or two R. 2 replace; R 1 R1a is selected from the following groups: H, C 1-4 -alkyl, C 1-4 -alkoxy, hydroxy-C 1-4 -alkyl, F 1-9 -Fluorine-C 1-4 -alkyl and F 1-8 -Fluorine-C 3-5 -cycloalkyl; R 2 R2a can be selected from the following groups: halogen, NH2, C 1-6 -alkyl, F 1-9 -Fluorine-C 1-6 -alkyl, C 1-4 -alkyl-O-H2CH2C-O-, F 1-9 -Fluorine-C 1-4 -alkoxy groups and CH3S(O); or R 2 R2b is selected from the following groups: phenyl, pyridinyl, isoxazolyl, oxazolyl, pyrazolyl, and thiophene; R2b can be arbitrarily and independently passed through one or two R... 3 replace; or R 2 R2c is selected from the following groups. and ; R 3 R3a is selected from the following: halogen, C 1-4 -alkyl, C 1-4 -alkoxy group, HO-C 1-4 -alkyl or CN; Or its salts, specifically its pharmaceutically acceptable salts.

[0031] Another embodiment of the present invention relates to a compound of formula (I), wherein A is A2, which is a 6-membered monoheteroaryl ring containing one or two heteroatom members selected from nitrogen; A can be independently selected from one or two R. 2 replace; And the substituent R 1 R 2 and R 3It is as defined in any of the aforementioned implementation schemes.

[0032] Another embodiment of the present invention relates to a compound of formula (I), wherein A is a group A3 consisting of: pyridinyl, pyrimidinyl, pyrazolo[1,5-b]pyridazinyl, 2H-[1,2,3]triazolo[4,5-b]pyridinyl, pyrazolo[3,4-b]pyridinyl, 1,2-dihydropyridin-2-one, imidazo[1,2-a]pyrimidinyl, [1,3]oxazolo[4,5-b]pyridinyl and furano[2,3-b]pyridinyl; And A can be independently selected via one or two R. 2 replace; And the substituent R 1 R 2 and R 3 It is as defined in any of the aforementioned implementation schemes.

[0033] Another embodiment of the present invention relates to a compound of formula (I), wherein A is selected from a group A4 consisting of the following components. , , , , , , , and ; And A can be independently selected via one or two R. 2 replace; And the substituent R 1 R 2 and R 3 It is as defined in any of the aforementioned implementation schemes.

[0034] Another embodiment of the present invention relates to a compound of formula (I), wherein A is selected from group A5 consisting of the following: , , and ; And A can be independently selected via one or two R. 2 replace; And the substituent R 1 R 2 and R 3 It is as defined in any of the aforementioned implementation schemes.

[0035] Another embodiment of the present invention relates to a compound of formula (I), wherein A is selected from a group A6 consisting of the following: , and ; And A can be independently selected via one or two R. 2 replace; And the substituent R 1 R 2 and R 3 It is as defined in any of the aforementioned implementation schemes.

[0036] R 1 R1b is selected from the following groups: H, C 1-4 -alkyl, C 1-4 -alkoxy, hydroxy-C 1-4 -alkyl, F 1-3 -Fluorine-C 1-4 -alkyl and F 1-3 -Fluorine-C 3-5 -cycloalkyl; And substituents A and R 2 and R 3 It is as defined in any of the aforementioned implementation schemes.

[0037] Another embodiment of the present invention relates to a compound of formula (I), wherein R 1 R1c is selected from the following groups: H, H3C-O-, F3C, HO-C(H3C)2- and ; And substituents A and R 2 and R 3 It is as defined in any of the aforementioned implementation schemes.

[0038] Another embodiment of the present invention relates to a compound of formula (I), wherein R 2 R2d is selected from the following groups: F, Cl, NH2, C 1-6 -alkyl, C 1-4 -alkyl-O-H2CH2C-O- and CH3S(O); or R 2 The R2e group is selected from the following groups: phenyl, pyridinyl, isoxazolyl, oxazolyl, pyrazolyl, and thiophene; R2e can be arbitrarily and independently passed through one or two R... 3 replace; or R 2 R2c is selected from the following groups. and ; And substituents A and R 1 and R 3 It is as defined in any of the aforementioned implementation schemes.

[0039] Another embodiment of the present invention relates to a compound of formula (I), wherein R 2 R2f can be selected from the following groups: F, Cl, NH2, H3C-O-H2CH2C-O- and CH3S(O); or R 2 The R2e group is selected from the following groups: phenyl, pyridinyl, isoxazolyl, oxazolyl, pyrazolyl, and thiophene; R2e can be arbitrarily and independently passed through one or two R... 3 replace; or R 2 R2c is selected from the following groups. and ; And substituents A and R 1 and R 3 It is as defined in any of the aforementioned implementation schemes.

[0040] Another embodiment of the present invention relates to a compound of formula (I), wherein R 2 R2f can be selected from the following groups: F, Cl, NH2, H3C-O-H2CH2C-O- and CH3S(O); or R 2 R2g is selected from the following groups. , , , , and ; R2g is arbitrarily and independently processed by one or two R... 3 replace; or R 2 R2c is selected from the following groups. and ; And substituents A and R 1 and R 3 It is as defined in any of the aforementioned implementation schemes.

[0041] Another embodiment of the present invention relates to a compound of formula (I), wherein R 2 R2f can be selected from the following groups: F, Cl, NH2, H3C-O-H2CH2C-O- and CH3S(O); or R2 R2h is selected from the following groups. , , , , , , , , , , , , , , , , , , , , and ; or R 2 R2c is selected from the following groups. and ; And substituents A and R 1 It is as defined in any of the aforementioned implementation schemes.

[0042] Another embodiment of the present invention relates to a compound of formula (I), wherein R 3 R3b is selected from the following: F, Cl, H3C, isopropyl, H3C-O-, HO-H2C or CN; And substituents A and R 1 and R 2 It is as defined in any of the aforementioned implementation schemes.

[0043] Another embodiment of the present invention relates to a compound of formula (I) having formula (Ia).

[0044] And substituents A and R 2 and R 3 It is as defined in any of the aforementioned implementation schemes.

[0045] Another embodiment of the invention relates to a compound of formula (I) having formula (Ib).

[0046] And substituents A and R 2 and R 3 It is as defined in any of the aforementioned implementation schemes.

[0047] Another embodiment of the present invention relates to a compound of formula (I) having formula (Ic).

[0048] Wherein the substituent R 1 It is as defined in any of the aforementioned implementation schemes.

[0049] Another embodiment of the present invention relates to a compound of formula (I) having formula (Id).

[0050] And the substituent R 1 R 2 and R 3 It is as defined in any of the aforementioned implementation schemes.

[0051] Another embodiment of the present invention relates to a compound of formula (I) having formula (Ie).

[0052] And the substituent R 1 R 2 and R 3 It is as defined in any of the aforementioned implementation schemes.

[0053] Another embodiment of the present invention relates to a compound of formula (I) having formula (If).

[0054] And the substituent R 1 R 2 and R 3 It is as defined in any of the aforementioned implementation schemes.

[0055] Particularly preferred are compounds of formula (I) with the following composition: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , and .

[0056] Particularly preferred are compounds of formula (I) with the following composition: Examples 1, 3, 4, 5, 26, 32, 38 and 40 described in the examples below.

[0057] Particularly preferred are compounds of formula (I) with the following composition: Examples 1, 2, 3, 4, 5, 7, 9, 12, 16, 24, 25, 26, 27, 28, 29, 30 and 31 described in the examples below.

[0058] The present invention provides a novel piperidinylpyridinyl carboxylate derivative of formula (I), which is surprisingly a potent QPCT / L inhibitor.

[0059] Another aspect of the invention relates to a compound of formula (I) which surprisingly exhibits potent inhibition of QPCT / L in cells associated with (but not limited to) lung disease or cancer.

[0060] Another aspect of the invention relates to compounds of formula (I), which are surprisingly potent cellular QPCT / L inhibitors with suitable membrane permeability and low in vitro efflux.

[0061] Another aspect of the invention relates to pharmaceutical compositions comprising at least one compound of formula (I) and optionally one or more inert carriers and / or diluents.

[0062] Another aspect of the present invention refers to the compound of formula (I) for the prevention and / or treatment of conditions associated with QPCT / L inhibition.

[0063] Another aspect of the present invention relates to a method for preparing the compounds of the present invention.

[0064] Other aspects of the invention will become apparent to those skilled in the art directly from the foregoing and the following description and examples.

[0065] Terms and definitions used General definition Terms not explicitly defined herein should be given the meanings that would be given to those skilled in the art based on the disclosure and context. However, unless the opposite meaning is specified, the following terms as used in the specification have the indicated meanings and follow the conventions outlined below.

[0066] In the group, radical, or moiety defined below, the number of carbon atoms is usually indicated before the group, for example, C. 1-6 Alkyl refers to an alkyl group having 1 to 6 carbon atoms. Typically, in groups such as HO, H2N, (O)S, (O)2S, NC (cyano), HOOC, and F3C, those skilled in the art can determine the group connection point to the molecule from the free valence of the group itself. For composite groups containing two or more daughter groups, the last named daughter group is the group connection point, for example, the substituent "aryl-C". 1-3 "-alkylene-" indicates bonding to C 1-3 An aryl group of an alkyl group, wherein the latter of the substituent is attached to the parent nucleus or to the group to which the substituent is attached.

[0067] If the compounds of this invention are described by chemical names or as chemical formulas, the chemical formula shall prevail in the event of any inconsistency. An asterisk may be used in the subsidiary formula to indicate bonds attached to the parent molecule as defined.

[0068] The atomic numbering of a substituent begins with the atom closest to the parent nucleus or the group to which the substituent is attached.

[0069] For example, the term "3-carboxypropyl-group" represents the following substituents:

[0070] The carboxyl group is attached to the third carbon atom of the propyl group. The terms "1-methylpropyl-", "2,2-dimethylpropyl-", or "cyclopropylmethyl-" represent the following groups:

[0071] An asterisk can be used in a subform to indicate a bond that is attached to the parent nucleus molecule as defined.

[0072] As used herein, the term "substituted" means that one or more hydrogen atoms on a specified atom are replaced by a group selected from a set of defined substituents, provided that the substitution does not exceed the normal valence of the specified atom and that the substitution produces a stable compound. Similarly, the term "substituted" may be used in conjunction with a chemical motif rather than a single atom, such as "substituted alkyl," "substituted aryl," etc.

[0073] Unless expressly indicated, throughout the specification and appended claims, the given chemical formulas and names shall cover tautomers and all stereo, optical and geometric isomers (e.g., enantiomers, diastereomers, E / Z isomers, etc.) and their racemates, as well as mixtures of individual enantiomers in different proportions, mixtures of diastereomers, or mixtures of any of the foregoing forms (if such isomers and enantiomers are present), and their solvates (e.g., hydrates).

[0074] Unless explicitly instructed otherwise, the term “pharmaceutically acceptable salt” as defined in more detail below should include its solvates (e.g., hydrates).

[0075] Generally, substantially pure stereoisomers can be obtained according to synthetic principles known to those skilled in the art, such as by separating the corresponding mixtures, by using stereochemically pure starting materials, and / or by stereoselective synthesis. Optically active forms are known in the art, for example by resolving racemic forms or by synthesis, for example, starting from optically active starting materials and / or using chiral reagents.

[0076] The enantiomeric compounds or intermediates of the present invention can be prepared via asymmetric synthesis, for example by the preparation and subsequent separation of suitable diastereomeric compounds or intermediates, which can be separated by known methods (e.g., by chromatographic separation or crystallization) and / or by using chiral reagents (e.g., chiral starting materials, chiral catalysts or chiral auxiliaries).

[0077] Furthermore, those skilled in the art know how to prepare enantiomerically pure compounds from the corresponding racemic mixture, for example by chromatographic separation of the corresponding racemic mixture on a chiral stationary phase; or by resolving the racemic mixture using a suitable resolving agent, for example by forming diastereomeric salts of the racemic compound using an optically active acid or base, followed by resolving the salt and releasing the desired compound from the salt; or by derivatizing the corresponding racemic compound using an optically active chiral auxiliary agent, followed by separation of the diastereomers and removal of the chiral auxiliary group; or by kinetic resolution of the racemic mixture (e.g., by enzymatic resolution); by enantioselective crystallization from clusters of enantiomeric crystals under suitable conditions; or by (fragmented) crystallization from a suitable solvent in the presence of an optically active chiral auxiliary agent.

[0078] The term “pharmaceutically acceptable” as used in this article refers to compounds, materials, compositions, and / or dosage forms that are suitable for use in human tissues within the limits of reasonable medical judgment and are free from excessive toxicity, irritation, allergic reactions, or other problems or complications, and that correspond to a reasonable benefit / risk ratio.

[0079] As used herein, "pharmaceutically acceptable salt" refers to a derivative of the disclosed compound in which the parent compound is modified by preparing its acidic or basic salt. Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues (e.g., amines), basic or organic salts of acidic residues (e.g., carboxylic acids), etc.

[0080] For example, such salts include those formed from benzenesulfonic acid, benzoic acid, citric acid, ethanesulfonic acid, fumaric acid, gentian acid, hydrobromic acid, hydrochloric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, 4-methylbenzenesulfonic acid, phosphoric acid, salicylic acid, succinic acid, sulfuric acid, and tartaric acid. Other pharmaceutically acceptable salts may be derived from ammonia, L-arginine, calcium, 2,2'-iminodiethanol, L-lysine, magnesium, etc. N It is formed by the cations of methyl-D-glucosamine, potassium, sodium and tris(hydroxymethyl)-aminomethane.

[0081] The pharmaceutically acceptable salts of the present invention can be synthesized by conventional chemical methods from parent compounds containing a basic or acidic moiety. Typically, such salts are prepared by reacting the free acidic or basic form of these compounds with a sufficient amount of a suitable base or acid in water or an organic diluent (e.g., diethyl ether, ethyl acetate, ethanol, isopropanol, or acetonitrile) or a mixture thereof.

[0082] Salts of acids other than those mentioned above (e.g., salts that can be used to purify or separate the compounds of the present invention) (e.g., trifluoroacetate) are also part of the present invention.

[0083] The term halogen refers to fluorine, chlorine, bromine, and iodine.

[0084] Term "C" 1-n -alkyl (where n is an integer selected from 2, 3, 4, 5 or 6 (preferably 4, 5 or 6), alone or in combination with other groups) represents an acyclic, saturated, branched or straight-chain hydrocarbon group having 1 to n C atoms. For example, the term C 1-5-Alkyl groups include H3C-, H3C-CH2-, H3C-CH2-CH2-, H3C-CH(CH3)-, H3C-CH2-CH2-CH2-, H3C-CH2-CH(CH3)-, H3C-CH(CH3)-CH2-, H3C-C(CH3)2-, H3C-CH2-CH2-CH2-CH2-, H3C-CH2-CH2-CH(CH3)-, H3C-CH2-CH(CH3)-CH2-, H3C-CH(CH3)-CH2-CH2-, H3C-CH2-C(CH3)2-, H3C-C(CH3)2-CH2-, H3C-CH(CH3)-CH(CH3)- and H3C-CH2-CH(CH2CH3)-.

[0085] Term "C" 2-m "-alkynyl" is used for groups in which m is an integer selected from 3, 4, 5 or 6 (preferably 4, 5 or 6). 2-m "-alkyl", provided that at least two carbon atoms of the group are bonded to each other by a triple bond.

[0086] Term "C" 3-k -cycloalkyl (where k is an integer selected from 3, 4, 5, 7 or 8 (preferably 4, 5 or 6), alone or in combination with other groups) represents a cyclic, saturated, unbranched hydrocarbon group having 3 to k carbon atoms. For example, the term C 3-7 -Cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.

[0087] The term "halogen," added to an "alkyl," "alkylene," or "cycloalkyl" group (saturated or unsaturated), defines an alkyl, alkylene, or cycloalkyl group in which one or more hydrogen atoms are replaced by a halogen atom selected from fluorine, chlorine, or bromine (preferably fluorine and chlorine, particularly fluorine). Examples include: H2FC-, HF2C-, F3C-.

[0088] The term "mono-heteroaryl ring" refers to a monocyclic aromatic ring system containing one or more heteroatoms selected from N, O or S and consisting of 5 to 6 ring atoms.

[0089] The term "monohyroaryl ring" is intended to include all possible isomers.

[0090] Therefore, the term "monohyaryl ring" includes the following exemplary structures (since each form can optionally be covalently linked to any atom as long as the appropriate chemical valence is maintained, it is not described as a group): .

[0091] The term "fused bicyclic heteroaryl ring" refers to a bicyclic aromatic ring system containing one or more heteroatoms selected from N, O, or S and consisting of 9 to 10 ring atoms. The term "fused bicyclic heteroaryl ring" is intended to include all possible isomers. Therefore, the term "bicyclic heteroaryl ring" includes the following exemplary structures (since each form can optionally be covalently bonded to any atom while maintaining appropriate valence, it is not described as a group): .

[0092] The term pyridyl refers to a group on the following ring: .

[0093] The term pyrimidinyl refers to a group on the following ring: .

[0094] The term pyrazolyl refers to a group on the following ring: .

[0095] The term thiophene group refers to a group that forms a ring with the following ring: .

[0096] The term oxazolyl refers to a group on the following ring: .

[0097] The term isoxazolyl refers to a group on the following ring: .

[0098] The term imidazo[1,2-a]pyrimidinyl refers to a group on the following ring: .

[0099] The term 2H-pyrazolo[3,4-b]pyridyl refers to a group on the following ring: .

[0100] The term 1H-[1,2,3]triazolo[4,5-b]pyridyl refers to a group on the following ring: .

[0101] The term imidazo[1,2-a]pyrimidinyl refers to a group on the following ring: .

[0102] The term pyrazolo[1,5-b]pyridazinyl refers to a group on the following ring: .

[0103] The aforementioned terms can be used independently and repeatedly in the formula or group and in the definition of each having one of the above meanings.

[0104] Biological assay Evaluation of the inhibitory activity of QPCT and QPCTL Assay A: Biochemical QPCT and QPCTL activity assays The activity of the compounds of this invention can be confirmed using the following biochemical enzyme activity assays: The QPCT or QPCTL-dependent conversion of the N-terminal glutamine to pyroglutamate of CD47 was monitored by MALDI-TOF MS. The test compound was dissolved in 100% DMSO and serially diluted to clear 1,536-well microtiter plates. The enzyme reaction was established in assay buffer containing 20 mM Tris pH 7.5, 0.1 mM TCEP, 0.01% BSA, and 0.001% Tween 20. 2.5 µL of 2x concentrated QPCTL (internal) or QPCT (Origine #TP700028) enzyme was added to each well in assay buffer (0.5 nM final concentration, columns 1-23) or ordinary assay buffer (column 24). The plates were incubated at 24°C in a humidified incubator for 10 min. Subsequently, 2.5 µL of CD47 peptide substrate substitute (…) was added to each well. 19 QLLFNKTKSVEFTFC 33 (Final concentration: 10 µM for QPCTL / 20 µM for QPCT). Mix the plate at 1,000 rpm for 30 sec and then incubate at 24 °C in a humidified incubator for 40 min. After incubation, add 1 µL of an internal standard peptide containing a stable isotope label. 19 [Pyr]LLFN(K)TKSVEFTFC 33 The enzyme reaction was terminated using reagents including (final concentration 4.0 µM) and SEN177 (final concentration 10 µM). The plate was sealed with foil, mixed at 1,000 rpm for 30 s, and stored at room temperature until the preparation of the MALDI target plate. The MALDI target plate was prepared as previously described. Mass spectrometry was obtained using a rapifleX MALDI-TOF / TOF instrument, which tracks the product (…). 19 [Pyr]LLFNKTKSVEFTFC 33 m / z 1,787.9037) and internal standard ( 19 [Pyr]LLFN(K)TKSVEFTFC 33 The signal of the peptide (m / z 1,795.9179) was measured. QPCT or QPCTL activity was monitored by calculating the ratio between the product signal and the internal standard signal, and then normalizing to high (100% activity) and low (0% activity) controls. Compound potency was determined by fitting dose-response data to a four-parameter logistic equation.

[0105] Table 2: Biological data of the compounds of the present invention obtained from determination A.

[0106] Examples Inhibition of QPCTL: IC50 50 [nM]]]> Inhibition of QPCT: IC50 50 [nM]]]> 1 2 3 2 1 1 3 1 4 4 1 1 5 4 9 7 8 12 9 2 4 10 8 30 11 13 48 12 3 4 13 8 24 14 3 11 15 15 58 16 1 1 17 5 5 18 2 1 19 1 1 20 2 3 21 2 1 22 2 2 23 4 4 24 14 8 25 1 6 26 3 2 27 4 3 28 1 1 29 2 1 30 2 1 31 3 3 32 1 1 33 1 2 Table 3: Biodata of prior art compounds obtained from determination A.

[0107] Prior art References Inhibition of QPCTL: IC50 50 [nM]]]> Inhibition of QPCT: IC50 50 [nM]]]> SEN177 J-S, NCB 2015; P, JBIC 2018 17 79 SEN180 J-S, NCB 2015 39 176 1094 WO 2018 / 178384 111 1707 1095 WO 2018 / 178384 13 260 3 WO 2022 / 086920 52 482 3 alternatives WO 2022 / 086920 837 4691 6 WO 2022 / 086920 1 1 21 CN 114874186 5493 1398 23 CN 114874186 4772 1845 Assay B: SIRPa signaling assay (using Raji or A549 cells) The activity of the compounds of the present invention can be confirmed using the following SIRPα signaling assay, which measures SIRPα linkage induced by CD47 via cell-cell interactions. Two cell types were used independently: the Raji cell line (a lymphoblastoid human cell line derived from B lymphocytes of a patient with Burkitt's lymphoma in 1963) and A549 cells (human alveolar basal epithelial cells of adenocarcinoma).

[0108] Dissolve the test compound in 100% DMSO and serially dilute to white 384-well microtiter cell culture plates (PerkinElmer #60076780 for Raji assays; Greiner #781945 for A549 assays using PDL-coated plates). Add 5000 Raji cells (ATCC #CC86) or 5000 A549 cells (ATCC #CCL-185) to each well in Whole Cell Spread Reagent 30 (DiscoverX 93-0563R30B). Incubate the plates at 37°C, 95% humidity, and 5% CO2 for 48 h. Add 15000 reporter cells (Jurkat PathHunter SIRPαV1, DiscoverX#93-1135C19) to each well and incubate for 5 h at 37°C, 95% humidity, and 5% CO2. Add Bioassay 1 of the PathHunter Bioassay Kit (DiscoverX 93-0001) to each well of the plate using a multichannel pipette, and then incubate at room temperature for 15 min. Then add Assay 2, and then incubate at room temperature for 60 min (in the dark).

[0109] Data analysis was performed using the luminescent signal generated by β-galactosidase in PathHunter reporter cell lines. Luminescence measurements were performed using a Pherastar multi-mode microplate reader. Dose-response curves and IC50 were calculated using a 4-parameter sigmoid dose-response model. 50 data.

[0110] Table 4: Biological data of the compounds of the present invention obtained by determination of B.

[0111] Examples Inhibition of SIRPa signaling induced by Raji cells: IC50 50 [nM]]]> Inhibition of SIRPa signaling induced by A549 cells: IC50 50 [nM]]]> 1 77 41 2 20 40 3 131 40 4 15 17 5 617 60 7 568 200 9 103 39 10 536 283 11 1167 419 12 150 45 13 596 358 14 673 202 15 296 410 16 27 32 17 274 71 18 44 16 19 20 6 20 83 29 21 16 11 22 117 51 23 158 46 24 981 184 25 150 78 26 148 101 27 106 52 28 70 53 29 64 27 30 87 17 31 54 31 32 72 21 33 53 30 Table 5: Biological data of prior art compounds obtained from determination of B.

[0112] Prior art References Inhibition of SIRPa signaling induced by Raji cells: IC50 50 [nM]]]> Inhibition of SIRPa signaling induced by A549 cells: IC50 50 [nM]]]> SEN177 J-S, NCB 2015; P, JBIC 2018 1365 214 SEN180 J-S, NCB 2015 2973 868 1094 WO 2018 / 178384 3240 3059 1095 WO 2018 / 178384 2203 161 3 WO 2022 / 086920 2736 921 3 alternatives WO 2022 / 086920 >10000 >10000 6 WO 2022 / 086920 8 12 21 CN 114874186 >10000 >10000 23 CN 114874186 >10000 >10000 Evaluation of penetration Determining the permeability of C:CACO-2 cells Caco-2 cells (1 - 2 x 10⁻⁶) 5 Cells / 1 cm 2 The inoculated areas were placed on filter inserts (CostarTranswell polycarbonate or PET filters, 0.4 μm pore size) and cultured (DMEM) for 10 to 25 days.

[0113] Dissolve the compound in a suitable solvent (e.g., DMSO, 1–20 mM stock solution). Dilute the stock solution with HTP-4 buffer (128.13 mM NaCl, 5.36 mM KCl, 1 mM MgSO4, 1.8 mM CaCl2, 4.17 mM NaHCO3, 1.19 mM Na2HPO4x7H2O, 0.41 mM NaH2PO4xH2O, 15 mM HEPES, 20 mM glucose, 0.25% BSA, pH 7.2) to prepare transport solutions (0.1–300 μM compound, final DMSO <= 0.5%). Apply the transport solution (TL) to the donor side, either at the top or outside the substrate, to measure AB or BA permeability (3 filters replicate). Collect samples from the donor side at the beginning and end of the experiment and also from the receiver side at different time intervals greater than 2 hours for concentration determination by HPLC-MS / MS or scintillation counting. Replace the sampled receiver volume with fresh receiver solution.

[0114] Expiratory Ratio (ER) = Permeability BA / Permeability AB Table 7: Biological data of the compounds of the present invention obtained from C.

[0115]

[0116] Table 8: Biological data of prior art compounds obtained from determination of C.

[0117]

[0118] Evaluation of microsomal clearance rate Microsomal clearance: Metabolic degradation of the test compound was determined using aggregated liver microsomes from different species at 37°C. At each time point, a final incubation volume of 60 μL contained 0.1 M TRIS buffer (pH 7.6), 5 mM magnesium chloride, microsomal protein (1 mg / mL for humans and dogs, 0.5 mg / mL for other species), and a final concentration of 1 μM of the test compound. A short pre-incubation was then performed at 37°C, initiated by the addition of β-nicotinamide adenine dinucleotide phosphate (reduced form) (NADPH, 1 mM), and terminated at different time points by transferring aliquots to the solvent. After centrifugation (10000 g, 5 min), aliquots of the supernatant were determined by LC-MS / MS to determine the amount of the parent compound. Half-life was determined by the slope of a semi-logarithmic plot of concentration-time characteristics.

[0119] The intrinsic clearance rate (CL_INTRINSIC) can be calculated by taking into account the amount of protein in culture: Intrinsic clearance rate [µl / min / mg protein] = (Ln 2 / (half-life [min] * protein content [mg / ml])) * 1000 Intrinsic clearance rate (CL_INTRINSIC_INVIVO) [ml / min / kg] = (Intrinsic clearance rate [µL / min / mg protein] × MPPGL [mg protein / g liver] × liver factor [g / kg body weight]) / 1000 Qh [%] = clearance rate [ml / min / kg] / liver blood flow [ml / min / kg] Human hepatocyte count: 120x10e 6 Cells / g liver Human liver factor: 25.7 g / kg body weight Human blood flow: 21 ml / (min x kg) Evaluation of hepatocyte clearance rate Hepatocyte clearance The metabolic degradation of the test compounds was determined in human hepatocyte suspensions. After recovery from cryopreservation, human hepatocytes were diluted in Dulbecco's modified Eagle medium (supplemented with 3.5 µg glucagon / 500 mL, 2.5 mg insulin / 500 mL, 3.75 mg hydrocortisone / 500 mL, and 5% human serum) to obtain 1.0 x 10⁻⁶ cells / mL. 6 Final cell density of cells / mL.

[0120] Subsequently, the cells were pre-incubated in a cell culture incubator (37°C, 10% CO2) for 30 minutes, and the test compound solution was added to the hepatocyte suspension to obtain a final test compound concentration of 1 µM and a final DMSO concentration of 0.05%.

[0121] Cell suspensions were incubated at 37°C (cell culture incubator, horizontal shaker) and samples were removed from the incubator after 0, 0.5, 1, 2, 4, and 6 hours. Samples were quenched with acetonitrile (containing an internal standard) and precipitated by centrifugation. The supernatant was transferred to 96-well deep-well plates for determination of the parent compound's descent by HPLC-MS / MS.

[0122] The percentage of remaining test compounds was calculated using the peak area ratio (test compound / internal standard) at each incubation time point relative to the peak area ratio at time 0. A curve was plotted of the logarithmically transformed data against incubation time, and the in vitro half-life (T1 / 2) was estimated using the absolute value of the slope obtained through linear regression analysis.

[0123] Autologous in vitro T1 / 2 calculation of in vitro intrinsic clearance (CLint) using 120x10 6 Hepatocyte count per cell / g liver, 25.7 g liver / kg of human liver per serving body weight, and in vitro culture parameters were scaled to whole liver using the following equation: Intrinsic clearance rate [mL / min / kg] = (Intrinsic clearance rate [µL / min / 10]) 6 [Number of cells] × Hepatocyte degree

[10] 6 [cells / g liver] × liver factor [g / kg body weight] / 1000 Based on a well-stirred liver model and considering a mean hepatic blood flow (QH) of 20.7 mL / min / kg, the hepatic blood clearance rate (CL) is predicted as follows: Clearance rate [mL / min / kg] = Intrinsic clearance rate [mL / min / kg] × Hepatic blood flow [mL / min / kg] / (Intrinsic clearance rate [mL / min / kg] + Hepatic blood flow [mL / min / kg]) The results are expressed as a percentage of hepatic blood flow: QH [%] = clearance rate [mL / min / kg] / hepatic blood flow [mL / min / kg] Evaluation of plasma protein binding The approximate in vitro fractional binding of test compounds to plasma proteins was determined using equilibrium dialysis technology and Dianorm Teflon dialysis cells (micro 0.2). Each dialysis cell consisted of a donor chamber and a recipient chamber, separated by an ultrathin semipermeable membrane with a 5 kDa molecular weight cutoff. A stock solution of each test compound was prepared in DMSO at 1 mM and serially diluted to obtain a final test concentration of 1 µM. A dialysis solution (supplemented with NaEDTA as an anticoagulant) was then prepared in plasma, and 200 µl of the test compound dialysis solution from an aliquot of the sample was allocated to the donor (plasma) chamber. 200 µl of dialysis buffer (100 mM potassium phosphate (pH 7.4), supplemented with up to 4.7% glucosamine) from an aliquot of the sample was allocated to the buffer (recipient) chamber. The mixture was incubated at 37°C with rotation for 2 hours to establish equilibrium.

[0124] At the end of the dialysis period, aliquots of the samples obtained from the donor and recipient chambers were transferred to reaction tubes and processed for HPLC-MS / MS analysis. The analyte concentrations in the aliquots were quantified using HPLC-MS / MS against a calibration curve.

[0125] Calculate the percentage of combination using the following formula: Binding % = (Plasma concentration - Buffer concentration / Plasma concentration) × 100 Evaluation of solubility A saturated solution is prepared on a well plate (the type of which depends on the robot) by adding an appropriate volume of a chosen aqueous medium (typically 0.25–1.5 ml) to each well containing a known amount of solid drug substance (typically in the range of 0.5–5.0 mg). The wells are agitated or stirred for a predetermined time period (typically in the range of 2–24 h) and then filtered using a suitable filter membrane (typically a PTFE membrane with a pore size of 0.45 µm). Membrane absorption is avoided by discarding the first few drops of filtrate. The amount of dissolved drug substance is determined by UV spectroscopy. Furthermore, the pH of the saturated aqueous solution is measured using a glass electrode pH meter.

[0126] Evaluation of human hepatocyte metabolism in vitro The metabolic pathways of the compounds were investigated using primary human hepatocytes in suspension. After recovery from cryopreservation, human hepatocytes were cultured in Duchenne modified Eagle medium containing 5% human serum supplemented with 3.5 µg glucagon / 500 ml, 2.5 mg insulin / 500 ml, and 3.75 mg / 500 ml hydrocortisone.

[0127] Subsequently, the cells were pre-cultured in a cell culture incubator (37°C, 10% CO2) for 30 minutes, and the test compound solution was added to the hepatocyte suspension to obtain 1.0*102 6 Up to 4.0*10 6 The final cell density was determined by the number of cells per ml (depending on the metabolic turnover rate of the compound observed using primary human hepatocytes), the final concentration of the test compound was 10 µM, and the final concentration of DMSO was 0.05%.

[0128] Cells were cultured for 6 hours in a cell culture incubator mounted on a horizontal shaker, and samples were removed from the culture at 0, 0.5, 1, 2, 4, or 6 hours (depending on the metabolic turnover rate). Samples were quenched with acetonitrile and precipitated by centrifugation. The supernatant was transferred to a 96-well deep-well plate, evaporated and resuspended under nitrogen, and then subjected to bioanalysis by liquid chromatography-high resolution mass spectrometry for identification of putative metabolites.

[0129] Based on Fourier transform MS n The data were initially structured. Metabolites were reported as a percentage of the parent cell with a threshold of ≥4% in human hepatocyte culture.

[0130] Evaluation of pharmacokinetic properties The test compound was administered intravenously or orally to each test species. Blood samples were collected at several time points after administration of the test compound for anticoagulation and centrifugation.

[0131] Quantify the concentration of the analyte—the administered compound and / or metabolite—in plasma samples. Calculate PK parameters using a compartmentless method. Normalize AUC and Cmax to a dose of 1 μmol / kg.

[0132] Treatment This invention relates to compounds of general formula (I) that can be used for the prevention and / or treatment of diseases and / or conditions associated with or regulated by QPCT / L activity, including but not limited to the treatment and / or prevention of cancer, fibrotic diseases, neurodegenerative diseases, atherosclerosis, infectious diseases, and chronic kidney disease.

[0133] Compounds of general formula (I) can be used for the prevention and / or treatment of the following diseases: (1) Pulmonary fibrotic diseases, such as pneumonia or interstitial pneumonia associated with collagen diseases (e.g., lupus erythematosus, systemic scleroderma, rheumatoid arthritis, polymyositis, and dermatomyositis); idiopathic interstitial pneumonia (e.g., pulmonary fibrosis (IPF)); nonspecific interstitial pneumonia; interstitial lung disease associated with respiratory bronchiolitis, desquamative interstitial pneumonia, cryptogenic organizing pneumonia, acute interstitial pneumonia, and lymphocytic interstitial pneumonia; lymphangioleiomyomatosis, pulmonary alveolar proteinosis, Langerhans cell histiocytosis, and pleural parenchymal elastic fiber hyperplasia; interstitial lung diseases of known etiology, such as those caused by occupational exposure (e.g., asbestosis, silicosis, miner's lung (coal dust), farmer's lung (hay and mold), pigeon fanciers' lung). Interstitial pneumonia caused by (birds) or other occupational airborne triggers (e.g., metal dust or mycobacteria) or by treatment (e.g., radiation, methotrexate, amiodarone, nitrofurantoin, or chemotherapy agents) or granulomatous diseases (e.g., granulomatous polyangiitis, Churg-Strauss syndrome, sarcoidosis, hypersensitivity pneumonitis); or by various causes (e.g., inhalation, inhalation of toxic gases, vapors, bronchitis, or pneumonia) or by heart failure, X-rays, radiation, chemotherapy, burke's sarcoma (M. boeck) or sarcoidosis, granulomatous disease, cystic fibrosis, or myxoviscosity or α-I-antitrypsin deficiency; (2) Other fibrotic diseases, such as hepatic bridging fibrosis, cirrhosis, non-alcoholic steatohepatitis (NASH), atrial fibrosis, endocardial fibrosis, old myocardial infarction, glial scars, arteriosclerosis, joint fibrosis, Dupuytren's contracture, keloids, scleroderma / systemic sclerosis, mediastinal fibrosis, myelofibrosis, Peyronie's disease, renal systemic fibrosis, retroperitoneal fibrosis, adhesive capsulitis; spontaneous acute exacerbations of pulmonary fibrosis and progressive pulmonary fibrosis or acute exacerbations of pulmonary fibrosis and progressive pulmonary fibrosis caused by infection, microaspiration, surgical lung biopsy, surgical resection, bronchoscopy (BAL, cryobiopsy), air pollution, previous exacerbations and drug-induced pulmonary fibrosis and progressive pulmonary fibrosis; (3) Leukemia, Acute Myeloid Leukemia (AML), Acute Promyelocytic Leukemia (APL), Chronic Myeloid Leukemia (CML), Acute Lymphoblastic Leukemia (ALL), T-cell Acute Lymphoblastic Leukemia (T-ALL), Lymphoma, B-cell Lymphoma, T-cell Lymphoma, Hodgkin's Disease, Non-Hodgkin's Lymphoma (NHL), Pilocytic Lymphoma, Burkitt Lymphoma, Multiple Myeloma (MM), Myelodysplastic Syndrome, Solid Tumors, Lung Cancer, Adenocarcinoma, Small Cell Lung Cancer (SCLC), Non-Small Cell Lung Cancer (NSCLC), Mediastinal Cancer, Peritoneal Cancer, Mesothelioma, Gastrointestinal Cancer, Gastric Cancer, Stomach Cancer, Intestinal Cancer, Small Intestinal Cancer, Large Intestinal Cancer, Colon Cancer, Colon Adenocarcinoma, Colon Adenoma Adenoma, rectal cancer, colorectal cancer, leiomyosarcoma, breast cancer, gynecological cancer, genitourinary cancer, ovarian cancer, endometrial cancer, cervical cancer, prostate cancer, testicular cancer, seminoma, teratoma, liver cancer, kidney cancer, bladder cancer, urethral cancer, bile duct cancer, pancreatic cancer, exocrine pancreatic cancer, esophageal cancer, nasopharyngeal cancer, head and neck squamous cell carcinoma (HNSCC), skin cancer, squamous cell carcinoma, Kaposi's sarcoma, melanoma, malignant melanoma, xeroderma pigmentosum, keratoacanthoma, bone cancer, bone sarcoma, osteosarcoma, rhabdomyosarcoma, fibrosarcoma, thyroid cancer, thyroid follicular carcinoma, adrenal cancer, nervous system cancer, brain cancer, astrocytoma, neuroblastoma, glioma, schwannoma, glioblastoma or sarcoma, gastrointestinal cancer, gastric cancer, stomach cancer. Cancer, esophageal cancer, head and neck squamous cell carcinoma (HNSCC), breast cancer, colorectal cancer, colon cancer, colon cancer, colon adenocarcinoma, colon adenoma, rectal cancer, ovarian cancer, pancreatic cancer, exocrine pancreatic cancer, leukemia, acute myeloid leukemia (AML), myelodysplastic syndrome, lymphoma, B-cell lymphoma, non-Hodgkin lymphoma (NHL), urothelial carcinoma or peritoneal cancer; (4) Inflammatory, autoimmune, or allergic diseases and conditions, such as asthma, childhood asthma, allergic bronchitis, alveolitis, hyperresponsive airways, allergic conjunctivitis, bronchiectasis, adult respiratory distress syndrome, bronchial and pulmonary edema, bronchitis or pneumonia, non-allergic asthma, chronic obstructive pulmonary disease (COPD), acute bronchitis, chronic bronchitis, emphysema; autoimmune diseases, such as rheumatoid arthritis, Graves' disease, Hugh Grant's syndrome, psoriatic arthritis, multiple sclerosis, systemic lupus erythematosus; inflammatory bowel diseases, such as Crohn's disease and ulcerative colitis, scleroderma; psoriasis (including T-cell mediated psoriasis) and inflammatory skin diseases (such as dermatitis, eczema, atopic dermatitis, allergic contact dermatitis, urticaria); vasculitis (such as necrotizing, cutaneous, and allergic vasculitis) or erythema nodosum; (5) Neurodegenerative diseases, such as amyotrophic lateral sclerosis, multiple sclerosis, Parkinson's disease, Alzheimer's disease, Huntington's disease, multiple system atrophy, or prion diseases.

[0134] Therefore, in another embodiment, the present invention relates to a compound of general formula (I) or a pharmaceutically acceptable salt thereof, which is used as a medicine.

[0135] Furthermore, the present invention relates to the use of compounds of general formula (I) for the prevention and / or treatment of diseases and / or conditions associated with or regulated by QPCT / L activity.

[0136] Furthermore, the present invention relates to the use of compounds of general formula (I) or pharmaceutically acceptable salts thereof or pharmaceutical compositions thereof for the treatment and / or prevention of cancer, fibrotic diseases, neurodegenerative diseases, atherosclerosis, infectious diseases, and chronic kidney disease.

[0137] Furthermore, the present invention relates to the use of compounds of general formula (I) or pharmaceutically acceptable salts thereof or pharmaceutical compositions thereof for the treatment and / or prevention of the following diseases: (1) pulmonary fibrosis diseases, such as pneumonia or interstitial pneumonia associated with collagen diseases (e.g., lupus erythematosus, systemic scleroderma, rheumatoid arthritis, polymyositis and dermatomyositis); idiopathic interstitial pneumonia (e.g. pulmonary fibrosis (IPF)); nonspecific interstitial pneumonia, interstitial lung disease associated with respiratory bronchiolitis, desquamative interstitial pneumonia, cryptogenic organizing pneumonia, acute interstitial pneumonia and lymphocytic interstitial pneumonia; lymphangioleiomyomatosis, pulmonary alveolar proteinosis, Langerhans cell histiocytosis, pleural parenchymal elastic fibers Interstitial lung disease of known etiology, such as interstitial pneumonia caused by occupational exposure (e.g., asbestosis, silicosis, miner's lung (coal dust), farmer's lung (hay and mold), pigeon fancier's lung (birds)) or other occupational airborne triggers (e.g., metal dust or mycobacteria) or by treatment (e.g., radiation, methotrexate, amiodarone, nitrofurantoin or chemotherapeutic agents) or granulomatous diseases (e.g., granulomatous polyangiitis, Churg-Strauss syndrome, sarcoidosis, allergic pneumonia); or interstitial pneumonia caused by various causes (e.g., inhalation of toxic gases, vapors, bronchitis or pneumonia) or by heart failure, X-rays, radiation, chemotherapy, Burke's sarcoma (M. boeck) or sarcoidosis, granulomatous disease, cystic fibrosis or myxoviscosity or α-I-antitrypsin deficiency; (2) Other fibrotic diseases, such as hepatic bridging fibrosis, cirrhosis, non-alcoholic steatohepatitis (NASH), atrial fibrosis, endocardial fibrosis, old myocardial infarction, glial scars, arteriosclerosis, joint fibrosis, Dupuytren's contracture, keloids, scleroderma / systemic sclerosis, mediastinal fibrosis, myelofibrosis, Peroni's disease, renal systemic fibrosis, retroperitoneal fibrosis, adhesive capsulitis; spontaneous acute exacerbations of pulmonary fibrosis and progressive pulmonary fibrosis or acute exacerbations of pulmonary fibrosis and progressive pulmonary fibrosis caused by infection, microaspiration, surgical lung biopsy, surgical resection, bronchoscopy (BAL, cryobiopsy), air pollution, previous exacerbations and drug-induced pulmonary fibrosis and progressive pulmonary fibrosis; (3) Leukemia, Acute Myeloid Leukemia (AML), Acute Promyelocytic Leukemia (APL), Chronic Myeloid Leukemia (CML), Acute Lymphoblastic Leukemia (ALL), T-cell Acute Lymphoblastic Leukemia (T-ALL), Lymphoma, B-cell Lymphoma, T-cell Lymphoma, Hodgkin's Disease, Non-Hodgkin's Lymphoma (NHL), Pilocytic Lymphoma, Burkitt Lymphoma, Multiple Myeloma (MM), Myelodysplastic Syndrome, Solid Tumors, Lung Cancer, Adenocarcinoma, Small Cell Lung Cancer (SCLC), Non-Small Cell Lung Cancer (NSCLC), Mediastinal Cancer, Peritoneal Cancer, Mesothelioma, Gastrointestinal Cancer, Gastric Cancer, Stomach Cancer, Intestinal Cancer, Small Intestinal Cancer, Large Intestinal Cancer, Colon Cancer, Colon Adenocarcinoma, Colon Adenoma Adenoma, rectal cancer, colorectal cancer, leiomyosarcoma, breast cancer, gynecological cancer, genitourinary cancer, ovarian cancer, endometrial cancer, cervical cancer, prostate cancer, testicular cancer, seminoma, teratoma, liver cancer, kidney cancer, bladder cancer, urethral cancer, bile duct cancer, pancreatic cancer, exocrine pancreatic cancer, esophageal cancer, nasopharyngeal cancer, head and neck squamous cell carcinoma (HNSCC), skin cancer, squamous cell carcinoma, Kaposi's sarcoma, melanoma, malignant melanoma, xeroderma pigmentosum, keratoacanthoma, bone cancer, bone sarcoma, osteosarcoma, rhabdomyosarcoma, fibrosarcoma, thyroid cancer, thyroid follicular carcinoma, adrenal cancer, nervous system cancer, brain cancer, astrocytoma, neuroblastoma, glioma, schwannoma, glioblastoma or sarcoma, gastrointestinal cancer, gastric cancer, stomach cancer. Cancer, esophageal cancer, head and neck squamous cell carcinoma (HNSCC), breast cancer, colorectal cancer, colon cancer, colon cancer, colon adenocarcinoma, colon adenoma, rectal cancer, ovarian cancer, pancreatic cancer, exocrine pancreatic cancer, leukemia, acute myeloid leukemia (AML), myelodysplastic syndrome, lymphoma, B-cell lymphoma, non-Hodgkin lymphoma (NHL), urothelial carcinoma or peritoneal cancer; (4) Inflammatory, autoimmune, or allergic diseases and conditions, such as asthma, childhood asthma, allergic bronchitis, alveolitis, hyperresponsive airways, allergic conjunctivitis, bronchiectasis, adult respiratory distress syndrome, bronchial and pulmonary edema, bronchitis or pneumonia, non-allergic asthma, chronic obstructive pulmonary disease (COPD), acute bronchitis, chronic bronchitis, emphysema; autoimmune diseases, such as rheumatoid arthritis, Graves' disease, Hugh Grant's syndrome, psoriatic arthritis, multiple sclerosis, systemic lupus erythematosus; inflammatory bowel diseases, such as Crohn's disease and ulcerative colitis, scleroderma; psoriasis (including T-cell mediated psoriasis) and inflammatory skin diseases (such as dermatitis, eczema, atopic dermatitis, allergic contact dermatitis, urticaria); vasculitis (such as necrotizing, cutaneous, and allergic vasculitis) or erythema nodosum; (5) Neurodegenerative diseases, such as amyotrophic lateral sclerosis, multiple sclerosis, Parkinson's disease, Alzheimer's disease, Huntington's disease, multiple system atrophy, or prion diseases.

[0138] In another aspect, the present invention relates to compounds of general formula (I) or pharmaceutically acceptable salts thereof or pharmaceutical compositions thereof for the treatment and / or prevention of the aforementioned diseases and conditions.

[0139] In another aspect, the present invention relates to compounds of general formula (I) or pharmaceutically acceptable salts thereof or pharmaceutical compositions thereof for the preparation of medicaments for the treatment and / or prevention of the aforementioned diseases and conditions.

[0140] In another aspect of the invention, the invention relates to a method for treating or preventing the aforementioned diseases and conditions, comprising administering to a person an effective amount of a compound of general formula (I) or a pharmaceutically acceptable salt thereof or a pharmaceutical composition thereof.

[0141] Combination therapy The compounds of the present invention may be further combined with one or more, preferably one, other therapeutic agents. According to one embodiment, the other therapeutic agents are selected from those that can be used to treat diseases or conditions as described above (specifically, diseases associated with cancer, fibrotic diseases, Alzheimer's disease, atherosclerosis, infectious diseases, chronic kidney disease, and autoimmune diseases).

[0142] Other therapeutic agents suitable for such combinations specifically include, for example, those that enhance the therapeutic effect of one or more active substances on one of the mentioned indications and / or allow for a reduction in the dosage of one or more active substances.

[0143] Therefore, the compounds of the present invention can be combined with one or more other therapeutic agents selected from: chemotherapy, targeted cancer therapy, cancer immunotherapy, radiation, antifibrotic agents, antitussives, anti-inflammatory agents, anti-atopic dermatitis agents, and bronchodilators.

[0144] Chemotherapy is a class of cancer treatments that use one or more chemical anticancer drugs (such as cell-inhibiting or cytotoxic substances, cell proliferation inhibitors, anti-angiogenic substances, etc.). Examples include folic acid (leucovorin), 5-fluorouracil, irinotecan, oxaliplatin, cisplatin, azacytidine, gemcitabine, alkylating agents, antimitotic agents, taxanes, and other existing or standard therapeutic compounds.

[0145] Targeted therapy is a class of cancer treatments that use drugs to target specific genes and proteins that help cancer cells survive and grow. Targeted therapies include drugs such as growth factors (e.g., platelet-derived growth factor (PDGF), fibroblast growth factor (FGF), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), insulin-like growth factor (IGF), human epidermal growth factor (HER, such as HER2, HER3, HER4, and hepatocyte growth factor), tyrosine kinases, KRAS, BRAF, BCR-ABL, mTOR, cyclin-dependent kinase inhibitors, or MDM2 inhibitors.

[0146] Cancer immunotherapy is a class of therapies that use substances to stimulate or suppress the immune system to help the body fight cancer. Cancer immunotherapy includes therapeutic antibodies, such as anti-Her2 antibodies, anti-EGFR antibodies, and anti-PDGFR antibodies; and anti-GD2 (ganglioside G2) antibodies. Examples include dinutuximab, olalatumab, trastuzumab, pertuzumab, ertumaxomab, cetuximab, necitumumab, nimotuzumab, panitumumab, or rituximab. Cancer immunotherapy also includes therapeutic antibodies that act as checkpoint inhibitors, such as anti-PD1, anti-PD-L1 antibodies, or CTLA4 inhibitors. Examples include atezolizumab, avelumab, durvalumab, ipilimumab, nivolumab, or pembrolizumab. Cancer immunotherapy also includes drugs that target (inhibit) the CD47-SIRPα signaling axis, such as drugs that bind to CD47 or SIRPα. Non-limiting examples include antibodies (e.g., anti-CD47 antibodies and anti-SIRPα antibodies) and recombinant Fc fusion proteins (e.g., CD47-Fc and SIRPα-Fc). Cancer immunotherapy also includes STING targets or T-cell connectors (e.g., belintoomeb).

[0147] Antifibrotic agents include, for example, nintedanib, pirfenidone, phosphodiesterase-IV (PDE4) inhibitors (e.g., roflumilast or specific PDE4b inhibitors such as BI1015550), autotaxin inhibitors (e.g., GLPG-1690 or BBT-877); connective tissue growth factor (CTGF) blocking antibodies (e.g., pamrevlumab); B-cell activating factor receptor (BAFF-R) blocking antibodies (e.g., lanalumab), α-V / β-6 blocking inhibitors (e.g., BG-00011 / STX-100), and recombinant pentamin-2. (PTX-2) (e.g., PRM-151); c-Jun-N-terminal kinase (JNK) inhibitors (e.g., CC-90001); galactolectin-3 inhibitors (e.g., TD-139); G protein-coupled receptor 84 (GPR84) inhibitors; G protein-coupled receptor 84 / G protein-coupled receptor 40 dual inhibitors (e.g., PBI-4050); Rho-associated coiled-coil protein kinase 2 (ROCK2) inhibitors (e.g., KD-025); heat shock protein 47 (HSP47) small interfering RNA. (e.g., BMS-986263 / ND-L02-s0201); Wnt pathway inhibitors (e.g., SM-04646); LD4 / PDE3 / 4 inhibitors (e.g., Tipelukast); recombinant immunoregulatory domains of histyl-tRNA synthetase (HARS) (e.g., ATYR-1923), prostaglandin synthase inhibitors (e.g., ZL-2102 / SAR-191801); 15-hydroxy-eicosapentaenoic acid (15-HEPE, e.g., DS-102); lysyl oxidase-like 2 (LOXL2) inhibitors (e.g., PAT-1251), PXS-5382 / PXS-5338; dual inhibitors of phosphatidylinositol 3-kinase (PI3K) / mammalian target of rapamycin (mTOR) (e.g., HEC-68498); calpases inhibitors (e.g., BLD-2660); mitogen-activated protein kinase kinase kinase (MAP3K19) inhibitors (e.g., MG-S-2525); chitosanase inhibitors (e.g., OATD-01); mitogen-activated protein kinase-activated protein kinase 2 (MAPKAPK2) inhibitors (e.g., MMI-0100); transforming growth factor β1 (TGF-β1) small interfering RNA (e.g., TRKZSO / BNC-1021); or lysophosphatidylcholine receptor antagonists (e.g., BMS986278).

[0148] The dosage of the combined partner mentioned above is typically 1 / 5 to 1 / 1 of the lowest recommended dose.

[0149] Therefore, in another aspect, the present invention relates to the use of the compounds of the invention in combination with one or more other therapeutic agents described above and below for the treatment of diseases or conditions that may be affected by or mediated by QPCT / L, particularly those as described above and below.

[0150] In another aspect, the present invention relates to a method for treating a disease or condition in a patient that is affected by QPCT / L inhibition, comprising the step of administering to a patient requiring such treatment a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof in combination with a therapeutically effective amount of one or more other therapeutic agents.

[0151] In another aspect, the present invention relates to the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in combination with one or more other therapeutic agents for the treatment of a disease or condition in patients in need of which is affected by QPCT / L inhibition.

[0152] In another aspect, the present invention relates to a method for treating a patient with a disease or condition mediated by QPCT / L activity, comprising the step of administering to a patient (preferred) requiring such treatment a therapeutically effective amount of the compound of the present invention in combination with a therapeutically effective amount of one or more other therapeutic agents described above and below.

[0153] The compounds of this invention can be used in combination with other therapeutic agents simultaneously or at staggered times.

[0154] The compounds of the present invention and one or more other therapeutic agents may be contained together in a single formulation (e.g., tablets or capsules) or separately in two identical or different formulations (e.g., in the form of so-called kits of parts).

[0155] Therefore, in another aspect, the present invention relates to pharmaceutical compositions comprising the compounds of the present invention and one or more other therapeutic agents described above and below, and optionally one or more inert carriers and / or diluents.

[0156] Other features and advantages of the invention will become apparent from the following more detailed embodiments, which illustrate the principles of the invention by way of example.

[0157] preparation The compounds and intermediates of the present invention can be obtained using synthetic methods known to those skilled in the art and described in the organic synthesis literature. Preferably, the compounds are obtained using a preparation method similar to that explained more fully below (specifically as described in the experimental section). In some cases, the order of the reaction steps may be varied. Variations of reaction methods known to those skilled in the art but not described in detail herein may also be used.

[0158] Those skilled in the art will understand the general methods for preparing the compounds of the present invention after studying the following schemes. Any functional group of the starting material or intermediate can be protected using conventional protecting groups. These protecting groups can be further cleaved at appropriate stages within the reaction sequence using methods well known to those skilled in the art.

[0159] The compounds of this invention are prepared by the synthetic methods described below, wherein the substituents of the general formula have the meanings given above. These methods are intended to illustrate the invention and not to limit its subject matter and the scope of the claimed compounds to these examples. If the preparation of a starting compound is not described, it is commercially available or can be prepared in a manner similar to that of the known compounds or methods described herein. The substances described in the literature are prepared according to the disclosed synthetic methods. Abbreviations are as defined in the Examples section.

[0160] The embodiments can be prepared as shown in Scheme I below.

[0161] Option I:

[0162] In Scheme I, N -Methyltriazolylpiperidine (Z=CH,CF) (A) undergoes nucleophilic aromatic substitution with a heteroaryl fluoride (X=Cl,Br) (B). This reaction is typically carried out at ambient temperature or at elevated temperatures (up to 110 °C) in the presence of a base (e.g., diisopropylethylamine). The intermediate (C) then undergoes Suzuki cross-coupling with a heteroarylboronic acid derivative in the presence of a suitable catalyst (e.g., Pd(dppf)Cl2) and a suitable base at elevated temperatures (e.g., 100 °C) to yield a compound of general formula (I).

[0163] Intermediate I can be prepared as shown in Scheme II below: Option II:

[0164] Compound (A) (where Z = CF) can be prepared from its corresponding piperidinyl ester (D) equipped with a suitable protecting group (PG, e.g., benzyl) by treatment with a suitable hydrazine source (e.g., N2H4*H2O) at an elevated temperature (e.g., 50 °C). The resulting hydrazine (E) is then activated with DMF / DMA at an elevated temperature (e.g., 50 °C) and subsequently treated with methylamine at an elevated temperature (e.g., 90 °C) to give the triazole derivative (F). Compound (A) can be obtained by cleaving the protecting group under suitable conditions (e.g., H2, Pd / C 10%, EtOH).

[0165] Intermediates II.2 to II.6 may be prepared as shown in Scheme III below: Option III:

[0166] In the case of R=H, the intermediate of formula (B) can be prepared starting from the corresponding carboxylic acid (G). In the case of R=Cl, the carboxylic acid (G) can be prepared from the corresponding pyridine (J) by deprotonation with a strong base (e.g., LDA) at low temperature (e.g., -70°C) and quenching with a suitable reagent (e.g., carbon dioxide). At ambient temperature, the carboxylic acid moiety is converted to the corresponding amide (H) using a suitable combination of reagents (e.g., 1,1'-carbonyldiimidazole and ammonia). Subsequently, (H) is treated at ambient temperature with a suitable dehydrating agent (e.g., Burgess reagent) to obtain the compound of formula (B). In the case of R=Me or CF3, the pyridine (J) is deprotonated at low temperature (e.g., -65°C) and quenched with DMF to give the corresponding aldehyde (K). In the case of R=CF3, the aldehyde (K) can be converted to the amide (H) and further to the nitrile (B) at ambient temperature using a suitable reagent (e.g., phenyltrimethylammonium tribromide), as described above for R=H.

[0167] Examples 1 and 2 can be prepared as shown in Scheme IV below: Option IV:

[0168] The pyridinium intermediate (B) (where R=Br and X=Cl) can be prepared according to Scheme III and can be further converted to an enol ether (L) at elevated temperatures (e.g., 70°C) using a suitable coupling agent (e.g., tributyl(1-ethoxyvinyl)stanane) and a suitable catalyst (e.g., Pd(dppf)Cl2). Following reaction with piperidine (A) at a suitable temperature (e.g., 15°C), followed by acidic post-treatment with a suitable acid (e.g., HCl / dioxane), a ketone (M) can be obtained. This ketone can be further converted to a tertiary alcohol (N) upon reaction with a suitable organometallic reagent (e.g., MeMgBr). The intermediate (N) is then subjected to Suzuki cross-coupling with a heteroarylboronic acid derivative at elevated temperatures (e.g., 100°C) in the presence of a suitable catalyst (e.g., Pd(dppf)Cl2) and a suitable base to obtain Example 1 or 2.

[0169] Example preparation The compounds and intermediates thereof of the present invention can be obtained using synthetic methods known to those skilled in the art and described in the organic synthesis literature, such as those described in "Comprehensive Organic Transformations," 2nd edition, Richard C. Larock, John Wiley & Sons, 2010, and "March's Advanced Organic Chemistry," 7th edition, Michael B. Smith, John Wiley & Sons, 2013. Preferably, the compounds are obtained by methods similar to those described more fully below, specifically as described in the Experimental Section. In some cases, the order in which the reaction scheme is carried out can be changed. Variations of these reactions known to those skilled in the art but not described in detail herein can also be used. Those skilled in the art will understand the general methods for preparing the compounds of the present invention after studying the schemes described below. The starting compounds are commercially available or can be prepared by methods described in the literature or herein, or can be prepared in a similar or analogous manner. Any corresponding functional groups in the starting compounds can be protected with conventional protecting groups before carrying out the reaction. These protecting groups can be cleaved again at appropriate stages within the reaction sequence using methods well known to those skilled in the art and described in the literature (e.g., in "Protecting Groups," 3rd edition, Philip J. Kocienski, Thieme, 2005, and "Protective Groups in Organic Synthesis," 4th edition, Peter GM Wuts, Theodora W. Greene, John Wiley & Sons, 2006). The terms "ambient temperature" and "room temperature" are used interchangeably and refer to a temperature of approximately 20°C, for example, between 19°C and 24°C.

[0170] abbreviation: ACN acetonitrile Aq. aqueous solution saline saturated NaCl aqueous solution ℃ degrees Celsius CyH / CH cyclohexane CO2 carbon dioxide conc. concentrated Cs2CO3 cesium carbonate DCM dichloromethane DIPA -diisopropylamine DIPEA -diisopropylethylamine DMF -dimethylformamide DMSO dimethyl sulfoxide ESI-MS electrospray ionization mass spectrometry EtOAc / EA ethyl acetate EtOH ethanol ex example eq equivalents FA formic acid h h [H2O] hour(s) H2O water HCl hydrochloric acid HPLC high performance liquid chromatography [K2CO3] Int. K3PO4 intermediate K2CO3 potassium carbonate KOAc potassium acetate L KOH potassium hydroxide L liter(s) LDA M lithium diisopropylamide LiOH lithium hydroxide M molar MgSO4 MeOH methanol MeTHF methyltetrahydrofuran MgSO4 magnesium sulfate min µL minute(s) [N2] mL milliliter(s) MTBE methyl-tert-butyl ether μL microliter(s) N2 Na2CO3 nitrogen NaHCO3 n-BuLi [CAT] n-butyllithium NH4CI NBS N-bromosuccinimide NCS Na2SO4 N-chlorosuccinimide [PdCl2(PPh3)2] Na2CO3 [Pd(dppf)Cl2] sodium carbonate [Pd(PPh3)4] NaHCO3 sodium bicarbonate NH3 ammonia NH4Cl ammonium chloride NaOH sodium hydroxide Na2SO4 sodium sulfate PdCl2(dppf)·CH2Cl2 PdCl2(dppf)·CH2Cl2 Pd(PPh3)4 SiO2 PE petroleum ether Prep. preparative RP reverse phase RT / rt room temperature (about 20 °C) sat. saturated SFC super critical fluid chromatography silica gel TEA triethylamine TFA trifluoroacetic acid TFAA trifluoroacetic anhydride THF tetrahydrofuran Xphos Pd G3 (2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'- biphenyl)]palladium(II) methanesulfonate Preparation of intermediates Synthesis of intermediate I.1

[0171] 4-Fluoro-4-(hydrazine carbonyl)piperidine-1-carboxylic acid tert-butyl ester In a round-bottom flask, 160 g (0.58 mol) of 4-ethyl 1-tert-butyl 4-fluoropiperidine-1,4-dicarboxylic acid was suspended in 640 mL of ethanol. At ambient temperature, hydrazine hydrate (70.6 mL, 1.16 mol) was added to the mixture. The reaction mixture was heated to 50 °C and stirred for 12 hours. After cooling to ambient temperature, the mixture was concentrated under reduced pressure to give tert-butyl 4-fluoro-4-(hydrazine carbonyl)piperidine-1-carboxylic acid, with a purity of 80%.

[0172] C 11 H 20 FN3O3 (M = 261.3 g / mol) ESI-MS: 284 [M+Na] + Rt (HPLC): 0.62 min (Method A) 4-Fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-carboxylic acid tert-butyl ester In a round-bottom flask, tert-butyl 4-fluoro-4-(hydrazine carbonyl)piperidin-1-carboxylate (135 g, 0.413 mol, 80% purity) was mixed with dioxane (945 mL). At ambient temperature, N , N Dimethylformamide dimethyl acetal (137 mL, 1.03 mol) was added to the mixture. The reaction mixture was heated to 50 °C and stirred for 1 hour. A solution of methylamine (299 g, 30% in EtOH, 2.89 mol) and acetic acid (165 mL, 2.89 mol) was added to the mixture. The resulting reaction mixture was heated to 90 °C and stirred for 11 hours. The mixture was concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, PE / EtOAc gradient 20:1 to 0:1) to give tert-butyl 4-fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-carboxylate.

[0173] C 13 H 21 FN4O2 (M=284.3 g / mol) ESI-MS: 285 [M+H] + Rt (HPLC): 0.77 min (Method A) Intermediate I.1: 4-Fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidine In a round-bottom flask, 90 g (0.32 mol) of 4-fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidine-1-carboxylic acid tert-butyl ester was combined with methanol (90 mL). A solution of HCl (4 M in MeOH, 450 mL, 1.8 mol) was slowly added at ambient temperature. The resulting reaction mixture was stirred at ambient temperature for 12 hours. The desired product was collected by filtration, washed with methanol, and dried to give 4-fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidine hydrochloride.

[0174] The hydrochloride (13.5 g) was added to a methanol solution of ammonia (7 M, 150 mL) and purified by column chromatography (BiotageSNAP Cartridge KP-NH 110 g, gradient DCM / MeOH 4:1 to 7:3) to give the title compound.

[0175] C8H 13 FN4 (M=184.2 g / mol) ESI-MS: 185 [M+H] + Rt (HPLC): 0.20 min (Method B) 4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidine (MFCD09055373, CAS: 297172-18-0) was obtained from a commercial supplier.

[0176] Synthesis of intermediates II.1 and II.2

[0177] 6-Bromo-2-chloro-3-fluoropyridine-4-carboxaldehyde Under an argon atmosphere, 6-bromo-2-chloro-3-fluoropyridine (6.80 g, 30.7 mmol) was added to THF (30 mL), and the resulting mixture was cooled to -75 °C. A solution of lithium diisopropylamino (1 M in THF, 30.7 mL, 30.7 mmol) was added dropwise, and the mixture was stirred at -75 °C for 1 hour. DMF (2.83 mL, 36.8 mmol) was added dropwise. The mixture was stirred at -78 °C for another 2 hours. The reaction mixture was quenched by adding acetic acid (2.64 mL) and diluted with water / salt water 1 / 1 and ethyl acetate, and then heated to ambient temperature. The organic phase was separated, dried over Na2SO4, and concentrated. The residue was purified by column chromatography (SiO2, CyH / EtOAc gradient 1:0 to 1:1) to give 6-bromo-2-chloro-3-fluoropyridine-4-carboxaldehyde.

[0178] C6H2BrClFNO (M=238.4 g / mol) ESI-MS: No quality detected Rt (HPLC): 0.44 min (Method B) 6-Bromo-2-chloro-3-fluoropyridine-4-carboxamide Ammonium acetate (12.0 g, 15.5 mmol) and 6-bromo-2-chloro-3-fluoropyridine-4-carboxaldehyde (3.70 g, 15.5 mmol) were mixed, and acetonitrile (50 mL) was added. Phenylenetrimethylammonium tribromide (12.0 g, 31.0 mmol) was added aliquots, and the resulting reaction mixture was stirred at ambient temperature for 16 hours. The mixture was filtered, and the residue was washed with acetonitrile and purified by column chromatography (dry loading, SiO2, CyH / EtOAc gradient 1:0 to 1:1) to obtain the desired product.

[0179] C6H3BrClFN2O (M=253.5 g / mol) ESI-MS: 251 / 253 [M−H] − Rt (HPLC): 0.41 min (Method B) Intermediate II.2: 6-bromo-2-chloro-3-fluoropyridine-4-carboxynitrile 6-Bromo-2-chloro-3-fluoropyridine-4-carboxamide (510 mg, 2.01 mmol) was suspended in dichloromethane (10 mL), and Burgess reagent (CAS: 29684-56-8, 742 mg, 3.02 mmol) was added at ambient temperature. The resulting reaction mixture was stirred for 16 hours and then purified directly by column chromatography (SiO2, CyH / EtOAc gradient 1:0 to 9:1) to give the title compound.

[0180] C6HBrClFN2 (M=235.4 g / mol) ESI-MS: No quality detected Rt (HPLC): 0.61 min (Method B) 1 H NMR (400 MHz, DMSO-d6) δ ppm = 8.45 (d, J = 3.8 Hz, 1H).

[0181] Intermediate II.1: 2-chloro-6-(1-ethoxyvinyl)-3-fluoropyridine-4-carboxynitrile Under an argon atmosphere, Int II.3 (50.0 mg, 0.21 mmol) and tributyl(1-ethoxyvinyl)stanane (87.9 µL, 0.23 mmol) were suspended in 1,4-dioxane (0.5 mL). Then, [1,1′-bis(diphenylphosphino)-ferrocene]-palladium(II) dichloride (Pd(dppf)Cl2, CAS: 72287-26-4) (15.5 mg, 0.02 mmol) was added, and the mixture was further degassed for 5 min. The reaction mixture was heated at 70 °C for 10 h. The mixture was concentrated, and the residue was purified by column chromatography (dry loading, SiO2, CyH / EtOAc gradient 1:0 to 9:1) to give the title compound.

[0182] C 10 H8ClFN2O (M=226.6 g / mol) ESI-MS: No quality detected Rt (HPLC): 0.80 min (Method B) 1 H NMR (400 MHz, DMSO-d6) δ ppm = 8.08 (d, J = 4.1 Hz, 1H), 5.31 (d, J =2.3 Hz, 1H), 4.62 (d, J = 2.4 Hz, 1H), 3.97 (d, J = 7.0 Hz, 2H), 1.38 (s, 3H).

[0183] Synthesis of intermediate II.4

[0184] 2,5-Dichloro-3-fluoropyridine-4-carboxylic acid At -70°C and under N2, n-BuLi (5.7 mL, 60.2 mmol) was added to a mixture of 2,5-dichloro-3-fluoropyridine (10.00 g, 60.2 mmol) and DIPA (7.5 mL, 66.3 mmol) contained in THF (100 mL). The mixture was stirred at -70°C for 2 hours. Freshly prepared pulverized solid CO2 (13.3 g, 301 mmol) was then added fractionally at -70°C. The resulting mixture was stirred at 25°C for 12 hours. The reaction mixture was quenched by adding NH4Cl (500 mL), diluted with H2O (500 mL), and extracted with EtOAc (2 x 500 mL). The combined organic layers were washed with brine (200 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, PE / EtOAc gradient 1:0 to 0:1) to obtain 2,5-dichloro-3-fluoropyridine-4-carboxylic acid.

[0185] C6H2Cl2FNO2 (M=210.0 g / mol) ESI-MS: 210 [M+H] + Rt (HPLC): 0.26 min (Method D) 2,5-Dichloro-3-fluoropyridine-4-carboxamide Oxaloyl chloride (21.8 g, 171 mmol) was added to a mixture of 2,5-dichloro-3-fluoropyridine-4-carboxylic acid (6.00 g, 28.6 mmol) contained in DCM (60 mL) under N2 conditions at 0 °C. After 2 hours, the reaction mixture was concentrated under reduced pressure, and the residue was suspended in DCM (60 mL). The reaction mixture was then slowly poured into NH3 / H2O (50 mL) and stirred at 25 °C for 12 hours. The reaction mixture was filtered and concentrated under reduced pressure to give 2,5-dichloro-3-fluoropyridine-4-carboxamide.

[0186] C6H3Cl2FN2O (M=209.0 g / mol) ESI-MS: No quality detected Rt (HPLC): 0.40 min (Method D) Intermediate II.4: 2,5-Dichloro-3-fluoropyridine-4-carboxynitrile Under N2 conditions at 0 °C, TEA (3.19 g, 31.6 mmol) and TFAA (3.32 g, 15.8 mmol) were added to a mixture containing 2,5-dichloro-3-fluoropyridine-4-carboxamide (3.00 g, 14.4 mmol) in DCM (30 mL). The mixture was stirred at 10 °C for 2 hours. The reaction mixture was diluted with H2O (100 mL) and extracted with EtOAc (2 x 100 mL). The combined organic layers were washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, PE / EtOAc gradient 1:0 to 3:1) to give the title compound.

[0187] C6HCl2FN2 (M=191.0 g / mol) ESI-MS: 190 [MH] Rt (GCMS): 6.40 min (Method E) Synthesis of Intermediate II.5

[0188] 2-Chloro-3-fluoro-6-(trifluoromethyl)pyridine-4-carboxaldehyde Under an argon atmosphere, 14.2 g (69.7 mmol) of 2-chloro-3-fluoro-6-(trifluoromethyl)pyridine was added to 330 mL of THF, and the resulting mixture was cooled to -75 °C. A solution of lithium diisopropylamino (1 M in THF, 77.0 mL, 77.0 mmol) was added dropwise over a 90-minute period, and the mixture was stirred at -75 °C for 60 minutes. 6.44 mL (83.7 mmol) of DMF was added dropwise. The mixture was stirred at -75 °C for another 30 minutes. The reaction was quenched by adding half of the concentrated acetic acid (40 mL) and diluted with water and EtOAc. The organic phase was separated, washed with brine, dried, and concentrated. The residue was purified by column chromatography (SiO2, CyH / EtOAc gradient 100:0 to 85:15) to obtain the desired compound.

[0189] C7H2ClF4NO (M=227.5 g / mol) EI-MS: 227 M*+ Rt (HPLC): 0.48 min (Method B) 2-Chloro-3-fluoro-6-(trifluoromethyl)pyridine-4-carboxamide Ammonium acetate (47.1 g, 611 mmol) and 2-chloro-3-fluoro-6-(trifluoromethyl)pyridine-4-carboxaldehyde (13.9 g, 61.1 mmol) were mixed, and acetonitrile (300 mL) was added. Phenylenetrimethylammonium tribromide (47.4 g, 122 mmol) was added aliquots, and the resulting reaction mixture was stirred at ambient temperature for 16 hours. The mixture was filtered through a diatomaceous earth filter, and the residue was washed with acetonitrile and purified by column chromatography (SiO2, CyH / EtOAc gradient 1:0 to 7:3) to obtain the desired product.

[0190] C7H3ClF4N2O (M=242.6 g / mol) ESI-MS: 241 [MH] − Rt (HPLC): 0.47 min (Method B) Intermediate II.6: 2-chloro-3-fluoro-6-(trifluoromethyl)pyridine-4-carboxynitrile The product of the previous step, 2-chloro-3-fluoro-6-(trifluoromethyl)pyridine-4-carboxamide (5.10 g, 21.0 mmol), was suspended in dichloromethane (400 mL), and Burgess reagent (CAS: 29684-56-8, 8.75 g, 35.6 mmol) was added at ambient temperature. The resulting reaction mixture was stirred for 40 hours and then directly purified by column chromatography (SiO2, CyH / EtOAc gradient 100:0 to 95:5) to give the title compound.

[0191] C7HClF4N2 (M=224.5 g / mol) ESI-MS: 225 [M+H] + Rt (HPLC): 0.57 min (Method F) Synthesis of intermediate II.6

[0192] 2-Bromo-3-fluoro-6-(trifluoromethyl)pyridine-4-carboxaldehyde Under an argon atmosphere, 2-bromo-3-fluoro-6-(trifluoromethyl)pyridine (3.08 g, 12.6 mmol) was added to THF (75 mL), and the resulting mixture was cooled to -70 °C. A solution of lithium diisopropylamino (1 M in THF, 13.9 mL, 13.9 mmol) was added dropwise, and the mixture was stirred at -70 °C for 90 min. DMF (1.17 mL, 15.1 mmol) was added dropwise. The mixture was stirred at -70 °C for another 30 min. The reaction was quenched by adding half of the concentrated acetic acid (800 µL) and diluted with water and EtOAc. The organic phase was separated, dried over MgSO4, and concentrated. The residue was purified by column chromatography (SiO2, CyH / EtOAc gradient 1:0 to 4:1) to give 2-bromo-3-fluoro-6-(trifluoromethyl)pyridine-4-carboxaldehyde.

[0193] C7H2BrF4NO (M=272.0 g / mol) ESI-MS: No quality detected Rt (HPLC): 0.51 min (Method B) 1 H NMR (400 MHz, DMSO- d 6) δ ppm 10.19 (s, 1 H), 8.26 (d, J =4.3 Hz, 1H).

[0194] 2-Bromo-3-fluoro-6-(trifluoromethyl)pyridine-4-carboxamide Ammonium acetate (5.53 g, 71.8 mmol) and 2-bromo-3-fluoro-6-(trifluoromethyl)pyridine-4-carboxaldehyde (2.17 g, 7.18 mmol) were mixed, and acetonitrile (44 mL) was added. Phenylenetrimethylammonium tribromide (5.57 g, 14.4 mmol) was added aliquots, and the resulting reaction mixture was stirred at ambient temperature for 72 hours. The mixture was filtered, and the residue was washed with acetonitrile and purified by column chromatography (using Celite®, SiO2 dry loading, CyH / EtOAc gradient 1:0 to 7:3) to obtain the desired product.

[0195] C7H3BrF4N2O (M=287.0 g / mol) ESI-MS: 285 / 287 [MH] − Rt (HPLC): 0.50 min (Method B) Intermediate II.6: 2-Bromo-3-fluoro-6-(trifluoromethyl)pyridine-4-carboxynitrile The product of the previous step, 2-bromo-3-fluoro-6-(trifluoromethyl)pyridine-4-carboxamide (975 mg, 3.40 mmol), was suspended in dichloromethane (80 mL), and Burgess reagent (CAS: 29684-56-8, 1.25 g, 5.10 mmol) was added at ambient temperature. The resulting reaction mixture was stirred for 40 hours and then directly purified by column chromatography (dry loading, SiO2, CyH / EtOAc gradient 1:0 to 9:1) to give the title compound.

[0196] C7HBrF4N2 (M=268.9 g / mol) ESI-MS: 268 / 270 [M+H] + Rt (HPLC): 0.59 min (Method C) Intermediate III.1: 6-acetyl-2-chloro-3-[4-fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]pyridin-4-carboxylonitrile

[0197] Int. II.1 (481 mg, 2.12 mmol) was suspended in DMSO (2.4 mL) and DIPEA (734 µL, 4.25 mmol). Int. I.1 (586 mg, 3.18 mmol) was added at 15 °C, and the resulting mixture was stirred at 15 °C for 18 hours. The mixture was diluted with water and the precipitate formed was filtered off. The residue was diluted with 1,4-dioxane and 4M HCl / 1,4-dioxane (2.0 mL) and stirred at room temperature for 3 hours. The reaction mixture was concentrated and ground with a semi-saturated aqueous solution of K₂CO₃. The precipitate formed was filtered off, washed with water and MTBE, and dried to give the title compound.

[0198] C 16 H 16 ClFN6O (M=362.8 g / mol) ESI-MS: 363 [M+H] + Rt (HPLC): 0.66 min (Method G) Intermediate III.2: 2-chloro-3-[4-fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]-6-(2-hydroxypropyl-2-yl)pyridin-4-carboxynitrile

[0199] Int. III.1 (265 mg, 0.73 mmol) was suspended in THF (4.0 mL) under an argon atmosphere. Methylmagnesium bromide (3.2 M in MeTHF, 274 µL, 0.88 mmol) was added dropwise at ambient temperature, and the reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was quenched by adding water (100 µL), concentrated, and purified by preparative HPLC (XBridge C18, acetonitrile / water gradient containing 0.1% TFA) to obtain the desired compound.

[0200] C 17 H 20 ClFN6O (M=378.8 g / mol) ESI-MS: 379 [M+H] + Rt (HPLC): 0.56 min (Method B) Intermediate III.3: 6-Bromo-2-chloro-3-[4-fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]pyridin-4-carboxylonitrile

[0201] Int. II.2 (1.00 g, 4.25 mmol) was suspended in DMSO (4.0 mL) and DIPEA (1.47 mL, 8.50 mmol). Int. I.1 (900 mg, 4.89 mmol) was added at 15 °C, and the resulting mixture was stirred at 15 °C for 2 hours. The mixture was diluted with ACN / water and purified by preparative HPLC (Sunfire C18, acetonitrile / water gradient containing 0.1% TFA) to obtain the desired compound.

[0202] C 14 H 13 BrClFN6 (M=399.6 g / mol) ESI-MS: 399 / 401 [M+H] + Rt (HPLC): 0.75 min (Method G) Intermediate III.4: 2-chloro-6-(2,2-difluorocyclopropyl)-3-[4-fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]pyridin-4-carboxynitrile

[0203] Int. III.3 (150 mg, 0.36 mmol, 95% purity) and (2,2-difluorocyclopropyl)boronic acid (200 mg, 1.59 mmol, 97% purity) were suspended in 1,4-dioxane (3 mL) under an argon atmosphere. K2CO3 (aq. 2 M, 535 µL, 1.07 mmol) was added, and the resulting mixture was degassed by passing argon through it for 10 min. [1,1′-bis(diphenylphosphino)-ferrocene]-palladium(II) dichloride (Pd(dppf)Cl2) (26.1 mg, 0.04 mmol) was added, and the mixture was degassed again for 5 min. The mixture was then heated to 70 °C for 12 h. After cooling to ambient temperature, the mixture was purified by preparative HPLC (Sunfire C18, ACN / water gradient containing 0.1% TFA) to give the title compound.

[0204] C 17 H 16 ClF3N6 (M=396.8 g / mol) ESI-MS: 397 [M+H] + Rt (HPLC): 0.56 min (Method F) Intermediates synthesized in a manner similar to those described for Int. III.3.

[0205]

[0206] Synthesis of Intermediate III.6

[0207] 2-Chloro-4-cyano-3-[4-fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]pyridin-1-onyl-1-olate Int. III.5 (500 mg, 1.56 mmol) was suspended in DCM (25 mL). H₂O₂ (30%, 530 mg, 4.68 mmol) and TFA (355 mg, 3.12 mmol) were added, and the reaction mixture was stirred at room temperature for 18 hours. The starting material remained. The same amounts of H₂O₂ and TFA were added three times, and the reaction mixture was stirred at room temperature for 90 hours. The reaction mixture was diluted with water (5 mL), and the DCM was concentrated. The residue was purified by preparative HPLC (XBridge C18, ACN / water gradient containing 0.1% TFA). The desired fractions were combined, the ACN was concentrated, and the aqueous residue was alkalized with K₂CO₃. The precipitate was filtered, washed with water, and dried to give the desired compound.

[0208] C 14 H 14 ClFN6O (M=336.8 g / mol) ESI-MS: 337 / 339 [M+H] + Rt (HPLC): 0.33 min (Method B) 2-Chloro-3-[4-fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]-6-hydroxypyridin-4-carboxynitrile 2-Chloro-4-cyano-3-[4-fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]pyridin-1-onthium-1-olate (100 mg, 0.30 mmol) was suspended in THF (4 mL). Trifluoroacetic anhydride (124 µL, 0.89 mmol) was added, and the reaction mixture was stirred at 60 °C for 18 h. The reaction mixture was quenched by adding saturated aqueous K₂CO₃. The THF was concentrated, and the aqueous residue was stirred at room temperature for 0.5 h. The reaction mixture was concentrated, and the residue was purified by preparative HPLC (XBridge C18, ACN / water gradient containing 0.1% NH₃) to give the desired compound.

[0209] C 14 H 14 ClFN6O (M=336.8 g / mol) ESI-MS: 337 [M+H] + Rt (HPLC): 0.22 min (Method B) Intermediate III.6: 2-chloro-3-[4-fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]-6-methoxypyridine-4-carboxynitrile 2-Chloro-3-[4-fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]-6-hydroxypyridin-4-carboxynitrile (63.0 mg, 0.19 mmol) was suspended in chloroform (5 mL), and silver carbonate (77.4 mg, 0.28 mmol) and methyl iodoform (115 mg, 0.81 mmol) were added. The reaction mixture was stirred at room temperature for 2 days. The reaction mixture was concentrated, diluted with ACN, filtered, and purified by preparative HPLC (XBridge C18, ACN / water gradient containing 0.1% NH3) to obtain the desired compound.

[0210] C 15 H16 ClFN6O (M=350.8 g / mol) ESI-MS: 351 / 353 [M+H] + Rt (HPLC): 0.56 min (Method B) Intermediate III.8: 2,5-Dichloro-3-[4-fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]pyridin-4-carboxynitrile

[0211] Int. I.1 (0.96 g, 5.21 mmol) and K₂CO₃ (2.40 g, 22.7 mmol) were added to Int. II.7 (0.17 g, 0.89 mmol) contained in DMF (5 mL). The mixture was stirred at 100 °C for 10 h. The reaction mixture was diluted with H₂O (100 mL) and extracted with EtOAc (2 x 200 mL). The combined organic layers were washed with brine (100 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO₂, PE / EtOAc gradient 1:0 to 0:1) and preparative HPLC (Agela DuraShell C18, ACN / water gradient containing 10 mM NH₄HCO₃) to give the title compound.

[0212] C 14 H 13 Cl2FN6 (M=355.2 g / mol) ESI-MS: 355 / 357 [M+H] + Rt (HPLC): 2.27 min (Method H) Synthesis of intermediate IV.1

[0213] 5-Bromo-2-tert-butyl-2H-pyrazolo[3,4-b]pyridine 5-Bromo-2H-pyrazolo[3,4-b]pyridine (8.00 g, 39.6 mmol) was suspended in toluene (50 mL) and tert-butyl acetate (53.1 mL, 396 mmol). Methanesulfonic acid (2.60 mL, 39.6 mmol) was added. The resulting reaction mixture was heated to 80 °C and stirred for 1 hour. After cooling to ambient temperature, additional methanesulfonic acid (2.60 mL, 39.6 mmol) was added, and the reaction mixture was heated to 80 °C and stirred for 1 hour. After cooling to ambient temperature, the reaction mixture was concentrated, and the residue was diluted with DCM. The mixture was alkalized by adding 1 M NaOH. The layers were separated, and the aqueous layer was extracted with DCM. The combined organic layers were washed with H₂O, dried over Na₂SO₄, filtered, and concentrated. The residue was purified by column chromatography (SiO₂, CyH / EtOAc gradient 9:1 to 0:10) to give the desired compound.

[0214] C 10 H 12 BrN3 (M=254.1 g / mol) ESI-MS: 254 / 256 [M+H] + Rt (HPLC): 0.85 min (Method C) 5-Bromo-2-tert-butyl-3-chloro-2H-pyrazolo[3,4-b]pyridine NCS (2.08 g, 15.6 mmol) was added to a mixture of 5-bromo-2-tert-butyl-2H-pyrazolo[3,4-b]pyridine (4.00 g, 14.2 mmol) contained in ACN (40 mL) under N2 conditions at 20 °C. The mixture was stirred at 85 °C for 12 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was diluted with H2O (20 mL) and extracted with EtOAc (3 x 20 mL). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, PE / EtOAc gradient 1:0 to 0:1) to give the desired compound.

[0215] C 10 H 11 BrClN3 (M=288.6 g / mol) ESI-MS: 288 / 290 [M+H] + Rt (HPLC): 0.55 min (Method I) Intermediate IV.1: {2-tert-butyl-3-chloro-2H-pyrazolo[3,4-b]pyridin-5-yl}boronic acid Under N2, 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)-1,3,2-dioxaborhexacyclopentane (3.09 g, 12.2 mmol), KOAc (2.75 g, 28.1 mmol), and Pd(dppf)Cl2 (0.68 g, 0.94 mmol) were added to a mixture containing 5-bromo-2-tert-butyl-3-chloro-2H-pyrazolo[3,4-b]pyridine (3.00 g, 9.36 mmol) in 1,4-dioxane (60 mL). The mixture was stirred at 100 °C under N2 for 12 hours. The reaction mixture was diluted with H2O (40 mL) and extracted with EtOAc (2 x 40 mL). The combined organic layers were washed with brine (20 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO₂, PE / EtOAc gradient 1:0 to 1:1) to give the title compound.

[0216] C 10 H 13 BClN3O3 (M=253.5 g / mol) ESI-MS: 254 / 256 [M+H] + Rt (HPLC): 0.33 min (Method I) Synthesis of intermediate IV.2

[0217] 5-Bromo-2-tert-butyl-2H-pyrazolo[3,4-b]pyridine 4.0 g (19.8 mmol) of 5-bromo-2H-pyrazolo[3,4-b]pyridine was suspended in toluene (23 mL), and tert-butyl acetate (26.6 mL, 198 mmol) was added. Methanesulfonic acid (1.3 mL, 19.8 mmol) was added slowly. The resulting reaction mixture was heated to 80 °C and stirred for 1 hour. After cooling to ambient temperature, additional methanesulfonic acid (1.3 mL, 19.8 mmol) was added, and the reaction mixture was heated to 80 °C and stirred for 1 hour. After cooling to ambient temperature, the reaction mixture was concentrated and purified by preparative HPLC (XBridge C18, ACN / water gradient containing 0.1% TFA) to give 5-bromo-2-tert-butyl-2H-pyrazolo[3,4-b]pyridine.

[0218] C 10 H 12 BrN3 (M=254.1 g / mol) ESI-MS: 254 / 256 [M+H] + Rt (HPLC): 0.50 min (Method F) Intermediate IV.2: {2-tert-butyl-2H-pyrazolo[3,4-b]pyridin-5-yl}boronic acid 5-Bromo-2-tert-butyl-2H-pyrazolo[3,4-b]pyridine (1.50 g, 3.87 mmol), bis(pinacol)diborane (1.21 g, 4.78 mmol), and potassium acetate (763 mg, 7.77 mmol) were added to 1,4-dioxane (15 mL), and the resulting mixture was degassed by passing argon through it for 10 minutes. [1,1′-bis(diphenylphosphino)-ferrocene]-palladium(II)dichloromethane complex (Pd(dppf)Cl2*CH2Cl2, CAS: 95464-05-4) (190 mg, 0.232 mmol) was added, and the mixture was degassed again for 3 minutes. The mixture was then heated to 110 °C and stirred at that temperature for 4 hours. After cooling to ambient temperature, the mixture was concentrated, and the residue was suspended in an ACN / water mixture, filtered, and purified by preparative HPLC (XBridge C18, ACN / water gradient containing 0.1% TFA) to obtain the title compound.

[0219] C 10 H 14 BN3O2 (M=219.1 g / mol) ESI-MS: 220 [M+H] + Rt (HPLC): 0.27 min (Method F) Intermediate IV.3: 2-nitro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)pyridine-3-amine

[0220] Under an argon atmosphere, 5-bromo-2-nitropyridine-3-amine (200 mg, 0.88 mmol, 96% purity), bis(pinacol)diborane (300 mg, 1.18 mmol), and potassium acetate (259 mg, 2.64 mmol) were added to 1,4-dioxane (4 mL), and the resulting mixture was degassed by passing argon gas through it for 15 min. PdCl₂(PPh₃)₂ (61.2 mg, 0.09 mmol) was added, and the mixture was heated to 60 °C and stirred at that temperature for 4.5 h. After cooling to ambient temperature, the mixture was diluted with EtOAc, filtered through a diatomaceous earth pad, and concentrated. The residue was ground with MTBE, the precipitate was filtered, washed with MTBE, and dried to give the title compound.

[0221] C 11 H 16 BN3O4 (M=265.1 g / mol) ESI-MS: 266 [M+H] + Rt (HPLC): 0.20 min (Method F) Synthesis of intermediate IV.4

[0222] 2-Benzyl-6-bromo-2H-[1,2,3]triazolo[4,5-b]pyridine 6-Bromo-1H-[1,2,3]triazolo[4,5-b]pyridine (600 mg, 2.96 mmol) was suspended in DMSO (5 mL) and DIPEA (1.00 mL, 5.91 mmol). Benzyl bromide (359 µL, 2.96 mmol) was added, and the resulting reaction mixture was heated to 90 °C and stirred for 16 hours. After cooling to ambient temperature, the reaction mixture was purified by preparative HPLC (XBridge C18, ACN / water gradient containing 0.1% NH3) to obtain the desired compound.

[0223] C 12 H9BrN4 (M=289.1 g / mol) ESI-MS: 289 / 291 [M+H] + Rt (HPLC): 0.59 min (Method F) Intermediate IV.4: {2-benzyl-2H-[1,2,3]triazolo[4,5-b]pyridin-6-yl}boronic acid 2-Benzyl-6-bromo-2H-[1,2,3]triazolo[4,5-b]pyridine (310 mg, 1.07 mmol), bis(pinacol)diborane (407 g, 1.61 mmol), and potassium acetate (315 mg, 3.22 mmol) were added to 1,4-dioxane (2 mL), and the resulting mixture was degassed by passing argon through it. PdCl2(PPh3)2 (75.3 mg, 0.11 mmol) was added, and the mixture was heated to 100 °C and stirred at that temperature for 3 hours. After cooling to ambient temperature, the mixture was diluted with EtOAc, filtered through a diatomaceous earth mat, and concentrated. The residue was suspended in an ACN / water mixture and purified by preparative HPLC (XBridge C18, ACN / water gradient containing 0.1% TFA) to give the title compound.

[0224] C 12 H 11 BN4O2 (M=254.1 g / mol) ESI-MS: 255 [M+H] + Rt (HPLC): 0.41 min (Method F) Intermediate IV.5: (1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-[1,2,3]triazolo[4,5-b]pyridin-6-yl)boronic acid

[0225] Under an argon atmosphere, 6-bromo-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-[1,2,3]triazolo[4,5-b]pyridine (100 mg, 0.30 mmol), bis(pinacol)diborane (231 mg, 0.91 mmol), and potassium acetate (116 mg, 1.18 mmol) were added to 1,4-dioxane (2 mL), and the resulting mixture was degassed by passing argon through it for 15 min. PdCl₂(PPh₃)₂ (21.3 mg, 0.03 mmol) was added, and the mixture was heated to 60 °C and stirred at that temperature for 16 h. After cooling to ambient temperature, the mixture was concentrated, and the residue was suspended in an ACN / water mixture, filtered, and purified by preparative HPLC (XBridge C18, ACN / water gradient containing 0.1% TFA) to give the title compound.

[0226] C 11 H 19 BN4O3Si (M=294.2 g / mol) ESI-MS: 295 [M+H] + Rt (HPLC): 0.51 min (Method F) Intermediate IV.6: {furano[2,3-b]pyridin-5-yl}boronic acid

[0227] Under an argon atmosphere, 5-bromofurano[2,3-b]pyridine (250 mg, 1.20 mmol, 95% purity), bis(pinacol)diborane (487 mg, 1.92 mmol), and potassium acetate (459 mg, 4.68 mmol) were added to 1,4-dioxane (5 mL), and the resulting mixture was degassed by passing argon gas through it for 15 min. PdCl₂(PPh₃)₂ (105 mg, 0.15 mmol) was added, and the mixture was heated to 60 °C and stirred at that temperature for 16 h. After cooling to ambient temperature, the mixture was diluted with EtOAc, filtered, and concentrated. The residue was suspended in an ACN / water mixture and purified by preparative HPLC (Sunfire C18, ACN / water gradient containing 0.1% TFA) to give the title compound.

[0228] C7H6BNO3 (M=162.9 g / mol) ESI-MS: 164 [M+H] + Rt (HPLC): 0.39 min (Method G) Synthesis of intermediate IV.7

[0229] 6-Bromo-2-tert-butyl-[1,3]oxazolo[4,5-b]pyridine 2-Amino-5-bromopyridin-3-ol (200 mg, 1.04 mmol) and neopentanoyl chloride (1.48 g, 12.1 mmol) were added to a microwave-safe vial, and the resulting reaction mixture was heated to 120 °C in a sealed vial and stirred at that temperature for 40 hours. After cooling to ambient temperature, the reaction mixture was diluted with DCM and washed with NaHCO3. The organic layer was dried, filtered, and concentrated to give the desired compound.

[0230] C 10 H 11 BrN₂O (M=255.1 g / mol) ESI-MS: 255 / 257 [M+H] + Rt (HPLC): 0.59 min (Method F) Intermediate IV.7: {2-tert-butyl-[1,3]oxazolo[4,5-b]pyridin-6-yl}boronic acid Under an argon atmosphere, 6-bromo-2-tert-butyl-[1,3]oxazolo[4,5-b]pyridine (210 mg, 0.82 mmol), bis(pinacol)diborane (334 mg, 1.32 mmol), and potassium acetate (315 mg, 3.21 mmol) were added to 1,4-dioxane (4 mL), and the resulting mixture was degassed by passing argon through it for 15 min. PdCl₂(PPh₃)₂ (72.2 mg, 0.10 mmol) was added, and the mixture was heated to 60 °C and stirred at that temperature for 90 h. After cooling to ambient temperature, the mixture was diluted with EtOAc, filtered, and concentrated. The residue was suspended in an ACN / water mixture and purified by preparative HPLC (Sunfire C18, ACN / water gradient containing 0.1% TFA) to give the title compound.

[0231] C 10 H 13 BN2O3 (M=220.0 g / mol) ESI-MS: 221 [M+H] + Rt (HPLC): 0.39 min (Method F) Synthesis of intermediate IV.8

[0232] methyl 6-methoxypyrazolo[1,5-b]pyridazine-3-carboxylate To a mixture containing hydroxylamine-O-sulfonic acid (17.0 g, 150 mmol) in H₂O (50 mL), 100 mL of aqueous NaHCO₃ solution was added to adjust the pH to 6 and the temperature was raised to 70 °C. Then, 3-methoxypyridazine (5.00 g, 45.4 mmol) was added to the mixture. The mixture was stirred at 70 °C for 2 hours. After cooling the reaction to 25 °C, the pH was adjusted to 8. A suspension containing methyl propionate-2-acetylacetate (6.30 g, 74.9 mmol) and KOH (10.0 g, 178 mmol) in DCM (50 mL) was added to the mixture. The resulting mixture was stirred at 25 °C for 12 hours. The reaction mixture was diluted with H₂O (200 mL) and extracted with EtOAc (2 x 200 mL). The combined organic layers were washed with brine (100 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, PE / EtOAc gradient 1:0 to 1:1) to obtain the desired compound.

[0233] C9H9N3O3 (M=207.2 g / mol) ESI-MS: 208 [M+H] + Rt (HPLC): 0.53 min (Method D) 6-Methoxypyrazolo[1,5-b]pyridazine-3-carboxylic acid LiOH (4.05 g, 96.5 mmol) and H₂O (10 mL) were added to a mixture containing methyl 6-methoxypyrazolo[1,5-b]pyridazine-3-carboxylate (10.0 g, 48.3 mmol) in THF (20 mL). The mixture was stirred at 25 °C for 12 hours. The reaction mixture was diluted with H₂O (100 mL) and extracted with EtOAc (2 x 200 mL). The aqueous phase was adjusted to pH 1, the precipitate was filtered, and the precipitate was concentrated to give the title compound.

[0234] C8H7N3O3 (M=193.2 g / mol) ESI-MS: 194 [M+H] + Rt (HPLC): 0.41 min (Method D) 3-Bromo-6-methoxypyrazolo[1,5-b]pyridazine NBS (14.7 g, 82.8 mmol) was added to a mixture containing 8.00 g (41.4 mmol) of 6-methoxypyrazolo[1,5-b]pyridazine-3-carboxylic acid in DMF (80 mL). The mixture was stirred at 25 °C for 16 hours. The reaction mixture was diluted with H₂O (200 mL) and extracted with EtOAc (2 x 200 mL). The combined organic layers were washed with brine (100 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO₂, PE / EtOAc gradient 1:0 to 3:1) to give the desired compound.

[0235] C7H6BrN3O (M=228.0 g / mol) ESI-MS: 228 / 230 [M+H] + Rt (HPLC): 0.63 min (Method D) 6-Methoxy-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)pyrazolo[1,5-b]pyridazine Under N2, bis(pinacol)diborane (11.4 g, 44.7 mmol), potassium acetate (17.8 g, 107 mmol), and Pd(PPh3)4 (3.45 g, 2.98 mmol) were added to a mixture containing 3-bromo-6-methoxy-pyrazolo[1,5-b]pyridazine (8.00 g, 29.8 mmol, 85% purity) in 1,4-dioxane (80 mL). The mixture was stirred at 100 °C for 12 hours. After cooling to ambient temperature, the reaction mixture was diluted with H2O (100 mL) and extracted with EtOAc (2 x 200 mL). The combined organic layers were washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, PE / EtOAc gradient 1:0 to 3:1) to give the desired compound.

[0236] C 13 H 18 BN3O3 (M=275.1 g / mol) ESI-MS: 276 [M+H] + Rt (HPLC): 0.73 min (Method D) Intermediate IV.8: {6-methoxypyrazolo[1,5-b]pyridazin-3-yl}boronic acid HCl (5 mL) was added to a mixture containing 7.00 g, 10.2 mmol, 40% purity, of 6-methoxy-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)pyrazolo[1,5-b]pyridazine (50 mL) in MeOH. The mixture was stirred at 25 °C for 5 hours. The reaction mixture was filtered and concentrated under reduced pressure to give the desired compound.

[0237] C7H8BN3O3 (M=193.0 g / mol) ESI-MS: 194 [M+H] + Rt (HPLC): 1.47 min (Method J) Synthesis of intermediate IV.9

[0238] 3-(3-Chlorophenyl)pyridazine Under N2 conditions, (3-chlorophenyl)boronic acid (29.9 g, 191 mmol), KOAc (43.3 g, 442 mmol), and Pd(dppf)Cl2*DCM (6.01 g, 7.36 mmol) were added to a mixture of 3-bromopyridazine (30.0 g, 147 mmol, 78% purity) contained in 1,4-dioxane (300 mL) and H2O (30 mL). The mixture was stirred at 100 °C for 12 hours. After cooling to ambient temperature, the reaction mixture was diluted with H2O (200 mL) and extracted with EtOAc (2 x 300 mL). The combined organic layers were washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, PE / EtOAc gradient 1:0 to 3:1) to give the desired compound.

[0239] C 10 H7ClN2 (M=190.6 g / mol) ESI-MS: 191 [M+H] + Rt (HPLC): 0.53 min (Method D) methyl 6-(3-chlorophenyl)pyrazolo[1,5-b]pyridazine-3-carboxylate To a mixture containing hydroxylamine-O-sulfonic acid (32.3 g, 285 mmol) in H₂O (200 mL), NaHCO₃ (200 mL) was added to adjust the pH to 6 and the temperature was raised to 70 °C. Then, 3-(3-chlorophenyl)pyridazine (35.0 g, 184 mmol) was added to the mixture. The mixture was stirred at 70 °C for 2 hours. After cooling the reaction to 25 °C, the pH was adjusted to 7. Methyl propionate-2-acetylacetic acid (4.00 g, 47.6 mmol) containing DCM (100 mL) was added to the mixture. The resulting mixture was stirred at 25 °C for 12 hours. The reaction mixture was diluted with H₂O (500 mL) and extracted with EtOAc (2 x 1000 mL). The combined organic layers were washed with brine (500 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, PE / EtOAc gradient 1:0 to 2:1) to obtain the desired compound.

[0240] C 14 H 10 ClN3O2 (M=287.7 g / mol) ESI-MS: 288 [M+H] + Rt (HPLC): 0.78 min (Method D) 6-(3-Chlorophenyl)pyrazolo[1,5-b]pyridazine-3-carboxylic acid LiOH (0.88 g, 20.9 mmol) and H₂O (30 mL) were added to a mixture containing methyl 6-(3-chlorophenyl)pyrazolo[1,5-b]pyridazine-3-carboxylate (5.00 g, 10.4 mmol, 60% purity) in THF (50 mL). The mixture was stirred at 25 °C for 5 hours. The reaction mixture was diluted with H₂O (100 mL) and extracted with EtOAc (2 x 200 mL). The aqueous phase was adjusted to pH 1. The aqueous phase was then extracted with EtOAc (2 x 200 mL). The combined organic layers were washed with brine (200 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to give the desired compound.

[0241] C 13 H8ClN3O2 (M=273.7 g / mol) ESI-MS: 274 [M+H] + Rt (HPLC): 0.64 min (Method D) 3-Bromo-6-(3-chlorophenyl)pyrazolo[1,5-b]pyridazine NBS (1.94 g, 10.9 mmol) was added to a mixture of 6-(3-chlorophenyl)pyrazolo[1,5-b]pyridazine-3-carboxylic acid (1.60 g, 5.44 mmol, 93% purity) contained in DMF (15 mL). The mixture was stirred at 25 °C for 12 hours. The reaction mixture was diluted with H₂O (50 mL) and extracted with EtOAc (2 x 200 mL). The combined organic layers were washed with brine (50 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO₂, PE / EtOAc gradient 1:0 to 1:1) to give the desired compound.

[0242] C 13 H8ClN3O2 (M=273.7 g / mol) ESI-MS: 274 [M+H] + Rt (HPLC): 0.64 min (Method D) 6-(3-Chlorophenyl)-3-(4,4,5,5-Tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)pyrazolo[1,5-b]pyridazine Under N2, bis(pinacol)diborane (3.95 g, 15.6 mmol), potassium acetate (0.91 g, 9.33 mmol), and Pd(PPh3)4 (0.60 g, 0.519 mmol) were added to a mixture containing 3-bromo-6-(3-chlorophenyl)pyrazolo[1,5-b]pyridazine (1.00 g, 2.59 mmol, 80% purity) in 10 mL of 1,4-dioxane. The mixture was stirred at 100 °C for 12 h. After cooling to ambient temperature, the reaction mixture was diluted with H2O (100 mL) and extracted with EtOAc (2 x 100 mL). The combined organic layers were washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, PE / EtOAc gradient 1:0 to 3:1) to give the desired compound.

[0243] C 18 H 19 BClN3O2 (M=355.6 g / mol) ESI-MS: 356 [M+H] + Rt (HPLC): 0.90 min (Method D) Intermediate IV.9: [6-(3-chlorophenyl)pyrazolo[1,5-b]pyridazin-3-yl]boronic acid 6-(3-chlorophenyl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)pyrazolo[1,5-b]pyridazine was purified by preparative HPLC (instrument: Shimadzu LC-8A preparative HPLC; column: Welch Xtimate C18250 * 70 mm, 10 µm; mobile phase: A was H2O (10 mM NH4HCO3) and B was ACN; gradient: B from 53% to 83% over 20 min; flow rate: 130 mL / min; wavelength: 220 and 254 nm) to give the title compound.

[0244] C 12 H9BClN3O2 (M=273.5 g / mol) ESI-MS: 274 [M+H] + Rt (HPLC): 2.57 min (Method K) Intermediate V.1: 5-chloro-6'-fluoro-3-[4-fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]-[2,3'-bipyridine]-4-carboxynitrile

[0245] Int. III.8 (20.0 mg, 0.06 mmol) and (6-fluoropyridin-3-yl)boronic acid (7.90 mg, 0.06 mmol) were suspended in 1,4-dioxane (3 mL) under an argon atmosphere. K2CO3 (aq. 2 M, 110 µL, 0.22 mmol) was added, and the resulting mixture was degassed by passing argon gas through it. [1,1′-bis(diphenylphosphino)-ferrocene]-palladium(II) dichloride (Pd(dppf)Cl2) (4.02 mg, 0.005 mmol) was added, and the mixture was degassed again. The mixture was then heated to 70 °C for 1 hour. After cooling to ambient temperature, the mixture was diluted with H2O, filtered, washed with DMF, and purified by preparative SFC (Torus_1AA; MeOH / CO2; 40 °C; BPR: 120 bar) to give the title compound.

[0246] C 19 H 16 ClF2N7 (M=415.8 g / mol) ESI-MS: 416 [M+H] + Rt (HPLC): 0.68 min (Method L) Synthesis of intermediate V.2

[0247] 3-[4-fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]-2-(1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-[1,2,3]triazolo[4,5-b]pyridin-6-yl)pyridin-4-carboxylonitrile Int. III.5 (96.4 mg, 0.30 mmol) and Int. IV.5 (88.4 mg, 0.30 mmol) were suspended in 1,4-dioxane (3 mL) under an argon atmosphere. K2CO3 (aq. 2 M, 300 µL, 0.60 mmol) was added, and the resulting mixture was degassed by passing argon gas through it. [1,1′-bis(diphenylphosphino)-ferrocene]-palladium(II) dichloride (Pd(dppf)Cl2) (22.0 mg, 0.03 mmol) was added, and the mixture was degassed again. The mixture was then heated to 100 °C for 3.5 h. After cooling to ambient temperature, the mixture was diluted with ACN, filtered, and purified by preparative HPLC (Sunfire C18, ACN / water gradient containing 0.1% TFA) to obtain the desired compound.

[0248] C 25 H 31 FN 10 OSi (M = 534.7 g / mol) ESI-MS: 535 [M+H] + Rt (HPLC): 0.83 min (Method G) Intermediate V.2: 3-[4-fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]-2-{1H-[1,2,3]triazolo[4,5-b]pyridin-6-yl}pyridin-4-carboxynitrile 3-[4-fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]-2-(1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-[1,2,3]triazolo[4,5-b]pyridin-6-yl)pyridin-4-carboxynitrile (129 mg, 0.24 mmol) was suspended in DCM (1.76 mL). TFA (0.88 mL, 11.4 mmol) was added, and the reaction mixture was stirred at room temperature for 5 hours. The reaction mixture was concentrated, and the residue was purified by preparative HPLC (Sunfire C18, ACN / water gradient containing 0.1% TFA) to obtain the desired compound.

[0249] C 19 H 17 FN 10 (M=404.4 g / mol) ESI-MS: 405 [M+H] + Rt (HPLC): 0.46 min (Method G) Synthesis of intermediate V.3

[0250] 3-[4-fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]-2-{6-methoxypyrazolo[1,5-b]pyridazin-3-yl}pyridin-4-carboxynitrile Int. III.5 (1.00 g, 3.12 mmol) and Int. IV.8 (663 mg, 3.27 mmol) were suspended in 1,4-dioxane (12 mL) under an argon atmosphere. Na₂CO₃ (aq. 2 M, 4.68 mL, 9.35 mmol) was added, and the resulting mixture was degassed by passing argon gas through it. Pd(dppf)Cl₂* DCM (50.9 mg, 0.06 mmol) was added, and the mixture was degassed again. The mixture was then heated to 100 °C for 2 hours. After cooling to ambient temperature, the mixture was diluted with water and extracted with DCM (2 x). The combined organic layers were dried over Na₂SO₄, filtered, and concentrated. The residue was purified by column chromatography (SiO₂, EtOAc / MeOH gradient 1:0 to 8:2) to give the title compound.

[0251] C 21 H 20 FN9O (M=433.4 g / mol) ESI-MS: 434 [M+H] + Rt (HPLC): 0.81 min (Method C) 3-[4-fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]-2-{6-hydroxypyrazolo[1,5-b]pyridazin-3-yl}pyridin-4-carboxynitrile 3-[4-fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]-2-{6-methoxypyrazolo[1,5-b]pyridazin-3-yl}pyridine-4-carboxynitrile (850 mg, 1.96 mmol) was suspended in DMF (8 mL). Sodium thiomethoxide (687 mg, 8.83 mmol, 90% purity) was added, and the reaction mixture was stirred at 130 °C for 1 hour. After cooling to ambient temperature, the reaction mixture was purified by column chromatography (SiO2, EtOAc / MeOH gradient 1:0 to 1:1) and by preparative HPLC (Sunfire C18, ACN / water gradient containing 0.1% TFA) to obtain the desired compound.

[0252] C 20 H 18 FN9O (M=419.4 g / mol) ESI-MS: 420 [M+H] + Rt (HPLC): 0.74 min (Method C) Intermediate V.3: 3-{4-cyano-3-[4-fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]pyridin-2-yl}pyrazolo[1,5-b]pyridazin-6-yl ester 3-[4-fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]-2-{6-hydroxypyrazolo[1,5-b]pyridazin-3-yl}pyridine-4-carboxynitrile (540 mg, 1.29 mmol) was suspended in DCM (20 mL) and TEA (0.29 mL, 2.06 mmol). Trifluoromethanesulfonic anhydride (1 M in DCM, 2.58 mL, 2.58 mmol) was added dropwise at -10 °C, and the reaction mixture was stirred at -10 °C for 1 hour. The mixture was quenched with water, allowed to warm to ambient temperature, and extracted with DCM. The organic layer was dried over Na₂SO₄, filtered, and concentrated. The residue was purified by column chromatography (SiO₂, EtOAc / MeOH gradient 10:0 to 9:1) to give the title compound.

[0253] C 21 H 17 F4N9O3S (M=551.5 g / mol) ESI-MS: 552 [M+H] + Rt (HPLC): 0.93 min (Method C) Preparation of the final compound Example 1

[0254] Potassium carbonate (2M in water, 92.4 µL, 0.19 mmol) was added to a mixture containing Int. III.2 (35.0 mg, 92 µmol) in 1,4-dioxane (1 mL) and 2-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)pyridine (30.9 mg, 0.14 mmol). The resulting mixture was purified by passing an argon stream through it. Pd(dppf)Cl2 (6.8 mg, 9.2 µmol) was added, and the mixture was purified again. The reaction mixture was stirred at 100 °C for 1.5 h. After cooling to ambient temperature, the mixture was diluted with an ACN / water mixture, filtered, and purified by preparative HPLC (XBridge C18 column with an ACN / water gradient containing 0.1% TFA) and purified again by preparative HPLC (XBridge C18 column with an ACN / water gradient containing 0.1% NH3) to obtain the title compound.

[0255] C 22 H 23 F2N7O (M=439.5 g / mol) ESI-MS: 440 [M+H] + Rt (HPLC): 0.72 min (Method F) 1 H NMR (400 MHz, DMSO- d 6) δ ppm 8.53 (d, J =2.3 Hz 1 H), 8.48 (s, 1 H), 8.27 (td, J =8.24, 2.5 Hz, 1 H), 7.89 (s, 1 H), 7.33 (dd, J =8.5, 2.7 Hz, 1H),5.41 (s, 1H), 3.74 (d, J=1.5 Hz, 3 H), 3.22 - 3.29 (m, 2 H), 3.12 - 3.19 (m, 2 H), 2.10 - 2.27 (m, 4 H), 1.46 (s, 6 H).

[0256] Similar to the synthesis of embodiments described in Example 1

[0257] Example 7

[0258] Int. V.1 (16.0 mg, 0.04 mmol) was suspended in MeOH (0.50 mL). Sodium methoxide (25% in MeOH, 26.4 µL, 0.12 mmol) was added, and the reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was diluted with water, filtered, and purified by preparative HPLC (Zorbax StableBond C18 column, ACN / water gradient containing 0.1% TFA) to give the title compound.

[0259] C 20 H 19 F2N7O (M=411.4 g / mol) ESI-MS: 412 [M+H] + Rt (HPLC): 0.77 min (Method N) Synthesis of Examples 13 and 14

[0260] Int. V. 2 x TFA (86.6 mg, 0.16 mmol) was suspended in DMF (1.0 mL). K₂CO₃ (45.3 mg, 0.33 mmol) and 5-(chloromethyl)-2-(trifluoromethyl)pyridine (32.7 mg, 0.16 mmol) were added at -10 °C, and the reaction mixture was stirred at room temperature for 28 hours. Residual starting material remained. More K₂CO₃ (20 mg, 0.14 mmol) and 5-(chloromethyl)-2-(trifluoromethyl)pyridine (15 mg, 0.08 mmol) were added, and the reaction mixture was stirred at room temperature for 60 hours. The reaction mixture was diluted with water and extracted with EtOAc. The organic layer was washed with brine, dried over MgSO₄, filtered, and concentrated. The residue was purified by preparative HPLC (Sunfire C18 column, ACN / water gradient containing 0.1% TFA) to give Examples 13 and 14.

[0261] Example 13 C 26 H 21 F4N 11 (M=563.5 g / mol) ESI-MS: 564 [M+H] + Rt (HPLC): 0.74 min (Method G) 1 H NMR (400 MHz, DMSO- d 6) δ ppm 9.06 (d, J =2.0 Hz, 1 H), 8.95 (d, J =1.7Hz, 1 H), 8.65 (d, J =2.0 Hz, 1 H), 8.59 (d, J =4.9 Hz, 1 H), 8.52 (s, 1 H), 8.19(dd, J =8.1, 1.7 Hz, 1 H), 7.98 (d, J =8.2 Hz, 1 H), 7.87 (d, J =4.9 Hz, 1 H), 6.28(s, 2 H), 3.70 (d, J =1.5 Hz, 3 H), 3.19 - 3.28 (m, 4 H), 2.05 - 2.21 (m, 4 H).

[0262] Example 14 C26 H 21 F4N 11 (M=563.5 g / mol) ESI-MS: 564 [M+H] + Rt (HPLC): 0.69 min (Method G) 1 H NMR (400 MHz, DMSO- d 6) δ ppm 8.99 (d, J =1.9 Hz, 1 H), 8.91 (d, J =1.7Hz, 1 H), 8.70 (d, J =1.9 Hz, 1 H), 8.57 - 8.60 (m, 1 H), 8.52 - 8.55 (m, 1 H), 8.03 (dd, J =8.1, 1.7 Hz, 1 H), 7.87 - 7.93 (m, 2 H), 6.25 (s, 2 H), 3.70 (d, J =1.5 Hz, 3 H), 3.10 - 3.23 (m, 4 H), 2.04 - 2.20 (m, 4 H).

[0263] Analysis data of the synthetic embodiment

[0264] Analytical HPLC methods Method A

[0265] Method B

[0266] Method C

[0267] Method D

[0268] Method E

[0269] Method F

[0270] Method G

[0271] Method H

[0272] Method I

[0273] Method J

[0274] Method K

[0275] Method L

[0276] Method M

[0277] Method N

[0278] Method O

[0279] Method P

[0280] Method Q

Claims

1. A compound of formula (I) or a salt thereof, specifically a pharmaceutically acceptable salt thereof. in A is A1a, which is a 5- or 6-membered monoheteroaryl ring containing one or two heteroatom members selected from nitrogen and oxygen; At least one of the heteroatom members is nitrogen; Or A is A1b, which is a 9- or 10-membered fused bicyclic heteroaryl ring containing one to four heteroatom members selected from nitrogen and oxygen; At least one of the heteroatom members is nitrogen; And A can be independently selected via one or two R. 2 replace; R 1 R1a is selected from the following groups: H, C 1-4 -alkyl, C 1-4 -alkoxy, hydroxy-C 1-4 -alkyl, F 1-9 -Fluorine-C 1-4 -alkyl and F 1-8 -Fluorine-C 3-5 -cycloalkyl; R 2 R2a can be selected from the following groups: halogen, NH2, C 1-6 -alkyl, F 1-9 -Fluorine-C 1-6 -alkyl, C 1-4 -alkyl-O-H2CH2C-O-, F 1-9 -Fluorine-C 1-4 -alkoxy groups and CH3S(O); or R 2 R2b is selected from the following groups: phenyl, pyridinyl, isoxazolyl, oxazolyl, pyrazolyl, and thiophene; R2b can be arbitrarily and independently passed through one or two R... 3 replace; or R 2 R2c is selected from the following groups. and ; R 3 R3a is selected from the following: halogen, C 1-4 -alkyl, C 1-4 -alkoxy group, HO-C 1-4 -alkyl or CN.

2. The compound of formula (I) according to claim 1, or a salt thereof, wherein A is a group A3 consisting of: pyridinyl, pyrimidinyl, pyrazolo[1,5-b]pyridazinyl, 2H-[1,2,3]triazolo[4,5-b]pyridinyl, pyrazolo[3,4-b]pyridinyl, 1,2-dihydropyridin-2-one, imidazo[1,2-a]pyrimidinyl, [1,3]oxazolo[4,5-b]pyridinyl and furano[2,3-b]pyridinyl; And A can be independently selected via one or two R. 2 replace.

3. The compound of formula (I) according to claim 1, or a salt thereof, wherein A is selected from a group A4 consisting of the following components. , , , , , , , and .

4. The compound of formula (I) or a salt thereof according to any one of claims 1 to 3, wherein R 1 R1b is selected from the following groups: H, C 1-4 -alkyl, C 1-4 -alkoxy, hydroxy-C 1-4 -alkyl, F 1-3 -Fluorine-C 1-4 -alkyl and F 1-3 -Fluorine-C 3-5 -Cycloalkyl.

5. The compound of formula (I) or a salt thereof according to any one of claims 1 to 4, wherein the compound has formula (Ia). 。 6. The compound of formula (I) according to claim 1 or claim 4, or a salt thereof, wherein the compound has formula (Ic). 。 7. The compound of formula (I) or a salt thereof according to claim 1 or claim 4, wherein the compound has formula (Id). 。 8. The compound of formula (I) according to claim 1 or claim 4, or a salt thereof, wherein the compound has formula (Ie). 。 9. The compound of formula (I) according to claim 1, or a salt thereof, wherein the compound is selected from the group consisting of: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 , and 。 10. A pharmaceutically acceptable salt of a compound according to one or more of claims 1 to 9.

11. A pharmaceutical composition comprising one or more compounds according to one or more of claims 1 to 9 or pharmaceutically acceptable salts thereof, and optionally one or more inert carriers and / or diluents.

12. A pharmaceutical composition comprising one or more compounds according to one or more of claims 1 to 9 or pharmaceutically acceptable salts thereof, and one or more other therapeutic agents, and optionally one or more inert carriers and / or diluents.

13. The pharmaceutical composition of claim 12, wherein the one or more other therapeutic agents are selected from anticancer agents and antifibrotic agents.

14. The compound or a pharmaceutically acceptable salt thereof according to one or more of claims 1 to 9, used as a medicine.

15. A method for treating a disease, such as cancer or fibrotic disease, and conditions associated with these diseases in a patient in need, the method being characterized by administering to the patient one or more compounds according to one or more of claims 1 to 9, or pharmaceutically acceptable salts thereof.

16. The compound or a pharmaceutically acceptable salt thereof according to one or more of claims 1 to 9, for use in methods of treating cancer, fibrotic diseases, neurodegenerative diseases, atherosclerosis, infectious diseases, or chronic kidney disease.

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