Pyrazole derivatives as PHD inhibitors

Abstract

A compound being substituted azine of formula (I), substituted pyrimidine of formula (IV) or pharmaceutically acceptable salt thereof

Description

Technical Field

[0001] This invention relates to a series of novel compounds and their use as inhibitors of the hypoxia-inducible factor prolyl hydroxylase domain (PHD inhibitors). Background Technology

[0002] Hypoxia-inducible factor (HIF) prolyl hydroxylase (PHD) is a target for therapeutic applications, including the treatment of anemia and other ischemia-related diseases, including cancer and inflammation. PHD is a Fe(II) and 2-oxoglutarate (2OG)-dependent oxygenase that catalyzes the hydroxylation of specific prolyl residues within the oxygen degradation domains (ODDs) of the HIF-α subunit. HIF-α levels increase with decreasing oxygen levels; HIF-α forms a dimer with HIF-β, and the α,β-HIF complex promotes the transcription of HIF target genes. However, it is likely that none of the currently "clinically" approved PHD inhibitors exhibit selectivity for PHD that is entirely superior to other human 2OG oxygenases. This lack of selectivity can lead to unpredictable off-target effects, meaning that treatment may be unsafe. Therefore, there is a need for more selective small-molecule PHD inhibitors.

[0003] Anemia, a reduction in the number of red blood cells (erythrocytes) in the bloodstream, is a leading cause of mortality and morbidity worldwide. A recent study suggests that up to one-third of the global population may be affected. Anemia can occur in various ways, including through infection, nutrition, chronic kidney disease, and iron deficiency.

[0004] Erythropoietin (EPO) deficiency is widely associated with anemia in a variety of chronic diseases, particularly those related to systemic inflammation. EPO production primarily occurs in the liver and kidneys, although the EPO gene is also expressed in many other organs. Studies have shown that EPO production is highly sensitive to changes in oxygen concentration in atmospheric blood.

[0005] Recombinant human erythropoietin (rhEPO) is currently used to treat anemia in patients; however, EPO cannot be administered orally. Numerous clinical studies have shown that higher doses of rhEPO are associated with an increased risk of cardiovascular events. This is related to the high dose of rhEPO, rather than an increase in hemoglobin levels. High plasma EPO concentrations are associated with vascular toxicity.

[0006] Therefore, there is a need to find alternative therapies for anemia that are easier to administer and do not carry the aforementioned risks.

[0007] Previous clinical studies using PHD inhibitors have demonstrated the achievement of therapeutically useful hemoglobin levels corresponding to normal physiological plasma EPO concentrations. HIF target genes have been reported to be involved in iron metabolism, transport, and absorption, potentially enhancing iron utilization in erythropoiesis. However, due to the widespread tissue distribution and complexity of the HIF system, predicting HIF's role beyond EPO regulation is challenging.

[0008] Hypoxia occurs in ischemic environments. Poor blood flow (organ ischemia) is a major clinical problem in modern times, occurring in circulatory and cardiovascular diseases and potentially during surgery and in the healing of damaged wounds. Poor blood flow commonly affects the kidneys, limbs, heart, and brain, and can be chronic or acute. It is envisioned that PHD inhibitors will stabilize HIF and produce protective and / or reparative responses to ischemic disease.

[0009] When designing PHD inhibitors and target selectivity, the goal is to find candidates that are highly effective and exhibit good properties suitable for use as drugs.

[0010] Currently available PHD inhibitors, including roxadustat, daprodustat, molidustat, desidustat, and vadadustat, exhibit only limited target selectivity for PHD. For example, inhibition of collagen prolyl hydroxylase (CPH), 2-oxoglutarate-containing iron-dependent oxygenase domain 1 (OFGOD1), and jumonji domain 6 (JMJD6) has been observed in one or more of these inhibitors. It is well known that the lack of selectivity of enzyme inhibitors can lead to unpredictable and undesirable off-target effects.

[0011] Therefore, there is a need for more specific PHD inhibitors with physical properties suitable for use as drugs. Summary of the Invention

[0012] This invention provides a series of novel compounds that have been shown to be highly efficient and selective inhibitors of human hypoxia-inducible factor (HIF) prolyl hydroxylase (PHD). Some compounds of this invention have been shown to have IC50 inhibitory effects on PHD2. 50 Less than 200 nM, which is a substantial improvement compared to known clinically used inhibitors (e.g., roxadustat in the PHD2 hydroxylation assay based on liquid chromatography). 50 (2.7 μM).

[0013] In addition to their potency, the compounds of the present invention have been found to have high selectivity for PHD, with a selectivity greater than 100-fold compared to other tested 2OG oxygenases.

[0014] In addition to these desirable biochemical properties, the compounds of the present invention also exhibit desirable physicochemical properties, including good solubility and permeability in cells.

[0015] These physicochemical properties mean that the compounds of the present invention have been found to stabilize HIF-α cells at concentrations within the nM potency range determined by enzyme analysis. Furthermore, in animal models, low doses of the compounds have shown to induce erythropoiesis.

[0016] Therefore, the compound has potential use in treating conditions that target HIF-PHD, including, for example, anemia and other ischemia-related diseases, inflammation, and other conditions mentioned below.

[0017] Therefore, the present invention provides compounds that are substituted azines of formula (I) or pharmaceutically acceptable salts thereof.

[0018]

[0019] in

[0020] X is CR 6 Or N;

[0021] R 0 It is H or unsubstituted or substituted C 1-6 alkyl;

[0022] R 1 It is H, unsubstituted or substituted C 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN, –N(R) t )C(O)N(R u )R v –C(O)OR w or –C(O)N(R) x )R 7 ;

[0023] R 2 It is H, –OR q C, whether substituted or not 1-6 Alkyl; and R 3 It is H, –OR 8 C, whether substituted or not 1-6 Alkyl; or R 2 It is –N= and R 3 =C(R) y ) – and R 2 and R 3 Together forming formula – N = C(R) y )– groups;

[0024] R 4 It is H, unsubstituted or substituted C 1-6 Alkyl, –OR 9 Or –C(O)OR 10 ;

[0025] R 5 It is H, unsubstituted or substituted C 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN, –N(R) t )C(O)N(R u )R v –C(O)OR w or –C(O)N(R) x )R 7 ;

[0026] R 6 It is H or unsubstituted or substituted C 1-6 alkyl;

[0027] R 7 It is –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar, –Ary–Ar, –CH(R 11 –Cyc, –CH2C≡CCH3, –Cyc, or –Ar, where Ar is an unsubstituted or substituted aryl or an unsubstituted or substituted heteroaryl, Ary is an unsubstituted or substituted arylene or an unsubstituted or substituted heteroaryl, and Cyc is an unsubstituted or substituted C 3-10 cycloalkyl, R 11 It is H, –C(O)OR z C, whether substituted or not 1-4 alkyl;

[0028] R 8 R 9 and R 10 Each is independently selected from H and unsubstituted or substituted C. 1-6 Alkyl groups; and

[0029] R t R u R v R w R x R y and R z Each is independently selected from H, unsubstituted or substituted C. 1-6 Alkyl groups and unsubstituted or substituted phenyl groups;

[0030] R q It is H, unsubstituted or substituted C 1-6Alkyl groups, or unsubstituted or substituted phenyl groups;

[0031] The condition is R 1 and R 5 One of them is –C(O)N(R) x )R 7 And R 1 and R 5 The other one is H, unsubstituted or substituted C. 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN, –N(R) t )C(O)N(R u )R v Or –C(O)OR w .

[0032] Preferred embodiments of the compounds are described below, which are substituted acridines of formula (I) or pharmaceutically acceptable salts thereof, including substituted acridines of formulas (Ia), (Ib), (Ic) and (Id) as defined below and pharmaceutically acceptable salts thereof.

[0033] In another aspect, the present invention provides a compound that is a substituted pyrimidine of formula (IV) or a pharmaceutically acceptable salt thereof.

[0034]

[0035] in

[0036] R 0 It is H or unsubstituted or substituted C 1-6 alkyl;

[0037] R 2 It is H, –OR q C, whether substituted or not 1-6 alkyl;

[0038] R 4 Yes – OR 9 ;

[0039] R 5 It is H, unsubstituted or substituted C 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN or –C(O)OR w ;

[0040] R 6 It is H or unsubstituted or substituted C 1-6 alkyl;

[0041] R 7 It is –CH(R) 11 –Ar、–CH(R) 11)–Ary–Ar, –Ary–Ar, –CH(R 11 –Cyc, –Cyc, or –Ar, where Ar is an unsubstituted or substituted aryl or an unsubstituted or substituted heteroaryl, Ary is an unsubstituted or substituted arylene or an unsubstituted or substituted heteroaryl, and Cyc is an unsubstituted or substituted C 3-10 cycloalkyl, R 11 It is H, –C(O)OR z C, whether substituted or not 1-4 alkyl;

[0042] R 9 Selected from H and unsubstituted or substituted C 1-6 alkyl;

[0043] R w and R z Each is independently selected from H, unsubstituted or substituted C. 1-4 Alkyl groups and unsubstituted or substituted phenyl groups; and

[0044] R q It is H, unsubstituted or substituted C 1-6 Alkyl, or unsubstituted or substituted phenyl.

[0045] The present invention also provides a pharmaceutical composition comprising a compound of the present invention as defined above and a pharmaceutically acceptable carrier or diluent. The pharmaceutical composition may also comprise one or more additional active agents, such as those described below.

[0046] In another aspect, the present invention provides compounds or pharmaceutical compositions of the present invention as defined above for therapeutic treatment of human or animal bodies.

[0047] The present invention also provides compounds or pharmaceutical compositions of the present invention as defined above, for use as regulators of hypoxia-inducible factor prolyl hydroxylase activity. Typically, these compounds or compositions are used as inhibitors of hypoxia-inducible factor prolyl hydroxylase activity.

[0048] This invention further provides compounds or pharmaceutical compositions of the invention as defined above for the treatment of PHD-related conditions, i.e., conditions that can be treated by modulating (e.g., inhibiting) hypoxia-inducible factor prolyl hydroxylase activity. As described below, such conditions include, but are not limited to, anemia, ischemia-related diseases, inflammation, non-fatty liver disease, irritable bowel disease, cardiovascular disease, heart failure, chronic kidney disease, renal insufficiency, Parkinson's disease, Alzheimer's disease, sickle cell anemia, and cancer. These conditions can be, for example, HIF-related conditions, EPO-related conditions, or VHL-related conditions, such as von Hippel-Lindau syndrome.

[0049] This invention also provides compounds or pharmaceutical compositions of the invention as defined above for the treatment of anemia, ischemia, inflammation, Parkinson's disease, Alzheimer's disease, non-fatty liver disease, irritable bowel disease, cardiovascular disease, heart failure, chronic kidney disease, sickle cell anemia, cancer, or renal insufficiency; or for skeletal muscle injury repair, increasing red blood cell count (RBC), increasing hemoglobin (HGB) production, increasing hematocrit (HCT) production, increasing erythropoietin (EPO) production, wound healing, angiogenesis, vascular regeneration, stem cell activation, or cardiac protection after myocardial infarction. Anemia can be renal anemia, such as anemia associated with chronic kidney disease, or anemia in dialysis patients, chemotherapy-induced anemia, sickle cell anemia (including through upregulation of fetal hemoglobin F), age-related anemia, or anemia caused by cancers such as leukemia, multiple myeloma, and pyogenic myeloma. Ischemia can be caused by circulatory or cardiovascular diseases, ischemia in myocardial infarction, ischemia during surgery, organ ischemia, ischemic diseases, diabetic limb ischemia, or sickle cell anemia.

[0050] The present invention also provides a method for treating a subject who has or is susceptible to PHD-related conditions, the method comprising administering to the subject an effective amount of a compound of the present invention as defined above or a pharmaceutical composition of the present invention.

[0051] The present invention also provides a method for treating a subject who has or is susceptible to anemia, ischemia, inflammation, Parkinson's disease, Alzheimer's disease, non-fatty liver disease, irritable bowel disease, cardiovascular disease, heart failure, chronic kidney disease, renal insufficiency, or skeletal muscle injury repair, the method comprising administering to the subject an effective amount of the compound of the present invention as defined above or the pharmaceutical composition of the present invention.

[0052] The present invention also provides a method for: increasing red blood cell count (RBC), increasing hemoglobin (HGB) production, increasing hematocrit (HCT) production, increasing erythropoietin (EPO) production, wound healing, angiogenesis, vascular regeneration, stem cell activation, or cardiac protection after myocardial infarction in a subject, the method comprising administering to the subject an effective amount of the compound of the present invention as defined above or the pharmaceutical composition of the present invention. Attached Figure Description

[0053] Figure 1 Immunoblots are shown of Hep3B cells treated with compounds 117, 119, 122, and 123 (Examples 93, 95, 98, and 99) for 3 hours with 100 μM (A) and 20 μM (B) PHD inhibitors. The blots show the protein levels of HIF1-α and β-actin at 3 hours post-treatment.

[0054] Figure 2Immunoblots are shown of HEK293 T cells treated with compound 68 (Example 46) at 0.5, 1, 5, 10, 20, 50, and 100 μM for 18 hours. The blots show the protein levels of HIF1-α and GAPDH at 18 hours post-treatment.

[0055] Figure 3 The figure (A) shows the red blood cell count (10) in seven C57BL / 6 mice treated with a mordant (1% methylcellulose) or a control (dapoxetine) compared to compound 68 of the present invention (Example 46) (x-axis). 6 (a) (μL) (y-axis); (b) Mouse hemoglobin levels (g / dL) in mice treated with the mordant (1% methylcellulose) or control (dapoxetine) compared to compound 68 of the present invention (Example 46) (x-axis) (y-axis); and (c) Mouse hematocrit percentage in mice treated with the mordant (1% methylcellulose) or control (dapoxetine) compared to compound 68 of the present invention (Example 46) (x-axis) (y-axis). Values ​​are shown before treatment, 4 days after treatment, and 8 days after treatment.

[0056] Figure 4 The IC50 of compound 68 of the present invention (Example 46) is shown when tested with off-target sites typically inhibited by existing PHD inhibitors. 50 . Detailed Implementation

[0057] definition

[0058] As used in this article, the term "alkyl" refers to a straight-chain or branched saturated hydrocarbon group. "C" n-m "Alkyl" refers to an alkyl group having n to m carbon atoms. Therefore, an alkyl group can be C 1-20 Alkyl, C 1-18 Alkyl, C 1-14 Alkyl, C 1-10 Alkyl, C 1-6 Alkyl or C 1-4 Alkyl group. C 1-10 Examples of alkyl groups are methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, or decyl. C 1-6 Examples of alkyl groups are methyl, ethyl, propyl, butyl, pentyl, or hexyl. C 1-4 Examples of alkyl groups are methyl, ethyl, isopropyl, n-propyl, tert-butyl, sec-butyl, or n-butyl. If the term "alkyl" is used anywhere herein without specifying a carbon number prefix, it has 1 to 6 carbons. For the avoidance of doubt, in the case of two alkyl moiety in a group, the alkyl moiety may be the same or different.

[0059] As used herein, the term "cycloalkyl" refers to a saturated or partially unsaturated cyclic hydrocarbon group. "Cn-m "Cycloalkyl" refers to a cycloalkyl group having n to m carbon atoms. Therefore, a cycloalkyl group can be C 3-20 cycloalkyl, C 3-10 cycloalkyl, C 3-8 cycloalkyl or C 3-6 Cycloalkyl. C 3-8 Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, cyclohex-1,3-dienyl, cycloheptyl, and cyclooctyl. 3-6 Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0060] As used herein, the term "alkenyl" refers to a straight-chain or branched hydrocarbon group containing one or more double bonds. "C" n- "m-alkenyl" refers to an alkenyl group having n to m carbon atoms. Therefore, an alkenyl group can be C 2-18 alkenyl, C 2-14 alkenyl, C 2-10 alkenyl, C 2-6 alkenyl or C 2-4 Alkenyl. C 2-10 Examples of alkenyl groups are vinyl (vinyl), propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl, or decenyl. 2-6 Examples of alkenyl groups are vinyl, propenyl, butenyl, pentenyl, or hexenyl. 2-4 Examples of alkenyl groups are vinyl, isopropenyl, n-propenyl, sec-butenyl, or n-butenyl. Alkenyl groups typically contain one or two double bonds.

[0061] As used herein, the term "alkynyl" refers to a straight-chain or branched hydrocarbon group containing one or more triple bonds. "C" n- "m-alkynyl" refers to an alkynyl group having n to m carbon atoms. Therefore, an alkynyl group can be C... 2-18 alkynyl group, C 2-14 alkynyl group, C 2-10 alkynyl group, C 2-6 alkynyl or C 2-4 Alkynyl group. C 2-10 Examples of alkynyl groups are ethynyl, propynyl, butynyl, penynyl, hexynyl, hepynyl, octyynyl, nonynyl, or decynyl. 1-6 Examples of alkynyl groups are ethynyl, propynyl, butynyl, pentynyl, or hexynyl. Alynyl groups typically contain one or two triple bonds.

[0062] C 3-20 A heterocyclic group is a monovalent moiety obtained by removing hydrogen atoms from the ring atoms of a heterocyclic compound. This moiety has 3 to 20 ring atoms (unless otherwise specified), of which 1 to 10 are cyclic heteroatoms. "C n-"m heterocyclic group" refers to a heterocyclic group having n to m ring atoms. Preferably, the ring has 3 to 7 ring atoms (i.e., it is C). 3-7 (Heterocyclic groups), of which 1 to 4 are cyclic heteroatoms.

[0063] Examples of 5- and 6-membered saturated heterocyclic groups include piperazine, piperidine, morpholine, 1,3-oxazine, pyrrolidine, imidazoline, and oxazolidinyl, including their quaternized derivatives, as defined herein. Examples of 5- and 6-membered partially saturated heterocyclic groups include tetrahydropyrazine, tetrahydropyridine, dihydro-1,4-oxazine, tetrahydropyrimidine, dihydro-1,3-oxazine, dihydropyrrole, dihydroimidazolium, and dihydrooxazole, including their quaternized derivatives, as defined herein. Therefore, heterocyclic groups include pyrazolylalkyl, piperidinyl, piperazinyl, thiomorpholinyl, S-oxo-thiomorpholinyl, S,S-dioxo-thiomorpholinyl, morpholinyl, pyrrolylalkyl, pyrrololinyl, imidazoylalkyl, imidazolinyl, 1,3-dioxopentyl, 1,4-dioxopentyl, and pyrazolylyl groups and moieties. Pyrazolyl, piperidinyl, piperazine, pyrazolylmorpholino, and imidazoalkyl groups and portions are typical examples.

[0064] Examples of 9- and 10-membered fused heterobicyclic groups include 9-membered fused heterobicyclic groups such as indoline, 2,3-dihydrobenzofuran, 2,3-dihydrobenzo[b]thiophene, 2,3-dihydro1H-benzo[d]imidazolium, 2,3-dihydrobenzo[d]oxazole, 2,3-dihydrobenzo[d]thiazole, benzo[d][1,3]dioxacyclopentene, 4,5,6,7-tetrahydrothiazo[5,4-c]pyridine, and 4,5,6,7-tetrahydrothiazo[4,5-c]pyridine, including their quaternized derivatives as defined herein; and 10-membered heterobicyclic groups such as 1,2,3, 4-Tetrahydroquinoline, 1,2,3,4-tetrahydroisoquinoline, chromium, isochromium, thiochromium, isothiochromium, 1,2,3,4-tetrahydroquinoxaline, 1,2,3,4-tetrahydroquinazoline, 1,4-dihydro-2H-benzo[d][1,3]oxazine, 3,4-dihydro-2H-benzo[b][1,4]oxazine, 3,4-dihydro-2H-benzo[b][1,4]thiazine, 1,4-dihydro-2H-benzo[d][1,3]thiazine, 4H-benzo[d][1,3]dioxin, and 2,3-dihydrobenzo[b][1,4]dioxin, including their quaternized derivatives. Preferably, the fused heterobicyclic group comprises 1, 2, or 3, preferably 1 or 2 nitrogen atoms.

[0065] For clarity, references to heterocyclic groups also include fused polycyclic systems, including, for example, fused bicyclic systems in which the heterocyclic group is fused with an aryl group. When the heterocyclic group is such a fused heterocyclic group, preferred examples are fused ring systems in which a 5- to 6-membered heterocyclic group is fused with a phenyl group. References to heterocyclic groups also include spirocyclic systems, such as 7-membered heterocyclic groups, such as 2,6-diazaspiro[3.3]heptane.

[0066] As used herein, the term "aryl" refers to a monocyclic, bicyclic, or polycyclic aromatic ring containing up to 14 carbon atoms in the ring moiety, typically 6 to 10 carbon atoms. Examples include phenyl, naphthyl, indenyl, and indenyl. Phenyl is preferred.

[0067] As used herein, the term "heteroaryl" refers to a monocyclic or bicyclic heteroaryl ring, which typically contains 5 to 10, for example, 6 to 10 atoms in its ring portion, including one or more heteroatoms. Heteroaryl rings are typically 5- or 6-membered rings containing at least one heteroatom selected from O, S, N, P, Se, and Si, more typically from O, S, and N. It may contain, for example, one, two, or three heteroatoms. Examples of heteroaryl rings include pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, furanyl, thiopheneyl, pyrazolylyl, pyrroleyl, oxazolyl, oxadiazolyl, isoxazolyl, thiadiazolyl, thiazolyl, isothiazolyl, imidazoleyl, pyrazolyl, pyridazolyl, quinolinyl, and isoquinolinyl. Furanyl, thiopheneyl, pyrazolyl, pyrazolyl, pyrimidinyl, and thiazolyl are typical examples.

[0068] As used herein, the terms "alkylene," "cycloalkylene," "heterocyclic," "alkenyl," "ynylene," "aryl," and "heteroaryl" refer to divalent groups obtained by removing a hydrogen atom from an alkyl, cycloalkyl, heterocyclic, alkenyl, ynyl, aryl, or heteroaryl group, respectively. Such divalent groups can be substituted or unsubstituted. Alkenylenes can be C10-20-3 ... 1-20 Alkylene, C 1-18 Alkylene, C 1-14 Alkylene, C 1-10 Alkylene, C 1-6 Alkylene or C 1-4 Alkylene. C 1-6 Examples of alkylene compounds are methylene, ethylene, propylene, butylene, pentylene, and hexylene. Cycloalkylene compounds can be C10, C20, C30, C40, C50, C60, C7 ... 3-10 Cycloalkylene, C 3-8 Cycloalkyl or C 3-6 Cycloalkylene. C 3-6 Examples of cycloalkylene groups include cyclopentylene and cyclohexylene. The alkenyl group can be C10. 2-18 imidene group, C 2-14 imidene group, C 2-10 imidene group, C 2-6 imide or C2-4 Alkenyl group. C 2-4 Examples of alkenyl groups include vinylidene, propenylidene, and butenylidene. Alkyne groups can be C16-26-3 ... 2-18 Ethyne group, C 2-14 Ethyne group, C 2-10 Ethyne group, C 2-6 Ethyne or C 2-4 etymynyl group. C 2-4 Examples of ynylenes include ethynylenes and propynylenes. Examples of arylenes include phenylenes, and examples of heteroarylenes include, for example, dieles derived from pyridine, dieles derived from thiophene, dieles derived from chromium, and dieles derived from chromium alcohols. For alkylenes, cycloalkylenes, alkenylenes, ynylenes, arylenes, and heteroarylenes, these groups can be bonded to other groups at any two positions on the group (in the case of heteroarylenes and heterocyclic groups, these positions are typically carbon atoms). Thus, propylene includes –CH2CH2CH2– and –CH2CH(CH3)–, and phenylenes include ortho, meta, and para-phenylenes.

[0069] As used herein in the context of substituted organic compounds and groups, the term "substituted" refers to having one or more groups selected from C10. 1-10 Alkyl, C 3-10 cycloalkyl, C 3-7 Heterocyclic, aryl, heteroaryl, cyano, amino, nitro, C 2-10 alkenyl, C 2-10 alkynyl group, C 1-10 Alkylamino, di(C) 1-10 )alkylamino, arylamino, diarylamino, aryl(C 1-10 Alkylamino, amino, amide, hydroxyl, oxo, halogen, carboxyl, ester, acyl, acyloxy, C 1-10 Alkyloxy, aryloxy, halogenated (C 1-10 )alkyl, sulfonic acid, thiol, C 1-10 Alkylthio, arylthio, sulfonyl, phosphoric acid, phosphate ester, phosphonic acid, phosphonate ester and SO3 - The substituent is an organic compound or group (e.g., alkyl, alkylene, cycloalkyl, heterocyclic, aryl, arylene, heteroaryl, or heteroaryl). Typically, one or more substituents are selected from cyano, amino, nitro, amide, amide, hydroxy, oxo, halogen, carboxyl, ester, acyl, acyloxy, sulfonic acid, thiol, sulfonyl, phosphoric acid, phosphate ester, phosphonic acid, phosphonate ester, and SO3. - When a compound or group is substituted, it typically has 1, 2, 3, or 4 substituents. For example, a substituted compound or group can have 1, 2, or 3 substituents, or, for example, 1 or 2 substituents.

[0070] However, when a group is substituted with a halogen, such as fluorine, the group can have 1, 2, 3, or 4 halogen substituents, or it can have more than 4 halogen substituents. In fact, the group can be perhalogenated, meaning all hydrogen atoms in the group can be replaced by halogen atoms. The group can, for example, be perfluorinated, meaning all hydrogen atoms in the group can be replaced by fluorine atoms. Therefore, in the context of substituted organic groups, such as substituted hydrocarbons, substituted alkyl groups, substituted cycloalkyl groups, substituted alkenyl groups, substituted alkynyl groups, substituted aryl groups, substituted alkylene groups, substituted cycloalkylene groups, substituted alkenylene groups, substituted alkynylene groups, and substituted arylene groups (including substituted heteroarylene groups), the term "substituted" as used herein includes perhalogenated groups, particularly perfluorinated groups. Thus, for example, the term "substituted C" as used herein... n-m "alkyl" includes C n-m Perfluoroalkyl, as used herein by the term "substituted C n-m "alkylene" includes C n-m Perfluoroalkylene, as used herein by the term "substituted C n-m "Hydrocarbon group" includes C n-m Perfluorocarbon groups, as used in this article, are referred to by the term "substituted C". n-m "Hydrocarbonyl" includes C n-m Perfluoroalkylene groups, as used in this article, are referred to by the term "substituted C". n-m "Alkoxy" includes C n-m Perfluoroalkoxy, etc.

[0071] As used in this article, oxy-representation refers to the group in the following formula: =O.

[0072] As used herein, the term acyl represents a group of the following formula: -C(=O)R, where R is an acyl substituent, such as a substituted or unsubstituted C. 1-20 Alkyl, substituted or unsubstituted C 3-20 Heterocyclic groups or substituted or unsubstituted aryl groups. Examples of acyl groups include, but are not limited to, -C(=O)CH3 (acetyl), -C(=O)CH2CH3 (propionyl), -C(=O)C(CH3)3 (tert-butyryl) and -C(=O)Ph (benzoyl, benzophenone).

[0073] As used herein, the term ester (or carboxylate, carboxylic acid ester, or oxycarbonyl) represents a group of the following formula: -C(=O)OR, where R is an ester substituent, such as a substituted or unsubstituted C. 1-20 Alkyl, substituted or unsubstituted C 3-20Heterocyclic groups, or substituted or unsubstituted aryl groups (usually phenyl). Examples of ester groups include, but are not limited to, -C(=O)OCH3, -C(=O)OCH2CH3, -C(=O)OC(CH3)3 and -C(=O)OPh.

[0074] As used herein, the term acyloxy (or trans ester) represents a group of the following formula: -OC(=O)R, where R is an acyloxy substituent, such as a substituted or unsubstituted C. 1-20 Alkyl, substituted or unsubstituted C 3-20 Heterocyclic group, or substituted or unsubstituted aryl group, usually C 1-6 Alkyl groups. Examples of acyl groups include, but are not limited to, -OC(=O)CH3 (acetoxy), -OC(=O)CH2CH3, -OC(=O)C(CH3)3, -OC(=O)Ph and -OC(=O)CH2Ph.

[0075] As used herein, the term phosphonic acid represents a group of the following formula: -P(=O)(OH)2. Those skilled in the art will understand that phosphonic acid groups can exist in both protonated and deprotonated forms (i.e., -P(=O)(OH)2, -P(=O)(O - )2 and -P(=O)(OH)(O - All of these fall within the scope of the term "phosphonic acid".

[0076] As used herein, the term phosphonate represents a salt of a phosphonic acid group. For example, a phosphonate can be of the formula -P(=O)(OH)(O - X + The group is ), where X is a monovalent cation. + It can be an alkali metal cation. For example, X + It could be Na + or K + .

[0077] As used herein, the term phosphonate represents a group of one of the following formulas: -P(=O)(OR)2 and -P(=O)(OR)O - Each R is independently a phosphonate substituent, such as -H, substituted or unsubstituted C. 1-20 Alkyl, substituted or unsubstituted C 3-20 Heterocyclic groups, with other C 3-20 C with heterocyclic substitution 3-20 Heterocyclic groups, substituted or unsubstituted C 1-20 Alkylene-C 3-20 Heterocyclic groups, substituted or unsubstituted C 3-25 cycloalkyl, substituted or unsubstituted C 1-20 Alkylene-C 3-25 Cycloalkyl, aryl, substituted or unsubstituted C1-20 Alkylene-aryl. Examples of phosphonate groups include, but are not limited to, -P(=O)(OCH3)2, -P(=O)(OCH2CH3)2, -P(=O)(Ot-Bu)2 and -P(=O)(OPh)2.

[0078] As used in this article, phosphoric acid represents the group in the following formula: -OP(=O)(OH)2.

[0079] As used in this article, the term phosphate ester represents a group of one of the following formulas: -OP(=O)(OR)2 and -OP(=O)(OR)O - Each R is independently a phosphate ester substituent, such as -H, substituted or unsubstituted C. 1-20 Alkyl, substituted or unsubstituted C 3-20 Heterocyclic group, with another C 3-20 C with heterocyclic substitution 3-20 Heterocyclic groups, substituted or unsubstituted C 1-20 Alkylene-C 3-20 Heterocyclic groups, substituted or unsubstituted C 3-25 cycloalkyl, substituted or unsubstituted C 1-20 Alkylene-C 3-25 Cycloalkyl, aryl, substituted or unsubstituted C 1-20 Alkylene-aryl. Examples of phosphate ester groups include, but are not limited to, -OP(=O)(OCH3)2, -OP(=O)(OCH2CH3)2, -OP(=O)(Ot-Bu)2, and -OP(=O)(OPh)2.

[0080] As used herein, the term amino represents a group with the formula -NH2. The term C1-C 10 Alkylamino groups are represented by the formula -NHR′, where R′ is a C 1-10 Alkyl, preferably C 1-6 Alkyl group, as previously defined. Term 2 (C 1-10 Alkylamino groups are represented by the formula -NR′R″, where R′ and R″ may be the same or different, representing C. 1-10 Alkyl, preferably C 1-6 Alkyl, as previously defined. The term arylamino represents a group of the formula -NHR′, wherein R′ is aryl, preferably phenyl, as previously defined. The term diarylamino represents a group of the formula -NR′R″, wherein R′ and R″ are the same or different and represent aryl, preferably phenyl, as previously defined. The term arylalkylamino represents a group of the formula -NR′R″, wherein R′ is C 1-10 Alkyl, preferably C 1-6 Alkyl group, R″ is aryl, preferably phenyl.

[0081] As used herein, the term amide represents a group of the following formula: -C(=O)NR'R", where R' and R" are independently amino substituents, such as p-di(C 1-10 Definition of alkylamino groups. Examples of amide groups include, but are not limited to, -C(=O)NH2, -C(=O)NHCH3, -C(=O)N(CH3)2, -C(=O)NHCH2CH3 and -C(=O)N(CH2CH3)2, and amide groups in which R' and R” together with the nitrogen atom to which they are attached form a heterocyclic structure, such as piperidine carbonyl, morpholine carbonyl, thiomorpholine carbonyl and piperazine carbonyl.

[0082] As used in this article, the term amide represents a group of the following formula: -NR 1 C(=O)R 2 , where R 1 It is an amide substituent, such as hydrogen, C 1-20 Alkyl, C 3-20 Heterocyclic or aryl groups, preferably hydrogen or C 1-20 Alkyl, R 2 It is an acyl substituent, such as C 1-20 Alkyl, C 3-20 Heterocyclic or aryl, preferably hydrogen or C 1-20 Alkyl groups. Examples of amide groups include, but are not limited to, -NHC(=O)CH3, -NHC(=O)CH2CH3, -NHC(=O)Ph, and -NHC(=O)C. 15 H 31 and -NHC(=O)C9H 19 Therefore, the replacement C 1-20 Alkyl groups may contain the following formula: -NHC(=O)-C 1-20 Alkyl groups are defined as amide substituents, such as -NHC(=O)C 15 H 31 or -NHC(=O)C9H 19 R 1 and R 2 They can form cyclic structures together, such as succinimide, maleimide, and phthalimide groups:

[0083]

[0084] C 1-10 The alkylthio group is the C group attached to the thio group. 1-10 Alkyl, preferably C 1-6 Alkyl group. Arylthio group is an aryl group attached to a thio group, preferably phenyl.

[0085] C 1-20 alkoxy groups are the substituted or unsubstituted C atoms attached to an oxygen atom. 1-20 Alkyl group. C1-6 alkoxy groups are the substituted or unsubstituted C atoms attached to an oxygen atom. 1-6 Alkyl group. C 1-4 An alkoxy group is a substituted or unsubstituted carbon atom attached to an oxygen atom. 1-4 Alkyl group. Substituted C 1-20 Alkoxy groups include C 1-20 Perfluoroalkoxy. C 1-20 Perfluoroalkoxy is a C atom bonded to an oxygen atom. 1-20 Perfluoroalkyl. C 1-20 An example of a perfluoroalkoxy group is tert-nonafluorobutoxy, -OC(CF3)3.

[0086] As defined herein, an aryloxy group is a substituted or unsubstituted aryl group attached to an oxygen atom. For example, it can be an unsubstituted or substituted phenoxy group. An example of an aryloxy group is -OPh (phenoxy).

[0087] As used herein in connection with any of the compounds described herein, the term "amino acid" refers to an amino acid residue. Amino acid residues are typically bonded to an atom in the compound described herein, either via their C-terminus or their N-terminus. For example, as understood by those skilled in the art, when an amino acid is allegedly bonded to a carbon atom of a carbonyl group in a compound described herein, the nitrogen atom at the N-terminus of the amino acid is typically bonded to that carbon atom. Similarly, when an amino acid is allegedly bonded to a nitrogen atom of an amine group in a compound described herein, the carbon atom at the C-terminus of the amino acid is typically bonded to that nitrogen atom. The carbon atom at the C-terminus of an amino acid may optionally be bonded to an oxygen atom in a compound described herein. The amino acids in any of the compounds described herein may be, for example, amino acid residues selected from arginine (Arg), histidine (His), lysine (Lys), aspartic acid (Asp), glutamic acid (Glu), serine (Ser), threonine (Thr), asparagine (Asn), glutamine (Gln), cysteine ​​(Cys), selenocysteine ​​(Sec), glycine (Gly), proline (Pro), alanine (Ala), valine (Val), isoleucine (Ile), leucine (Leu), methionine (Met), phenylalanine (Phe), tyrosine (Tyr), and tryptophan (Trp).

[0088] As used herein, acridine refers to a heterocyclic compound containing a 6-membered aromatic ring in which one or more ring carbon atoms are replaced by nitrogen atoms. For example, pyridine is an acridine, as are pyridazine.

[0089] Unless otherwise stated, the above includes the well-known ionic, salt, solvate, and protected forms of these substituents. For example, references to carboxylic acids or carboxyl groups (-COOH) also include the anionic (carboxylate) form (-COO). -), its salts or solvates, and conventional protected forms. Similarly, references to amino groups include the protonated form (-N). + HR 1 R 2 ), amino salts or solvates (e.g., hydrochlorides), and the conventional protected forms of amino groups. Similarly, references to hydroxyl groups also include the anionic form (-O). - ), its salts or solvates, and conventional forms of protection.

[0090] The compounds of the present invention can exist in a variety of tautomer forms, and it should be understood that the present invention includes all such tautomer forms.

[0091] In some compounds of this invention, chiral carbon atoms may be present depending on the nature of the substituents, and therefore these compounds may exist in stereoisomers. This invention extends to all optical isomers, such as stereoisomers of the compounds of this invention, including enantiomers, diastereomers, and mixtures thereof, such as racemates. Different stereoisomers can be separated or resolved from each other by conventional methods, or any given isomer can be obtained by conventional stereoselective or stereospecific synthesis.

[0092] It should also be understood that any atom present in the compounds of this invention can be in any available naturally occurring isotopic form. For example, a carbon atom can be... 12 C or 13 C. A hydrogen atom can be 1 H or 2 H (deuterium).

[0093] As used herein, the term “treatment” refers to therapeutic procedures and preventative or preventive measures aimed at preventing or mitigating undesirable physiological changes or conditions, such as the development or spread of a disease. “Treatment” can also refer to the extension of life compared to the expected lifespan without treatment. Those who require treatment include those who already have the condition, those who are predisposed to it, or those who need to prevent it.

[0094] The phrase “pharmaceutically acceptable” means that a substance or composition must be chemically and / or toxicologically compatible with the other components of the formulation and / or the patients treated with it.

[0095] The compounds of the present invention

[0096] This invention relates to a series of novel compounds and their use as inhibitors of the hypoxia-inducible factor prolyl hydroxylase domain (PHD inhibitors). Therefore, these compounds have potential use in treating conditions targeting HIF-PHD, including, for example, anemia and other ischemia-related diseases, and other conditions mentioned below.

[0097] Therefore, the present invention provides compounds that are substituted azines of formula (I) or pharmaceutically acceptable salts thereof.

[0098]

[0099] In equation (I) above, X is C(R) 6 X is C(R) or N. Preferably, X is C(R). 6 That is, the preferred X is the same as R. 6 Bonded cyclic carbon atoms (in this case, the substituted acridine is a substituted pyridine). However, usually X is N (in this case, the substituted acridine is a substituted pyridazine).

[0100] R 0 It is H or unsubstituted or substituted C 1-6 Alkyl group. Typically, R 0 Is it H or unsubstituted C? 1-6 Alkyl group. Typically, R 0 Selected from H, methyl, and ethyl. Typically, R... 0 It is H or methyl.

[0101] R 1 It is H, unsubstituted or substituted C 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN, –N(R) t )C(O)N(R u )R v –C(O)OR w or –C(O)N(R) x )R 7 .

[0102] Typically, R 1 It is –C(O)N(R) x )R 7 When R 5 Not –C(O)N(R) x )R 7 This is especially typical at times. Therefore, when R 5 It is H, unsubstituted or substituted C 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN, –N(R) t )C(O)N(R u )R v Or –C(O)OR w At that time, R 1 It is –C(O)N(R) x )R 7 This is particularly typical.

[0103] Typically, R1 It can also be H, unsubstituted or substituted C. 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN, –N(R) t )C(O)N(R u )R v Or –C(O)OR w When R 5 –C(O)N(R) x )R 7 This is especially typical at times. For example, R 1 It can be H, unsubstituted or substituted C. 1-6 Alkyl, –CN or –C(O)OR w H, –CN, or –C(O)OR are preferred. w When R 5 It is –C(O)N(R) x )R 7 This situation usually occurs at that time.

[0104] Therefore, R 1 It can be selected from H, unsubstituted or substituted C. 1-6 Alkyl, –CN, –C(O)OR w and –C(O)N(R) x )R 7 R 1 It can be selected from, for example, H, –CN, –C(O)OR w and –C(O)N(R) x )R 7 R 1 It can be selected, for example, from H, unsubstituted or substituted C. 1-6 Alkyl, –CN and –C(O)OR w Or, for example, selected from H, –CN, and –C(O)OR w .

[0105] R 2 It is H, –OR q C, either unsubstituted or substituted 1-6 Alkyl group. Or, R 2 It is –N=, in this case, R 3 =C(R) y )–, and R 2 and R 3 Together forming formula – N = C(R) y )– groups.

[0106] R 2 For example, it could be H, –OR q C, whether substituted or not 1-6 Alkyl group. Typically, in this case, R2 It is H or unsubstituted or substituted C 1-6 Alkyl group. For example, R 2 It can be H or unsubstituted C 1-6 Alkyl groups, such as R 2 It can be H, ethyl, or methyl. R 2 For example, it could be H or methyl.

[0107] R 3 It is H, –OR 8 C, whether substituted or not 1-6 Alkyl group. Or, R 3 =C(R) y –, in this case, R 2 It is –N=, and R 2 and R 3 Together forming formula – N = C(R) y )– groups.

[0108] R 3 Examples could be H, –OR 8 C, whether substituted or not 1-6 Alkyl group. Typically, in this case, R 3 Is it H or –OR 8 R 3 It can also typically be selected from H or unsubstituted or substituted C. 1-6 Alkyl group. However, R 3 Usually –OR 8 .

[0109] In some implementation schemes, R 2 It is –N=, and R 3 =C(R) y )–, and R 2 and R 3 Together forming formula – N = C(R) y )– groups.

[0110] R 4 It is H, unsubstituted or substituted C 1-6 Alkyl, –OR 9 Or –C(O)OR 10 R 4 Examples could be H or unsubstituted C. 1-6 Alkyl, –OR 9 Or –C(O)OR 10 However, R 4 Usually H, –OR 9 Or –C(O)OR 10 Typically, R 4 Selected from –OR 9 and –C(O)OR10 , or R 4 Yes - OR 9 Or, R 4 It can be selected from H or unsubstituted or substituted C. 1-6 Alkyl groups, such as R 4 It could be H. However, R 4 It is usually C(O)OH or OH. For example, R4 is usually OH.

[0111] R 5 It is H, unsubstituted or substituted C 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN, –N(R) t )C(O)N(R u )R v –C(O)OR w or –C(O)N(R) x )R 7 .

[0112] R 5 Typically, it consists of H, unsubstituted or substituted C. 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN, –N(R) t )C(O)N(R u )R v Or –C(O)OR w When R 1 It is –C(O)N(R) x )R 7 This is especially typical at times. For example, R 5 It can be H, unsubstituted or substituted C. 1-6 Alkyl, –CN or –C(O)OR w H, –CN, or –C(O)OR are preferred. w When R 1 It is –C(O)N(R) x )R 7 This situation usually occurs at that time.

[0113] Typically, R 5 It can also be –C(O)N(R) x )R 7 When R 1 Not –C(O)N(R) x )R 7 This is especially typical at times. Therefore, when R 1 It is H, unsubstituted or substituted C 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN, –N(R) t )C(O)N(Ru )R v Or –C(O)OR w At that time, R 5 It is –C(O)N(R) x )R 7 This is particularly typical.

[0114] Typically, R 5 It is H, unsubstituted or substituted C 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN, –C(O)OR w or –C(O)N(R) x )R 7 Typically, R 5 Selected from H, unsubstituted or substituted C 1-6 Alkyl, –CN, –C(O)OR w and –C(O)N(R) x )R 7 R 5 It can be selected from, for example, H, –CN, –C(O)OR w and –C(O)N(R) x )R 7 R 5 It can be selected, for example, from H, unsubstituted or substituted C. 1-6 Alkyl, –CN and –C(O)OR w Or, for example, selected from H, –CN, and –C(O)OR w R 5 For example, it could be –C(O)OR w R 5 It is usually –C(O)OH.

[0115] R 6 It is H or unsubstituted or substituted C 1-6 Alkyl group. Typically R 6 Is it H or unsubstituted C? 1-6 Alkyl group. Typically, R 6 It is H.

[0116] R 7 It is –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar, –Ary–Ar, –CH(R 11 –Cyc, –CH2C≡CCH3, –Cyc, or –Ar. R 7 For example, it could be –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar, –Ary–Ar or –CH(R 11 )–Cyc.

[0117] Ar is an unsubstituted or substituted aryl group or an unsubstituted or substituted heteroaryl group. For example, Ar can be an unsubstituted or substituted aryl group or an unsubstituted heteroaryl group. For example, Ar can be an unsubstituted or substituted phenyl group or an unsubstituted heteroaryl group. Ar can be selected, for example, from unsubstituted phenyl groups, unsubstituted pyrimidinyl groups, unsubstituted benzothiazole groups, and phenyl groups substituted with –C(O)OH, –C(O)OMe, –C(O)OEt, –C(O)NH2, –C(O)N(H)Me, –OMe, or N-morpholinoyl.

[0118] Ary is an unsubstituted or substituted arylene, or an unsubstituted or substituted heteroarylene. Ary can be, for example, an unsubstituted arylene or an unsubstituted heteroarylene. Typically, Ary is an unsubstituted phenylene or an unsubstituted pyridylene.

[0119] Cyc is unsubstituted or substituted C 3-10 Cycloalkyl. Typically, Cyc is an unsubstituted or substituted cyclohexyl group. For example, Cyc can be an unsubstituted cyclohexyl group or a cyclohexyl group substituted with –CF3 or –OCF3.

[0120] R 11 It is H, –C(O)OR z C, whether substituted or not 1-4 Alkyl group. R 11 Examples could be H, –C(O)OR z or unreplaced C 1-4 Alkyl group. Typically, R 11 It is H, –C(O)OR z Or methyl.

[0121] R 8 R 9 and R 10 Each is independently selected from H and unsubstituted or substituted C. 1-6 alkyl.

[0122] Therefore R 8 It can be H or unsubstituted or substituted C. 1-6 Alkyl group. Typically, R 8 It's H.

[0123] R 9 It can also be selected from H or unsubstituted or substituted C. 1-6 Alkyl group. Typically, R 9 It's H.

[0124] R 10 It can also be selected from H or unsubstituted or substituted C. 1-6 Alkyl group. Typically, R 10 It's H.

[0125] Typically, R 8 R 9 and R 10 They can be the same or different, and they are each independently selected from H and unsubstituted C. 1-6 Alkyl groups and those with phenyl groups or –OC(O)R 99 Replacement C 1-6 Alkyl group. R 99 It is phenyl, unsubstituted C 1-6 Alkyl, –N(R) a (R) b –C(O)R c –OR d Or amino acids. R a R b R c and R d Each is independently selected from H, unsubstituted or substituted C. 1-6 Alkyl groups and amino acids.

[0126] Therefore, R 8 Usually selected from H, unsubstituted C 1-6 Alkyl groups and those with phenyl groups or -OC(O)R 99 Replacement C 1-6 Alkyl, wherein R 99 As defined above.

[0127] Typically, R 9 Selected from H, unsubstituted C 1-6 Alkyl groups and those with phenyl groups or -OC(O)R 99 Replacement C 1-6 Alkyl, wherein R 99 As defined above.

[0128] R 10 Usually selected from H, unsubstituted C 1-6 Alkyl groups and those with phenyl groups or -OC(O)R 99 Replacement C 1-6 Alkyl, wherein R 99 As defined above.

[0129] Where R 8 R 9 or R 10 Structures of formula (I) that are not H include prodrug compounds. Specifically, where R... 8 R 9 or R 10 Is it unsubstituted or substituted C? 1-6 Alkyl groups, especially substituted C groups 1-6 Alkyl-substituted azazides of formula (I) include prodrug compounds. For example, where R 8 R 9or R 10 It is a phenyl or -OC(O)R 99 (where R) 99 As defined above) replaces C 1-6 Alkyl-substituted azazons of formula (I) include prodrug compounds. R 8 R 9 or R 10 For example, it can be a replacement for C. 1-6 Alkyl, wherein C 1-6 Substituents on the alkyl group, or one of them, of the formula -OC(O)R 99 The group, wherein R 99 It is phenyl, unsubstituted C 1-6 Alkyl, –N(R) a (R) b ), –C(O)R c –OR d Or amino acids, and in which R a R b R c and R d Each is independently selected from H, unsubstituted or substituted C. 1-6 Alkyl groups and amino acids. Such compounds include prodrugs. Therefore, R 8 It is usually -OC(O)R 99 Replacement C 1-6 Alkyl, wherein R 99 As defined above. Similarly, R 9 It can be -OC(O)R 99 Replacement C 1-6 Alkyl, wherein R 99 As defined above. R 10 It can be -OC(O)R 99 Replacement C 1-6 Alkyl, wherein R 99 As defined above.

[0130] Typically, R a R b R c and R d Each is independently selected from H or unsubstituted or substituted C. 1-6 Alkyl group. Typically, R a R b R c and R d Each is independently selected from H, methyl, or ethyl.

[0131] R t R u R v R w R x Ry and R z Each is independently selected from H, unsubstituted or substituted C. 1-6 Alkyl groups and unsubstituted or substituted phenyl groups.

[0132] R t Is it unsubstituted or substituted C? 1-4 Alkyl or H. Typically, R t It is unreplaced C 1-4 Alkyl or H. Typically, R... t It's H.

[0133] R u Is it unsubstituted or substituted C? 1-4 Alkyl or H. Typically, R u It is unreplaced C 1-4 Alkyl or H. Typically, R... u It's H.

[0134] R v Is it unsubstituted or substituted C? 1-4 Alkyl or H. Typically, R v It is unreplaced C 1-4 Alkyl or H. Typically, R... v It's H.

[0135] Therefore, R t R u and R v It's usually H.

[0136] R x It is usually H.

[0137] Typically, R y Is it H or unsubstituted C? 1-6 Alkyl groups, such as R y It can be H or methyl.

[0138] R z It is usually H.

[0139] R w It is H, unsubstituted or substituted C 1-6 Alkyl group, or unsubstituted or substituted phenyl group. R w It can be H or unsubstituted or substituted C. 1-6 Alkyl group. Typically, for example, R w It is H, unsubstituted C 1-6 Alkyl, or phenyl or –OC(O)R ww Replacement C 1-6 Alkyl, wherein R ww It is phenyl, unsubstituted C 1-6 Alkyl, –N(R) a (R)b ), –C(O)R c –OR d Or amino acids, of which R a R b R c and R d Each is independently selected from H, unsubstituted or substituted C. 1-6 Alkyl groups and amino acids.

[0140] Where R w Substituted azazons of formula (I) that are not H include prodrug compounds. Specifically, where R... w Is it unsubstituted or substituted C? 1-6 Compounds of formula (I) containing alkyl or unsubstituted or substituted phenyl groups include prodrug compounds. For example, where R... w It is a phenyl or –OC(O)R ww (where R) ww As defined above) replaces C 1-6 Alkyl compounds of formula (I) include prodrug compounds. R w For example, it can be a replacement for C. 1-6 Alkyl, wherein C 1-6 Substituents on the alkyl group, or one of them, of the formula -OC(O)R ww The group, wherein R ww It is phenyl, unsubstituted C 1-6 Alkyl, –N(R) a (R) b ), –C(O)R c –OR d Or amino acids, of which R a R b R c and R d Each is independently selected from H, unsubstituted or substituted C. 1-6 Alkyl groups and amino acids. Such compounds include prodrugs. Therefore, R w It is usually -OC(O)R ww Replacement C 1-6 Alkyl, wherein R ww As defined above. Typically, R a R b R c and R d Each is independently selected from H or unsubstituted or substituted C. 1-6 Alkyl group. Typically, R a R b R c and R d Each is independently selected from H, methyl, or ethyl. R w It can be H or unsubstituted C. 1-6Alkyl groups or phenyl groups or -OC(O)R ww Replacement C 1-6 Alkyl, wherein R ww It is a phenyl or unsubstituted C 1-6 alkyl.

[0141] R w It can be H.

[0142] R q It is H, unsubstituted or substituted C 1-6 Alkyl, or unsubstituted or substituted phenyl. Typically, R q It's H.

[0143] Typically, in equation (I), R 1 and R 5 One of them is –C(O)N(R) x )R 7 And R 1 and R 5 The other one is not –C(O)N(R) x )R 7 When R 1 and R 5 One of them is not –C(O)N(R) x )R 7 When, it can be R specified in this article. 1 or R 5 Any other definition of it. Therefore, it can be H, unsubstituted or substituted C. 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN, –N(R) t )C(O)N(R u )R v Or –C(O)OR w Any one of them.

[0144] Typically, in equation (I), R 1 and R 5 One of them is –C(O)N(R) x )R 7 And R 1 and R 5 The other one is H, unsubstituted or substituted C. 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN, –N(R) t )C(O)N(R u )R v Or –C(O)OR w Typically, R 1 and R 5 One of them is –C(O)N(R) x )R7 And R 1 and R 5 The other one is H, unsubstituted or substituted C. 1-6 Alkyl, –CN or –C(O)OR w For example, R 1 and R 5 One of them could be –C(O)N(R) x )R 7 And R 1 and R 5 The other one can be H, –CN, or –C(O)OR w .

[0145] In some embodiments of the substituted azazine of formula (I):

[0146] R 0 Is it H or unsubstituted C? 1-6 alkyl;

[0147] R 1 It is H, –CN, –C(O)OR w or –C(O)N(R) x )R 7 ;

[0148] R 2 Is it H or unsubstituted C? 1-6 Alkyl; and R 3 Is it H or –OR 8 Or R 2 It is –N= and R 3 =C(R) y ) – and R 2 and R 3 Together forming formula – N = C(R) y The group )–. In such an embodiment, R 4 It can be H or –OR 9 Or –C(O)OR 10 ;R 5 It can be H, –CN, –C(O)OR w Or –C(O)N(R) x )R 7 And R 6 It is H. Furthermore, typically, R 7 It is –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar, –Ary–Ar or –CH(R 11 )–Cyc, where Ar is an unsubstituted or substituted aryl or an unsubstituted or substituted heteroaryl, Ary is an unsubstituted arylene or an unsubstituted heteroaryl, and Cyc is an unsubstituted or substituted C3-10 cycloalkyl, R 11 It is H, –C(O)OR z or unreplaced C 1-4 alkyl.

[0149] Typically in such implementation schemes, R 8 R 9 and R 10 Each is independently selected from H and unsubstituted or substituted C. 1-6 Alkyl group. Typically, R x It is H, R z It is H, R y Is it H or unsubstituted C? 1-6 Alkyl, R w It is H, unsubstituted C 1-6 Alkyl or phenyl or –OC(O)R ww Replacement C 1-6 Alkyl, wherein R ww It is phenyl, unsubstituted C 1-6 Alkyl, –N(R) a (R) b ), –C(O)R c –OR d Or amino acids, of which R a R b R c and R d Each is independently selected from H, unsubstituted or substituted C. 1-6 Alkyl groups and amino acids. Typically, R a R b R c and R d Each is independently selected from H or unsubstituted or substituted C. 1-6 Alkyl group. Typically, R a R b R c and R d Each is independently selected from H, methyl, or ethyl.

[0150] Typically, compounds are provided in which R 1 and R 5 One of them is –C(O)N(R) x )R 7 R 1 and R 5 The other one is H, –CN, or –C(O)OR. w .

[0151] Therefore, typically, R 0 Is it H or unsubstituted C? 1-6 Alkyl; R 1It is H, –CN, –C(O)OR w or –C(O)N(R) x )R 7 ;R 2 It is H or an unsubstituted C1-6 alkyl; and R 3 Is it H or –OR 8 Or R 2 It is –N= and R 3 =C(R) y )–, and R 2 and R 3 Together forming formula – N = C(R) y ) group; R 4 It is H, –OR 9 Or –C(O)OR 10 ;R 5 It is H, –CN, –C(O)OR w or –C(O)N(R) x )R 7 ;R 6 It is H; R 7 It is –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar, –Ary–Ar or –CH(R 11 )–Cyc, where Ar is an unsubstituted or substituted aryl or an unsubstituted or substituted heteroaryl, Ary is an unsubstituted arylene or an unsubstituted heteroaryl, and Cyc is an unsubstituted or substituted C 3-10 Cycloalkyl, and R 11 It is H, –C(O)OR z or unreplaced C 1-4 Alkyl; R 8 R 9 and R 10 Each is independently selected from H and unsubstituted or substituted C. 1-6 Alkyl; and R x It is H, R z It is H, R y Is it H or unsubstituted C? 1-6 Alkyl, and R w It is H, unsubstituted C 1-6 Alkyl or phenyl or –OC(O)R ww Replacement C 1-6 Alkyl, wherein R ww It is phenyl, unsubstituted C 1-6 Alkyl, –N(R) a (R) b ), –C(O)R c –OR d Or amino acids, of which R a Rb R c and R d Each is independently selected from H, unsubstituted or substituted C. 1-6 Alkyl groups and amino acids; condition R 1 and R 5 One of them is –C(O)N(R) x )R 7 And R 1 and R 5 The other one is H, –CN, or –C(O)OR. w .

[0152] In some embodiments of the substituted azazine of formula (I), R 0 It is H or methyl; R 1 It is H, –CN, –C(O)OR w 、or –C(O)N(R) x )R 7 ;R 2 It is H or methyl; and R 3 Is it H or –OR 8 Or R 2 It is –N= and R 3 =C(R) y )–, and R 2 and R 3 Together forming formula – N = C(R) y )– groups.

[0153] Typically, in such compounds, R 4 It is H, –OR 9 Or –C(O)OR 10 ;R 5 It is H, –CN, –C(O)OR w , or –C(O)N(R x )R 7 ;R 6 It is H; and R 7 It is –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar, –Ary–Ar or –CH(R 11 –Cyc; where Ar is an unsubstituted phenyl, unsubstituted pyrimidinyl, unsubstituted benzothiazole, or a phenyl substituted with –C(O)OH, –C(O)OMe, –C(O)OEt, –C(O)NH2, –C(O)N(H)Me, –OMe, or N-morpholino; Ary is an unsubstituted phenylene or unsubstituted pyridylene; Cyc is an unsubstituted cyclohexyl or a cyclohexyl substituted with –CF3 or –OCF3; and R 11 It is H, –C(O)ORz Or methyl.

[0154] Typically, R 8 R 9 and R 10 Each is independently selected from H and unsubstituted C. 1-6 Alkyl, or phenyl or –OC(O)R 99 Replacement C 1-6 Alkyl, wherein R 99 It is a phenyl group, with an unsubstituted C. 1-6 Alkyl, –N(R) a (R) b –C(O)R c –OR d Or amino acids, of which R a R b R c and R d Each is independently selected from H, unsubstituted or substituted C. 1-6 Alkyl groups and amino acids. Typically, R a R b R c and R d Each is independently selected from H and unsubstituted or substituted C. 1-6 Alkyl group. Typically, R a R b R c and R d Each is independently selected from H, methyl, or ethyl.

[0155] Typically, R x It is H; R z It is H; R w It is H, unsubstituted C 1-6 Alkyl groups or phenyl groups or -OC(O)R ww Replacement C 1-6 Alkyl, wherein R ww It is a phenyl or unsubstituted C 1-6 Alkyl; and R y It is H or methyl.

[0156] For compounds like those of the present invention, the typical form is R. 1 and R 5 One of them is –C(O)N(R) x )R 7 And R 1 and R 5 The other one is H, –CN, or –C(O)OR. w .

[0157] Therefore, typically, R 0 It is H or methyl; R 1It is H, –CN, –C(O)OR w or –C(O)N(R) x )R 7 ;R 2 It is H or methyl; and R 3 Is it H or –OR 8 Or R 2 It is –N=, and R 3 =C(R) y )–, and R 2 and R 3 Together forming formula – N = C(R) y )– group; R 4 It is H, –OR 9 Or –C(O)OR 10 ;R 5 It is H, –CN, –C(O)OR w or –C(O)N(R) x )R 7 ;R 6 It is H; R 7 It is –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar, –Ary–Ar or –CH(R 11 Ar is an unsubstituted phenyl, unsubstituted pyrimidinyl, unsubstituted benzothiazole, or a phenyl substituted with –C(O)OH, –C(O)OMe, –C(O)OEt, –C(O)NH2, –C(O)N(H)Me, –OMe, or N-morpholino; Ary is an unsubstituted phenylene or unsubstituted pyridylene; Cyc is an unsubstituted cyclohexyl or a cyclohexyl substituted with –CF3 or –OCF3; and R 11 It is H, –C(O)OR z or methyl; R 8 R 9 and R 10 Each is independently selected from H and unsubstituted C. 1-6 Alkyl groups and those with phenyl groups or –OC(O)R 99 Replacement C 1-6 Alkyl, wherein R 99 It is phenyl, unsubstituted C 1-6 Alkyl, –N(R) a (R) b –C(O)R c –OR d Or amino acids, of which R a R b R c and R d Each is independently selected from H, unsubstituted or substituted C. 1-6Alkyl groups and amino acids; and R x It is H; R z It is H; R w It is H, unsubstituted C 1-6 Alkyl groups or phenyl groups or -OC(O)R ww Replacement C 1-6 Alkyl, wherein R ww It is a phenyl or unsubstituted C 1-6 Alkyl; and R y It is H or methyl; the condition is R. 1 and R 5 One of them is –C(O)N(R) x )R 7 And R 1 and R 5 The other one is H, –CN, or –C(O)OR. w .

[0158] Typically, in substituted azazides of formula (I), the situation is (a)R 5 It is –C(O)N(R) x )R 7 and (b)R 3 Yes – OR 8 or R 4 Yes – OR 9 In these implementation schemes, R x R 7 R 8 and R 9 The compounds of this invention may be defined as described anywhere herein.

[0159] In another typical case, in the substituted azazine of formula (I), (a)R 1 It is –C(O)N(R) x )R 7 , and (b)R 4 Yes – OR 9 Or –C(O)OR 10 , or R 5 It is –C(O)OR w In these implementation schemes, R x R 7 R 8 and R 9 The compounds of this invention may be defined as described anywhere herein.

[0160] Therefore, typically, among the substituted azazines of formula (I), one of the following is true:

[0161] (1)(a)R 5 It is –C(O)N(R) x )R 7, and (b)R 3 Is – OR 8 or R 4 Is – OR 9 ;or

[0162] (2)(a)R 1 It is –C(O)N(R) x )R 7 , and (b)R 4 Yes – OR 9 Or –C(O)OR 10 , or R 5 It is –C(O)OR w .

[0163] R x R 7 R 8 and R 9 It can be further defined as described herein. In some implementations, for example, R x It is H, R 8 It is H, R 9 It is H, and R 7 As defined anywhere in this article. For example, R 7 It can be –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar, –Ary–Ar or –CH(R 11 )–Cyc, where Ar is an unsubstituted or substituted aryl or an unsubstituted or substituted heteroaryl, Ary is an unsubstituted arylene or an unsubstituted heteroaryl, and Cyc is an unsubstituted or substituted C 3-10 cycloalkyl, R 11 It is H, –C(O)OR z or unreplaced C 1-4 Alkyl group. R z It can be defined as anywhere in this document, but it is usually H.R. 7 For example, it could be –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar, –Ary–Ar or –CH(R 11 –Cyc; where Ar is an unsubstituted phenyl, unsubstituted pyrimidinyl, unsubstituted benzothiazole, or a phenyl substituted with –C(O)OH, –C(O)OMe, –C(O)OEt, –C(O)NH2, –C(O)N(H)Me, –OMe, or N-morpholino; Ary is an unsubstituted phenylene or unsubstituted pyridylene; Cyc is an unsubstituted cyclohexyl or a cyclohexyl substituted with –CF3 or –OCF3; and R 11 It is H, –C(O)OR z Or methyl. Rz It can be as defined anywhere in this document, but it is usually H.

[0164] In the compounds of the present invention, the substituted acridine may have the following formula (Ia). Therefore, in some embodiments, the present invention relates to substituted acridine compounds of formula (Ia) or pharmaceutically acceptable salts thereof:

[0165]

[0166] R in equation (Ia) 0 R 9 X, R x and R 7 Each can be defined as described anywhere in this document for equation (Ia). R in equation (Ia) 1 It can be H, unsubstituted or substituted C. 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN, –N(R) t )C(O)N(R u )R v Or –C(O)OR w , where R t R u R v and R w Equation (I) can be defined as anywhere in this document. R in equation (Ia) 2 It is H, –OR q C, whether substituted or not 1-6 Alkyl, wherein R q It is H, unsubstituted or substituted C 1-6 Alkyl, or unsubstituted or substituted phenyl. R in formula (Ia) 3 It is H or unsubstituted or substituted C 1-6 alkyl.

[0167] Therefore, when the substituted azazine of the compound of the present invention has formula (Ia), typically, X is C(R) 6 X is C(R) or N. Preferably, X is C(R). 6 However, alternatively, X can be N.

[0168] Typically, R 0 Is it H or unsubstituted or substituted C? 1-6 Alkyl group. However, R in formula (Ia) 0 Equation (I) can be further defined as anywhere in this document.

[0169] R 1 Typically, it consists of H, unsubstituted or substituted C. 1-6Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN or –C(O)OR w However, R in equation (Ia) 1 Equation (I) can be further defined as anywhere in this document.

[0170] R 2 It can be H or –OR q C, whether substituted or not 1-6 Alkyl, and R 3 It can be H or unsubstituted or substituted C. 1-6 Alkyl group. However, R in formula (Ia) 2 and R 3 Equation (I) can be further defined as anywhere in this document.

[0171] Typically, R 6 It is H or unsubstituted or substituted C 1-6 Alkyl group. However, R in formula (Ia) 6 Equation (I) can be further defined as anywhere in this document.

[0172] R 7 Usually –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar, –Ary–Ar, –CH(R 11 –Cyc, –CH2C≡CCH3, –Cyc, or –Ar, where Ar is an unsubstituted or substituted aryl group, or an unsubstituted or substituted heteroaryl group, Ary is an unsubstituted or substituted arylene group, or an unsubstituted or substituted heteroarylene group, and Cyc is an unsubstituted or substituted C group. 3-10 Cycloalkyl, and R 11 Usually H, –C(O)OR z C, whether substituted or not 1-4 Alkyl group. However, R in formula (Ia) 7 and R 11 Equation (I) can be further defined as anywhere in this document.

[0173] In embodiments where the substituted azazine has the formula (Ia), R 9 Typically, it is H or unsubstituted or substituted C. 1-6 Alkyl group. R w R x and R z Each can be independently selected from H, unsubstituted or substituted C. 1-4 Alkyl groups and unsubstituted or substituted phenyl groups. Typically, R q It is H, unsubstituted or substituted C 1-6Alkyl, or unsubstituted or substituted phenyl. However, R in formula (Ia) 9 R w R x R z and R q Equation (I) can be further defined as anywhere in this document.

[0174] In some preferred embodiments, the substituted azazine has the formula (Ia), R 9 It is H.

[0175] The compounds of the present invention may be substituted azazines selected from any of the following structures of formula (Ia) or pharmaceutically acceptable salts thereof:

[0176]

[0177]

[0178]

[0179]

[0180] The numbers in parentheses next to the above structures match the compound numbers given in the Examples section below.

[0181] In the compounds of the present invention, the substituted acridine may have the following formula (Ib). Therefore, in some embodiments, the present invention relates to substituted acridine compounds of formula (Ib) or pharmaceutically acceptable salts thereof:

[0182]

[0183] R in equation (Ib) 0 R 4 R 6 R 7 R 8 and R x Each can be defined as it is anywhere in this document for equation (I). R in equation (Ib) 1 It can be H, unsubstituted or substituted C. 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN, –N(R) t )C(O)N(R u )R v Or –C(O)OR w , where R t R u R v and R w Equation (I) can be defined as anywhere in this document. R in equation (Ib)2 It is H, –OR q C, whether substituted or not 1-6 Alkyl, wherein R q It is H, unsubstituted or substituted C 1-6 Alkyl, or unsubstituted or substituted phenyl.

[0184] Therefore, when the substituted azazine compounds of the present invention have formula (Ib), typically, R 0 It is H or unsubstituted or substituted C 1-6 Alkyl group. However, R in formula (Ib) 0 Equation (I) can be further defined as anywhere in this document.

[0185] R 1 It can be H, unsubstituted or substituted C. 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN or –C(O)OR w However, R in equation (Ib) 1 Equation (I) can be further defined as anywhere in this document.

[0186] Typically, R 2 It is H, –OR q C, either unsubstituted or substituted 1-6 Alkyl group. However, R in formula (Ib) 2 Equation (I) can be further defined as anywhere in this document.

[0187] Typically, R in equation (Ib) 4 It is H or unsubstituted or substituted C 1-6 Alkyl group. R 6 It can be H or unsubstituted or substituted C. 1-6 Alkyl group. For example, R 4 It can be H or unsubstituted C 1-6 Alkyl groups, and typically H. R in formula (Ib) 4 Equation (I) can be further defined as anywhere in this document.

[0188] Typically, R 6 It is H or unsubstituted or substituted C 1-6 Alkyl group. For example, R 6 It can be H or unsubstituted C 1-6 Alkyl groups, and typically H. R in formula (Ib) 6 Equation (I) can be further defined as anywhere in this document.

[0189] Typically, R 7 It is –CH(R) 11 –Ar、–CH(R)11 )–Ary–Ar, –Ary–Ar, –CH(R 11 –Cyc, –Cyc, or –Ar, where Ar is an unsubstituted or substituted aryl group, or an unsubstituted or substituted heteroaryl group, Ary is an unsubstituted or substituted arylene group, or an unsubstituted or substituted heteroaryl group, and Cyc is an unsubstituted or substituted C group. 3-10 Cycloalkyl, and R 11 It is H, –C(O)OR z C, whether substituted or not 1-4 Alkyl group. However, R in formula (Ib) 7 and R 11 Equation (I) can be further defined as anywhere in this document.

[0190] Typically, for compounds of formula (Ib) or their pharmaceutically acceptable salts, R 8 It is H or unsubstituted or substituted C 1-6 Alkyl group. However, R in formula (Ib) 8 Equation (I) can be further defined as anywhere in this document.

[0191] Typically, R w R x and R z Each is independently selected from H, unsubstituted or substituted C. 1-4 Alkyl groups and unsubstituted or substituted phenyl groups. Typically, R q It is H, unsubstituted or substituted C 1-6 Alkyl groups, or unsubstituted or substituted phenyl groups. For example, R q Typically, it is H. However, R in equation (Ib) w R x R z and R q Equation (I) can be further defined as anywhere in this document.

[0192] In some preferred substituted azazines of formula (Ib), R 8 It's H.

[0193] The compounds of the present invention may be substituted azazines selected from any of the following structures of formula (Ib) or pharmaceutically acceptable salts thereof:

[0194]

[0195]

[0196] The numbers in parentheses next to the above structures match the compound numbers given in the Examples section below.

[0197] In the compounds of the present invention, the substituted acridine may have the following formula (Ic). Therefore, in some embodiments, the present invention relates to substituted acridine compounds of formula (Ic) or pharmaceutically acceptable salts thereof:

[0198]

[0199] R in equation (Ic) 0 R 4 R 6 R x and R 7 Each can be defined as described anywhere in this document for equation (I). R in equation (Ic) 5 It can be H, unsubstituted or substituted C. 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN, –N(R) t )C(O)N(R u )R v Or –C(O)OR w , where R t R u R v and R w Equation (I) can be defined anywhere in this document. R in equation (Ic) 2 It is H, –OR q C, whether substituted or not 1-6 Alkyl, wherein R q It is H, unsubstituted or substituted C 1-6 Alkyl, or unsubstituted or substituted phenyl. R in formula (Ic) 3 It is H, –OR 8 C, whether substituted or not 1-6 Alkyl, wherein R 8 Selected from H and unsubstituted or substituted C 1-6 alkyl.

[0200] Therefore, when the substituted azazine compounds of the present invention have formula (Ic), typically, R 0 It is H or unsubstituted or substituted C 1-6 Alkyl group. However, R in formula (Ic) 0 Further definitions of equation (I) can be found anywhere in this document. Typically, R... 2 It is H, –OR q C, either unsubstituted or substituted 1-6 Alkyl group. However, R in formula (Ic) 2 Equation (I) can be further defined as anywhere in this document. 3 It can be H or –OR 8 C, either unsubstituted or substituted1-6 Alkyl group. However, R in formula (Ic) 3 As can be further defined anywhere in this text for equation (I), R 8 Similarly. R in equation (Ic) 3 Usually H or –OR 8 R 8 For example, it could be unsubstituted C. 1-6 alkyl.

[0201] For substituted azazons of formula (Ic), R is usually... 4 It is H, unsubstituted or substituted C 1-6 Alkyl, –OR 9 Or –C(O)OR 10 , where R 9 and R 10 As defined anywhere in this document, equation (I) is used. However, R in equation (Ic) 4 Equation (I) can be further defined as anywhere in this document. 4 It can be selected from, for example, H, –OR 9 Or –C(O)OR 10 R 4 It can be selected, for example, from H, –OH or –C(O)OH.

[0202] R in equation (Ic) 5 It can be H, unsubstituted or substituted C. 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN or –C(O)OR w However, R in equation (Ic) 5 As can be further defined anywhere in this text for equation (I), R w That's also possible.

[0203] Typically, R 6 It is H or unsubstituted or substituted C 1-6 Alkyl group. However, R in formula (Ic) 6 Equation (I) can be further defined as anywhere in this document.

[0204] R 7 It can be –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar, –Ary–Ar, –CH(R 11 –Cyc, –Cyc, or –Ar, where Ar is an unsubstituted or substituted aryl group, or an unsubstituted or substituted heteroaryl group, Ary is an unsubstituted or substituted arylene group, or an unsubstituted or substituted heteroaryl group, and Cyc is an unsubstituted or substituted C group. 3-10 Cycloalkyl, and R11 It is H, –C(O)OR z C, whether substituted or not 1-4 Alkyl group. However, R in formula (Ic) 7 and R 11 Equation (I) can be further defined as anywhere in this document.

[0205] In embodiments where the substituted azazine has the formula (Ic), R 8 R 9 and R 10 Each is independently selected from H and unsubstituted or substituted C. 1-6 Alkyl group. However, R in formula (Ic) 8 R 9 and R 10 Each can be further defined as in any place in this document as Equation (I) is.

[0206] Typically, R w R x and R z Each is independently selected from H, unsubstituted or substituted C. 1-4 Alkyl groups and unsubstituted or substituted phenyl groups. However, R in formula (Ic) w R x and R z Each can be further defined as in any place in this document as Equation (I) is.

[0207] R q It can be H, unsubstituted or substituted C. 1-6 Alkyl, or unsubstituted or substituted phenyl. However, R in formula (Ic) q Equation (I) can be further defined as anywhere in this document.

[0208] Typically, in substituted azazides of formula (Ic), R 4 It is OH or C(O)OH. For example, in substituted azazides of formula (Ic), the usual case is R. 4 It is OH or C(O)OH, R 5 It is H. For example, R 4 It can be OH, R 5 It can be H. Similarly, typically, R 4 It is C(O)OH, R 5 It is H. Similarly, typically, R 4 It is OH or C(O)OH, and R5 is CN. For example, R 4 It can be OH, R 5 It can be CN.

[0209] Similarly, in substituted azazines of formula (Ic), R5 It is C(O)OH. For example, in substituted azazides of formula (Ic), the usual case is R. 5 It is C(O)OH, R 4 It is H. Similarly, typically, R 5 It is C(O)OH, R 4 It is OH.

[0210] Therefore, in some preferred embodiments of the substituted azazine of formula (Ic): (i)R 4 It is OH or C(O)OH, and / or (ii)R 5 It is C(O)OH.

[0211] In fact, preferably, in the substituted azazine of formula (Ic): (a) R 4 It is OH; or (b)R 4 It is C(O)OH; or (c)R 5 It is C(O)OH; or (d)R 4 It is OH, and R 5 It is C(O)OH.

[0212] The compounds of the present invention may be substituted azazines selected from any of the following structures of formula (Ic) or pharmaceutically acceptable salts thereof:

[0213]

[0214]

[0215]

[0216] The numbers in parentheses next to the above structures match the compound numbers given in the Examples section below.

[0217] In the compounds of the present invention, the substituted acridine may have the following formula (Id). Therefore, in some embodiments, the present invention relates to substituted acridine compounds of formula (Id) or pharmaceutically acceptable salts thereof:

[0218] R in equation (Id) 0 R 1 R 6 R 9 R y R x and R 7 Each can be defined as shown anywhere in this document for equation (I). However, R in equation (Id) 1 Typically, it consists of H, unsubstituted or substituted C. 1-6Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN, –N(R) t )C(O)N(R u )R v Or –C(O)OR w , where R t R u R v and R w Equation (I) can be defined as it is anywhere in this document.

[0219] Therefore, when the substituted azazine of the compounds of the present invention has the formula (Id), typically, R 0 It is H or unsubstituted or substituted C 1-6 Alkyl group. However, R in formula (Id) 0 Equation (I) can be further defined anywhere in this document. R in equation (Id) 0 It is usually H or methyl. Typically, it is methyl.

[0220] Typically, R in equation (Id) 1 It is H, unsubstituted or substituted C 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN or –C(O)OR w However, R in equation (Id) 1 Equation (I) can be further defined anywhere in this document. R in equation (Id) 1 H is preferred.

[0221] R in equation (Id) w Selected from H, unsubstituted or substituted C 1-4 Alkyl groups, and unsubstituted or substituted phenyl groups. However, R in formula (Id) w Equation (I) can be further defined as anywhere in this document.

[0222] Typically, R y Selected from H, unsubstituted or substituted C 1-4 Alkyl groups, and unsubstituted or substituted phenyl groups. R in formula (Id) y Equation (I) can be further defined as anywhere in this document. However, R in equation (Id) y It is usually H or methyl.

[0223] Typically, in equation (Id), R y and R 0 Both are methyl groups. R y and R 0 They can all be hydrogen.

[0224] Typically, R 6It is H or unsubstituted or substituted C 1-6 Alkyl group. R in formula (Id) 6 Further definitions of equation (I) are possible anywhere in this document. However, generally, R in equation (Id) 6 It's H.

[0225] For compounds of formula (Id) or their pharmaceutically acceptable salts, R 7 Usually –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar, –Ary–Ar, –CH(R 11 –Cyc, –Cyc, or –Ar, where Ar is an unsubstituted or substituted aryl group, or an unsubstituted or substituted heteroaryl group, Ary is an unsubstituted or substituted arylene group, or an unsubstituted or substituted heteroaryl group, and Cyc is an unsubstituted or substituted C group. 3-10 Cycloalkyl, and typically, R 11 It is H, –C(O)OR z C, whether substituted or not 1-4 Alkyl group. However, R in formula (Id) 7 and R 11 Equation (I) can be further defined as anywhere in this document. z Selected from H, unsubstituted or substituted C 1-4 Alkyl groups, and unsubstituted or substituted phenyl groups. However, R in formula (Id) z Equation (I) can also be further defined as anywhere in this document.

[0226] Typically, R 9 It is H or unsubstituted or substituted C 1-6 alkyl.

[0227] R in equation (Id) x Selected from H, unsubstituted or substituted C 1-4 Alkyl groups, and unsubstituted or substituted phenyl groups. However, R in formula (Id) x Further definitions of equation (I) can be found anywhere in this document. Typically, R in equation (Id) x It's H.

[0228] In some preferred embodiments, the substituted azine has the formula (Id), R 9 It's H.

[0229] The compounds of the present invention may be substituted azazides selected from any of the following structures of formula (Id) or pharmaceutically acceptable salts thereof:

[0230]

[0231]

[0232]

[0233] The numbers in parentheses next to the structures listed above correspond to the compound numbers given in the Examples section below.

[0234] In another embodiment, the substituted azine of formula (I) has any one of the following structures. Therefore, the present invention provides a compound or a pharmaceutically acceptable salt thereof, the compound being a substituted azine having any one of the following structures:

[0235]

[0236] The numbers in parentheses next to the structures listed above correspond to the compound numbers given in the Examples section below.

[0237] The present invention also provides substituted azazine compounds of formula (Ia) as defined herein, or pharmaceutically acceptable salts thereof, wherein R in formula (Ia) 9 Not H (and where X, R) 0 R 1 R 2 R 3 R 7 and R x As defined in this document for formula Ia). Such compounds include prodrugs. Typically, R in this embodiment... 9 Is it unsubstituted or substituted C? 1-6 Alkyl group. In one aspect of this embodiment, R 9 It is the replacement of C 1-6 Alkyl group. R 9 It can be, for example, phenyl or –OC(O)R 99 Replacement C 1-6 Alkyl, wherein R 99 As defined above. Therefore, R 99 It can be phenyl, unsubstituted C 1-6 Alkyl, –N(R) a (R) b –C(O)R c –OR d Or amino acids, of which R a R b R c and R d Each is independently selected from H, unsubstituted or substituted C. 1-6 Alkyl groups and amino acids. Typically, R a R b R c and Rd Each is independently selected from H or unsubstituted or substituted C. 1-6 Alkyl group. Typically, R a R b R c and R d Each is independently selected from H-methyl or ethyl. R 9 It can be, for example, by -OC(O)R 99 Replacement C 1-6 Alkyl, wherein R 99 As defined above. In another aspect of this implementation scheme, R... 9 It is unreplaced C 1-6 Alkyl group. R 9 It can be, for example, methyl. The substituted azazine of formula (Ia) can be selected, for example, from any of the following structures:

[0238]

[0239]

[0240]

[0241]

[0242] The numbers in parentheses next to the structures listed above correspond to the compound numbers given in the Examples section below.

[0243] The present invention also provides substituted azazine compounds of formula (Ib) as defined herein, or pharmaceutically acceptable salts thereof, wherein R in formula (Ib) 8 Not H (and where R) 0 R 1 R 2 R 4 R 6 R 7 and R x As defined in this document for formula Ib). Such compounds include prodrugs. Typically, R in this embodiment... 8 Is it unsubstituted or substituted C? 1-6 Alkyl group. In one aspect of this embodiment, R 8 It is the replacement of C 1-6 Alkyl group. R 8 It can be, for example, phenyl or –OC(O)R 99 Replacement C 1-6 Alkyl, wherein R 99 As defined above. Therefore, R 99 It can be phenyl, unsubstituted C 1-6 Alkyl, –N(R) a (R) b–C(O)R c –OR d Or amino acids, of which R a R b R c and R d Each is independently selected from H, unsubstituted or substituted C. 1-6 Alkyl groups and amino acids. Typically, R a R b R c and R d Each is independently selected from H or unsubstituted or substituted C. 1-6 Alkyl group. Typically, R a R b R c and R d Each is independently selected from H, methyl, or ethyl. R 8 It can be, for example, by -OC(O)R 99 Replacement C 1-6 Alkyl, wherein R 99 As defined above. In another aspect of this implementation scheme, R... 8 It is unreplaced C 1-6 Alkyl group. R 8 It can be, for example, methyl. The substituted azazine of formula (Ib) can be selected, for example, from any of the following structures:

[0244]

[0245]

[0246] The numbers in parentheses next to the structures listed above correspond to the compound numbers given in the Examples section below.

[0247] The present invention also provides substituted azazine compounds of formula (Ic) as defined herein, or pharmaceutically acceptable salts thereof, wherein R 4 Yes – OR 9 Or –C(O)OR 10 , and / or R 5 It is –C(O)OR w , where R 9 R 10 and R w Not H (and where R) 0 R 2 R 3 R 4 R 5 R 6 R 7 and R xAs defined in this article for formula Ic). Such compounds include prodrugs. Typically, in substituted azazides of formula (Ic): (a) R 4 Yes – OR 9 ; or (b)R 4 It is –C(O)OR 10 ; or (c)R 5 It is –C(O)OR w ; or (d)R 4 Yes – OR 9 R 5 It is –C(O)OR w R 9 R 10 and R w Same or different, is C that is unsubstituted or substituted. 1-6 alkyl.

[0248] In one aspect of this implementation plan, R 9 R 10 and R w It is the replacement of C 1-6 Alkyl group. R 9 and R 10 It can be, for example, phenyl or –OC(O)R 99 Replacement C 1-6 Alkyl, wherein R 99 As defined above. Therefore, R 99 It can be phenyl, unsubstituted C 1-6 Alkyl, –N(R) a (R) b ), –C(O)R c –OR d Or amino acids, of which R a R b R c and R d Each is independently selected from H, unsubstituted or substituted C. 1-6 Alkyl groups and amino acids. Typically, R a R b R c and R d Each is independently selected from H or unsubstituted or substituted C. 1-6 Alkyl group. Typically, R a R b R c and R d Each is independently selected from H, methyl, or ethyl. R 9 and R 10 It can be, for example, by -OC(O)R 99 Replacement C 1-6 Alkyl, wherein R 99As defined above. Similarly, in this aspect of the implementation scheme, R w It can be a substitute for C 1-6 Alkyl group. R w It can be, for example, phenyl or –OC(O)R ww Replacement C 1-6 Alkyl, wherein R ww As defined above. Therefore, R ww It can be phenyl, unsubstituted C 1-6 Alkyl, –N(R) a (R) b ), –C(O)R c –OR d Or amino acids, wherein Ra, Rb, Rc, and Rd are each independently selected from H, unsubstituted or substituted C. 1-6 Alkyl groups and amino acids. Typically, R a R b R c and R d Each is independently selected from H or unsubstituted or substituted C. 1-6 Alkyl group. Typically, R a R b R c and R d Each is independently selected from H, methyl, or ethyl. R w It can be, for example, by -OC(O)R ww Replacement C 1-6 Alkyl, wherein R ww As defined above.

[0249] In another aspect of this implementation scheme, R 9 R 10 and R w It can be the same or different, and is an unsubstituted C. 1-6 Alkyl group. R 9 R 10 and R w It can be selected, for example, from methyl and ethyl.

[0250] The substituted azazine of formula (Ic) can be selected, for example, from any of the following structures:

[0251]

[0252]

[0253] The numbers in parentheses next to the structures listed above correspond to the compound numbers given in the Examples section below.

[0254] The present invention also provides substituted azazine compounds of formula (Id) as defined herein, wherein R9 Not H (and where R) 0 R 1 R y R 6 R 7 and R x (As defined in this document for formula Id). Such compounds include prodrugs. Typically, R in this embodiment... 9 Is it unsubstituted or substituted C? 1-6 Alkyl group. In one aspect of this embodiment, R 9 It is the replacement of C 1-6 Alkyl group. R 9 It can be, for example, phenyl or –OC(O)R 99 Replacement C 1-6 Alkyl, wherein R 99 As defined above. Therefore, R 99 It can be phenyl, unsubstituted C 1-6 Alkyl, –N(R) a (R) b ), –C(O)R c –OR d Or amino acids, of which R a R b R c and R d Each is independently selected from H, unsubstituted or substituted C. 1-6 Alkyl groups and amino acids. Typically, R a R b R c and R d Each is independently selected from H or unsubstituted or substituted C. 1-6 Alkyl group. Typically, R a R b R c and R d Each is independently selected from H, methyl, or ethyl. R 9 It can be, for example, by -OC(O)R 99 Replacement C 1-6 Alkyl, wherein R 99 As defined above. In another aspect of this implementation scheme, R... 9 It is unreplaced C 1-6 Alkyl group. R 9 It can be, for example, methyl. Substituted azazines of formula (Id) can, for example, have the following structures.

[0255]

[0256] The numbers in parentheses next to the structures listed above correspond to the compound numbers given in the Examples section below.

[0257] Compounds of formulas (Ia), (Ib), (Ic) and (Id) above (where typically, R) 8 R 9 R 10 or R w Is it unsubstituted or substituted C? 1-6 Alkyl groups, as prodrug structures of compounds of formulas (Ia), (Ib), (Ic), or (Id), have surprising advantages; they are potent HIF-PHD inhibitors. In particular, compounds having the structures described above have surprisingly shown enhanced inhibitory efficacy in cell assays, even when they themselves do not possess high potency as HIF-PHD inhibitors. The reduced potency and increased activity in cell assays suggest that these compounds have the potential to provide targeted inhibition and reduce off-target effects.

[0258] The present invention also provides a compound which is a substituted pyrimidine of formula (IV) or a pharmaceutically acceptable salt thereof.

[0259]

[0260] in

[0261] R 0 It is H or unsubstituted or substituted C 1-6 alkyl;

[0262] R 2 It is H, –OR q C, whether substituted or not 1-6 alkyl;

[0263] R 4 Yes – OR 9 , where R 9 Selected from H and unsubstituted or substituted C 1-6 alkyl;

[0264] R 5 It is H, unsubstituted or substituted C 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN or –C(O)OR w ;

[0265] R 6 It is H or unsubstituted or substituted C 1-6 alkyl;

[0266] R 7 It is –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar, –Ary–Ar, –CH(R 11–Cyc, –Cyc, or –Ar, where Ar is an unsubstituted or substituted aryl group, or an unsubstituted or substituted heteroaryl group, Ary is an unsubstituted or substituted arylene group, or an unsubstituted or substituted heteroaryl group, and Cyc is an unsubstituted or substituted C group. 3-10 Cycloalkyl, and R 11 It is H, –C(O)OR z C, whether substituted or not 1-4 Alkyl; R x It is H, unsubstituted or substituted C 1-4 Alkyl groups, or unsubstituted or substituted phenyl groups;

[0267] R w and R z Each is independently selected from H, unsubstituted or substituted C. 1-4 Alkyl groups and unsubstituted or substituted phenyl groups; and

[0268] R q It is H, unsubstituted or substituted C 1-6 Alkyl, or unsubstituted or substituted phenyl.

[0269] Typically, in equation (IV), R 0 Is it H or unsubstituted C? 1-6 Alkyl groups, such as H or methyl. Typically R 0 It's H.

[0270] Typically, in equation (IV), R 2 Is it H or unsubstituted C? 1-6 Alkyl groups, such as H or methyl. Typically R 2 It's H.

[0271] Typically, in equation (IV), R 0 It is H, R 2 It is H

[0272] Typically, in equation (IV), R 5 Yes –CN.

[0273] In equation (IV), R 6 Typically H or unsubstituted C 1-6 Alkyl groups, such as H or methyl. Typically, R 6 In equation (IV), it is H.

[0274] R 4 In equation (IV), it is –OR 9 And R 9 Selected from H and unsubstituted or substituted C 1-6 Alkyl group. However, in formula (IV), R 9 Equation (I) can be defined as it is anywhere in this document.

[0275] Typically, in equation (IV), R 9 It is H. When R 9 When it is H (i.e., when R) 4 When it is OH), then typically, R 5 Yes –CN.

[0276] However, in some implementations, R in formula (IV) 9 C can be unsubstituted or substituted. 1-6 Alkyl group. Such embodiments include prodrugs. In one aspect of this embodiment, R in formula (IV) 9 It is the replacement of C 1-6 Alkyl group. R 9 It can be, for example, phenyl or –OC(O)R 99 Replacement C 1-6 Alkyl, wherein R 99 As defined above. Therefore, R 99 It can be phenyl, unsubstituted C 1-6 Alkyl, –N(R) a (R) b –C(O)R c –OR d Or amino acids, of which R a R b R c and R d Each is independently selected from H, unsubstituted or substituted C. 1-6 Alkyl groups and amino acids. Typically, R a R b R c and R d Each is independently selected from H or unsubstituted or substituted C. 1-6 Alkyl group. Typically, R a R b R c and R d Each is independently selected from H, methyl, or ethyl. R in formula (IV) 9 It can be, for example, by -OC(O)R 99 Replacement C 1-6 Alkyl, wherein R 99 As defined above. In another aspect of this implementation, R in equation (IV) 9 It is unreplaced C 1-6 Alkyl group. R 9 For example, it can be methyl.

[0277] In equation (IV), typically, R x Is it H or unsubstituted C? 1-4 Alkyl, such as H or methyl. Typically, R in formula (IV) is...x It's H.

[0278] In equation (IV), R w It is H, unsubstituted or substituted C 1-6 Alkyl, or unsubstituted or substituted phenyl. R w It can be H or unsubstituted or substituted C. 1-6 Alkyl group. Typically, for example, R w It is H, unsubstituted C 1-6 Alkyl, or phenyl or –OC(O)R ww Replacement C 1-6 Alkyl, wherein R ww It is phenyl, unsubstituted C 1-6 Alkyl, –N(R) a (R) b ), –C(O)R c –OR d Or amino acids, of which R a R b R c and R d Each is independently selected from H, unsubstituted or substituted C. 1-6 Alkyl groups and amino acids. Typically, R in formula (IV) is... w It's H.

[0279] In equation (IV), typically, R z Is it H or unsubstituted C? 1-4 Alkyl, such as H or methyl. Typically, R in formula (IV) is... z It's H.

[0280] In equation (IV), typically, R q Is it H or unsubstituted C? 1-6 Alkyl, or unsubstituted phenyl. It is typically, for example, H or unsubstituted C. 1-4 Alkyl, such as H or methyl. Typically, R in formula (IV) q It's H.

[0281] Typically, R in equation (IV) 7 It is –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar, or –CH(R 11 )–Cyc, where Ar is an unsubstituted or substituted aryl group, or an unsubstituted or substituted heteroaryl group, Ary is an unsubstituted or substituted arylene group, or an unsubstituted or substituted heteroaryl group, and Cyc is an unsubstituted or substituted C group. 3-10 Cycloalkyl, and R 11 It is H, –C(O)OR z C, whether substituted or not1-4 Alkyl group. Typically, R 11 Is it H or unsubstituted C? 1-4 Alkyl groups, such as H or methyl. Typically R 11 It is H.

[0282] R in equation (IV) 7 For example, it could be –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar, or –CH(R 11 –Cyc; where Ar is an unsubstituted phenyl, unsubstituted pyrimidinyl, unsubstituted benzothiazole, or a phenyl substituted with –C(O)OH, –C(O)OMe, –C(O)OEt, –C(O)NH2, –C(O)N(H)Me, –OMe, or N-morpholino; Ary is an unsubstituted phenylene or unsubstituted pyridylene; Cyc is an unsubstituted cyclohexyl or a cyclohexyl substituted with –CF3 or –OCF3; and R 11 As defined above, typically, it is H. Usually, R in equation (IV) 7 It is –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar or –CH(R 11 Ar is an unsubstituted phenyl group or a phenyl group substituted with –C(O)OH or –C(O)OMe; Ary is an unsubstituted phenylene group or an unsubstituted pyridylene group; Cyc is an unsubstituted cyclohexyl group or a cyclohexyl group substituted with –CF3; and R 11 As defined above, typically, it is H.

[0283] Compounds of formula (IV) may be represented by one of the following structures or a pharmaceutically acceptable salt thereof:

[0284] The numbers in parentheses next to the structures listed above correspond to the compound numbers given in the Examples section below.

[0285] General Synthesis Method

[0286] The compounds of this invention can be prepared by any suitable method. Detailed general synthetic routes for the compounds of this invention are set forth below and in the examples.

[0287] The substituted azazine of formula (I) and the substituted pyrimidine of formula (IV) can be synthesized, for example, using the methods listed below in the Examples section titled "General Method A," "General Method B," "General Method C," and "General Method D." The production of the substituted azazine of formula (I) using these general methods is shown and described below with reference to schemes 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12, with specific synthetic examples described in the Examples section below. Similarly, the production of the substituted pyrimidine of formula (IV) using these general methods is shown and described below with reference to scheme 11.

[0288] The substituted azazine of formula (I) and the substituted pyrimidine of formula (IV) can be synthesized using an amide coupling method (general method A or B), which can be used by activating an amide group (e.g., -C(O)N(R)). x )R 7 An amide group is introduced by coupling the amide group to a carboxylic acid or ester group in the precursor compound. The amide group introduced in the preceding step can then be further modified using a Pd-catalyzed coupling method (General Method C) to obtain the desired C(O)N(R) group in the final compound. x )R 7 Then, the alkoxy dealkylation step (General Method D) can be used as a final step to provide an OH group in the final compound.

[0289] For example, Scheme 10 below illustrates how to produce a substituted azazine of formula (Ia) using general methods B, C, and D provided in the Examples section. The same general methods B, C, and D can also be used to produce a substituted azazine of formula (Ib). Scheme 6 below illustrates how to produce a substituted azazine of formula (Ic) using general methods B and D described in the Examples section. Schemes 7, 8, and 12 below illustrate the synthesis of a pyrazolo[1,5-A]pyrido[3,2-e]pyrimidine-7-acylamino structure of formula (Id) using general methods A and D in the Examples. Furthermore, Scheme 11 illustrates how to produce a substituted pyrimidine of formula (IV) using general method B. Schemes 1 and 2 are also provided below to illustrate general methods for preparing some of the reference examples described herein. As those skilled in the art will understand, alternative precursor compounds having substituents different from those shown in the schemes below can be used in the same methods to achieve variations within the range of formulas (I), (Ia), (Ib), (Ic), (Id), and (IV) herein.

[0290] Therefore, the compounds described in this article can be prepared according to the following reaction scheme:

[0291]

[0292] Option 1

[0293] Step (i) of Scheme 1 can be performed using any suitable peptide coupling agent. Typically, step (i) involves treatment with an acid anhydride (e.g., propanephosphonic anhydride) in the presence of a base (such as N,N-diisopropylethylamine (DIPEA)). Typically, the reaction occurs in a solvent. This solvent can be a polar solvent. Typically, the solvent can be a polar aprotic solvent. This polar aprotic solvent can be dimethylacetamide (DMAc). Typically, step (i) occurs at standard atmospheric temperature and pressure (SATP), which is approximately 25°C and 1 atmosphere (approximately 100,000 Pa). This step can last from 1 to 24 hours, for example, approximately 16 hours.

[0294] Step (ii) of Scheme 1 includes treatment in the presence of a catalyst. The catalyst may be a palladium catalyst. Typically, the catalyst may be Pd. t BuXPhos. This step can also be carried out in the presence of a base. The base can be a carbonate. Typically, the base is cesium carbonate (Cs₂CO₃). The reaction usually occurs in a solvent. The solvent can be a mixture of solvents. Typically, the solvent can be a mixture of one or more polar solvents, usually a polar aprotic solvent. Typically, the reaction occurs in a mixture of dimethylacetamide (DMAc) and tetrahydrofuran (THF). This mixture can be a 1:1 mixture (1:1). Typically, step (ii) occurs at temperatures above room temperature. For example, typically, step (ii) occurs between 60°C and 100°C. Typically, this step occurs at around 80°C. This step can last from 1 to 24 hours, for example, about 16 hours.

[0295] Step (iii) of Scheme 1 is a deprotection step, which may include treatment with any suitable reagent known to a person skilled in the art. In some cases, silyl halides are used. Typically, trimethylsilyl iodide (TMS-I) is used. Typically, this step is carried out in the presence of a solvent. Typically, the solvent is a polar solvent, and more typically, a polar aprotic solvent. The solvent may be dichloromethane (CH2Cl2). Typically, step (iii) occurs at a temperature above room temperature. For example, typically, step (iii) occurs between 40°C and 80°C.

[0296] This step typically occurs at around 60°C. It can last from 1 to 24 hours, for example, about 8 hours.

[0297]

[0298] Option 2

[0299] Step (i) of Scheme 2 may include treatment with any suitable amide coupling agent. Typically, step (i) involves treatment with an acid anhydride (e.g., propanephosphonic anhydride (T3P)) in the presence of a base (such as N,N-diisopropylethylamine (DIPEA)). Typically, this reaction occurs in a solvent. The solvent can be a polar solvent. Typically, the solvent can be a polar aprotic solvent. A polar aprotic solvent can be dimethylacetamide (DMAc). Typically, step (i) occurs under SATP. This step can last from 1 to 24 hours, for example, about 16 hours.

[0300] Step (ii) of Scheme 2 includes treatment in the presence of a catalyst. The catalyst may be a palladium catalyst. Typically, the catalyst is Pd. t BuXPhos. This step can also be carried out in the presence of a base. The base can be a carbonate. Typically, the base is Cs₂CO₃. Typically, the reaction occurs in a solvent. The solvent can be a mixture of solvents. Typically, the solvent can be a mixture of one or more polar solvents, usually a polar aprotic solvent. Typically, the reaction occurs in a mixture of DMAc and tetrahydrofuran (THF). This mixture can be approximately 1:1 (1:1). Typically, step (ii) occurs at temperatures above room temperature. For example, typically, step (ii) occurs between 80°C and 120°C. Typically, this step occurs at around 100°C. This step can last from 30 minutes to 6 hours, for example, about 1 hour.

[0301]

[0302] Option 3

[0303] Scheme 3 above illustrates how to produce certain substituted azazines of formula (I) using the general synthetic methods described in the Examples section below (see general methods A, B, C and D in the Examples).

[0304] Step (i) of Scheme 3 may include treatment with any suitable peptide coupling agent. Typically, step (i) includes treatment with an acid anhydride (e.g., propanephosphonic anhydride) in the presence of a base (such as N,N-diisopropylethylamine (DIPEA)). Typically, this reaction occurs in a solvent. The solvent can be a polar solvent. Typically, the solvent can be a polar aprotic solvent. A polar aprotic solvent can be dimethylacetamide (DMAc). Typically, step (i) occurs under SATP. This step can last from 1 to 24 hours, for example, about 16 hours.

[0305] Step (ii) of Scheme 3 includes treatment in the presence of a catalyst. The catalyst may be a palladium catalyst. Typically, the catalyst may be Pd. tBuXPhos. This step can also be carried out in the presence of a base. The base can be a carbonate. Typically, the base is Cs₂CO₃, Na₂CO₃, or K₂CO₃. Usually, the base is Cs₂CO₃. Typically, the reaction occurs in a solvent. The solvent can be a mixture of solvents. Typically, the solvent can be a mixture of one or more polar solvents, usually a polar aprotic solvent. Typically, the reaction occurs in a mixture of DMAc and tetrahydrofuran (THF). The mixture can be a 1:1 mixture (1:1). Typically, step (ii) occurs at temperatures above room temperature. For example, typically, step (ii) occurs between 80°C and 120°C. Typically, the step occurs at around 100°C. The step can last from 30 minutes to 6 hours, for example, about 1 hour.

[0306] Step (iii) can be carried out using any suitable ester hydrolysis reagent. Typically, the reaction can be carried out in the presence of a hydroxide base (usually lithium hydroxide). The reaction is typically carried out in the presence of a solvent. The solvent can be a mixture of one or more solvents (typically one or more polar solvents). The one or more polar solvents can be protic or aprotic solvents. For example, the solvent can be a mixture of THF and H₂O. Typically, the mixture is approximately 1:1. Typically, step (iv) occurs under SATP.

[0307] This step can last from 1 to 24 hours, for example, about 16 hours.

[0308]

[0309] Option 4

[0310] Scheme 4 above illustrates how to produce the 4-hydroxypyridine / pyridinone structure of formula (Ia) using the general synthetic method described in the Examples section below (see General Methods A and D in the Examples).

[0311] Typically, step (i) of Scheme 4 involves treatment with R-NH2. Step (i) of Scheme 4 may include treatment with any suitable amide coupling agent. Typically, step (i) involves treatment with an acid anhydride (e.g., propanephosphonic anhydride) in the presence of a base (such as N,N-diisopropylethylamine (DIPEA)). Typically, the reaction occurs in a solvent, which may be a polar solvent. Typically, the solvent may be a polar aprotic solvent. The polar aprotic solvent may be dimethylacetamide (DMAc). Typically, step (i) occurs under SATP. This step may last from 1 to 24 hours, for example, about 16 hours.

[0312] Step (ii) of Scheme 4 may include treatment with other reagents, such as pyrazoles and substituted pyrazoles. Step (ii) of Scheme 4 includes treatment in the presence of a catalyst. The catalyst may be a palladium catalyst. Typically, the catalyst may be Pd. t BuxPhos G3. This step can also be carried out in the presence of a base. The base can be a carbonate. Typically, the base is cesium carbonate (Cs₂CO₃). Typically, the reaction occurs in a solvent. The solvent can be a mixture of solvents. Typically, the solvent can be a mixture of one or more polar solvents (usually polar aprotic solvents). Typically, the reaction occurs in tert-butanol or 1,4-dioxane. Typically, step (ii) occurs at temperatures above room temperature. For example, typically, step (ii) occurs between 40°C and 80°C. Typically, this step occurs at around 60°C. This step can last from 1 hour to 48 hours, for example, about 16 hours.

[0313] Step (iii) of Scheme 4 is a deprotection step, which may include treatment with any suitable reagent known to a person skilled in the art. In some cases, lithium chloride is used. Typically, this step is carried out in the presence of a solvent. Typically, the solvent is a polar solvent, more typically, a polar aprotic solvent. The solvent may be DMAc. Typically, step (v) occurs at a temperature above room temperature. For example, step (v) typically occurs between 80°C and 120°C. Usually, this step occurs at around 100°C. This step can last from 30 minutes to 6 hours, for example, about 2 hours.

[0314] Step (iv) of Scheme 4 can be carried out using any suitable ester hydrolysis reagent, if desired. Typically, the reaction can be carried out in the presence of a hydroxide (usually lithium hydroxide). The reaction is generally carried out in the presence of a solvent. The solvent can be a mixture of one or more solvents (usually one or more polar solvents). The one or more polar solvents can be proton or aprotic solvents. For example, the solvent can be a mixture of THF and H₂O. Typically, the mixture is a 1:1 mixture. Step (ii) typically occurs under SATP. This step can last from 1 to 24 hours, for example, about 16 hours.

[0315]

[0316] Option 5

[0317] Scheme 5 above illustrates how to produce the 4-hydroxypyridine / pyridinone structure of formula (Ic) using the general synthetic method described in the Examples section below (see General Methods A and D in the Examples).

[0318] Step (i) of Scheme 5 may include treatment with any suitable amide coupling agent. Typically, step (i) includes treatment with an acid anhydride (e.g., propanephosphonic anhydride) in the presence of a base (such as N,N-diisopropylethylamine (DIPEA)). Typically, the reaction occurs in a solvent. The solvent may be a polar solvent. Typically, the solvent may be a polar aprotic solvent. A polar aprotic solvent may be dimethylacetamide (DMAc). Typically, step (i) occurs under SATP. This step may last from 1 to 24 hours, for example, about 16 hours.

[0319] Step (ii) of Scheme 5 includes treatment in the presence of a catalyst. The catalyst may be a palladium catalyst. Typically, the catalyst may be Pd. t BuxPhos G3. This step can also be carried out in the presence of a base. The base can be a carbonate. Typically, the base is cesium carbonate (Cs₂CO₃). Typically, the reaction occurs in a solvent. The solvent can be a mixture of solvents. Typically, the solvent can be a mixture of one or more polar solvents (usually polar aprotic solvents). Typically, the reaction occurs in tert-butanol or 1,4-dioxane. Typically, step (ii) occurs at temperatures above room temperature. For example, typically, step (ii) occurs between 40°C and 80°C. Typically, this step occurs at around 60°C. This step can last from 1 hour to 48 hours, for example, about 16 hours. Step (ii) may also involve treatment with other reagents, such as pyrazoles and substituted pyrazoles.

[0320] Step (iii) of Scheme 5 is a deprotection step, which may include treatment with any suitable reagent known to a person skilled in the art. In some cases, lithium chloride is used. Typically, this step is carried out in the presence of a solvent. Typically, the solvent is a polar solvent, more typically, a polar aprotic solvent. The solvent may be DMAc. Typically, step (v) occurs at a temperature above room temperature. For example, typically, step (v) occurs between 80°C and 120°C. Typically, this step occurs at around 100°C. This step can last from 30 minutes to 6 hours, for example, about 2 hours.

[0321]

[0322] Option 6

[0323] Scheme 6 above illustrates how to produce substituted azazine of formula (Ic) using the general synthesis methods described in the Examples section below (see General Methods B and D in the Examples).

[0324] Step (i) of Scheme 6 includes treatment in the presence of a catalyst. The catalyst can be a palladium catalyst. Typically, the catalyst can be RockPhos Pd G3. This step can also be carried out in the presence of a base. The base can be a carbonate. Typically, the base is cesium carbonate (Cs2CO3). Typically, the reaction occurs in a solvent. The solvent can be a nonpolar solvent. Typically, the solvent is a nonpolar protic solvent. Usually, the solvent is tert-butanol ( t BuOH). Typically, step (i) occurs at temperatures above room temperature. For example, step (i) typically occurs between 60°C and 100°C. Usually, this step occurs at around 80°C. This step can last from 1 hour to 24 hours, for example, about 16 hours.

[0325] Step (ii) can be carried out using any suitable ester hydrolysis reagent. Typically, the reaction can be carried out in the presence of a hydroxide base (usually lithium hydroxide). The reaction is typically carried out in the presence of a solvent. The solvent can be a mixture of one or more solvents (typically one or more polar solvents). The one or more polar solvents can be protic or aprotic solvents. For example, the solvent can be a mixture of THF and H2O. Typically, the mixture is a 1:1 mixture. Typically, step (ii) occurs under SATP. This step can last from 1 to 24 hours, for example, about 16 hours.

[0326] In step (iii) of Scheme 6, the starting material is treated with the R-NH2 group. For step (iii), any suitable peptide coupling agent can be used. Typically, step (iii) of Scheme 6 involves treatment with an acid anhydride (e.g., propanephosphonic anhydride) in the presence of a base (such as N,N-diisopropylethylamine (DIPEA)). Typically, this reaction occurs in a solvent. The solvent can be a polar solvent. Typically, the solvent can be a polar aprotic solvent. A polar aprotic solvent can be dimethylacetamide (DMAc). Typically, step (iii) occurs under SATP. This step can last from 1 to 24 hours, for example, about 16 hours.

[0327] Step (iv) of Scheme 6 is a deprotection step, which may include treatment with any suitable reagent known to a person skilled in the art. In some cases, lithium chloride is used. Typically, this step is carried out in the presence of a solvent. Typically, the solvent is a polar solvent, more typically, a polar aprotic solvent. The solvent may be DMAc. Typically, step (iv) occurs at a temperature above room temperature. For example, typically, step (iv) occurs between 80°C and 120°C. Typically, this step occurs at around 100°C. This step can last from 30 minutes to 6 hours, for example, about 2 hours.

[0328]

[0329] Option 7

[0330] Scheme 7 above illustrates how to produce the pyrazolo[1,5-a]pyrido[3,2-e]pyrimidine-7-acylamino structure of formula (Id) using the general synthetic method described above.

[0331] Step (i) of Scheme 7 is a Michael addition reaction, which can be performed using any suitable reagent known to a person skilled in the art. In some cases, sodium ethoxide is used. Typically, this step is carried out in the presence of a solvent. Typically, the solvent is a polar solvent, more typically a polar aprotic solvent. The solvent can be EtOH. Typically, step (i) occurs at a temperature above room temperature. Typically, this step occurs between 80°C and 120°C. Usually, this step occurs around 100°C. This step can last from 1 hour to 48 hours, for example, about 2 hours.

[0332] Step (ii) of Scheme 7 is intramolecular cyclization, which may include treatment with any suitable reagent known to a person skilled in the art. Typically, this step is carried out in the presence of a solvent. Typically, the solvent is a high-boiling point solvent. The solvent may be a diphenyl ether. Typically, step (ii) occurs at a temperature above room temperature. Typically, the temperature is between 140°C and 250°C. Generally, this step occurs at around 250°C. This step can last from 10 minutes to 8 hours, for example, about 30 minutes.

[0333] Step (iii) of Scheme 7 involves direct amide coupling of ethyl ester and includes treatment in the presence of a catalyst. Typically, the catalyst can be DABCO-(AlMe3)2. The reaction typically occurs in a solvent. The solvent can be a mixture of solvents. Typically, the reaction occurs in tetrahydrofuran. Step (iii) typically occurs at temperatures above room temperature. For example, step (iii) typically occurs between 40°C and 150°C. Typically, the step occurs at around 130°C. The step can last from 10 minutes to 12 hours, for example, about 1 hour.

[0334]

[0335] Option 8

[0336] Scheme 8 above illustrates how to produce the pyrazolo[1,5-a]pyrido[3,2-e]pyrimidine-7-acylamino structure of formula (Id) using the general synthetic methods described in the Examples section below (see General Methods A and D in the Examples).

[0337] In step (i) of scheme 8, the starting material is treated with the R-NH2 group. For step (i), any suitable amide coupling agent can be used. Typically, step (i) of scheme 4 involves treatment with an acid anhydride (e.g., propanephosphonic anhydride) in the presence of a base (such as N,N-diisopropylethylamine (DIPEA)). Typically, this reaction occurs in a solvent. The solvent can be a polar solvent. Typically, the solvent is a polar aprotic solvent. A polar aprotic solvent can be dimethylacetamide (DMAc). Typically, step (i) occurs under SATP. This step can last from 1 to 24 hours, for example, about 16 hours.

[0338] Step (ii) of Scheme 8 involves treatment with triethyl orthoformate. Typically, this reaction is carried out under solvent-free conditions. Typically, step (ii) occurs at temperatures above room temperature. For example, step (ii) can occur between 100°C and 140°C. Generally, this step occurs at around 120°C. This step can last from 30 minutes to 6 hours, for example, about 2 hours.

[0339] Step (iii) of Scheme 8 involves heating the reagent at a temperature above room temperature. Typically, the reagent is heated to above 200°C, typically to around 240°C. This step can last from 10 minutes to 2 hours, for example, around 30 minutes. This step can also be carried out in the presence of other reagents, such as diphenyl ether.

[0340]

[0341] Option 9

[0342] Scheme 9 above illustrates how to produce the 3-hydroxypyridine structure of formula (Ib) using the general synthetic methods described in the Examples section below (see General Methods A and D in the Examples).

[0343] Typically, step (i) of Scheme 9 includes treatment with R-NH2. Step (i) of Scheme 9 may include treatment with any suitable amide coupling agent. Typically, step (i) includes treatment with an acid anhydride (e.g., propanephosphonic anhydride) in the presence of a base (such as N,N-diisopropylethylamine (DIPEA)). Typically, the reaction occurs in a solvent, which may be a polar solvent. Typically, the solvent may be a polar aprotic solvent. The polar aprotic solvent may be dimethylacetamide (DMAc). Typically, step (i) occurs under SATP. This step may last from 1 to 24 hours, for example, about 16 hours.

[0344] Step (ii) of Scheme 9 involves treatment with other reagents, such as pyrazoles and substituted pyrazoles. Step (ii) of Scheme 9 also includes treatment in the presence of a catalyst. The catalyst may be a palladium catalyst. Typically, the catalyst may be Pd. tBuxPhos G3. This step can also be carried out in the presence of a base. The base can be a carbonate. Typically, the base is cesium carbonate (Cs₂CO₃). Typically, the reaction occurs in a solvent. The solvent can be a mixture of solvents. Typically, the solvent can be a mixture of one or more polar solvents (usually polar aprotic solvents). Typically, the reaction occurs in tert-butanol or 1,4-dioxane. Typically, step (ii) occurs at temperatures above room temperature. For example, typically, step (ii) occurs between 40°C and 80°C. Typically, this step occurs at around 60°C. This step can last from 1 hour to 48 hours, for example, about 16 hours. Step (ii) may also involve treatment with other reagents, such as pyrazoles and substituted pyrazoles.

[0345] Step (iii) of Scheme 9 is a deprotection step, which may include treatment with any suitable reagent known to a person skilled in the art. In some cases, lithium chloride is used. Typically, this step is carried out in the presence of a solvent, typically a polar solvent, and more typically a polar aprotic solvent. The solvent may be DMAc. Typically, step (v) occurs at a temperature above room temperature. For example, step (v) typically occurs between 80°C and 120°C. Usually, this step occurs at around 100°C. This step can last from 30 minutes to 6 hours, for example, about 2 hours.

[0346] Step (iv) of Scheme 9 can be carried out using any suitable ester hydrolysis reagent, if desired. Typically, the reaction can be carried out in the presence of a hydroxide (usually lithium hydroxide). The reaction is generally carried out in the presence of a solvent. The solvent can be a mixture of one or more solvents (typically one or more polar solvents). The one or more polar solvents can be proton or aprotic solvents. For example, the solvent can be a mixture of THF and H₂O. Typically, the mixture is a 1:1 mixture. Step (ii) typically occurs under SATP. This step can last from 1 to 24 hours, for example, about 16 hours.

[0347]

[0348] Option 10

[0349] Scheme 10 above illustrates how to produce substituted azazine of formula (Ia) using the general synthesis methods described in the Examples section below (see general methods B, C and D in the Examples).

[0350] Step (i) of Scheme 10 can be carried out using any suitable esterification reagent known to those skilled in the art. In some cases, N'-ethylcarboimide hydrochloride (EDC.HCl) is used. Typically, a catalyst is also present. The catalyst is usually an organic catalyst. The catalyst can be 4-dimethylaminopyridine (DMAP). A base may also be present in step (i) of Scheme 10. The base can be N,N-diisopropylethylamine (DIPEA). Typically, step (i) of Scheme 10 is carried out in the presence of a solvent. The solvent can be a mixture of two solvents. Typically, the solvent is a mixture of two polar solvents. Usually, the solvent is a mixture of a polar protic solvent and a polar aprotic solvent. Thus, the solvent can be a mixture of dimethylformamide (DMF) and ethanol. Typically, step (i) occurs under SATP. This step can last from 1 hour to 24 hours, for example, about 16 hours.

[0351] In step (ii) of scheme 10, the product of step (i) is treated with pyrazole. Step (ii) includes treatment in the presence of a catalyst. The catalyst may be a palladium catalyst. Typically, the catalyst may be Pd. t BuxPhos G3. This step can also be carried out in the presence of a base. The base can be a carbonate. Typically, the base is cesium carbonate (Cs₂CO₃). Typically, step (ii) occurs at temperatures above room temperature. For example, step (ii) typically occurs between 40°C and 80°C. Typically, this step occurs at around 60°C. This step can last from 1 hour to 24 hours, for example, about 16 hours.

[0352] Step (iii) can be carried out using any suitable ester hydrolysis reagent. Typically, the reaction can be carried out in the presence of a hydroxide (usually lithium hydroxide). The reaction is typically carried out in the presence of a solvent. The solvent can be a mixture of one or more solvents (typically one or more polar solvents). The one or more polar solvents can be protic or aprotic solvents. For example, the solvent can be a mixture of THF and H2O. Typically, the mixture is approximately 1:1. Typically, step (iii) occurs under SATP. This step can last from 1 to 24 hours, for example, about 16 hours.

[0353] In step (iv) of Scheme 10, the product of step (iii) is treated with the group R-NH2. For step (iv), any suitable peptide coupling agent can be used. Typically, step (iv) of Scheme 10 involves treatment with HATU in the presence of a base (such as DIPEA). Typically, this reaction occurs in a solvent. The solvent can be a polar solvent. Typically, the solvent can be a polar aprotic solvent. A polar aprotic solvent can be N,N-dimethylacetamide (DMAc). Typically, step (iv) occurs under SATP. This step can last from 1 to 24 hours, for example, about 16 hours.

[0354] Step (v) of Scheme 10 is a deprotection step, which may include treatment with any suitable reagent known to a person skilled in the art. In some cases, lithium chloride is used. Typically, this step is carried out in the presence of a solvent. Typically, the solvent is a polar solvent, more typically, a polar aprotic solvent. The solvent may be DMAc. Typically, step (v) occurs at a temperature above room temperature. For example, step (v) typically occurs between 80°C and 120°C. Usually, this step occurs at around 100°C. This step can last from 30 minutes to 6 hours, for example, about 2 hours.

[0355] Step (vi) of Scheme 10 involves a reaction in the presence of a catalyst. The catalyst may be a palladium catalyst. Typically, the catalyst may be PdAmPhos. This step may also be carried out in the presence of a base. The base may be a carbonate. Typically, the base is cesium carbonate (Cs₂CO₃). In step (vi), the product of step (iv) is typically treated with an organoborane compound containing the group -R. Typically, this compound is a compound of the formula RB(OH)₂ or an RB-pinacol ester. Typically, step (vi) occurs at a temperature above room temperature. For example, typically, step (vi) occurs between 80°C and 120°C. Typically, this step occurs at around 100°C. This step may last from 30 minutes to 6 hours, for example, about 2 hours. Typically, this step is carried out in the presence of a solvent. Typically, the solvent is a nonpolar solvent, more typically, a nonpolar aprotic solvent. The solvent may be 1,4-dioxane.

[0356]

[0357] Option 11

[0358] Scheme 11 above illustrates how to produce substituted pyrimidines of formula (IV) using the general synthetic method described in the Examples section below (see General Method B in the Examples).

[0359] Typically, step (i) of scheme 11 involves treatment with an acid. The acid is usually a protic acid, such as HCl. For example, 4M HCl in 1,4-dioxane can be used. Typically, step (i) occurs at a temperature above room temperature. For example, step (i) typically occurs between 80°C and 120°C. Typically, the step occurs at around 100°C. The step can last from 1 hour to 24 hours, for example, about 16 hours.

[0360] Typically, step (ii) of scheme 11 involves treatment in the presence of a base. Any suitable base can be used. Typically, the base is a carbonate. K₂CO₃ can be used. Typically, step (ii) is carried out in a solvent. The solvent can be a polar solvent, typically a polar protic solvent. Usually, the solvent is methanol. Typically, step (ii) occurs under SATP. This step can last from 1 to 24 hours, for example, about 16 hours.

[0361] Step (iii) can be carried out using any suitable ester hydrolysis reagent. Typically, the reaction can be carried out in the presence of a hydroxide base (usually lithium hydroxide). The reaction is typically carried out in the presence of a solvent. The solvent can be a mixture of one or more solvents (typically one or more polar solvents). The one or more polar solvents can both be protic solvents. For example, the solvent can be a mixture of methanol and H₂O. Typically, the mixture is a 1:1 mixture. Typically, step (iii) occurs under SATP. This step can last from 1 to 24 hours, for example, about 16 hours.

[0362] In step (iv) of Scheme 11, the product of step (iii) is treated with the group R-NH2. For step (iv), any suitable peptide coupling agent can be used. Typically, step (iv) of Scheme 11 involves treatment with T3P in the presence of a base (such as N,N-diisopropylethylamine (DIPEA)). Typically, this reaction occurs in a solvent. The solvent can be a polar solvent. Typically, the solvent can be a polar aprotic solvent. A polar aprotic solvent can be N,N-dimethylacetamide (DMAc). Typically, step (iv) occurs under SATP. This step can last from 1 to 24 hours, for example, about 16 hours.

[0363]

[0364] Option 12

[0365] Scheme 12 above illustrates how to produce a pyrazolo[1,5-a]pyrido[3,2-e]pyrimidine-7-acylamino structure of formula (Id) using the general synthetic methods described in the Examples section below (see General Methods A and D in the Examples).

[0366] In scheme 12, step (i) typically involves heating the raw materials together. Typically, heating occurs at a temperature of 100°C to 200°C, more typically at 140°C to 160°C. Heating can occur at around 150°C. This step can last from 30 minutes to 6 hours, for example, about 1 hour.

[0367] Step (ii) of Scheme 12 involves heating the product of step (i) with diethyl ethoxymethylene malonate. Typically, step (ii) is carried out in a solvent. The solvent can be a polar solvent or a nonpolar protic solvent. Typically, the solvent is toluene. Heating typically occurs at temperatures between 80°C and 160°C, more typically between 100°C and 140°C. Heating can occur at approximately 120°C. Step (ii) typically lasts between approximately 24 and 72 hours. For example, step (ii) can last approximately 48 hours.

[0368] Typically, step (iii) of Scheme 12 involves treatment with a reagent to promote ring formation. Any suitable reagent can be used. Typically, Eaton's reagent (10 wt% phosphorus pentoxide in methanesulfonic acid solution) is used. Typically, step (iii) occurs at a temperature above room temperature. For example, typically, step (iii) occurs between 50°C and 90°C. Usually, this step occurs at around 70°C. This step can last from 16 hours to 30 hours, for example, about 24 hours.

[0369] Typically, step (iv) of Scheme 12 involves treatment with a chlorinating agent. Any suitable chlorinating agent known to a person skilled in the art can be used. Phosphoryl chloride (POCl3) is commonly used. Typically, step (iv) occurs at a temperature above room temperature. For example, step (iv) typically occurs between 50°C and 90°C. Typically, this step occurs at around 70°C. This step can last from 1 hour to 6 hours, for example, about 3 hours.

[0370] Typically, step (v) of scheme 12 involves treatment with sodium methoxide (NaOMe). Step (v) is typically carried out in a solvent. The solvent can be a polar solvent, typically a polar protic solvent. Typically, the solvent is methanol. Step (v) typically occurs under SATP. This step can last from 1 to 10 hours, for example, about 4 hours.

[0371] Typically, step (vi) of scheme 12 involves treatment with R-NH2 in the presence of a reagent that promotes amide formation. The reagent may be an organoaluminum reagent, typically bis(trimethylaluminum)-1,4-diazabicyclo[2.2.2]octane adduct (DABAL-AlMe3). Typically, step (vi) is carried out in the presence of a solvent. The solvent is typically a nonpolar solvent and is typically a nonpolar aprotic solvent, such as THF. Typically, step (vi) is carried out at a temperature above room temperature. For example, step (vi) typically occurs between 100°C and 140°C. Typically, the step occurs at around 120°C. The step can last from 30 minutes to 6 hours, for example, about 3 hours.

[0372] Step (vii) of Scheme 12 may include treatment with any reagent suitable for converting the ether to a hydroxyl group. Typically, lithium chloride can be used. Typically, the reaction occurs in a solvent. The solvent can be a polar solvent. Typically, the solvent can be a polar aprotic solvent. A polar aprotic solvent can be N,N-dimethylacetamide (DMAc). Typically, step (vii) occurs at a temperature above room temperature. For example, step (vii) typically occurs between 80°C and 120°C. Typically, this step occurs at around 100°C. This step can last from 30 minutes to 6 hours, for example, about 2 hours.

[0373] The compounds of the present invention containing one or more chiral centers can be used in enantiomerically pure or diastereomerically pure form, or in the form of a mixture of isomers. For clarity, the compounds of the present invention can be used as solvates if desired. Furthermore, for clarity, the compounds of the present invention can be used in any tautomer form.

[0374] As used herein, a pharmaceutically acceptable salt is a salt formed with a pharmaceutically acceptable acid or base. Pharmaceutically acceptable acids include inorganic acids (such as hydrochloric acid, sulfuric acid, phosphoric acid, diphosphoric acid, hydrobromic acid, or nitric acid) and organic acids (such as citric acid, fumaric acid, maleic acid, malic acid, ascorbic acid, succinic acid, tartaric acid, benzoic acid, acetic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, or p-toluenesulfonic acid). Pharmaceutically acceptable bases include hydroxides of alkali metals (such as sodium or potassium) and alkaline earth metals (such as calcium or magnesium) and organic bases (such as alkylamines, aralkylamines, and heterocyclic amines). Substituted azathenes of formula (I), (Ia), (Ib), (Ic), or (Id) or substituted pyrimidines of formula (IV) can be converted into pharmaceutically acceptable salts by conventional methods, and the salts can be converted into free compounds.

[0375] Pharmaceutical Composition

[0376] The present invention also provides a pharmaceutical composition comprising the compound of the present invention as defined anywhere herein and a pharmaceutically acceptable carrier or diluent.

[0377] Typically, the composition contains up to 85 wt% of the compound of the present invention. More typically, it contains up to 50 wt% of the compound of the present invention. Preferred pharmaceutical compositions are sterile and pyrogen-free. Furthermore, when the pharmaceutical compositions provided by the present invention contain optically active compounds of the present invention, the compounds of the present invention are typically substantially pure optical isomers.

[0378] The compositions of the present invention may be provided in the form of a kit, which includes instructions for use as described herein or details about which subjects the composition may be used on.

[0379] Typically, the compositions of the present invention are formulated for administration together with a pharmaceutically acceptable carrier or diluent. For example, the solid oral form may contain the active compound and diluent (e.g., lactose, glucose, sucrose, cellulose, corn starch, or potato starch); lubricant (e.g., silica, talc, stearic acid, magnesium stearate, or calcium stearate and / or polyethylene glycol); binder (e.g., starch, gum arabic, gelatin, methylcellulose, carboxymethylcellulose, or polyvinylpyrrolidone); depolymerizing agent (e.g., starch, alginate, alginate, or sodium carboxymethyl starch); effervescent mixture; dye; sweetener; humectant (e.g., lecithin, polysorbate, dodecyl sulfate); and non-toxic and pharmacologically inactive substances commonly used in pharmaceutical formulations. Such pharmaceutical formulations can be manufactured in known ways, for example by mixing, granulation, tableting, sugar coating, or film coating processes.

[0380] The compositions of the present invention can be formulated as solutions or suspensions for inhalation (nebulization) administration. The compounds or combinations of the present invention can be administered via metered-dose inhalers (MDIs) or nebulizers (such as electronic or jet nebulizers). Alternatively, the compounds or combinations of the present invention can be formulated as powdered medications for inhalation administration, such formulations being administered via dry powder inhalers (DPIs). When formulated for inhalation administration, the compounds or combinations of the present invention can be delivered in particulate form, wherein the mass median aerodynamic diameter (MMAD) of the particulates is 1 to 100 μm, preferably 1 to 50 μm, more preferably 1 to 20 μm, for example 3 to 10 μm, for example 4 to 6 μm. When the compounds or combinations of the present invention are delivered as a nebulized aerosol, the mentioned particulate diameter defines the MMAD of the aerosol droplet. MMAD can be measured by any suitable technique, such as laser diffraction.

[0381] Orally administered liquid dispersions can be syrups, emulsions, and suspensions. Syrups may contain a carrier, such as sucrose or sucrose with glycerol and / or mannitol and / or sorbitol.

[0382] Suspensions and emulsions may contain carriers such as natural gums, agar, sodium alginate, pectin, methylcellulose, carboxymethylcellulose, or polyvinyl alcohol. Suspensions or solutions intended for intramuscular injection or inhalation may contain active compounds and pharmaceutically acceptable carriers such as sterile water, olive oil, ethyl oleate, glycols (e.g., propylene glycol), and, if desired, an appropriate amount of lidocaine hydrochloride.

[0383] Solutions intended for inhalation, injection, or infusion may contain a carrier, such as sterile water, or preferably they may be in the form of sterile, aqueous, isotonic saline solutions. Pharmaceutical compositions suitable for needle-free injection, such as transdermal delivery, may also be used.

[0384] The compositions of the present invention may also contain one or more other active agents. These other active agents may be selected from ACE inhibitors, angiotensin II receptor agonists, β-blockers, calcium channel blockers, PDE inhibitors, mineralocorticoid receptor antagonists, diuretics, aspirin, iron supplements, vitamin B12 and folic acid supplements, statins, digitalis (digoxin) derivatives, chemotherapy drugs for cancer, and antibiotics.

[0385] Therapeutic uses

[0386] The compounds of this invention have shown high efficiency and specificity as inhibitors of hypoxia-inducible factor (HIF) prolyl hydroxylase (PHD). For example, some compounds of this invention have been shown to have an IC50 response to PHD2. 50 Less than 200 nM, which is a significant improvement compared to known inhibitors (e.g., roxadustat in the LCMS PHD2 hydroxylation assay used in this paper, IC50). 50 (2.7 μM). In addition to their potency, the compounds of the present invention have been found to have high selectivity for PHD, with a selectivity greater than 100-fold compared to other tested 2OG oxygenases.

[0387] In addition to these desirable biochemical properties, the compounds of the present invention also exhibit desirable physical properties, including good solubility and permeability in cells. These physical properties mean that the compounds of the present invention have been found to effectively stabilize HIF-1α cells. Furthermore, in animal models, low in vivo doses of this compound have been shown to induce erythropoiesis.

[0388] Therefore, the compounds and pharmaceutical compositions of the present invention have potential use in treating conditions targeting PHD.

[0389] Therefore, this invention provides compounds of the invention as defined herein or pharmaceutical compositions of the invention as defined herein for therapeutic treatment of a human or animal body. As discussed above, the term "treatment" herein refers to therapeutic treatments and preventative or preventative measures.

[0390] This invention also provides compounds or pharmaceutical compositions as defined herein for use as regulators of hypoxia-inducible factor prolyl hydroxylase activity. Typically, these compounds or pharmaceutical compositions are used as inhibitors of hypoxia-inducible factor prolyl hydroxylase activity. Therefore, this invention also provides compounds or pharmaceutical compositions as defined herein for use as PHD inhibitors.

[0391] The present invention also provides compounds of the present invention as defined herein or pharmaceutical compositions of the present invention as defined herein for the treatment of PHD-related conditions.

[0392] As used herein, "PHD-related syndromes" refer to syndromes that can be treated by modulating hypoxia-inducible factor prolyl hydroxylase activity. Typically, PHD-related syndromes are those that can be treated by inhibiting hypoxia-inducible factor prolyl hydroxylase activity. Technicians can readily identify PHD-related syndromes experimentally.

[0393] PHD-related conditions include, but are not limited to, anemia, ischemia, inflammation, Parkinson's disease, sickle cell anemia (including through upregulation of fetal hemoglobin F), Alzheimer's disease, non-fatty liver disease, irritable bowel disease, cardiovascular disease, heart failure, chronic kidney disease, cancer, and renal insufficiency. Therefore, the compounds or pharmaceutical compositions of the present invention can be used to treat any of the above-mentioned conditions.

[0394] Similarly, the compounds or pharmaceutical compositions of the present invention can be used for skeletal muscle injury repair, increasing red blood cell count (RBC), increasing hemoglobin (HGB) production, increasing hematocrit (HCT) production, increasing erythropoietin (EPO) production, wound healing, angiogenesis, vascular regeneration, stem cell activation, or cardiac protection after myocardial infarction.

[0395] Therefore, this invention provides compounds or pharmaceutical compositions as defined herein for the treatment of anemia, ischemia, inflammation, Parkinson's disease, Alzheimer's disease, non-fatty liver disease, irritable bowel disease, sickle cell anemia, cancer, cardiovascular disease, heart failure, chronic kidney disease or renal insufficiency; or for skeletal muscle injury repair, increasing red blood cell count (RBC), increasing hemoglobin (HGB) production, increasing hematocrit (HCT) production, increasing erythropoietin (EPO) production, wound healing, angiogenesis, vascular regeneration, stem cell activation, or cardiac protection after myocardial infarction. Sickle cell anemia may be caused by the upregulation of fetal hemoglobin F.

[0396] Typically, the compounds or pharmaceutical compositions of this invention are used to treat anemia. Anemia can be renal anemia, such as anemia associated with chronic kidney disease, or anemia in dialysis patients, anemia caused by chemotherapy, cancer-related anemia, age-related anemia, or anemia caused by cancer (such as leukemia, multiple myeloma, and pyogenic myeloma). Anemia can also be sickle cell anemia, such as sickle cell anemia caused by the upregulation of fetal hemoglobin F.

[0397] Typically, the compounds or pharmaceutical compositions of the present invention are used to increase red blood cell count (RBC), increase hemoglobin (HGB) production, increase hematocrit (HCT) production, or increase erythropoietin (EPO) production.

[0398] Furthermore, the compounds or pharmaceutical compositions of the present invention are typically used to treat ischemia. Therefore, ischemia-related diseases can be treated with the compounds or pharmaceutical compositions of the present invention. Thus, the compounds or pharmaceutical compositions of the present invention can be used to treat ischemia in circulatory or cardiovascular diseases, myocardial infarction, ischemia during surgery, organ ischemia, ischemic diseases, or diabetic limb ischemia or sickle cell anemia. The compounds or pharmaceutical compositions of the present invention can be used to provide cardiac protection after myocardial infarction.

[0399] This invention also provides a method for treating a subject with or susceptible to PHD-related conditions, the method comprising administering to the subject an effective amount of a compound or pharmaceutical composition of the invention as defined herein. PHD-related conditions may include, for example, anemia, ischemia, sickle cell anemia, cancer, inflammation, Parkinson's disease, Alzheimer's disease, non-fatty liver disease, irritable bowel disease, cardiovascular disease, heart failure, chronic kidney disease, or renal insufficiency. Conditions may also include, for example, HIF-related conditions, EPO-related conditions, or VHL-related conditions, such as von Hippel-Lindau (VHL) syndrome.

[0400] The present invention also provides a method for repairing skeletal muscle injury, increasing red blood cell count (RBC), increasing hemoglobin (HGB) production, increasing hematocrit (HCT) production, increasing erythropoietin (EPO) production, wound healing, angiogenesis, vascular regeneration, stem cell activation, or cardiac protection after myocardial infarction in a subject, the method comprising administering to the subject an effective amount of the compound or pharmaceutical composition of the present invention as defined herein.

[0401] Subjects are typically mammals, usually humans. However, they may not be human. Preferred non-human animals include, but are not limited to, primates such as marmosets or monkeys, commercially farmed animals such as horses, cattle, sheep, or pigs, and pets such as dogs, cats, mice, rats, guinea pigs, ferrets, gerbils, or hamsters.

[0402] Diseases such as anemia, ischemia, and inflammation may be associated with hypoxic states, and they have been shown to be closely related to erythropoietin (EPO) deficiency. Previous studies have shown that the role of EPO can be improved by activating the HIF pathway. As previously mentioned, PHD is an oxygenase that catalyzes the hydroxylation of specific prolyl residues within the oxygen degradation domain of the hypoxia-inducible factor α (HIF-α) subunit. Therefore, the compounds and compositions of the present invention are potent PHD inhibitors that can be used to treat HIF pathway-related diseases and EPO-related diseases (such as anemia, ischemia, and inflammation).

[0403] Increased EPO production also leads to an increase in red blood cell count (RBC) and hemoglobin (HGB) production. In turn, increased red blood cell count can promote wound healing, angiogenesis, blood vessel regeneration, or stem cell activation.

[0404] Both Parkinson's disease and Alzheimer's disease are associated with neuronal hypoxia. Specifically, HIF stabilization in Parkinson's patients is thought to increase dopamine synthesis and dopaminergic neuron growth. In Alzheimer's patients, hypoxia is known to stimulate the production of amyloid-β peptide, which can disrupt the membrane localization of glucose transporters (GLUTs), thereby affecting glucose levels in the brain. Stabilizing HIF can upregulate neuronal glucose transporters (such as GLUT-1 and GLUT-3), which may mitigate this effect.

[0405] Therefore, the compounds or pharmaceutical compositions of the present invention are typically used to treat anemia.

[0406] Anemia, as mentioned in this article, can be any form of anemia. For example, anemia includes anemia related to impaired blood cell production, such as iron deficiency anemia, vitamin deficiency anemia, inflammatory anemia, aplastic anemia (such as pure red cell aplasia and Fanconi anemia), anemia related to bone marrow disorders, hemolytic anemia, sickle cell anemia, thalassemia, renal anemia (such as anemia associated with chronic kidney disease or anemia in dialysis patients), anemia due to endocrine disorders, megaloblastic anemia (such as pernicious anemia and folic acid deficiency anemia), anemia in premature infants, congenital erythropoiesis-related anemia, as well as myeloid tuberculosis anemia and myelodysplastic syndrome.

[0407] Anemia can also be anemia associated with increased red blood cell destruction (i.e., hemolytic anemia). Hemolytic anemia can be caused by intrinsic abnormalities (such as hereditary spherocytosis, hereditary elliptocytosis, alipoproteinemia, enzyme deficiency, and sickle cell anemia), extrinsic abnormalities (such as antibody-mediated anemia, including rhesus monkey disease and transfusion reactions, or mechanical damage to red blood cells, including cardiac surgery, hemodialysis, and infection), or parasites (such as Trypanosoma congo).

[0408] Anemia can also be blood loss-related, such as premature birth, trauma or surgery, gastrointestinal diseases, gynecological diseases, menstruation, and iatrogenic anemia.

[0409] Other causes of anemia can include fluid overload and intestinal inflammation (e.g., caused by Helicobacter pylori infection, gluten-related diseases such as celiac disease, or inflammatory bowel disease).

[0410] Typically, anemia is renal anemia (such as anemia associated with chronic kidney disease or anemia in dialysis patients), chemotherapy-induced anemia, sickle cell anemia, cancer-related anemia, age-related anemia, or anemia caused by cancer (such as leukemia, multiple myeloma, and pyogenic myeloma).

[0411] Anemia can often be renal anemia, such as anemia associated with chronic kidney disease or anemia in dialysis patients.

[0412] The compounds or compositions of this invention can also be used to treat ischemia.

[0413] In this article, ischemia can refer to any type of ischemia. Ischemia includes ischemia of the heart or circulatory system (such as ischemia in circulatory or cardiovascular diseases, myocardial infarction, coronary artery ischemia, coronary artery disease, myocardial ischemia, and ischemic heart disease).

[0414] Ischemia also includes organ ischemia, such as intestinal ischemia (e.g., intestinal ischemia, which includes ischemic colitis and mesenteric ischemia); cerebral ischemia (including acute ischemia, such as ischemic stroke and transient ischemic attack, as well as chronic ischemia that can lead to vascular dementia); and renal ischemia.

[0415] Ischemia can also be limb ischemia (such as acute limb ischemia, chronic limb-threatening ischemia, and diabetic limb ischemia).

[0416] Ischemia can also be associated with ischemic diseases. Ischemia can occur during surgery. It can also be skin ischemia (such as cyanosis or gangrene).

[0417] Typically, ischemia is ischemia in circulatory or cardiovascular diseases, myocardial infarction, ischemia during surgery, organ ischemia, ischemic diseases, or diabetic limb ischemia.

[0418] Administering a therapeutically effective amount of the compound of the present invention to a subject, as used herein, means a therapeutically or preventively effective amount. Similarly, compositions comprising a therapeutically effective amount of the compound of the present invention may be administered to a subject. The dosage may be determined based on a variety of parameters, particularly based on the compound used; the age, weight, and condition of the subject to be treated; the route of administration; and the desired treatment regimen. Likewise, the physician will be able to determine the route of administration and dosage required for any particular subject. Typical daily doses are from about 0.01 to 100 mg / kg body weight, preferably from about 0.1 mg / kg body weight to 50 mg / kg body weight, for example from about 1 to 10 mg / kg body weight, depending on the activity of the specific inhibitor, the age, weight, and condition of the subject to be treated, the type and severity of the disease, and the frequency and route of administration. Preferably, the daily dose level is from 1 mg to 2 g.

[0419] The present invention will be further described in the following embodiments and reference embodiments:

[0420] Example

[0421] Synthesis Examples

[0422] General Method

[0423] All reactions involving moisture-sensitive reagents were performed under a nitrogen atmosphere using standard vacuum pipeline techniques. Glassware was dried and cooled under nitrogen before use. Commercially available anhydrous solvents and HPLC-grade solvents used in the reactions were used for post-processing and chromatographic analysis. Aqueous solutions were prepared using deionized water and purified using an Elix UV-10 system. Thin-layer chromatography (TLC) was performed using Merck (Darmstadt, Germany) silica gel 60F254 TLC plates. TLC visualization was performed under UV light and stained with one of three staining agents: ninhydrin, potassium permanganate, or anisaldehyde. (Uppsala, Sweden) IsoleraOne or SP4 rapid purification system, using Chromatographic analysis was performed using a pre-packed SNAP column. An Agilent (Cheshire, UK) 1200 series 6120 quadrupole LC-MS system was used, with Merck... The reaction was monitored using a Performance RP-18 HPLC column. The deuterated solvent was obtained from Sigma-Aldrich. 1 H NMR spectroscopy is used with H NMR spectroscopy. 13C cryopreservation probes were obtained using a Bruker AVANCE AVIII HD 400 nanobay (400 MHz) or Bruker AV500 (500 MHz) instrument. All signals are expressed in δppm, and multiplies are indicated by the abbreviations s, d, t, q, and m for singlet, doublet, triplet, quadruplet, and multiplies, respectively. Chemical shifts in the presented NMR spectra are referenced to the residual solvent peak, and the coupling constant J is reported in Hertz (Hz) with an accuracy of 0.5 Hz. For high-resolution mass spectrometry (HR-MS), a Bruker MicroTOF instrument with an ESI source and a time-of-flight (TOF) analyzer was used. MS data are expressed as mass-to-charge ratio (m / z) in Daltons. Fourier transform infrared (FT-IR) spectra were obtained using a Bruker Tensor 27 instrument. Spectro-grade solvent and a Perkin Elmer 241 polarimeter were used to obtain optical rotation.

[0424] All chemicals, reagents, and solvents were purchased from Sigma-Aldrich (Dorset, UK) and were ready for use without further purification. HPLC-grade solvents were used for reactions, chromatography, and post-processing.

[0425] General Method A

[0426] Ethyl ester amide coupling: Add the relevant ethyl ester (1 equivalent), the relevant amine (1 equivalent), and DABACO-(AlMe3)2 (1.0 equivalent); rinse the microwave vial with N2 and remove N2 under vacuum (3 times) before adding anhydrous THF. Then heat the reaction mixture at 130°C for 8 minutes using biotage microwave irradiation (unless otherwise specified). Dilute the reaction mixture with a mixture of CH3Cl:IPA (3:1, 20 ml), followed by the addition of KNaC4H4O6·4H2O. aq (50 ml). Stir the resulting mixture for 1 hour. Then separate the phases, wash the organic phase with water and brine, and dry with Na2SO4. Remove the solvent under vacuum. Purify the crude compound using rapid column chromatography (20 column volumes, according to the conditions described for each reaction) to obtain the desired compound.

[0427] General Method B:

[0428] Amide coupling: Carboxylic acid (1 equivalent) and DIPEA (2.5 equivalent) were dissolved in DMF. Then, T3P (1.5 equivalent, 50% DMF solution) or HATU (2 equivalent) was added. The resulting reaction mixture was stirred at room temperature for 30 minutes, then an amine (1.2 equivalent) was added. The resulting mixture was stirred overnight at room temperature. Ethyl acetate (20 mL) and H2O (100 mL) were added to the reaction mixture. The organic and aqueous phases were separated. The aqueous layer was extracted twice more with EtOAc (30 mL). The organic fractions were combined, washed with brine, and dried over anhydrous Na2SO4. The crude compound was then purified by rapid column chromatography (20 column volumes) to obtain the desired compound.

[0429] General Method C:

[0430] Pd-catalyzed amination: An aryl halide (1 equivalent), an amine (1.2 equivalent), Cs₂CO₃ (2 equivalent), and a Pd-ligand conjugate (0.1 equivalent) were placed under an anhydrous N₂ atmosphere, followed by the addition of tert-butanol. The resulting mixture was heated at 80 °C for 16 hours. The reaction mixture was then allowed to cool to room temperature. Ethyl acetate (20 ml) and H₂O (100 ml) were added to the reaction mixture. The organic and aqueous phases were separated. The aqueous layer was extracted twice more with EtOAc (30 ml). The organic fractions were combined, washed with brine, and dried over anhydrous Na₂SO₄. The crude compound was then purified by rapid column chromatography (0-100% EtOAc in cyclohexane solution) at 20 column volumes to obtain the desired compound.

[0431] General Method D:

[0432] C-4 methoxy demethylation: The methoxy starting material (1 equivalent) was dissolved in DMAc; then LiCl·H2O (10 equivalents) was added. The resulting mixture was heated under microwave radiation at 120 °C for 2 hours (unless otherwise specified). The resulting mixture was diluted with water (100 ml) and extracted with EtOAc (3 × 20 ml). The organic phases were combined, washed with water and brine, and dried over anhydrous Na2SO4. The eluent was then evaporated under vacuum and purified by rapid column chromatography (100%–95% CH2Cl2, 0%–20% MeOH) at 15 column volumes (unless otherwise specified) to obtain the desired compound.

[0433] Reference Example 1 – Synthesis of N-([1,1'-biphenyl]-4-ylmethyl)-4-chloro-2-methoxybenzamide (14)

[0434]

[0435] 14 (451 mg, 1.24 mmol, 95%) was obtained by general method B: 4-chloro-2-methoxybenzoic acid (250 mg, 1.34 mmol), 4-phenylbenzylamine (294 mg, 1.6 mmol), T3P (1.06 g, 3.36 mmol), and DIPEA (412 mg, 3.36 mmol).

[0436] 1 H NMR (400MHz, chloroform-d) δ8.21(d,J=8.5Hz,1H),8.12(t,J=6.0Hz,1H),7.60–7.56(m,4H),7.46–7.41(m,4H),7 .37–7.30(m,1H),7.08(dd,J=8.5,2.0Hz,1H),6.97(d,J=2.0Hz,1H),4.71(d,J=6.0Hz,2H),3.93(s,3H).

[0437] HRMS (ESI-TOF) is calculated as C 21 H 19 O2N 35 Cl[M+H] + :352.1098, actual value: 352.1098.

[0438] Reference Example 2 – Synthesis of N-([1,1'-biphenyl]-4-ylmethyl)-2-methoxy-4-(1H-pyrazol-1-yl)benzamide (15)

[0439]

[0440] According to the general method, C:14 (100 mg, 0.284 mmol), Pd t BuXPhos G3 (20 mg, 0.028 mmol), Cs2CO3 (185 mg, 0.568 mmol), and pyrazole (23 mg, 0.34 mmol) yielded 15 (28 mg, 0.073 mmol, 26%).

[0441] 1 H NMR (400MHz, DMSO-d6) δ8.77(t,J=6.0Hz,1H),8.66(d,J=2.5Hz,1H),7.80(d,J=1.5Hz, 1H),7.69–7.32(m,12H),6.60(dd,J=2.5,1.5Hz,1H),4.56(d,J=6.0Hz,2H),4.01(s,3H)

[0442] HRMS (ESI-TOF) is calculated as C 24 H22 O2N3[M+H] + :384.1704, actual value: 384.1704.

[0443] Reference Example 3 – Synthesis of N-([1,1'-biphenyl]-4-ylmethyl)-2-hydroxy-4-(1H-pyrazol-1-yl)benzamide (16)

[0444]

[0445] TMS-I (39 mg, 0.195 mmol) was added to a solution of 15 (25 mg, 0.0652 mmol) and CH2Cl2 (2.5 mL). The resulting mixture was refluxed at 90 °C for 8 hours, then cooled to room temperature, and HCl was added. aq (1.5 ml, 1 M), then extracted with CH2Cl2 (3 × 10 ml). The organic fractions were combined, dried over anhydrous Na2SO4, concentrated under vacuum, and then purified by rapid column chromatography (0%–5% MeOH, CH2Cl2, 1% NH3) at 20 column volumes to give 16 (7.5 mg, 0.020 mmol, 31%).

[0446] 1 H NMR (400MHz, DMSO-d6) δ12.96 (s, 1H), 9.43 (t, J = 6.0Hz, 1H), 8.61 (dd, J = 2.5, 1.0Hz, 1H), 8.05 (d, J = 8. 0Hz,1H),7.79(d,J=1.5Hz,1H),7.70–7.31(m,11H),6.58(dd,J=2.5,1.5Hz,1H),4.57(d,J=6.0Hz,2H).

[0447] HRMS (ESI-TOF) is calculated as C 23 H 20 O2N3[M+H] + 370.1548, actual value: 370.1548.

[0448] Reference Example 4 – Synthesis of N-([1,1'-biphenyl]-4-ylmethyl)-2-chloropyrimidine-5-carboxamide (17)

[0449]

[0450] 17 (285 mg, 0.88 mmol, 47%) was obtained by general method B: 2-chloro-5-carboxypyrimidine (300 mg, 1.89 mmol), 4-phenylbenzylamine (370 mg, 2.26 mmol), T3P (819 mg, 2.83 mmol) and DIPEA (365 mg, 2.83 mmol).

[0451] 1 H NMR (400MHz, DMSO-d6) δ9.46 (t, J = 6.0 Hz, 1H), 9.18 (s, 2H), 7.87–6.86 (m, 9H), 4.56 (d, J = 6.0 Hz, 2H).

[0452] HRMS (ESI-TOF) is calculated as C 18 H 15 35 ClN3O[M+H] + 324.0898, actual value: 324.0899.

[0453] Reference Example 5 – Synthesis of 2-chloro-N-(4-phenoxybenzyl)pyrimidine-5-carboxamide (18)

[0454]

[0455] 18 (336 mg, 0.99 mmol, 52%) was obtained by general method B: 2-chloro-5-carboxypyrimidine (300 mg, 1.89 mmol), 4-phenoxybenzylamine (410 μl, 2.26 mmol) and T3P (1.5 g, 4.72 mmol).

[0456] 1 H NMR (400MHz, DMSO-d6) δ9.41 (t, J = 6.0 Hz, 1H), 9.16 (s, 2H), 7.53–7.27 (m, 4H), 7.19–6.78 (m, 5H), 4.50 (d, J = 6.0Hz, 2H).

[0457] HRMS (ESI-TOF) is calculated as C 18 H 14 35 ClN3O2[M+H] + 340.0847, actual value: 340.1327.

[0458] Reference Example 6 – Synthesis of N-([1,1'-biphenyl]-4-ylmethyl)-6-chloronicotinamide (19)

[0459]

[0460] 19 (154 mg, 0.48 mmol, 91%) was obtained by general method A: 2-chloroethyl nicotinate (83 μl, 0.53 mmol), 4-phenyl-benzylamine (97 mg, 0.53 mmol) and DABCO-(AlMe3)2 (108 mg, 0.424 mmol).

[0461] 1 H NMR (400MHz, DMSO-d6) δ9.34(t,J=6.0Hz,1H),8.90(d,J=2.5Hz,1H),8.30(dd,J=8.5,2.5Hz,1H),7.87–7.19(m,10H),4.54(d,J=6.0Hz,2H).

[0462] HRMS (ESI-TOF) is calculated as C 19 H 15 ClN2O[M+H] + 322.0873, actual value: 322.0821.

[0463] Reference Example 7 – Synthesis of N-([1,1'-biphenyl]-4-ylmethyl)-2-(1H-pyrazol-1-yl)pyrimidine-5-carboxamide (20)

[0464]

[0465] According to the general method, C:17 (57 mg, 0.176 mmol), Pd t BuXPhos G3 (15 mg, 0.0176 mmol), Cs2CO3 (201 mg, 0.619 mmol), and pyrazole (42 mg, 0.619 mmol) yielded 20 (10 mg, 0.0281 mmol, 16%).

[0466] 1 H NMR (400MHz, DMSO-d6) δ9.44(t,J=6.0Hz,1H),9.27(s,2H),8.73(d,J=3.0Hz,1H),7.93(d,J=1.5Hz,1H),7 .74–7.60(m,4H),7.51–7.40(m,4H),7.39–7.31(m,1H),6.65(dd,J=3.0,1.5Hz,1H),4.58(d,J=5.8Hz,2H).

[0467] HRMS (ESI-TOF) is calculated as C 21 H 18 ON5[M+H] +:356.1505, actual value: 356.1504.

[0468] Example 8 Synthesis of –N-(4-phenoxybenzyl)-2-(1H-pyrazol-1-yl)pyrimidine-5-carboxamide (21)

[0469]

[0470] According to the general method C:18 (100 mg, 0.294 mmol), pyrazole (40 mg, 0.589 mmol), Pd t BuXPhos G3 (23 mg, 0.0294 mmol), Cs2CO3 (238 mg, 0.735 mmol) yielded 21 (64 mg, 0.172 mmol, 58%).

[0471] 1 H NMR(400MHz,DMSO-d6)δ8.96(d,J=3.0Hz,1H),8.68(d,J=3.0Hz,1H),8.51–8.37(m,1 H), 8.03–7.98 (m, 1H), 7.48–6.90 (m, 10H), 6.62–6.61 (m, 1H), 4.51 (d, J = 6.0Hz, 2H).

[0472] HRMS (ESI-TOF) is calculated as C 21 H 17 N5O2[MH] - 370.1382, actual value: 370.1309.

[0473] Example 9 – Synthesis of N-([1,1'-biphenyl]-4-ylmethyl)-6-(1H-pyrazol-1-yl)nicotinamide (22)

[0474]

[0475] According to method C: 19 (100 mg, 0.31 mmol), pyrazole (40 mg, 0.589 mmol), Pd t BuXPhos G3 (25 mg, 0.031 mmol) and Cs2CO3 (238 mg, 0.735 mmol) yielded 22 (53 mg, 0.028 mmol, 48%).

[0476] 1H NMR (400MHz, DMSO-d6) δ9.32(t,J=6.0Hz,1H),8.97(d,J=2.0Hz,1H),8.69(d,J=2.5Hz,1H),8.47(dd,J=9.0,2.0Hz,1H), 8.30(dd,J=9.0,2.0Hz,1H),7.90(d,J=1.5Hz,1H),7.74–7.22(m,9H),6.63(dd,J=2.5,1.5Hz,1H),4.57(d,J=6.0Hz,2H).

[0477] HRMS (ESI-TOF) is calculated as C 22 H 19 ON4[M+H] + :355.1553, actual result: 355.1551.

[0478] Reference Example 10 – Synthesis of N-benzyl-6-chloronicotinamide (28)

[0479]

[0480] 28 (220 mg, 0.897 mmol, 83%) was obtained by general method A: 2-chloro-ethyl nicotinate (200 mg, 1.081 mmol), benzylamine (116 mg, 1.08 mmol) and DABACO-AlMe3 (221 mg, 0.864 mmol).

[0481] Solvent system used for purification: 0%-100% EtOAc in cyclohexane solution.

[0482] 1 H NMR(400MHz,DMSO-d6)δ9.32(t,J=6.0Hz,1H),8.89(dd,J=2.5,1.0Hz,1H),8.29(d d,J=8.5,2.5Hz,1H),7.87–7.57(m,1H),7.50–6.86(m,5H),4.51(d,J=6.0Hz,2H).

[0483] HRMS (ESI-TOF) is calculated as C 13 H 10 ON2 35 Cl[MH] - :245.0487, actual value: 245.0482.

[0484] Example 11 – Synthesis of 1-(5-(([1,1'-biphenyl]-4-ylmethyl)carbamoyl)pyridin-2-yl)-1H-pyrazole-4-carboxylic acid ethyl ester (29)

[0485]

[0486] According to the general method, C:19 (125 mg, 0.388 mmol), pyrazole-4-carboxylic acid ethyl ester (65 mg, 0.465 mmol), PdtBuxPhos G3 (31 mg, 0.0388 mmol), Cs2CO3 (252 mg, 0.776 mmol) and t BuOH (3 ml) yielded 29 (97 mg, 0.227 mmol, 59%).

[0487] 1 H NMR (400MHz, DMSO-d6) δ9.38(t,J=6.0Hz,1H),9.04(s,1H),9.02(d,J=2.5Hz,1H),8.51(dd,J=8.5,2.5Hz,1H),8.27( s,1H),8.07(d,J=8.5Hz,1H),7.69–7.30(m,9H),4.54(d,J=6.0Hz,2H),4.21(q,J=7.0Hz,2H),1.31(t,J=7.0Hz,3H).

[0488] HRMS (ESI-TOF) is calculated as C 25 H 21 O3N4[MH] - :425.1619, actual value: 425.1622.

[0489] Example 12 – Synthesis of 1-(5-(benzylcarbamoyl)pyridin-2-yl)-1H-pyrazole-4-carboxylic acid ethyl ester (30)

[0490]

[0491] According to the general method C: 28 (80mg, 0.325mmol), Pd t BuXPhos G3 (25 mg, 0.0325 mmol), Cs2CO3 (317 mg, 0.97 mmol), pyrazole-4-carboxylic acid ethyl ester (67 mg, 0.48 mmol) and t BuOH (3 ml) yielded 30 (41 mg, 0.117 mmol, 36%).

[0492] 1H NMR (400MHz, DMSO-d6) δ9.33(t,J=6.0Hz,1H),9.04(d,J=1.0Hz,1H),8.99(dd,J=2.5,1.0Hz,1H),8.49(dd,J=8.5,2.5Hz,1H),8.27( d,J=1.0Hz,1H,),8.06(dd,J=8.5,1.0Hz,1H),7.38–7.32(m,5H),4.53(d,J=6.0Hz,2H),4.28(q,J=7.0Hz,2H),1.31(t,J=7.0Hz,3H).

[0493] HRMS (ESI-TOF) is calculated as C 19 H 17 O3N4[MH] - 349.1306, actual value: 349.1302.

[0494] Example 13 – Synthesis of 1-(5-(([1,1'-biphenyl]-4-ylmethyl)carbamoyl)pyridin-2-yl)-1H-pyrazole-4-carboxylic acid (31)

[0495]

[0496] 29 mg (63 mg, 0.147 mmol) was dissolved in a mixture of THF and water (10 mL (10:1)), and then LiOH monohydrate (19 mg, 0.45 mmol) was added. The resulting mixture was stirred for 16 hours, and the reaction was confirmed by TLC. HCl was then added to the reaction mixture. aq (10 ml, 1 M), the resulting mixture was extracted with EtOAc (3 × 20 ml). The organic fractions were combined and washed with brine, dried with Na2SO4, and purified by rapid column chromatography (CH2Cl2, MeOH 0-5%, formic acid 1%) with 20 column volumes to give 31 (15 mg, 0.037 mmol, 26%).

[0497] 1 H NMR (400MHz, DMSO-d6) δ9.37–9.34(m,1H),9.01(d,J=2.5Hz,1H),8.92(d,J=2.5Hz,1H),8.53–8.47(m,1H ),8.09–8.01(m,1H),7.70–7.62(m,4H),7.46(t,J=7.8Hz,4H),7.39–7.30(m,2H),4.56(d,J=6.0Hz,2H).

[0498] HRMS (ESI-TOF) is calculated as C23 H 17 O3N4[MH] - :397.1306, actual value: 397.1310.

[0499] Example 14 – Synthesis of 1-(5-(benzylcarbamoyl)pyridin-2-yl)-1H-pyrazole-4-carboxylic acid (32)

[0500]

[0501] 30 (20 mg, 0.043 mmol) was dissolved in THF (2 mL). MeOH (2 mL) and water (0.5 mL) were added to the reaction mixture, followed by LiOH monohydrate (16 mg, 0.40 mmol). The resulting mixture was stirred overnight at room temperature. The reaction mixture was acidified to pH 3 with HCl (1 M) solution, extracted with EtOAc (3 × 25 mL), washed with brine, and dried over anhydrous Na₂SO₄. The crude compound was then purified by rapid column chromatography (CH₂Cl₂, MeOH 0.5%, formic acid 1%) at 20 column volumes to give 32 (13 mg, 0.040 mmol, 94%).

[0502] 1 H NMR (400MHz, DMSO-d6) δ9.34(t,J=6.0Hz,1H),8.99(dd,J=2.5,1.0Hz,1H),8.96(s,1H),8.48(d d,J=8.5,2.5Hz,1H),8.19(s,1H),8.08–8.02(m,1H),7.48–7.21(m,5H),4.54(d,J=6.0Hz,2H).

[0503] HRMS (ESI-TOF) is calculated as C 17 H 13 O3N4[MH] - 321.0993, actual value: 321.0994.

[0504] Example 15 – Synthesis of N-([1,1'-biphenyl]-4-ylmethyl)-6-chloro-4-methoxynicotinamide (33)

[0505]

[0506] According to general method B: 6-chloro-4-methoxy-nicotinic acid (300 mg, 1.60 mmol), 4-phenylbenzylamine (439 mg, 2.4 mmol), T3P (1.27 g, 4 mmol) and DIPEA (825 μL, 4.8 mmol), 33 (432 mg, 1.22 mmol, 76%) was obtained.

[0507] 1 H NMR (400MHz, DMSO-d6) δ8.82(t,J=6.0Hz,1H),8.51(s,1H),7.68–7.60(m,4H),7.57–7.27(m,6H),4.53(d,J=6.0Hz,2H),3.99(s,3H).

[0508] HRMS (ESI-TOF) is calculated as C 20 H 18 O2N2 35 Cl[M+H] + :353.1051, actual value: 353.1048.

[0509] Synthesis of 16–N-([1,1'-biphenyl]-4-yl)-6-chloro-4-methoxynicotinamide (34)

[0510]

[0511] According to general method B: 6-chloro-4-methoxy-nicotinic acid (500 mg, 2.67 mmol), 4-aminobiphenyl (540 mg, 3.2 mmol), T3P (2.12 g, 6.68 mmol) and DIPEA (846 μl, 6.68 mmol) were used to obtain 34 (311 mg, 0.92 mmol, 34%).

[0512] 1 H NMR (400MHz, DMSO-d6) δ10.36(s,1H),8.46(s,1H),7.87–7.77(m,2H),7.71–7.62(m,4H),7.52–7.30(m,4H),3.99(s,3H).

[0513] HRMS (ESI-TOF) is calculated as C 19 H 16 O2N2 35 Cl[M+H] + :339.0894, actual value: 339.0893.

[0514] Reference Example 17 – Synthesis of 6-chloro-4-methoxy-N-(3-(trifluoromethyl)benzyl)nicotinamide (35)

[0515]

[0516] According to general method B: 6-chloro-4-methoxy-nicotinic acid (500 mg, 2.67 mmol), 3-trifluoromethylbenzylamine (566 mg, 3.2 mmol), T3P (2.12 g, 6.68 mmol) and DIPEA (846 μL, 6.68 mmol) were used to obtain 35 (518 mg, 1.505 mmol, 56%).

[0517] 1 H NMR (400MHz, DMSO-d6) δ8.91(t,J=6.0Hz,1H),8.48(s,1H),7.75–7.53(m,4H),7.35(s,1H),4.57(d,J=6.0Hz,2H),3.98(s,3H).

[0518] HRMS (ESI-TOF) is calculated as C 15 H 13 O2N2 35 ClF3[M+H] + 345.0612, actual value: 345.0613.

[0519] Reference Example 18 - 6-chloro-N-(cyclohexylmethyl)-4-methoxynicotinamide (36)

[0520]

[0521] According to general method B: 6-chloro-4-methoxy-nicotinic acid (500 mg, 2.67 mmol), cyclohexane-methylamine (361 mg, 3.2 mmol), T3P (2.12 g, 6.68 mmol) and DIPEA (846 μl, 6.68 mmol) were used to obtain 36 (438 mg, 1.55 mmol, 58%).

[0522] 1 H NMR (400MHz, DMSO-d6) δ8.40(s,1H),8.17(t,J=6.0Hz,1H),7.30(s,1H),3.95(s,3H,),3.12–3.06(m,2H),1.87–0.75(m,11H).

[0523] HRMS (ESI-TOF) is calculated as C 14 H 20 O2N2 35 Cl[M+H]+ :283.1207, actual value: 283.1208.

[0524] Example 19 -N-([1,1'-biphenyl]-4-ylmethyl)-4-methoxy-6-(1H-pyrazol-1-yl)nicotinamide (37)

[0525]

[0526] According to the general method C:33 (50mg, 0.142mmol), Pd t BuXPhos G3 (11.2 mg, 0.0142 mmol), Cs2CO3 (138 mg, 0.426 mmol), and pyrazole (24 mg, 0.355 mmol) yielded 37 (30 mg, 0.078 mmol, 55%).

[0527] 1 H NMR(400MHz,DMSO-d6)δ8.79(t,J=6.0Hz,1H),8.68–8.65(m,2H),7.88(dd,J=1.5,1.0Hz,1H),7.6 7–7.46(m,9H),7.39–7.33(m,1H),6.61(dd,J=2.5,1.5Hz,1H),4.55(d,J=6.0Hz,2H),4.08(s,3H).

[0528] HRMS (ESI-TOF) is calculated as C 23 H 21 O2N4[M+H] + :385.1659, actual value: 385.1658.

[0529] Example 20: Synthesis of N-([1,1'-biphenyl]-4-yl)-4-methoxy-6-(1H-pyrazol-1-yl)nicotinamide (38)

[0530]

[0531] According to the general method C:34 (150mg, 0.43mmol), Pd t BuXPhos G3 (34 mg, 0.043 mmol), Cs2CO3 (354 mg, 1.09 mmol), and pyrazole (74 mg, 1.09 mmol) yielded 38 (67 mg, 0.181 mmol, 40%).

[0532] 1H NMR (400MHz, DMSO-d6) δ8.68(d,J=2.5Hz,1H),8.59(s,1H),7.90(d,J=1.5Hz,1H) ,7.70–7.63(m,6H),7.52–7.42(m,4H),6.63(dd,J=2.5,1.5Hz,1H),4.08(s,3H).

[0533] HRMS (ESI-TOF) is calculated as C 22 H 19 O2N4[M+H] + :371.1502, actual value: 371.1501.

[0534] Example 21 – Synthesis of 6-chloro-4-methoxy-N-(3-(trifluoromethyl)benzyl)nicotinamide (39)

[0535]

[0536] According to the general method, C: 35 (150 mg, 0.43 mmol), Pd t BuXPhos G3 (34 mg, 0.043 mmol), Cs2CO3 (354 mg, 1.09 mmol), and pyrazole (74 mg, 1.09 mmol) yielded 39 (72 mg, 0.191 mmol, 44%).

[0537] 1 H NMR(400MHz, DMSO-d6)δ9.26(t,J=6.0Hz,1H),8.66(dd,J=2.5,1.0Hz,1H),8.62(s,1H),8.33(d,J=2.5Hz,1H), 7.88(dd,J=1.5,1.0Hz,1H),7.68–7.57(m,4H),6.61(dd,J=2.5,1.5Hz,1H),4.59(d,J=6.0Hz,2H),4.07(s,3H).

[0538] HRMS (ESI-TOF) is calculated as C 18 H 16 O2N4F3[M+H] + :377.1219, actual value: 377.1220.

[0539] Example 22 – Synthesis of N-(cyclohexylmethyl)-4-methoxy-6-(1H-pyrazol-1-yl)nicotinamide (40)

[0540]

[0541] According to the general method C: 36 (150 mg, 0.43 mmol), Pd t BuXPhos G3 (34 mg, 0.043 mmol), Cs2CO3 (354 mg, 1.09 mmol), and pyrazole (74 mg, 1.09 mmol) yielded 40 (67 mg, 0.21 mmol, 40%).

[0542] 1 H NMR (400MHz, DMSO-d6) δ8.68(dd,J=2.5,1.0Hz,1H),8.56(s,1H),7.90(dd,J=1.5,1.0Hz,1H),7.57(s,1H),6.64 (dd,J=2.5,1.5Hz,1H),3.95(s,3H),3.06–3.03(m,2H),1.79–1.58(m,5H),1.28–1.09(m,4H),1.00–0.83(m,2H).

[0543] HRMS (ESI-TOF) is calculated as C 17 H 23 O2N4[M+H] + :315.1815, actual value: 315.1816.

[0544] Example 23 – Synthesis of N-([1,1'-biphenyl]-4-ylmethyl)-4-hydroxy-6-(1H-pyrazol-1-yl)nicotinamide (41)

[0545]

[0546] TMS-I (17 mg, 0.0858 mmol) was added to a mixture of 37 (11 mg, 0.0286 mmol) and CH2Cl2 (1 ml). The resulting mixture was heated at 90 °C for 90 minutes, then cooled to room temperature, and HCl was added. aq (1.5 ml, 1 M) was then extracted with CH2Cl2 (3 × 10 ml). The organic fractions were combined, dried over anhydrous Na2SO4, concentrated under vacuum, and then purified by rapid column chromatography (0%–5% MeOH, CH2Cl2, 1% NH3) at 20 column volumes to give 41 (2 mg, 0.0054 mmol, 20%).

[0547] 1H NMR (400MHz, DMSO-d6) δ8.76(s,1H),8.64(d,J=2.5Hz,1H),7.86(d,J=1.5Hz,1H),7.7 0–7.60(m,5H),7.52–7.29(m,6H),6.60(dd,J=2.5,1.5Hz,1H),4.59(d,J=6.0Hz,2H).

[0548] HRMS (ESI-TOF) is calculated as C 22 H 19 O2N4[M+H] + :371.1502, Actual value: 371.1502.

[0549] Example 24 – Synthesis of N-([1,1'-biphenyl]-4-yl)-4-hydroxy-6-(1H-pyrazol-1-yl)nicotinamide (42)

[0550]

[0551] TMS-I (66 mg, 0.33 mmol) was added to a reaction mixture of 38 (41 mg, 0.11 mmol) and CH2Cl2 (2.5 mL). The resulting mixture was heated at 90 °C for 90 minutes, then cooled to room temperature, and HCl was added. aq (1.5 ml, 1 M), then extracted with CH2Cl2 (3 × 10 ml). The organic fractions were combined, dried over anhydrous Na2SO4, concentrated under vacuum, and then purified by rapid column chromatography (0%–5% MeOH, CH2Cl2, 1% NH3) at 20 column volumes to give 42 (7 mg, 0.0196 mmol, 18%).

[0552] 1 H NMR (400MHz, DMSO-d6) δ8.70(d,J=2.0Hz,1H),8.64(s,1H),7.92(d,J=2.0Hz,1H),7.72–7.62(m,5H),7.55–7.27(m,6H),6.65(dd,J=2.0Hz,1H).

[0553] HRMS (ESI-TOF) is calculated as C 21 H 17 O2N4[M+H] + :357.1346, actual value: 357.1343.

[0554] Example 25: Synthesis of 4-hydroxy-6-(1H-pyrazol-1-yl)-N-(3-(trifluoromethyl)benzyl)nicotinamide (43)

[0555]

[0556] TMS-I (59 mg, 0.295 mmol) was added to a reaction mixture of 39 (37 mg, 0.0986 mmol) and CH2Cl2 (2.5 mL). The resulting mixture was heated at 90 °C for 90 minutes, then cooled to room temperature, and HCl was added. aq (1.5 ml, 1 M) was then extracted with CH2Cl2 (3 × 10 ml). The organic fractions were combined, dried over anhydrous Na2SO4, concentrated under vacuum, and then purified by rapid column chromatography (0%-5% MeOH, CH2Cl2, 1% NH3) at 20 column volumes to give 43 (20 mg, 0.055 mmol, 57%).

[0557] 1 H NMR (400MHz, DMSO-d6) δ8.66(s,1H),8.59(dd,J=2.5,1.0Hz,1H),7.82(dd,J=1.5,1.0Hz ,1H),7.71–7.49(m,4H),7.26(s,1H),6.55(dd,J=2.5,1.5Hz,1H),4.59(d,J=6.0Hz,2H).

[0558] HRMS (ESI-TOF) is calculated as C 17 H 14 O2N4F3[M+H] + :363.1063, actual value: 363.1067.

[0559] Example 26 – Synthesis of N-(cyclohexylmethyl)-4-hydroxy-6-(1H-pyrazol-1-yl)nicotinamide (44)

[0560]

[0561] TMS-I (128 mg, 0.64 mmol) was added to a reaction mixture of 40 (67 mg, 0.214 mmol) and CH2Cl2 (2.5 mL). The resulting mixture was heated at 90 °C for 90 minutes, then cooled to room temperature, and HCl was added. aq (1.5 ml, 1 M) was then extracted with CH2Cl2 (3 × 10 ml). The organic fractions were combined, dried over anhydrous Na2SO4, concentrated under vacuum, and then purified by rapid column chromatography (0%–5% MeOH, CH2Cl2, 1% NH3) at 20 column volumes to give 44 (17 mg, 0.056 mmol, 26%).

[0562] 1 H NMR (400MHz, DMSO-d6) δ8.63(d,J=2.5Hz,1H),8.47(s,1H),8.29(d,J=2.5Hz,1H),8.21(s,1H),6 .65–6.63(m,1H),3.05(t,J=6.0Hz,2H),1.75–1.44(m,5H),1.27–1.10(m,3H),1.05–0.80(m,2H).

[0563] HRMS (ESI-TOF) is calculated as C 16 H 21 O2N4[M+H] + :301.1659, actual value: 301.1656.

[0564] Reference Example 27 – Synthesis of 6-chloro-4-methoxy-N-(4-(trifluoromethyl)benzyl)nicotinamide (45)

[0565]

[0566] 45 (437 mg, 1.27 mmol, 66%) was obtained by general method B: 6-chloro-5-methoxy-nicotinic acid (350 mg, 1.87 mmol), 4-trifluoromethylbenzylamine (448 mg, 2.56 mmol), T3P (1.48 g, 4.67 mmol) and DIPEA (803 μl, 4.67 mmol).

[0567] 1 ¹H NMR (400MHz, chloroform-d) δ 9.02 (s, 1H), 7.81 (t, J = 6.0Hz, 1H), 7.58 (d, J = 8.0Hz, 2H), 7.43 (d, J = 8.0Hz, 2H), 6.93 (s, 1H), 4.69 (d, J = 6.0Hz, 2H), 4.01 (s, 3H).

[0568] HRMS (ESI-TOF) is calculated as C 15 H 13 O2N2 35 ClF3[M+H] + 345.0612, actual value: 345.0608.

[0569] Reference Example 28 – Synthesis of N-([1,1'-biphenyl]-3-ylmethyl)-6-chloro-4-methoxynicotinamide (46)

[0570]

[0571] 46 (570 mg, 1.61 mmol, 76%) was obtained by general method B: 6-chloro-5-methoxy-nicotinic acid (400 mg, 2.13 mmol), 3-phenylbenzylamine (469 mg, 2.56 mmol), T3P (1.62 g, 5.12 mmol) and DIPEA (880 μl, 5.12 mmol).

[0572] 1 H NMR (400MHz, DMSO-d6) δ8.85(t,J=6.0Hz,1H),8.48(s,1H),7.70–7.30(m,10H),4.56(d,J=6.0Hz,2H),3.96(s,3H).

[0573] HRMS (ESI-TOF) is calculated as C 20 H 18 O2N2 35 Cl[M+H] + :353.1051, actual value: 353.1053.

[0574] Reference Example 29 – Synthesis of N-([1,1'-biphenyl]-3-yl)-6-chloro-4-methoxynicotinamide (47)

[0575]

[0576] 47 (421 mg, 1.24 mmol, 66%) was obtained by general method B: 6-chloro-5-methoxy-nicotinic acid (350 mg, 1.87 mmol), 3-aminobiphenyl (411 mg, 2.43 mmol), T3P (1.48 g, 4.67 mmol) and DIPEA (803 μl, 4.67 mmol).

[0577] 1 ¹H NMR (400MHz, chloroform-d) δ 9.24 (s, 1H), 9.05 (s, 1H), 7.90–7.87 (m, 1H), 7.66–7.30 (m, 8H), 6.91 (s, 1H), 4.06 (s, 3H).

[0578] HRMS (ESI-TOF) is calculated as C 19 H 16 O2N2 35 Cl[M+H] + :339.0894, actual value: 339.0894.

[0579] Reference Example 30 – Synthesis of 6-chloro-4-methoxy-N-(4-(trifluoromethoxy)benzyl)nicotinamide (48)

[0580]

[0581] 48 (434 mg, 1.20 mmol, 75%) was obtained by general method B: 6-chloro-5-methoxy-nicotinic acid (300 mg, 1.60 mmol), 4-trifluoromethoxybenzylamine (367 mg, 1.92 mmol), T3P (1.01 g, 3.2 mmol) and DIPEA (803 μl, 4.67 mmol).

[0582] 1 ¹H NMR (400MHz, chloroform-d) δ 8.97 (s, 1H), 7.78 (t, J = 6.0Hz, 1H), 7.37–7.29 (m, 2H), 7.17–7.11 (m, 2H), 6.90 (s, 1H), 4.61 (d, J = 6.0Hz, 2H), 3.98 (s, 3H).

[0583] HRMS (ESI-TOF) is calculated as C 15 H 13 O3N2 35 ClF3[M+H] + :361.0561, actual result: 361.0563.

[0584] Reference Example 31 – Synthesis of 6-chloro-4-methoxy-N-((4-(trifluoromethyl)cyclohexyl)methyl)nicotinamide (49)

[0585]

[0586] 49 (402 mg, 1.14 mmol, 86%) was obtained by general method B: 6-chloro-5-methoxy-nicotinic acid (250 mg, 1.33 mmol), C-(4-trifluoromethyl-cyclohexylamine (255 mg, 1.59 mmol), T3P (827 mg, 2.6 mmol) and DIPEA (803 μl, 4.67 mmol).

[0587] 1 H NMR (400MHz, CDCl3) δ8.89(s,1H),7.31(s,1H),6.82(s,1H),3.94(s,3H),3.23(t,J=6.2Hz,2H), 1.95–1.74(m,5H),1.57–1.42(m,1H),1.21(qd,J=12.7,2.9Hz,2H),0.93(qd,J=12.8,3.1Hz,2H).

[0588] HRMS (ESI-TOF) is calculated as C 15 H19 O2N2 35 ClF3[M+H] + :351.1081, actual value: 351.1078.

[0589] Reference Example 32 – Synthesis of ((6-chloropyridin-3-yl)methyl)tert-butyl carbamate (50)

[0590]

[0591] 6-Chloropyridin-3-yl)methylamine (4.0 g, 0.0281 mol) was dissolved in CH2Cl2 (50 mL), and then DIPEA (36.9 mL, 0.049 mol) was added. Di-tert-butyl dicarbonate (7.6 g, 0.035 mol) was slowly added to the reaction mixture, and the resulting mixture was stirred at room temperature for 16 hours. The reaction mixture was then washed with water (3 × 50 mL) and brine (50 mL), and dried over Na2SO4. The organic phase was concentrated under vacuum and then purified by rapid column chromatography (cyclohexane 100%–50%, EtOAc 0%–50%) using 20 column volumes to give 50 (6.08 g, 0.0251 mmol, 89%).

[0592] 1 H NMR (400MHz, chloroform-d) δ8.25(dd,J=2.5,1.0Hz,1H),7.58(dd,J=8.5,2.5Hz,1H,),7.24(d,J=1.0Hz,1H),5.08(s,1H),4.26(d,J=6.0Hz,2H),1.41(s,9H).

[0593] HRMS (ESI-TOF) is calculated as C 11 H 16 O2N2 35 Cl[M+H] + :243.0894, actual value: 243.0895.

[0594] Synthesis of tert-butyl carbamate (51) as described in Example 33

[0595]

[0596] 50 (500 mg, 2.06 mmol), phenylboronic acid (249 mg, 2.06 mmol), tetrapalladium (118 mg, 0.103 mmol), and Cs₂CO₃ (1.33 g, 4.12 mmol) were dissolved in anhydrous dioxane (5 mL). The resulting mixture was heated at 100 °C for 30 min under microwave irradiation. The reaction mixture was filtered through a diatomaceous earth mat, and water (25 mL) was added to the reaction mixture, followed by extraction with CH₂Cl₂ (3 × 25 mL). The organic fractions were combined, washed with water (3 × 50 mL) and brine (50 mL), and dried over Na₂SO₄. The organic phase was concentrated under vacuum and then purified by rapid column chromatography (cyclohexane 100%–50%, EtOAc 0%–50%) using 20 column volumes to give 51 (520 mg, 1.83 mmol, 89%).

[0597] 1 ¹H NMR (400MHz, chloroform-d) δ 8.58 (t, J = 1.5Hz, 1H), 7.98–7.93 (m, 2H), 7.67 (d, J = 1.5Hz, 2H), 7.48–7.38 (m, 3H), 5.08 (s, 1H), 4.34 (d, J = 6.0Hz, 2H), 1.46 (s, 9H).

[0598] HRMS (ESI-TOF) is calculated as C 17 H 21 O2N2[M+H] + :285.1597, actual value: 285.1594.

[0599] Reference Example 34 — Synthesis of (6-phenylpyridin-3-yl)methylamine (52)

[0600]

[0601] 51 (500 mg, 1.76 mmol) was dissolved in CH2Cl2 (5 mL), and HCl ((2 M) diethyl ether solution (3 mL)) was added to the solution. The resulting mixture was placed under vacuum and washed with N2; this was repeated 3 times. The resulting mixture was stirred at room temperature for 16 hours. The evaporation was evaporated under vacuum to give 52 (312 mg, 1.69 mmol, 96%).

[0602] 1 H NMR (400MHz, DMSO-d6) δ8.85(s,2H),8.79(dd,J=2.5,1.0Hz,1H),8.15–8.07(m,3H,),8.02(dd,J=8.0,1.0Hz,1H),7.55–7.41(m,3H),4.08(s,2H).

[0603] HRMS (ESI-TOF) is calculated as C 12 H 13 N2[M+H] + :185.1073, Actual value: 185.1073.

[0604] Synthesis of 35–6-chloro-4-methoxy-N-((6-phenylpyridin-3-yl)methyl)nicotinamide (53)

[0605]

[0606] 53 (144 mg, 0.41 mmol, 90%) was obtained by general method B: 52 (100 mg, 0.54 mmol), 6-chloro-4-methoxy-nicotinic acid (85 mg, 0.45 mmol) and T3P (358 mg, 1.125 mmol).

[0607] 1 H NMR (400MHz, DMSO-d6) δ 8.91 (t, J = 6.0Hz, 1H), 8.51 (s, 1H), 8.13–7.79 (m, 4H), 7.63–7.37 (m, 4H), 7.32 (s, 1H), 4.54 (d, J = 6.0Hz, 2H), 3.98 (s, 3H). HRMS (ESI-TOF) calculated as C 19 H 17 O2N3 35 Cl[M+H] + :354.1003, actual value: 354.1002.

[0608] Example 36 – Synthesis of 4-methoxy-6-(1H-pyrazol-1-yl)-N-(4-(trifluoromethyl)benzyl)nicotinamide (56)

[0609]

[0610] According to the general method, C: 45 (200 mg, 0.58 mmol), Pd t BuXPhos G3 (46 mg, 0.058 mmol), Cs2CO3 (377 mg, 1.16 mmol), pyrazole (42 mg, 0.63 mmol) and t BuOH (2 ml) yielded 56 (69 mg, 0.185 mmol, 32%).

[0611] 1H NMR (400MHz, DMSO-d6) δ8.88(t,J=6.0Hz,1H),8.66(dd,J=2.5,1.0Hz,1H),8.64(s,1H),7.89(t,J=1.5,1.0Hz,1H),7.7 2(d,J=7.0Hz,2H),7.60(s,1H),7.56(d,J=7.0Hz,2H),6.62(dd,J=2.5,1.5Hz,1H),4.59(d,J=6.0Hz,2H),4.08(s,3H).

[0612] HRMS (ESI-TOF) is calculated as C 18 H 16 O2N4F3[M+H] + :377.1221, Actual value: 377.1221.

[0613] Example 37 – Synthesis of N-([1,1'-biphenyl]-3-ylmethyl)-4-methoxy-6-(1H-pyrazol-1-yl)nicotinamide (57)

[0614]

[0615] According to the general method C: 46 (150 mg, 0.42 mmol), Pd t BuXPhos G3 (33 mg, 0.0426 mmol), Cs2CO3 (341 mg, 1.05 mmol), pyrazole (42 mg, 0.63 mmol) and t BuOH (2 ml) yielded 57 (78 mg, 0.203 mmol, 48%).

[0616] 1 ¹H NMR (400MHz, chloroform-d) δ 9.03 (d, J = 2.0 Hz, 1H), 8.53 (d, J = 2.5 Hz, 1H), 7.85–7.76 (m, 1H), 7.70–7.63 (m, 1H), 7.56–7.20 (m, 9H), 7.20–7.15 (m, 1H), 6.40 (dd, J = 2.5, 2.0 Hz, 1H), 4.66 (d, J = 6.0 Hz, 2H), 3.97 (s, 3H).

[0617] HRMS (ESI-TOF) is calculated as C 23 H 21 O2N4[M+H] + 385.1659, actual value: 385.1656.

[0618] Example 38 – Synthesis of N-([1,1'-biphenyl]-3-yl)-4-methoxy-6-(1H-pyrazol-1-yl)nicotinamide (58)

[0619]

[0620] According to the general method C: 47 (100mg, 0.295mmol), Pd t BuXPhos G3 (20 mg, 0.028 mmol), Cs2CO3 (185 mg, 0.568 mmol), pyrazole (23 mg, 0.34 mmol) and t BuOH (2 ml) yielded 58 (50 mg, 0.13 mmol, 46%).

[0621] 1 ¹H NMR (400MHz, chloroform-d) δ 9.42 (s, 1H), 9.18 (s, 1H), 8.64 (dd, J = 2.5, 1.0 Hz, 1H), 7.91 (m, 1H), 7.78 (dd, J = 1.5, 1.0 Hz, 1H), 7.69–7.33 (m, 9H), 6.51 (dd, J = 2.5, 1.5 Hz, 1H), 4.23 (s, 3H).

[0622] HRMS (ESI-TOF) is calculated as C 22 H 19 O2N4[M+H] + :371.1502, actual value: 371.1497.

[0623] Example 39 – Synthesis of 4-methoxy-6-(1H-pyrazol-1-yl)-N-(4-(trifluoromethoxy)benzyl)nicotinamide (59)

[0624]

[0625] According to the general method C: 48 (100mg, 0.277mmol), Pd t BuXPhos G3 (19 mg, 0.027 mmol), Cs2CO3 (180 mg, 0.554 mmol), pyrazole (23 mg, 0.33 mmol) and t BuOH (2 ml) yielded 59 (79 mg, 0.201 mmol, 73%).

[0626] 1H NMR (400MHz, chloroform-d) δ9.12(s,1H),8.63(dd,J=2.5,1.0Hz,1H),7.76(t,J=1.5,1.0Hz,1H),7.61(s,1H),7.4 0(d,J=7.5Hz,2H),7.20(d,J=7.5Hz,2H),6.50(dd,J=2.5,1.5Hz,1H,),4.68(d,J=6.0Hz,2H),4.10(s,3H).

[0627] HRMS (ESI-TOF) is calculated as C 18 H 16 O3N4F3[M+H] + :393.1169, actual value: 393.1163.

[0628] Example 40 – Synthesis of 4-methoxy-6-(1H-pyrazol-1-yl)-N-((4-(trifluoromethyl)cyclohexyl)methyl)nicotinamide (60)

[0629]

[0630] According to the general method C: 49 (100mg, 0.277mmol), Pd t BuXPhos G3 (19 mg, 0.027 mmol), Cs2CO3 (180 mg, 0.554 mmol), pyrazole (23 mg, 0.33 mmol) and t BuOH (2 ml) yielded 60 (49 mg, 0.127 mmol, 46%).

[0631] 1 H NMR (400MHz, CDCl3) δ9.07 (s, 1H), 8.62 (dd, J = 2.6, 0.7Hz, 1H), 7.75 (d, J = 1.6Hz, 1H), 7.60 (s, 1H), 7.54 (d, J = 6.2Hz, 1H), 6.49 (dd, J = 2. 7,1.7Hz,1H),4.13(s,3H),3.35(t,J=6.4Hz,2H),2.06–1.90(m,5H),1.69–1.57(m,1H),1.43–1.20(m,2H),1.05(qd,J=12.8,2.6Hz,2H).

[0632] HRMS (ESI-TOF) is calculated as C 18 H 22 O2N4F3[M+H] + :383.1689, actual value: 383.1689.

[0633] Example 41 – Synthesis of 4-methoxy-N-((6-phenylpyridin-3-yl)methyl)-6-(1H-pyrazol-1-yl)nicotinamide (61)

[0634]

[0635] According to the general method C: 53 (80mg, 0.226mmol), Pd t BuXPhos G1 (14 mg, 0.0226 mmol), Cs2CO3 (145 mg, 0.452 mmol), pyrazole (15 mg, 0.226 mmol) and t BuOH (2 ml) yielded 61 (16 mg, 0.043 mmol, 19%).

[0636] 1 H NMR (400MHz, chloroform-d) δ9.11(s,1H),8.69(dd,J=2.5,1.0Hz,1H),8.62(dd,J=2.5,1.0Hz,1H),8.01– 7.68(m,7H),7.52–7.38(m,3H),6.48(dd,J=2.5,1.5Hz,1H),4.73(d,J=6.0Hz,2H),4.10(s,3H).

[0637] HRMS (ESI-TOF) is calculated as C 22 H 20 O2N4[M+H] + :386.1611, actual value: 386.1604.

[0638] Example 42 – Synthesis of 4-hydroxy-6-(1H-pyrazol-1-yl)-N-(4-(trifluoromethyl)benzyl)nicotinamide (64)

[0639]

[0640] According to the general method, 64 (6 mg, 0.016 mmol, 52%) was obtained from a DMAc solution (2 ml) of D: 56 (12 mg, 0.0319 mmol) and LiCl (19 mg, 0.319 mmol).

[0641] 1H NMR(400MHz,DMSO-d6)δ10.12(s,1H),8.68(s,1H),8.64–8.58(m,1H),7.81(s,1H,),7.71(d ,J=8.0Hz,2H),7.56(d,J=8.0Hz,2H),7.20(m,1H),6.60–6.53(m,1H),4.61(d,J=6.0Hz,2H).

[0642] HRMS (ESI-TOF) is calculated as C 17 H 12 O2N4F3[MH] - :361.0917, Actual value: 361.0917.

[0643] Example 43 – Synthesis of N-([1,1'-biphenyl]-3-ylmethyl)-4-hydroxy-6-(1H-pyrazol-1-yl)nicotinamide (65)

[0644]

[0645] According to the general method, 65 (30 mg, 0.081 mmol, 63%) was obtained from a DMAc solution (5 ml) of D: 57 (50 mg, 0.129 mmol) and LiCl (78 mg, 1.29 mmol).

[0646] 1 H NMR (400MHz, DMSO-d6) δ13.21(s,1H),9.26(t,J=6.0Hz,1H),8.69(dd,J=2.5,1.0Hz,1H),8.64(d,J=2.5Hz,1H),8.28 (d,J=2.5Hz,1H),7.91(dd,J=1.5,1.0Hz,1H),7.59–7.29(m,9H),6.64(dd,J=2.5,1.5Hz,1H),4.63(d,J=6.0Hz,2H,).

[0647] HRMS (ESI-TOF) is calculated as C 22 H 19 O2N4[M+H] + :371.1502, Actual value: 371.1507

[0648] Example 44 – Synthesis of N-([1,1'-biphenyl]-3-yl)-4-hydroxy-6-(1H-pyrazol-1-yl)nicotinamide (66)

[0649]

[0650] According to the general method, 66 (19 mg, 0.053 mmol, 50%) was obtained from a DMAc solution (2 ml) of D: 58 (40 mg, 0.107 mmol) and LiCl (45 mg, 1.07 mmol).

[0651] 1 H NMR (400MHz, DMSO-d6) δ8.74–8.64(m,2H),8.04(s,1H),7.93(s,1H),7.76–7.63(m,3H),7.56–7.29(m,7H),6.68–6.63(m,1H).

[0652] HRMS (ESI-TOF) is calculated as C 21 H 17 O2N4[M+H] + :357.1346, actual value: 357.1345.

[0653] Example 45 – Synthesis of 4-hydroxy-6-(1H-pyrazol-1-yl)-N-(4-(trifluoromethoxy)benzyl)nicotinamide (67)

[0654]

[0655] According to the general method, 67 (43 mg, 0.114 mmol, 64%) was obtained from a DMAc solution (2 ml) of D: 59 (70 mg, 0.178 mmol) and LiCl (74 mg, 1.78 mmol).

[0656] 1 H NMR(400MHz,DMSO-d6)δ13.16(s,1H),9.41(s,1H),8.76(s,1H),8.67–8.61(m,1H),7.89–7 .84(m,1H),7.51–7.45(m,2H),7.35–7.31(m,3H),6.61–6.58(m,1H),4.57(d,J=6.0Hz,2H).

[0657] HRMS (ESI-TOF) is calculated as C 17 H 12 O3N4F3[MH] - 377.0867, actual value: 377.865.

[0658] Example 46: Synthesis of 4-hydroxy-6-(1H-pyrazol-1-yl)-N-((4-(trifluoromethyl)cyclohexyl)methyl)nicotinamide (68)

[0659]

[0660] According to the general method, 68 (8.5 mg, 0.023 mmol, 30%) was obtained by preparing a DMAc solution (2 ml) of D:60 (30 mg, 0.078 mmol) and LiCl (33 mg, 0.78 mmol).

[0661] 1 H NMR (400MHz, DMSO) δ13.33(s,1H),9.10(s,1H),8.72(s,1H),8.63(d,J=2.6Hz,1H),7.86(d,J=1.6Hz,1H),7.28(s,1H),6.6 0(dd,J=2.7,1.7Hz,1H),3.20(t,J=6.3Hz,2H),2.29–2.13(m,1H),1.96–1.78(m,4H),1.65–1.47(m,1H),1.32–0.96(m,4H).

[0662] HRMS (ESI-TOF) is calculated as C 17 H 20 O2N4F3[M+H] + :369.1532, actual value: 369.1533.

[0663] Example 47 – Synthesis of 4-hydroxy-N-((6-phenylpyridin-3-yl)methyl)-6-(1H-pyrazol-1-yl)nicotinamide (69)

[0664]

[0665] 69 (6 mg, 0.0161 mmol, 40%) was obtained by using a DMAc solution (2 ml) of D:61 (16 mg, 0.041 mmol) and lithium chloride (17 mg, 0.41 mmol) according to the general method.

[0666] 1 H NMR (400MHz, DMSO-d6) δ 8.80–8.62 (m, 3H), 8.15–7.86 (m, 6H), 7.56–7.43 (m, 4H), 6.62–6.59 (m, 1H), 4.62 (d, J = 6.0Hz, 2H).

[0667] HRMS (ESI-TOF) is calculated as C 21 H 18 O2N5[M+H] + :372.1455, actual value: 372.1447.

[0668] Synthesis of N-([1,1'-biphenyl]-4-ylmethyl)-6-chloro-5-methoxynicotinamide (72) as described in Example 48

[0669]

[0670] According to general method B: 6-chloro-5-methoxy-nicotinic acid (850 mg, 4.55 mmol), 4-phenylbenzylamine (1 g, 5.46 mmol), T3P (4.32 g, 13.6 mmol) and DIPEA (2.2 ml, 6.68 mmol) were used to obtain 72 (502 mg, 1.42 mmol, 31%).

[0671] 1 H NMR (400MHz, DMSO-d6) δ 9.32 (t, J = 6.0Hz, 1H), 8.50 (d, J = 2.0Hz, 1H), 7.96 (d, J = 2.0Hz, 1H), 7.74–7.31 (m, 9H), 4.56 (d, J = 6.0Hz, 2H), 3.96 (s, 3H). HRMS (ESI-TOF) calculated as C 20 H 18 O2N2 35 Cl[M+H] + :353.1051, actual value: 353.1049.

[0672] Example 49 – Synthesis of N-([1,1'-biphenyl]-4-ylmethyl)-5-methoxy-6-(1H-pyrazol-1-yl)nicotinamide (73)

[0673]

[0674] According to the general method, C:72 (100 mg, 0.284 mmol), Pd t BuXPhos G3 (22 mg, 0.0284 mmol), Cs2CO3 (279 mg, 0.852 mmol), pyrazole (38 mg, 0.568 mmol) and t BuOH (2 ml) yielded 73 (23 mg, 0.0598 mmol, 20%).

[0675] 1H NMR (400MHz, chloroform-d) δ8.46(d,J=2.0Hz,1H),8.30(dd,J=2.5,1.0Hz,1H),7.97(d,J=2.0Hz,1H),7.76(dd ,J=1.5,1.0Hz,1H),7.64–7.32(m,9H),6.46(dd,J=2.5,1.5Hz,1H),4.67(d,J=6.0Hz,2H),3.93(s,3H).

[0676] HRMS (ESI-TOF) is calculated as C 23 H 21 O2N4[M+H] + 385.1659, actual value: 385.1661.

[0677] Example 50 – Synthesis of N-([1,1'-biphenyl]-3-ylmethyl)-5-methoxy-6-(1H-pyrazol-1-yl)nicotinamide (74)

[0678]

[0679] [N-([1,1'-biphenyl]-3-ylmethyl)-6-chloro-5-methoxynicotinamide] was obtained by general method B: 6-chloro-5-methoxynicotinic acid (250 mg, 1.33 mmol), 3-phenylbenzylamine (285 mg, 1.56 mmol), T3P (1.08 g, 3.4 mmol), and DIPEA (574 μl, 3.34 mmol). The crude substance was not purified and proceeded to the next step according to general method C: N-([1,1'-biphenyl]-3-ylmethyl)-6-chloro-5-methoxynicotinamide (100 mg, 0.284 mmol), Pd... t BuXPhos G3 (20 mg, 0.028 mmol), Cs2CO3 (185 mg, 0.568 mmol), and pyrazole (23 mg, 0.34 mmol) yielded 74 (54 mg, 0.14 mmol, 49%).

[0680] 1 H NMR (400MHz, DMSO-d6) δ9.37(t,J=6.0Hz,1H),8.62(d,J=2.0Hz,1H),8.28(dd,J=2.5,1.0Hz,1H),8.10(d,J=2.0Hz, 1H),7.77(t,J=1.5,1.0Hz,1H),7.69–7.32(m,9H),6.53(dd,J=2.5,1.5Hz,1H),4.62(d,J=6.0Hz,2H),3.94(s,3H).

[0681] HRMS (ESI-TOF) is calculated as C 23 H 21 O2N4[M+H] + 385.1659, actual value: 385.1661.

[0682] Example 51 – Synthesis of N-([1,1'-biphenyl]-4-ylmethyl)-5-hydroxy-6-(1H-pyrazol-1-yl)nicotinamide (75)

[0683]

[0684] 75 (4 mg, 0.01 mmol, 35%) was obtained by preparing a DMAc solution (2 ml) of 73 (12 mg, 0.031 mmol) and LiCl (19 mg, 0.31 mmol) according to the general method.

[0685] 1 H NMR (500MHz, DMSO-d6) δ11.68(s,1H),9.28(t,J=6.0Hz,1H),8.76(d,J=2.5Hz,1H),8.52(d,J=2.0Hz,1H),7.96(d ,J=2.0Hz,1H),7.72–7.60(m,4H),7.48–7.41(m,4H),7.39–7.30(m,1H),6.74–6.72(m,1H),4.55(d,J=6.0Hz,2H).

[0686] HRMS (ESI-TOF) is calculated as C 22 H 19 O2N4[M+H] + :371.1502, actual value: 371.1503.

[0687] Example 52 – Synthesis of N-([1,1'-biphenyl]-3-ylmethyl)-5-hydroxy-6-(1H-pyrazol-1-yl)nicotinamide (76)

[0688]

[0689] 76 (14 mg, 0.037 mmol, 32%) was obtained by preparing a DMAc solution (3.5 ml) of D:74 (45 mg, 0.117 mmol) and LiCl (71 mg, 1.17 mmol) according to the general method.

[0690] 1H NMR (400MHz, DMSO-d6) δ11.68(s,1H),9.28(t,J=6.0Hz,1H),8.76(d,J=2.5Hz,1H),8.52(d,J=2 .0Hz,1H),7.97(d,J=2.0Hz,1H),7.71–7.32(m,10H),6.75–6.71(m,1H),4.59(d,J=6.0Hz,2H).

[0691] HRMS (ESI-TOF) is calculated as C 22 H 19 O2N4[M+H] + :371.1502, Actual value: 371.1502.

[0692] Example 53 – Synthesis of 1-(5-(([1,1'-biphenyl]-4-ylmethyl)carbamoyl)-4-methoxypyridin-2-yl)-1H-pyrazole-4-carboxylic acid ethyl ester (77)

[0693]

[0694] According to the general method, C:33 (150 mg, 0.43 mmol), Pd t BuXPhos G3 (34 mg, 0.043 mmol), Cs2CO3 (354 mg, 1.09 mmol), pyrazole-4-carboxylic acid ethyl ester (89 mg, 0.649 mmol) and t BuOH (2 ml) yielded 77 (93 mg, 0.20 mmol, 48%).

[0695] 1 H NMR(400MHz,DMSO-d6)δ9.37(s,1H),9.00(s,1H),8.85(s,1H),8.24(s,1H),7.69–7.62(m,4H),7. 54–7.29(m,6H),4.59(d,J=6.0Hz,2H),4.28(q,J=7.0Hz,2H),4.09(s,3H),1.31(t,J=7.0Hz,3H).

[0696] HRMS (ESI-TOF) is calculated as C 26 H 25 O4N4[M+H] + :457.1870, actual value: 457.1876.

[0697] Example 54 – Synthesis of N-([1,1'-biphenyl]-4-ylmethyl)-6-(4-cyano-1H-pyrazol-1-yl)-4-methoxynicotinamide (78)

[0698]

[0699] According to the general method, C:33 (150 mg, 0.426 mmol), Pd t BuXPhos G3 (16 mg, 0.0213 mmol), Cs2CO3 (276 mg, 0.852 mmol), pyrazole-4-onitrile (47 mg, 0.511 mmol) and t BuOH (4 ml) yielded 78 (9 mg, 0.022 mmol, 5%).

[0700] 1 H NMR (400MHz, DMSO-d6) δ9.47 (d, J=.0 1Hz, 1H), 8.87 (t, J = 6.0Hz, 1H), 8.66 (s, 1H), 8.48 (d, J = 1.0Hz, 1H), 7.70–7.31 (m, 10H), 4.55 (d, J = 6.0Hz, 2H), 4.09 (s, 3H).

[0701] HRMS (ESI-TOF) is calculated as C 24 H 18 O2N5[MH] - :408.1466, actual value: 408.1467.

[0702] Example 55 – Synthesis of 1-(5-(([1,1'-biphenyl]-4-ylmethyl)carbamoyl)-3-methoxypyridin-2-yl)-1H-pyrazole-4-carboxylic acid ethyl ester (79)

[0703]

[0704] According to the general method, C:72 (150 mg, 0.43 mmol), Pd t BuXPhos G3 (34 mg, 0.043 mmol), Cs2CO3 (354 mg, 1.09 mmol), pyrazole-4-carboxylic acid ethyl ester (89 mg, 0.649 mmol) and t BuOH (4 ml) yielded 79 (44 mg, 0.096 mmol, 23%).

[0705] 1H NMR (400MHz, DMSO-d6) δ9.41(d,J=6.0Hz,1H),8.78–8.75(m,1H),8.64(d,J=2.5Hz,1H),8.15(d,J=2.5Hz,1H),7.67–7.61(m,5H ),7.53–7.42(m,4H),7.39–7.34(m,1H),4.59(d,J=6.0Hz,2H),4.27(q,J=7.0Hz,2H),4.00–3.94(s,3H),1.30(t,J=7.0Hz,3H).

[0706] HRMS (ESI-TOF) is calculated as C 26 H 25 O4N4[M+H] + :457.1870, actual value: 457.1874.

[0707] Example 56 – Synthesis of 1-(4-methoxy-5-((4-(trifluoromethyl)benzyl)carbamoyl)pyridin-2-yl)-1H-pyrazole-4-carboxylic acid ethyl ester (80)

[0708]

[0709] According to the general method, C:45 (100 mg, 0.29 mmol), RockPhos Pd G3 (20 mg, 0.029 mmol), Cs2CO3 (188 mg, 0.58 mmol), 4-ethylpyrazole (24 mg, 0.63 mmol) and t BuOH (4 ml) yielded 80 (86 mg, 0.192 mmol, 66%).

[0710] 1 H NMR (400MHz, chloroform-d) δ9.12(s,1H),9.07(d,J=1.0Hz,1H),8.11(d,J=1.0Hz,1H),7.89(t,J=6.0Hz,1H),7.63(s,1H),7. 62–7.58(m,2H),7.50–7.45(m,2H),4.73(d,J=6.0Hz,2H),4.35(q,J=7.0Hz,2H),4.11(s,3H),1.37(t,J=7.0Hz,3H).

[0711] HRMS (ESI-TOF) is calculated as C 21 H 18 O4N4F3[MH] - :447.1285, actual value: 447.1281.

[0712] Example 57 – Synthesis of 6-(4-cyano-1H-pyrazol-1-yl)-4-methoxy-N-(4-(trifluoromethyl)benzyl)nicotinamide (81)

[0713]

[0714] According to the general method, C:45 (100 mg, 0.29 mmol), RockPhos Pd G3 (24 mg, 0.029 mmol), Cs2CO3 (180 mg, 0.554 mmol), pyrazole-4-onitrile (24 mg, 0.35 mmol) and t BuOH (4 ml) yielded 81 (23 mg, 0.057 mmol, 20%).

[0715] 1 ¹H NMR (400MHz, chloroform-d) δ 9.13 (s, 1H), 9.04 (d, J = 1.0 Hz, 1H), 7.99 (d, J = 1.0 Hz, 1H), 7.88 (t, J = 6.0 Hz, 1H), 7.67–7.56 (m, 3H), 7.51–7.42 (m, 2H), 4.74 (d, J = 6.0 Hz, 2H), 4.03 (s, 3H).

[0716] HRMS (ESI-TOF) is calculated as C 18 H 15 O4N4F3[M+H] + :402.1172, actual value: 402.1172.

[0717] Example 58 – Synthesis of 1-(4-methoxy-5-(((6-phenylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)-1H-pyrazole-4-carboxylic acid ethyl ester (82)

[0718]

[0719] According to the general method, C:53 (80 mg, 0.226 mmol), RockPhosPd G3 (19 mg, 0.0226 mmol), Cs2CO3 (145 mg, 0.452 mmol), 4-ethylpyrazole (31 mg, 0.226 mmol) and t BuOH (2.5 ml) yielded 82 (11 mg, 0.024 mmol, 11%).

[0720] 1¹H NMR (400MHz, chloroform-d) δ 9.13 (s, 1H), 9.08 (s, 1H), 8.70–8.68 (m, 1H), 8.12 (s, 1H), 8.01–7.94 (m, 2H), 7.90 (t, J = 6.0Hz, 1H), 7.84–7.37 (m, 6H), 4.73 (d, J = 6.0Hz, 2H), 4.32 (q, J = 7.0Hz, 2H), 4.11 (s, 3H), 1.37 (t, J = 7.0Hz, 3H).

[0721] HRMS (ESI-TOF) is calculated as C 25 H 24 O4N5[M+H] + :458.1822, actual value: 458.1816.

[0722] Example 59 – Synthesis of 1-(5-(([1,1'-biphenyl]-4-ylmethyl)carbamoyl)-4-hydroxypyridin-2-yl)-1H-pyrazole-4-carboxylic acid ethyl ester (83)

[0723]

[0724] 83 (70 mg, 0.158 mmol, 90%) was obtained by using a DMAc solution (35 ml) of D:80 (82 mg, 0.179 mmol) and LiCl (1.79 mg, 1.79 mmol) according to the general method.

[0725] 1 H NMR(400MHz,DMSO-d6)δ9.42(s,1H),9.00(s,1H),8.85(s,1H),8.24(s,1H),7.72–7.58(m,4H),7.54– 7.39(m,5H),7.38–7.29(m,1H),4.59(d,J=6.0Hz,2H,),4.27(q,J=7.0Hz,2H),1.30(t,J=7.0Hz,3H).

[0726] HRMS (ESI-TOF) is calculated as C 25 H 23 O4N4[M+H] + :443.1713, Actual value: 443.1713.

[0727] Example 60 – Synthesis of 1-(5-(([1,1'-biphenyl]-4-ylmethyl)carbamoyl)-4-hydroxypyridin-2-yl)-1H-pyrazole-4-carboxylic acid (84)

[0728]

[0729] Dissolve 83 (65 mg, 0.147 mmol) in THF (5 ml), add water (0.5 ml) to the reaction mixture, and then add LiOH monohydrate (16 mg, 0.40 mmol). Stir the resulting mixture overnight at room temperature. Then, use HCl. aq (1M) The reaction mixture was acidified to pH 3, extracted with EtOAc (3 × 25 ml), washed with brine, and dried over anhydrous Na2SO4. The crude compound was then purified by rapid column chromatography (CH2Cl2, MeOH 0-20%, formic acid 1%) at 20 column volumes to give 84 (52 mg, 0.125 mmol, 85%) as a grayish-white solid.

[0730] 1 H NMR (400MHz, DMSO-d6) δ13.34(s,1H),9.42(s,1H),9.00(s,1H),8.96(s,1H),8.19(s,1H),7.71–7.31(m,11H),4.59(d,J=6.0Hz,2H).

[0731] HRMS (ESI-TOF) is calculated as C 23 H 19 O4N4[M+H] + :415.1400, actual value: 415.1401.

[0732] Example 61 – Synthesis of N-([1,1'-biphenyl]-4-ylmethyl)-6-(4-cyano-1H-pyrazol-1-yl)-4-hydroxynicotinamide (85)

[0733]

[0734] 78 (6 mg, 0.0146 mmol) was dissolved in DMAc (1.5 mL) in a microwave-safe vial, followed by the addition of Cs₂CO₃ (14.3 mg, 0.044 mmol). The resulting reaction mixture was heated under microwave irradiation at 130 °C for 1 hour. H₂O (50 mL) was added, and the resulting mixture was extracted with ethyl acetate (3 × 15 mL). The combined organic fractions were washed with brine, dried over Na₂SO₄, and the solvent was removed under vacuum. The crude compound was then purified by rapid column chromatography (CH₂Cl₂:MeOH 0-10%) at 20 column volumes to give 85 (2.5 mg, 0.006 mmol, 44%).

[0735] 1H NMR (400MHz, DMSO-d6) δ9.45 (s, 1H), 8.87 (d, J = 2.0Hz, 1H), 8.44 (s, 1H), 7.73–7.42 (m, 11H), 4.60 (d, J = 6.0Hz, 2H).

[0736] HRMS (ESI-TOF) is calculated as C 23 H 16 O2N5[MH] - :394.1309, actual value: 394.1309.

[0737] Example 62 – Synthesis of 1-(5-(([1,1'-biphenyl]-4-ylmethyl)carbamoyl)-3-hydroxypyridin-2-yl)-1H-pyrazole-4-carboxylic acid (86)

[0738]

[0739] Dissolve 79 (20 mg, 0.043 mmol) in THF (2 ml), add MeOH (2 ml) and water (0.5 ml) to the reaction mixture, then add LiOH monohydrate (16 mg, 0.40 mmol). Stir the resulting mixture overnight at room temperature. Then, use HCl... aq The reaction mixture was acidified to pH 3 with (1M) solution, extracted with EtOAc (3 × 25 ml), washed with brine, and dried over anhydrous Na2SO4. The crude compound was then purified by rapid column chromatography (CH2Cl2:MeOH 0-20%, formic acid 1%) at 20 column volumes to give 86 (8 mg, 0.019 mmol, 45%).

[0740] 1 H NMR (400MHz, DMSO-d6) δ9.09 (s, 1H), 8.90 (d, J = 6.0Hz, 1H), 7.92–7.09 (m, 12H), 4.47 (d, J = 6.0Hz, 2H).

[0741] HRMS (ESI-TOF) is calculated as C 23 H 17 O4N4[MH] - :413.1255, actual value: 413.1252.

[0742] Example 63 – Synthesis of 1-(4-hydroxy-5-((4-(trifluoromethyl)benzyl)carbamoyl)pyridin-2-yl)-1H-pyrazole-4-carboxylic acid ethyl ester (87)

[0743]

[0744] 87 (48 mg, 0.110 mmol, 73%) was obtained by preparing a DMAc solution (3.5 ml) of D:80 (68 mg, 0.151 mmol) and LiCl (91 mg, 1.51 mmol) according to the general method.

[0745] 1 H NMR(400MHz,THF-d8)δ13.46(s,1H),8.99(d,J=1.0Hz,1H),8.93–8.85(m,1H),8.74(s,1H),8.07(d,J=1.0Hz,1H),7.65( d, J=7.0Hz, 2H), 7.58 (d, J=7.0Hz, 2H), 7.48 (s, 1H), 4.71 (d, J=6.0Hz, 2H), 4.29 (q, J=7.0Hz, 2H), 1.33 (t, J=7.0Hz, 3H).

[0746] HRMS (ESI-TOF) is calculated as C 20 H 16 O4N4F3[MH] - :433.1129, actual value: 433.1125.

[0747] Example 64 – Synthesis of 1-(4-hydroxy-5-(((6-phenylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)-1H-pyrazole-4-carboxylic acid ethyl ester (88)

[0748]

[0749] 88 (3.5 mg, 0.0079 mmol, 53%) was obtained by using a DMAc solution (2 ml) of D:82 (7 mg, 0.015 mmol) and LiCl (9 mg, 0.15 mmol) according to the general method.

[0750] 1 H NMR (400MHz, DMSO-d6) δ9.00(s,1H),8.80(s,1H),8.67(d,J=2.0Hz,1H),8.24(s,1H),8.07(dd,J=8.0,2.0Hz,2H),7.95(d,J=8 .0Hz,1H),7.85(dd,J=8.0,2.0Hz,1H),7.54–7.39(m,5H),4.61(d,J=6.0Hz,2H),4.26(q,J=7.0Hz,2H),1.30(t,J=7.0Hz,3H).

[0751] HRMS (ESI-TOF) is calculated as C24 H 22 O4N5[M+H] + :444.1666, actual value: 444.1659.

[0752] Example 65 – Synthesis of 6-(4-cyano-1H-pyrazol-1-yl)-4-hydroxy-N-(4-(trifluoromethyl)benzyl)nicotinamide (89)

[0753]

[0754] 89 (5 mg, 0.012 mmol, 26%) was obtained by preparing a DMAc solution (2 ml) of 81 (20 mg, 0.049 mmol) and LiCl (30 mg, 0.50 mmol) according to the general method.

[0755] 1 H NMR (400MHz, DMSO-d6) δ9.43(d,J=1.0Hz,1H),8.81(s,1H),8.43(d,J=1.0Hz,1H) ,7.71(d,J=8.0Hz,2H),7.57(d,J=8.0Hz,2H),7.42(s,1H),4.63(d,J=6.0Hz,2H).

[0756] HRMS (ESI-TOF) is calculated as C 18 H 11 O4N4F3[MH] - 386.0870, actual value: 386.0867.

[0757] Example 66 – Synthesis of 1-(4-hydroxy-5-((4-(trifluoromethyl)benzyl)carbamoyl)pyridin-2-yl)-1H-pyrazole-4-carboxylic acid (90)

[0758]

[0759] Dissolve 87 (40 mg, 0.092 mmol) in THF (2 ml), add MeOH (2 ml) and water (0.5 ml) to the reaction mixture, then add LiOH monohydrate (19 mg, 0.46 mmol). Stir the resulting mixture overnight at room temperature. Then, use HCl. aq (1M) The reaction mixture was acidified to pH 3, extracted with EtOAc (3 × 25 ml), washed with brine, and dried over anhydrous Na2SO4. The crude compound was then purified by rapid column chromatography (CH2Cl2:MeOH 0-20%, formic acid 1%) at 20 column volumes to give 90 (17 mg, 0.042 mmol, 46%).

[0760] 1 H NMR(400MHz,DMSO-d6)δ9.56(s,1H),8.94(s,1H),8.78(s,1H),8.17(s,1H),7. 70 (d, J = 8.0 Hz, 2H), 7.56 (d, J = 8.0 Hz, 2H), 7.38 (s, 1H), 4.63 (d, J = 6.0 Hz, 2H).

[0761] HRMS (ESI-TOF) is calculated as C 18 H 12 O4N4F3[MH] - :405.0816, actual value: 405.0815.

[0762] Example 67 – Synthesis of 1-(4-hydroxy-5-(((6-phenylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)-1H-pyrazole-4-carboxylic acid (91)

[0763]

[0764] Dissolve 88 (3 mg, 0.0072 mmol) in THF (0.8 mL), add MeOH (0.15 mL) and water (0.05 mL) to the reaction mixture, then add LiOH monohydrate (1 mg, 0.023 mmol). Stir the resulting mixture overnight at room temperature. Then, use HCl... aq (1M) The reaction mixture was acidified to pH 3 and 91 (1 mg, 0.0024 mmol, 33%) was obtained by preparative HPLC.

[0765] 1 H NMR(500MHz,DMSO-d6)δ8.94(s,1H),8.68(s,1H),8.19(s,1H),8.13–8.05(m,2H),7. 96(d,J=8.5Hz,1H),7.87(d,J=8.5Hz,1H),7.55–7.39(m,6H),4.62(d,J=6.0Hz,2H).

[0766] HRMS (ESI-TOF) is calculated as C 22 H 18 O4N5[M+H] + :416.1353, actual value: 416.1352.

[0767] Synthesis of N-([1,1'-biphenyl]-4-ylmethyl)-1H-pyrazole-4-carboxamide (92) as described in Example 68

[0768]

[0769] According to the general method B: 1H-pyrazole-4-carboxylic acid (1 g, 8.9 mmol), 4-phenylbenzylamine (2.44 g, 13.35 mmol), T3P (7.05 g, 22.2 mmol) and DIPEA (4.6 g, 35.7 mmol), 92 (462 mg, 1.66 mmol, 19%) was obtained.

[0770] 1 H NMR (400MHz, DMSO-d6) δ8.66(t,J=6.0Hz,1H),8.09(s,2H),7.70–7.58(m,4H),7.50–7.30(m,5H,),4.47(d,J=6.0Hz,2H).

[0771] HRMS (ESI-TOF) is calculated as C 17 H 16 ON3[M+H] + :278.1287, actual value: 278.1290.

[0772] Example 69 – Synthesis of N-([1,1'-biphenyl]-4-ylmethyl)-1-(5-cyano-4-methoxypyridin-2-yl)-1H-pyrazole-4-carboxamide (93)

[0773]

[0774] Universal Method C: 6-Chloro-4-methoxynicotinonitrile (40 mg, 0.238 mmol), 92 (65 mg, 0.238 mmol), Pd t BuXPhos Pd G3 (19 mg, 0.023 mmol) and Cs2CO3 (232 mg, 0.714 mmol) yielded 93 (14 mg, 0.034 mmol, 12%).

[0775] 1 H NMR (400MHz, DMSO-d6) δ 9.03 (t, J = 6.0Hz, 1H), 8.82 (s, 1H), 8.30–8.29 (m, 1H), 7.70–7.59 (m, 5H), 7.50–7.31 (m, 6H), 4.50 (d, J = 6.0Hz, 2H), 4.12 (s, 3H). HRMS (ESI-TOF) calculated as C 24 H 20 O2N5[M+H] + :410.1622, actual value: 410.1611.

[0776] Example 70 – Synthesis of 6-(4-(([1,1'-biphenyl]-4-ylmethyl)carbamoyl)-1H-pyrazol-1-yl)-4-methoxynicotinic acid ethyl ester (94)

[0777]

[0778] According to the general method, C: ethyl 6-chloro-4-methoxynicotinic acid (39 mg, 0.18 mmol), 92 (50 mg, 0.18 mmol), Pd t BuXPhos Pd G3 (25 mg, 0.009 mmol) and dioxane (3 ml) yielded 94, a clear oily substance (23 mg, 0.05 mmol, 28%).

[0779] 1 H NMR(400MHz,THF-d8)δ9.08(d,J=1.0Hz,1H),8.68(s,1H),8.13(d,J=1.0Hz,1H),8.05(t,J=6.0Hz,1H),7.69 (s,1H),7.64–7.23(m,9H),4.59(d,J=6.0Hz,2H),4.30(q,J=7.0Hz,2H),4.03(s,3H),1.34(t,J=7.0Hz,3H).

[0780] HRMS (ESI-TOF) is calculated as C 26 H 25 O4N4[M+H] + :457.1870, actual value: 457.1868.

[0781] Example 71 – Synthesis of N-([1,1'-biphenyl]-4-ylmethyl)-1-(5-cyano-4-hydroxypyridin-2-yl)-1H-pyrazole-4-carboxamide (95)

[0782]

[0783] 95 (1.9 mg, 0.0048 mmol, 35%) was obtained by preparing a DMAc solution (1 ml) of 93 (6 mg, 0.014 mmol) and LiCl (8.5 mg, 0.14 mmol) according to the general method.

[0784] 1H NMR (400MHz, DMSO-d6) δ9.14 (s, 1H), 8.95 (t, J = 6.0Hz, 1H), 8.45 (s, 1H), 8.16 (s ,1H),7.70–7.59(m,4H),7.53–7.29(m,5H),7.16(s,1H),4.48(d,J=6.0Hz,2H).

[0785] HRMS (ESI-TOF) is calculated as C 23 H 18 O2N5[M+H] + :396.1466, actual value: 396.1456.

[0786] Example 72 – Synthesis of 6-(4-(([1,1'-biphenyl]-4-ylmethyl)carbamoyl)-1H-pyrazol-1-yl)-4-hydroxynicotinic acid (96)

[0787]

[0788] 96 was obtained by general method using a DMAc solution (3 ml) of D:94 (23 mg, 0.0504 mmol) and LiCl (30 mg, 0.504 mmol), as a yellow oily substance (9 mg, 0.0216 mmol, 43%).

[0789] 1 H NMR (600MHz, DMSO-d6) δ9.21(s,1H),8.96(t,J=6.0Hz,1H),8.71(s,1H),8.22(s,1H),7.64(m,5H),7.51–7.32(m,5H),4.49(d,J=6.0Hz,2H).

[0790] HRMS (ESI-TOF) is calculated as C 23 H 19 O4N4[M+H] + :415.1400, actual value: 415.1401.

[0791] Example 73 – Synthesis of N-([1,1'-biphenyl]-4-ylmethyl)-1-(5-cyanopyridin-2-yl)-1H-pyrazole-4-carboxamide (97)

[0792]

[0793] 97 (6 mg, 0.015 mmol, 16%) was obtained by general method using C: 6-chloro-nicotinonitrile (30 mg, 0.216 mmol), 92 (30 mg, 0.108 mmol), PdtBuXPhosG3 (17 mg, 0.0216 mmol), and Cs2CO3 (175 mg, 0.432 mmol).

[0794] 1 H NMR(400MHz,DMSO-d6)δ9.27(d,J=1.0Hz,1H),9.10–8.96(m,2H),8.51(dd,J=8.5,2.0Hz,1H),8.29(d, J=1.0Hz,1H),8.10(dd,J=8.5,1.0Hz,1H),7.67–7.59(m,4H),7.51–7.31(m,5H),4.50(d,J=6.0Hz,2H).

[0795] HRMS (ESI-TOF) is calculated as C 23 H 16 ON5[MH] - 378.1360, actual value: 378.1354.

[0796] Example 74 – Synthesis of N-([1,1'-biphenyl]-4-ylmethyl)-1-(4-methoxypyridin-2-yl)-1H-pyrazole-4-carboxamide (98)

[0797]

[0798] 98 (28 mg, 0.072 mmol, 63%) was obtained by general methods using C: 2-chloro-4-methoxypyridine (16 mg, 0.114 mmol), 92 (31 mg, 0.114 mmol), RockPhos Pd G3 (9.5 mg, 0.0114 mmol) and Cs2CO3 (92 mg, 0.285 mmol).

[0799] 1 H NMR (400MHz, DMSO-d6) δ9.18(d,J=1.0Hz,1H),8.95(t,J=6.0Hz,1H),8.32(d,J=6.0Hz,1H),8.19(d ,J=1.0Hz,1H),7.69–7.31(m,10H),7.00(dd,J=6.0,2.5Hz,1H),4.49(d,J=6.0Hz,2H),3.93(s,3H).

[0800] HRMS (ESI-TOF) is calculated as C23 H 21 O2N4[M+H] + 385.1690, actual value: 385.1659.

[0801] Example 75 – Synthesis of N-([1,1'-biphenyl]-4-ylmethyl)-1-(4-hydroxypyridin-2-yl)-1H-pyrazole-4-carboxamide (99)

[0802]

[0803] 99 (3 mg, 0.008 mmol, 45%) was obtained by following the general method: D: 92 (7 mg, 0.018 mmol) and LiCl (11 mg, 0.18 mmol) dissolved in DMAc (1 ml).

[0804] 1 H NMR(400MHz, DMSO-d6)δ9.14(d,J=1.0Hz,1H),8.89(t,J=6.0Hz,1H),8.73–8.66(m,1H),8.33–8.2 9(m,1H),7.91–7.89(m,1H),7.50–7.28(m,9H),6.77(dd,J=6.0,2.5Hz,1H),4.48(d,J=6.0Hz,2H).

[0805] HRMS (ESI-TOF) is calculated as C 22 H 19 O2N4[M+H] + :371.1506, Actual value: 371.1506.

[0806] Example 76 – Synthesis of 6-(4-(([1,1'-biphenyl]-4-ylmethyl)carbamoyl)-1H-pyrazol-1-yl)nicotinic acid ethyl ester (100)

[0807]

[0808] Following the general method, a dioxane solution (5 ml) of C: 6-chloronicotinic acid ethyl ester (42 mg, 0.23 mmol), 92 (30 mg, 0.108 mmol), PdtBuXPhos G3 (18 mg, 0.023 mmol), and Cs2CO3 (149 mg, 0.46 mmol) was prepared to yield 100 (10 mg, 0.023 mmol, 21%).

[0809] 1H NMR(400MHz,THF-d8)δ9.09(d,J=1.0Hz,1H),9.00(dd,J=2.0,1.0Hz,1H),8.70–8.60(m,1H),8.14–8.10(m,1H), 7.96(t,J=6.0Hz,1H),7.67–7.35(m,10H,),4.59(d,J=6.0Hz,2H),4.40(q,J=7.0Hz,2H),1.38(t,J=7.0Hz,3H).

[0810] HRMS (ESI-TOF) is calculated as C 25 H 23 O3N4[M+H] + :427.1766, Actual value: 427.1766.

[0811] Example 77 – Synthesis of 6-(4-(([1,1'-biphenyl]-4-ylmethyl)carbamoyl)-1H-pyrazol-1-yl)nicotinic acid (101)

[0812]

[0813] 100 (8 mg, 0.0187 mmol) was dissolved in a mixture of THF and water (3 mL (10:1)). Lithium hydroxide monohydrate (1.5 mg, 0.0374 mmol) was added to the reaction mixture, and the resulting mixture was stirred at room temperature for 16 hours. The reaction was confirmed to be complete by TLC, and HCl was added to the reaction mixture. aq (5 ml, 1 M). The mixture was extracted with EtOAc (3 × 10 ml), washed with brine, dried over anhydrous Na2SO4, and purified by rapid column chromatography (CH2Cl2, MeOH 0-5%, 1% formic acid) to give 101 (2 mg, 0.005 mmol, 27%).

[0814] 1 H NMR(400MHz, DMSO-d6)δ9.26(d,J=1.0Hz,1H),8.97(t,J=6.0Hz,1H),8.87(s,1H),8.35(d,J=2.0Hz,1H), 8.19(d,J=7.0Hz,1H),7.89(d,J=7.0Hz,1H),7.68–7.55(m,4H),7.52–7.29(m,5H),4.49(d,J=6.0Hz,2H).

[0815] HRMS (ESI-TOF) is calculated as C 23 H 17 O3N4[MH]- :397.1304, actual value: 397.1304.

[0816] Example 78 – Synthesis of 2-(4-(([1,1'-biphenyl]-4-ylmethyl)carbamoyl)-1H-pyrazol-1-yl)isonicotinic acid methyl ester (102)

[0817]

[0818] According to the general method, C: methyl 2-chloroisonicotinic acid (37 mg, 0.216 mmol), 92 (30 mg, 0.108 mmol), t A dioxane solution (3 ml) of BuXPhos Pd G3 (17 mg, 0.0216 mmol) and Cs2CO3 (175 mg, 0.54 mmol) yielded 102 (6 mg, 0.014 mmol, 13%).

[0819] 1 H NMR (400MHz, DMSO-d6) δ9.24(d,J=1.0Hz,1H),8.99(t,J=6.0Hz,1H),8.73(dd,J=5.0,1.0Hz,1H),8.34(dd,J=1.5,1.0Hz,1H) ,8.26(d,J=1.0Hz,1H),7.85(dd,J=5.0,1.5Hz,1H),7.66–7.61(m,4H),7.51–7.29(m,5H),4.50(d,J=6.0Hz,2H),3.95(s,3H).

[0820] HRMS (ESI-TOF) is calculated as C 24 H 21 O3N4[M+H] + :413.1608, actual value: 413.1608.

[0821] Example 79 – Synthesis of 2-(4-(([1,1'-biphenyl]-4-ylmethyl)carbamoyl)-1H-pyrazol-1-yl)isonicotinic acid (103)

[0822]

[0823] 102 (4 mg, 0.0097 mmol) was dissolved in a mixture of THF and water (3 mL (10:1)). Lithium hydroxide monohydrate (1 mg, 0.0194 mmol) was added to the reaction mixture, and the resulting mixture was stirred at room temperature for 16 hours. The reaction was confirmed to be complete by LCMS, and HCl was added to the reaction mixture. aq(5 ml, 1 M). The mixture was extracted with EtOAc (3 × 10 ml), washed with brine, dried over anhydrous Na2SO4, and purified by rapid column chromatography (CH2Cl2, MeOH 0-5%, 1% formic acid) at 20 column volumes to give 103 (3 mg, 0.0075 mmol, 78%).

[0824] 1 H NMR (400MHz, DMSO-d6) δ9.24(s,1H),8.98(t,J=6.0Hz,1H),8.68(d,J=5.0Hz,1H),8.33(t,J=1.5Hz,1H) ,8.24(s,1H),7.81(dd,J=5.0,1.5Hz,1H),7.69–7.61(m,4H),7.50–7.37(m,5H),4.50(d,J=6.0Hz,2H).

[0825] HRMS (ESI-TOF) is calculated as C 23 H 17 O3N4[MH] - :397.1304, actual value: 397.1304.

[0826] Synthesis of 1-(4-methoxypyridin-2-yl)-1H-pyrazole-4-carboxylic acid ethyl ester (104) from Example 80

[0827]

[0828] According to the general method, C: 4-methoxy-2-chloropyridine (500 mg, 3.49 mmol), 4-ethylpyrazole (725 mg, 5.2 mmol), RockPhos Pd G3 (244 mg, 0.349 mmol), Cs2CO3 (2.8 g, 8.7 mmol) and t BuOH (10 ml) yielded 104 (190 mg, 0.769 mmol, 22%).

[0829] 1 H NMR(400MHz,THF-d8)δ8.23(d,J=5.5Hz,1H),8.13(d,J=5.5Hz,1H),7.57(d,J=2.0Hz, 1H), 6.94 (d, J = 2.0Hz, 2H), 4.29 (q, J = 7.0Hz, 2H), 3.94 (s, 3H), 1.33 (t, J = 7.0Hz, 3H).

[0830] HRMS (ESI-TOF) is calculated as C 12 H 14O3N3[M+H] + :248.1029, actual value: 248.1033.

[0831] Reference Example 81 – Synthesis of 1-(4-methoxypyridin-2-yl)-1H-pyrazole-4-carboxylic acid (105)

[0832]

[0833] 104 (190 mg, 0.769 mmol) was dissolved in a mixture of THF and water (11 mL (10:1)). Lithium hydroxide monohydrate (78 mg, 2.19 mmol) was added to the reaction mixture, and the resulting mixture was stirred at room temperature for 16 hours. The reaction was confirmed to be complete by LCMS, and HCl (1 M, 5 mL) was added to the reaction mixture. The resulting mixture was extracted with EtOAc (3 × 10 mL), washed with brine, and dried over Na2SO4. The solvent was removed under vacuum to give 105 (73 mg, 0.33 mmol, 44%).

[0834] 1 H NMR (400MHz, DMSO-d6) δ 8.90 (s, 1H), 8.34 (d, J = 6.0 Hz, 1H), 8.15 (s, 1H), 7.45 (d, J = 2.5 Hz, 1H), 7.03 (dd, J = 6.0, 2.5 Hz, 1H), 3.93 (s, 3H).

[0835] HRMS (ESI-TOF) is calculated as C 10 H8O3N3[MH] - :218.0569, actual value: 218.0571.

[0836] Example 82 – Synthesis of N-([1,1'-biphenyl]-4-yl)-1-(4-methoxypyridin-2-yl)-1H-pyrazole-4-carboxamide (106)

[0837]

[0838] 106 (27 mg, 0.072 mmol, 54%) was obtained by general method B: 105 (30 mg, 0.136 mmol), 4-aminobiphenyl (35 mg, 0.204 mmol), T3P (108 mg, 0.34 mmol) and DIPEA (116 μl, 0.68 mmol).

[0839] 1H NMR(400MHz,THF-d8)δ9.29(s,1H),9.18(d,J=1.0Hz,1H),8.24(d,J=6.0Hz,1H),8.17(d ,J=1.0Hz,1H),7.91–7.84(m,2H),7.68–7.21(m,8H),6.87(d,J=6.0Hz,1H),3.95(s,3H).

[0840] HRMS (ESI-TOF) is calculated as C 22 H 19 O2N4[M+H] + 371.1502, actual value: 371.1396.

[0841] Example 83 – Synthesis of N-([1,1'-biphenyl]-3-ylmethyl)-1-(4-methoxypyridin-2-yl)-1H-pyrazole-4-carboxamide (107)

[0842]

[0843] 107 (32 mg, 0.083 mmol, 61%) was obtained by general method B: 105 (30 mg, 0.136 mmol), 3-aminomethylbiphenyl (36 mg, 0.204 mmol), T3P (108 mg, 0.34 mmol) and DIPEA (116 μl, 0.68 mmol).

[0844] 1 ¹H NMR (400MHz, chloroform-d) δ 8.97 (d, J = 1.0 Hz, 1H), 8.15 (d, J = 6.0 Hz, 1H), 8.07 (d, J = 1.0 Hz, 1H), 7.58–7.28 (m, 10H), 6.73 (d, J = 6.0 Hz, 1H), 6.59 (t, J = 6.0 Hz, 1H), 4.64 (d, J = 6.0 Hz, 2H), 3.90 (s, 3H).

[0845] HRMS (ESI-TOF) is calculated as C 23 H 21 O2N4[M+H] + 385.1659, actual value: 385.1656.

[0846] Example 84 – Synthesis of N-([1,1'-biphenyl]-4-yl)-1-(4-hydroxypyridin-2-yl)-1H-pyrazole-4-carboxamide (108)

[0847]

[0848] Following the general method, a DMAc solution (2 ml) of D:106 (27 mg, 0.073 mmol) and LiCl (29 mg, 0.73 mmol) was reacted for 8 hours to obtain 108 (15 mg, 0.0421 mmol, 57%).

[0849] 1 H NMR(400MHz,DMSO-d6)δ10.18(s,1H),9.39(s,1H),8.32–8.18(m,2H),7.91–7 .82(m,2H),7.70–7.63(m,4H),7.50–7.31(m,4H),6.82(dd,J=5.5,2.5Hz,1H).

[0850] HRMS (ESI-TOF) is calculated as C 21 H 17 O2N4[M+H] + 357.1346, actual value: 357.1348.

[0851] Example 85: Synthesis of N-([1,1'-biphenyl]-3-ylmethyl)-1-(4-hydroxypyridin-2-yl)-1H-pyrazole-4-carboxamide (109)

[0852]

[0853] 109 (11 mg, 0.029 mmol, 36%) was obtained by general method using a DMAc solution (2 ml) of D: 107 (32 mg, 0.083 mmol) and LiCl (35 mg, 0.83 mmol).

[0854] 1 H NMR (400MHz, DMSO-d6) δ11.23(s,1H),9.14(d,J=1.0Hz,1H),8.93(t,J=6.0Hz,1H),8.19(d,J=5.5Hz ,1H),8.15(d,J=1.0Hz,1H),7.70–7.27(m,10H),6.78(dd,J=5.5,2.5Hz,1H),4.52(d,J=6.0Hz,2H).

[0855] HRMS (ESI-TOF) is calculated as C 22 H 19 O2N4[M+H] + :371.1502, actual value: 371.1497.

[0856] Reference Example 86 – Synthesis of 1-(5-cyano-4-methoxypyridin-2-yl)-1H-pyrazole-4-carboxylic acid ethyl ester (110)

[0857]

[0858] According to the general method, C: 6-chloro-4-methoxynicotinonitrile (400 mg, 2.38 mmol), pyrazole-4-carboxylic acid ethyl ester (500 mg, 3.57 mmol), t A dioxane solution (10 ml) of BuxPhos Pd G3 (188 mg, 0.238 mmol) and Cs2CO3 (2.3 g, 7.14 mmol) yielded 110 (110 mg, 0.40 mmol, 17%).

[0859] 1 H NMR (400MHz, DMSO-d6) δ9.02 (s, 1H), 8.83 (s, 1H), 8.32 (s, 1H), 7.69 (s, 1H), 4.28 (q, J = 7.0Hz, 2H), 4.13 (s, 3H), 1.30 (t, J = 7.0Hz, 3H).

[0860] HRMS (ESI-TOF) is calculated as C 13 H 13 O3N4[M+H] + :273.0978, Actual value: 273.0978.

[0861] Reference Example 87 – Synthesis of 1-(5-cyano-4-methoxypyridin-2-yl)-1H-pyrazole-4-carboxylic acid (111)

[0862]

[0863] 110 (95 mg, 0.399 mmol) was dissolved in a mixture of THF and water (11 mL (10:1)). Lithium hydroxide monohydrate (14 mg, 0.399 mmol) was added to the reaction mixture, and the resulting mixture was stirred at room temperature for 16 hours. The reaction was confirmed to be complete by TLC, and HCl (10 mL, 1 M) was added to the reaction mixture. The resulting mixture was extracted with EtOAc (3 × 10 mL), washed with brine, dried over anhydrous Na₂SO₄, and concentrated under vacuum to give 111 (90 mg, 0.368 mmol, 93%).

[0864] 1H NMR (400MHz, DMSO-d6) δ8.89 (d, J = 1.0 Hz, 1H), 8.76 (s, 1H), 8.20 (d, J = 1.0 Hz, 1H), 7.63 (s, 1H), 4.08 (s, 3H).

[0865] HRMS (ESI-TOF) is calculated as C 11 H7O3N4[MH] - :243.0523, actual value: 243.0520.

[0866] Example 88 – Synthesis of 1-(5-cyano-4-methoxypyridin-2-yl)-N-((6-phenylpyridin-3-yl)methyl)-1H-pyrazole-4-carboxamide (112)

[0867]

[0868] 112 (28 mg, 0.068 mmol, 35%) was obtained according to general method B: 111 (40 mg, 0.164 mmol), 52 (56 mg, 0.306 mmol), T3P (130 mg, 0.409 mmol) and DIPEA (116 μl, 0.68 mmol).

[0869] 1 H NMR(400MHz,THF-d8)δ9.06(d,J=1.0Hz,1H),8.57(s,1H),8.34(dd,J=8.5,1.5Hz,1H),8.18(t,J=6.0Hz,1H), 8.12–8.04(m,2H),7.81(d,J=1.5Hz,2H),7.73(s,1H),7.44–7.31(m,4H),4.59(d,J=6.0Hz,2H),4.12(s,3H).

[0870] HRMS (ESI-TOF) is calculated as C 23 H 19 O2N6[M+H] + :411.1564, actual value: 411.1555.

[0871] Example 89 – Synthesis of 1-(5-cyano-4-methoxypyridin-2-yl)-N-(4-(trifluoromethyl)benzyl)-1H-pyrazole-4-carboxamide 113

[0872]

[0873] 113 (38 mg, 0.0947 mmol, 47%) was obtained by general method B: 111 (40 mg, 0.164 mmol), 4-trifluoromethylbenzylamine (54 mg, 0.306 mmol), T3P (130 mg, 0.409 mmol) and DIPEA (116 μl, 0.68 mmol).

[0874] 1 H NMR(400MHz,THF-d8)δ8.82(s,1H),8.41(dd,J=4.5,1.5Hz,1H),8.11(dd,J=8.5,1.5Hz,1H) ,7.50(s,1H),7.39–7.29(m,4H),7.23(t,J=6.0Hz,1H),4.39(d,J=6.0Hz,2H),3.89(s,3H).

[0875] HRMS (ESI-TOF) is calculated as C 19 H 13 O2N5F3[MH] - :400.1026, actual value: 400.1022.

[0876] Example 90 – Synthesis of 1-(5-cyano-4-hydroxypyridin-2-yl)-N-((6-phenylpyridin-3-yl)methyl)-1H-pyrazole-4-carboxamide (114)

[0877]

[0878] 114 (8 mg, 0.020 mmol, 42%) was obtained by general method using a DMAc solution (2 ml) of D:112 (20 mg, 0.0487 mmol) and LiCl (20.5 mg, 0.487 mmol).

[0879] 1 H NMR(400MHz, DMSO-d6)δ9.11(d,J=1.0Hz,1H),9.10–9.04(m,1H),8.64–8.61(m,1H),8.37(s,1H),8.16(s,1H),8.09–8.0 3(m,2H),7.93(dd,J=8.0,1.0Hz,1H),7.81(dd,J=8.0,2.5Hz,1H),7.53–7.38(m,3H),7.15(s,1H),4.49(d,J=6.0Hz,2H).

[0880] HRMS (ESI-TOF) is calculated as C 22 H 17 O2N6[M+H]+ :397.1407, Actual value: 397.1407.

[0881] Example 91 – Synthesis of 1-(5-cyano-4-hydroxypyridin-2-yl)-N-(4-(trifluoromethyl)benzyl)-1H-pyrazole-4-carboxamide 115

[0882]

[0883] 115 (5 mg, 0.0129 mmol, 21%) was obtained by general method D: 113 (25 mg, 0.062 mmol) and DMAc solution (2 ml) of lithium chloride (26 mg, 0.62 mmol).

[0884] 1 H NMR(400MHz,DMSO-d6)δ9.06(s,1H),8.96(t,J=6.0Hz,1H),8.13(s,1H),8.08(s,1H ), 7.70 (d, J = 8.0Hz, 2H), 7.53 (d, J = 8.0Hz, 2H), 6.78 (s, 1H), 4.51 (d, J = 6.0Hz, 2H).

[0885] HRMS (ESI-TOF) is calculated as C 18 H 11 O2N5F3[MH] - :386.0864, ​​Actual yield: 386.0867. Reference Example 92– Synthesis of 6-hydroxypyrazolo[1,5-a]pyrido[3,2-e]pyrimidine-7-carboxylic acid ethyl ester (116)

[0886]

[0887] Pyrazolo[1,5-a]pyrimidin-7-amine (250 mg, 1.86 mmol) and diethyl ethoxymethylene malonate (402 mg, 1.86 mmol) were added to a microwave-safe vial and dissolved in anhydrous EtOH (5 ml), followed by the addition of NaOEt (189 mg, 2.79 mmol). The reaction mixture was heated at 100 °C for 2 hours under microwave irradiation, then cooled to room temperature. The reaction mixture was then filtered, dried, and diphenyl ether (2 ml) was added. The resulting mixture was then heated to 240 °C and held for 30 minutes, then cooled to room temperature. The reaction mixture was then filtered, washed with Et2O (4 × 50 ml), and dried to give 116 (210 mg, 0.81 mmol, 43%).

[0888] 1H NMR (400MHz, DMSO-d6) δ 8.97 (s, 1H), 8.62 (s, 1H), 8.15 (d, J = 2.0Hz, 1H), 6.68 (d, J = 2.0Hz, 1H), 4.20 (q, J = 7.0Hz, 2H), 1.28 (t, J = 7.0Hz, 3H).

[0889] HRMS (ESI-TOF) is calculated as C 12 H 11 O3N4[M+H] + :259.0825, actual value: 259.0826.

[0890] Example 93 – Synthesis of N-([1,1'-biphenyl]-4-ylmethyl)-6-hydroxypyrazolo[1,5-a]pyrido[3,2-e]pyrimidine-7-carboxamide (117)

[0891]

[0892] General Method A: 116 (100 mg, 0.38 mmol), 4-phenylbenzylamine (70 mg, 0.38 mmol), DABCO-(AlMe3)2 (100 mg, 0.38 mmol) yielded 117 (4 mg, 0.01 mmol, 3%).

[0893] Solvent system used for purification: 0%-5% MeOH in CH3Cl solution.

[0894] 1 H NMR (400MHz, DMSO-d6) δ9.04 (s, 1H), 8.72 (s, 1H), 8.26 (d, J = 2.0Hz, 1H), 7.74 –7.59(m,4H),7.54–7.25(m,5H),6.78(d,J=2.0Hz,1H),4.60(d,J=6.0Hz,2H).

[0895] HRMS (ESI-TOF) is calculated as C 23 H 16 O2N5[MH] - 394.1309, actual value: 394.1303.

[0896] Reference Example 94 – Synthesis of methyl 3-([1,1'-biphenyl]-4-ylamino)-3-oxopropionate (118)

[0897]

[0898] 118 (230 mg, 0.855 mmol, 24%) was obtained by general method B: methyl malonate (585 mg, 4.97 mmol), 4-aminobiphenyl (600 mg, 3.55 mmol), T3P (3.16 g, 9.94 mmol) and DIPEA (1.8 g, 14.2 mmol).

[0899] 1 ¹H NMR (400MHz, chloroform-d) δ 9.27 (s, 1H), 7.68–7.30 (m, 9H), 3.82 (s, 3H), 3.52 (s, 2H).

[0900] HRMS (ESI-TOF) is calculated as C 16 H 16 O3N[M+H] + :270.1124, actual value: 270.1122.

[0901] Example 95: Synthesis of N-([1,1'-biphenyl]-4-yl)-6-hydroxypyrazolo[1,5-a]pyrido[3,2-e]pyrimidine-7-carboxamide (119)

[0902]

[0903] 118 (100 mg, 0.37 mmol) was added to a microwave-safe vial, washed with N2 and the N2 was removed under vacuum (3 times), followed by the addition of anhydrous ethyl orthoformate (3 ml). The reaction mixture was heated at 120 °C for 3 hours. Excess ethyl orthoformate was removed under vacuum, and the resulting mixture was dissolved in diphenyl ether (5 ml). Pyrazolo[1,5-a]pyrimidine-7-amine (75 mg, 0.55 mmol) was added, and the resulting mixture was heated to 240 °C and held for 30 minutes. After cooling to room temperature, it was directly purified by 20 column volumes of rapid column chromatography (CH2Cl2, MeOH 0-10%, 1% HCOOH) to give 119 (5.5 mg, 0.015 mmol, 4%).

[0904] 1 H NMR (400MHz, DMSO-d6) δ13.31(s,1H),9.11(s,1H),8.95(s,1H),8.11(d,J=2.0Hz,1H),7.85–7.27(m,9H),6.68(d,J=2.0Hz,1H).

[0905] HRMS (ESI-TOF) is calculated as C 22 H 14 O2N5[MH] -:380.1153, actual value: 380.1150.

[0906] Reference Example 96 – Synthesis of methyl 3-(([1,1'-biphenyl]-4-ylmethyl)amino)-3-oxopropionate (120)

[0907]

[0908] 120 (1.5 g, 5.3 mmol, 60.0%) was obtained by general method B: methyl malonate (1.00 g, 8.9 mmol), 4-aminomethylbiphenyl (1.83 g, 10.0 mmol), T3P (6.7 g, 21.1 mmol) and DIPEA (2.7 g, 21.1 mmol).

[0909] 1 H NMR (400MHz, THF-d8) δ7.71 (s, 1H), 7.41–7.03 (m, 9H), 4.23 (d, J = 6.0Hz, 2H), 3.46 (s, 3H), 3.10 (s, 2H).

[0910] HRMS (ESI-TOF) is calculated as C 17 H 18 O3N[M+H] + :284.1281, actual value: 284.1281.

[0911] Reference Example 97 – Synthesis of methyl 3-oxo-3-((4-(trifluoromethyl)benzyl)amino)propionate (121)

[0912]

[0913] 121 (736 mg, 2.67 mmol, 30%) was obtained by general method B: methyl malonate (1.00 g, 8.9 mmol), 4-trifluorobenzylamine (1.75 g, 10.0 mmol), T3P (6.7 g, 21.1 mmol) and DIPEA (2.7 g, 21.1 mmol).

[0914] 1 ¹H NMR (400MHz, chloroform-d) δ 7.86 (s, 1H), 7.52 (d, J = 8.0Hz, 2H), 7.35 (d, J = 8.0Hz, 2H), 4.46 (d, J = 6.0Hz, 2H), 3.68 (s, 3H), 3.33 (s, 2H).

[0915] HRMS (ESI-TOF) is calculated as C 12 H 13O3NF3[M+H] + :276.0842, actual value: 276.0843.

[0916] Example 98 – Synthesis of N-([1,1'-biphenyl]-4-ylmethyl)-6-hydroxy-2,5-dimethylpyrazolo[1,5-a]pyridolo[3,2-e]pyrimidine-7-carboxamide (122)

[0917]

[0918] 120 (200 mg, 0.70 mmol) was added to a microwave-safe vial, washed with N2 and the N2 was removed under vacuum (3 times), then anhydrous ethyl orthoformate (5 ml) was added. The reaction mixture was heated at 120 °C for 3 hours under microwave irradiation. Excess ethyl orthoformate was removed under vacuum, and the resulting mixture was dissolved in diphenyl ether (5 ml). 2,5-Dimethylpyrazolo[1,5-a]pyrimidine-7-amine (114 mg, 0.70 mmol) was added, and the resulting mixture was heated to 240 °C and held for 30 minutes. After cooling to room temperature, it was directly purified by 20 column volumes of rapid column chromatography (CH2Cl2:MeOH 0-10%, 1% formic acid) to give 122 (10 mg, 0.024 mmol, 3%).

[0919] 1 H NMR (400MHz, DMSO-d6) δ10.54(s,1H),8.59(s,1H),7.69–7.30(m,9H),6.44(s,1H),4.59(d,J=6.0Hz,2H),2.88(s,3H),2.45(s,3H).

[0920] HRMS (ESI-TOF) is calculated as C 25 H 22 O2N5[M+H] + :424.1768, actual value: 424.1768.

[0921] Example 99 – Synthesis of 6-hydroxy-2,5-dimethyl-N-(4-(trifluoromethyl)benzyl)pyrazolo[1,5-a]pyrido[3,2-e]pyrimidine-7-carboxamide (123)

[0922]

[0923] 121 (200 mg, 0.70 mmol) was added to a microwave-safe vial, washed with N2 and the N2 was removed under vacuum three times, then anhydrous ethyl orthoformate (5 ml) was added. The reaction mixture was heated at 120 °C for 3 hours under microwave irradiation. Excess ethyl orthoformate was removed under vacuum, and the resulting mixture was dissolved in diphenyl ether (5 ml). 2,5-Dimethylpyrazolo[1,5-α]pyrimidine-7-amine (114 mg, 0.70 mmol) was added, and the resulting mixture was heated to 240 °C and held for 30 minutes. After cooling to room temperature, it was directly purified by 20 column volumes of rapid column chromatography (CH2Cl2:MeOH 0-10%, 1% formic acid) to give 123 (8 mg, 0.020 mmol, 3%).

[0924] 1 H NMR (400MHz, DMSO-d6) δ 8.52 (s, 1H), 7.71 (d, J = 8.1Hz, 2H), 7.55 (d, J = 8.0Hz, 2H), 6.50 (s, 1H), 4.64 (d, J = 6.0Hz, 2H), 2.89 (s, 3H), 2.47 (s, 3H).

[0925] HRMS (ESI-TOF) is calculated as C 20 H 15 O2N5F3[MH] - :414.1183, actual value: 414.1182.

[0926] Example 100: Synthesis of N-([1,1'-biphenyl]-4-ylmethyl)-6-(4-cyano-1H-pyrazol-1-yl)-5-methoxynicotinamide (124)

[0927]

[0928] The title compound (56 mg, 0.137 mmol, 33%) was obtained by conventional method using C:72 (147 mg, 0.42 mmol), PdtBuXPhos G3 (33 mg, 0.04 mmol), cesium carbonate (273 mg, 0.84 mmol) and 1H-pyrazole-4-onitrile (70 mg, 0.5 mmol).

[0929] 1H NMR (400MHz, DMSO) δ9.41(t,J=6.0Hz,1H),9.11(d,J=1.0Hz,1H),8.65(d,J=2.0Hz,1H),8.36(d,J=1.0Hz,1H),8. 18(d,J=2.0Hz,1H),7.67–7.62(m,4H),7.49–7.42(m,4H),7.40–7.32(m,1H),4.59(d,J=6.0Hz,2H),3.97(s,3H).

[0930] Example 101: Synthesis of N-([1,1'-biphenyl]-4-ylmethyl)-6-(4-cyano-1H-pyrazol-1-yl)-5-hydroxynicotinamide (125)

[0931]

[0932] Following general method D: 125 (56 mg, 0.14 mmol) and lithium chloride (58 mg, 1.37 mmol) were mixed, and then N,N-dimethylacetamide (3 ml) was added. The resulting mixture was microwaved at 150 °C for 8 hours. The crude substance was then purified by rapid reversed-phase chromatography (H₂O + 0.1% formic acid (0-100% ACN, 0.1% formic acid)) to give the title compound (3 mg, 0.007 mmol, 5%).

[0933] 1 H NMR (400MHz, DMSO) δ9.35(t,J=5.9Hz,1H),9.24(s,1H),8.53(d,J=1.9Hz,1H),8.42(s,1H),7.95(d,J =1.9Hz,1H),7.67–7.62(m,4H),7.45(qd,J=7.1,1.9Hz,4H),7.38–7.32(m,1H),4.55(d,J=6.0Hz,2H).

[0934] Reference Example 102 – Synthesis of 6-chloro-5-methoxy-N-((4-(trifluoromethyl)cyclohexyl)methyl)nicotinamide (126)

[0935]

[0936] The title compound (1625 mg, 4.64 mol, 86%) was obtained by general method B: 6-chloro-5-methoxynicotinic acid (1000 mg, 5.35 mmol), HATU (4064 mg, 10.70 mmol), C-4 trifluorocyclohexylamine (1209 μL, 8.02 mmol) and DIPEA (2759 μL, 16.04 mmol).

[0937] LRMS m / z is calculated as C 15 H 19 ClF3N2O2[M+H] + :351.1, Actual value: 351.1

[0938] Example 103 – Synthesis of 1-(3-methoxy-5-(((4-(trifluoromethyl)cyclohexyl)methyl)carbamoyl)pyridin-2-yl)-1H-pyrazole-4-carboxylic acid ethyl ester (127)

[0939]

[0940] The target compound (92 mg, 0.20 mmol, 23%) was obtained by using the general method with C:127 (300 mg, 0.86 mmol), PdtBuXPhos G3 (68 mg, 0.09 mmol), 1H-pyrazole-4-carboxylic acid ethyl ester (180 mg, 1.29 mmol) and Cs2CO3 (835 mg, 2.57 mmol).

[0941] 1 H NMR (400MHz, DMSO) δ8.80(t,J=6.0Hz,1H),8.74(d,J=0.7Hz,1H),8.57(d,J=1.8Hz,1H),8.14(d,J=0.6Hz,1H),8.07(d,J=1.8Hz,1H),4.27(q,J=7 .0Hz,2H),3.96(s,3H),3.19(t,J=6.0Hz,2H),2.35–2.16(m,1H),1.94–1 .82(m,4H),1.63–1.50(m,1H),1.30(t,J=7.0Hz,3H),1.27–0.98(m,4H).

[0942] LRMS m / z is calculated as C 21 H 26 F3N4O4[M+H] + :455.19, Actual Winnings: 455.30

[0943] Example 104 – Synthesis of 1-(3-hydroxy-5-(((4-(trifluoromethyl)cyclohexyl)methyl)carbamoyl)pyridin-2-yl)-1H-pyrazole-4-carboxylic acid ethyl ester (128)

[0944]

[0945] The title compound (20 mg, 0.04 mmol, 20%) was obtained by conventional method using D:128 (92 mg, 0.20 mmol), LiCl (85 mg, 2.03 mmol), and DMSO (2 ml).

[0946] 1 H NMR (400MHz, DMSO) δ8.95(s,1H),8.69(t,J=6.0Hz,1H),8.42(d,J=2.0Hz,1H),8.24(s,1H),7.87(d,J=2.0Hz,1H),4.28(q,J=7.0Hz,2H),3.15(t, J=6.3Hz,2H),1.92–1.79(m,5H),1.55(dp,J=18.8,7.4,5.7Hz,1H),1.29 (t,J=7.0Hz,3H),1.25–1.16(m,2H),1.02(qd,J=13.8,13.1,3.9Hz,2H).

[0947] Example 105 – Synthesis of 1-(3-hydroxy-5-(((4-(trifluoromethyl)cyclohexyl)methyl)carbamoyl)pyridin-2-yl)-1H-pyrazole-4-carboxylic acid (129)

[0948]

[0949] 129 (20 mg, 0.04 mmol) was dissolved in THF (10 mL), and then LiOH aqueous solution (500 μL, 2 M) was added. The resulting mixture was stirred at room temperature for 16 hours. The reaction mixture was neutralized with formic acid, and then diatomaceous earth was added. The solvent was then removed under vacuum, and the crude mixture was purified by rapid reversed-phase column chromatography (0%-100% ACN (0.1% formic acid) in H2O solution (0.1% formic acid)) to give the title compound (6 mg, 0.014 mmol, 36%).

[0950] 1 H NMR (400MHz, DMSO) δ8.82(s,1H),8.65(t,J=6.0Hz,1H),8.40(d,J=2.0Hz,1H),8.09(s,1H),7.88(d,J=2.0Hz,1H),3.14( t,J=6.0Hz,2H),2.28–2.15(m,1H),1.92–1.80(m,5H),1.21(qt,J=13.9,6.7Hz,3H),1.02(qd,J=13.7,12.9,4.0Hz,2H).

[0951] LRMS m / z is calculated as C 18 H18 F3N4O4[MH] - :411.13, Actual value: 411.0

[0952] Reference Example 106 – Synthesis of 6-chloro-4-methoxynicotinic acid ethyl ester (130)

[0953]

[0954] 6-Chloro-4-methoxynicotinic acid (5 g, 0.03 mol), EDC.HCl (6.22 g, 0.04 mol), and DMAP (0.05 g, 1.2 mmol) were dissolved in DMF (50 mL). DIPEA (4.60 mL, 0.03 mmol) and EtOH (5 mL) were then added to the reaction mixture. The resulting mixture was stirred at room temperature for 16 hours. Ethyl acetate (50 mL), 1 M HCl (50 mL), and H₂O (100 mL) were added to the reaction mixture. The organic and aqueous layers were separated. This process was repeated at least twice. The organic layers were combined, washed with brine, and dried over anhydrous Na₂SO₄. The crude compound was then purified by rapid column chromatography (15 column volumes, cyclohexane 100%–50%, EtOAc 0%–50%) to give the desired compound (2.02 g, 9.39 mmol, 31%).

[0955] LRMS m / z calculation is C9H 11 ClNO3[M+H] + :216.0, actual value: 216.2.

[0956] Reference Example 107 – Synthesis of 4-methoxy-6-(1H-pyrazol-1-yl)nicotinic acid ethyl ester (131)

[0957]

[0958] The title compound (1.9 g, 7.65 mmol, 87%) was obtained by means of C:130 (1.90 g, 0.088 mol), PdtBuXPhos G3 (0.35 g, 0.00044 mol), cesium carbonate (4.30 g, 0.0132 mol), pyrazole (0.90 g, 0.0132 mol), and anhydrous 1,4-dioxane (50 ml) according to the general method.

[0959] LRMS m / z is calculated as C 12 H 14 N3O3[M+H] + 248.10, Actual amount received: 248.10

[0960] Synthesis of 108–4-methoxy-6-(1H-pyrazol-1-yl)nicotinic acid (132)

[0961]

[0962] 131 (1.9 g, 7.65 mmol) was dissolved in a mixture of THF (65 ml), MeOH (35 ml), and H₂O (10 ml), and then lithium hydroxide monohydrate (3.7 g, 90.5 mmol) was added. The resulting mixture was stirred at room temperature for 16 hours. The reaction was confirmed to be complete by TLC. The reaction mixture was acidified, cooled to 5 °C, and the precipitate was collected by filtration. The precipitate was washed with diethyl ether to give the title compound (1.4 g, 6.39 mmol, 84%).

[0963] LRMS m / z is calculated as C 10 H 10 N3O4[M+H] + :220.07, actual value: 220.2.

[0964] Example 109: Synthesis of N-(but-2-yn-1-yl)-4-methoxy-6-(1H-pyrazol-1-yl)nicotinamide (133)

[0965]

[0966] The title compound (148 mg, 0.54 mmol, 63%) was obtained by general method B: 132 (187 mg, 0.85 mmol), HATU (646 mg, 1.7 mmol), but-2-yn-1-amine hydrogen chloride (135 mg, 1.27 mmol) and DIPEA (438 μL, 2.55 mmol).

[0967] LRMS m / z is calculated as C 14 H 15 N4O2[M+H] + 271.1, Actual value: 271.2

[0968] Example 110 Synthesis of N-(but-2-yn-1-yl)-4-hydroxy-6-(1H-pyrazol-1-yl)nicotinamide (134)

[0969]

[0970] The title compound (44 mg, 0.171 mmol, 33%) was obtained by conventional method using D:133 (143 mg, 0.53 mmol) and lithium chloride (111 mg, 2.64 mmol).

[0971] 1 H NMR (400MHz, DMSO) δ11.38(t,J=5.4Hz,1H),8.48–8.39(m,2H),7.73–7.62(m,1H),6.73(d ,J=3.1Hz,1H),6.42(q,J=2.1Hz,1H),4.01(dq,J=5.2,2.5Hz,2H),1.78(t,J=2.7Hz,3H).

[0972] LRMS m / z is calculated as C 13 H 13 N4O2[M+H] + 257.10, Actual Winnings: 257.30

[0973] Example 111 Synthesis of 135-N-((6-chloropyridin-3-yl)methyl)-4-methoxy-6-(1H-pyrazol-1-yl)nicotinamide (135)

[0974]

[0975] The title compound (1.315 g, 3.83 mmol, 93%) was obtained by general method B: 132 (900 mg, 4.11 mmol), HATU (3123 mg, 8.22 mmol), (6-chloropyridin-3-yl)methylamine (700 mg, 4.93 mmol) and DIPEA (2.1 ml, 12.33 mmol).

[0976] LRMS m / z is calculated as C 16 H 15 ClN5O2[M+H] + 344.1, Actual value: 344.2

[0977] Example 112: Synthesis of ethyl 4-(5-((4-methoxy-6-(1H-pyrazol-1-yl)nicotinamide)methyl)pyridin-2-yl)benzoate (136)

[0978]

[0979] The title compound (50 mg, 0.109 mmol, 54%) was obtained by conventional method C:135 (44 mg, 0.128 mmol), (4-ethoxycarbonyl)phenyl)boronic acid (42 mg, 0.22 mmol), palladium aminophosphine (5.16 mg, 0.01 mmol) and cesium carbonate (142.13 mg, 0.44 mmol).

[0980] LRMS m / z is calculated as C 25 H 24N5O4[M+H] + :458.18, actual result: 458.3, [M+2H] / 2 observed 229.7.

[0981] Example 1. Synthesis of 13–4-methoxy-6-(1H-pyrazol-1-yl)-N-((6-(pyrimidin-5-yl)pyridin-3-yl)methyl)-nicotinamide (137)

[0982]

[0983] The title compound (9 mg, 0.012 mmol, 15%) was obtained by conventional method C: 135 (27 mg, 0.078 mmol), pyrimidine-5-ylboronic acid (27 mg, 0.22 mmol), palladium aminophosphine (5.16 mg, 0.01 mmol) and cesium carbonate (142.13 mg, 0.44 mmol).

[0984] LRMS m / z is calculated as C 20 H 18 N7O2[M+H] + :388.15, Actual value: 388.00.[M+2H] / 2 Observed value: 194.8

[0985] Example 114– Synthesis of 4-methoxy-N-((6-(4-(methylcarbamoyl)phenyl)pyridin-3-yl)methyl)-6-(1H-pyrazol-1-yl)nicotinamide (138)

[0986]

[0987] The title compound (39 mg, 0.088 mmol, 59%) was obtained by conventional method C: 135 (50 mg, 0.15 mmol), (4-(methylcarbamoyl)phenyl)boronic acid (39 mg, 0.22 mmol), palladium aminophosphine (5.16 mg, 0.01 mmol) and cesium carbonate (142.13 mg, 0.44 mmol).

[0988] LRMS m / z is calculated as C 24 H 23 N6O3[M+H] + :443.18, actual result: 443.1, [M+2H] / 2 observed 222.20.

[0989] Example 115 Synthesis of N-((6-(4-carbamoylphenyl)pyridin-3-yl)methyl)-4-methoxy-6-(1H-pyrazol-1-yl)nicotinamide (139)

[0990]

[0991] The title compound (22 mg, 0.051 mmol, 34%) was obtained by conventional method C: 135 (50 mg, 0.15 mmol), (4-(carbamoyl)phenyl)boronic acid (36 mg, 0.22 mmol), palladium aminophosphine (5.16 mg, 0.01 mmol) and cesium carbonate (142.13 mg, 0.44 mmol).

[0992] LRMS m / z is calculated as C 23 H 21 N6O3[M+H] + :429.17, Actual amount received: 429.2

[0993] Example 1. Synthesis of 116–4-methoxy-N-((6-(4-morpholinophenyl)pyridin-3-yl)methyl)-6-(1H-pyrazol-1-yl)nicotinamide (140)

[0994]

[0995] The title compound (50 mg, 0.106 mmol, 71%) was obtained by conventional method C:135 (50 mg, 0.15 mmol), (4-morpholinophenyl)boronic acid (59 mg, 0.22 mmol), palladium aminophosphine (5.16 mg, 0.01 mmol) and cesium carbonate (142.13 mg, 0.44 mmol).

[0996] LRMS m / z is calculated as C 26 H 27 N6O3[M+H] + :471.21, Actual result: 471.20, [M+2H] / 2 observed 236.20.

[0997] Example 117 – Synthesis of N-((6-(benzo[d]thiazolyl)pyridin-3-yl)methyl)-4-methoxy-6-(1H-pyrazol-1-yl)nicotinamide (141)

[0998]

[0999] The title compound (22 mg, 0.0497 mmol, 33%) was obtained by conventional method C: 135 (50 mg, 0.15 mmol), (5-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)benzo[d]thiazole (57.07 mg, 0.22 mmol), palladium aminophosphine (5.16 mg, 0.01 mmol) and cesium carbonate (142.13 mg, 0.44 mmol).

[1000] LRMS m / z is calculated as C 23 H 19 N6O2S[M+H] + :443.13, measured [M+2H] 2+ / 2:222.0

[1001] Example 118: Synthesis of ethyl 4-(5-((4-hydroxy-6-(1H-pyrazol-1-yl)nicotinamide)methyl)pyridin-2-yl)benzoate (142)

[1002]

[1003] The title compound (10 mg, 0.022 mmol, 22%) was obtained by conventional method D:140 (50 mg, 0.11 mmol) and lithium chloride (46 mg, 1.09 mmol), and purified by reversed-phase rapid chromatography (0-100% CAN (0.1% formic acid) in H2O solution (0.1% formic acid)) to give the title compound (10 mg, 0.0225 mmol, 22%).

[1004] LRMS m / z is calculated as C 24 H 22 N5O4[M+H] + :444.10, Actual value: 444.2, [M+2H] 2+ / 2 observed 222.7.

[1005] Example 119 – Synthesis of 4-(5-((4-hydroxy-6-(1H-pyrazol-1-yl)nicotinamide)methyl)pyridin-2-yl)benzoic acid (143)

[1006]

[1007] 143 (6 mg, 0.0135 mmol) was dissolved in a mixture of MeOH (5 mL) and H₂O (1 mL), and then lithium hydroxide (2.84 mg, 0.07 mmol) was added. The resulting mixture was stirred at room temperature for 16 hours. 1 M HCl (1 mL) was added to the reaction mixture, the resulting precipitate was filtered, washed with Et₂O, and dried to give the title compound (2 mg, 0.0048 mmol, 35%).

[1008] 1H NMR (400MHz, DMSO) δ11.88(t,J=5.8Hz,1H),8.65(d,J=2.3Hz,1H),8.48(s,1H),8.44(s,1H),8.15(d,J=8.2Hz,2H),8.01(dd,J=8. 3, 6.3Hz, 3H), 7.82 (dd, J = 8.0, 2.5Hz, 1H), 7.65 (d, J = 1.5Hz, 1H), 6.68 (s, 1H), 6.41 (dd, J = 2.1, 2.1Hz, 1H), 4.55 (d, J = 5.8Hz, 2H).

[1009] LRMS m / z is calculated as C 22 H 18 N5O4[M+H] + :416.14, Actual value: 416.20, [M+2H] + / 2 observed 208.6

[1010] Example 1: Synthesis of 20–4-hydroxy-6-(1H-pyrazol-1-yl)-N-((6-(pyrimidin-5-yl)pyridin-3-yl)methyl)-nicotinamide (144)

[1011]

[1012] The title compound (6 mg, 0.0160 mmol, 81%) was obtained by conventional method D:137 (9 mg, 0.02 mmol) and lithium chloride (9.77 mg, 0.23 mmol).

[1013] 1 H NMR (400MHz, DMSO) δ9.43(d,J=1.9Hz,2H),9.24(s,1H),8.75(d,J=2.2Hz,1H),8.64(d,J=2.7Hz,1H ),8.15–8.10(m,1H),7.97–7.85(m,2H),7.35(s,1H),6.61(q,J=2.1Hz,1H),4.63(d,J=5.9Hz,2H).

[1014] LRMS m / z is calculated as C 19 H 16 N7O2[M+H] + :374.14, Actual value: 374.00, [M+2H] + / 2 = 187.5

[1015] Example 121: Synthesis of 4-hydroxy-N-((6-(4-(methylcarbamoyl)phenyl)pyridin-3-yl)methyl)-6-(1H-pyrazol-1-yl)nicotinamide (145)

[1016]

[1017] The title compound (6 mg, 0.0160 mmol, 81%) was obtained by conventional method D:138 (35 mg, 0.09 mmol) and lithium chloride (37 mg, 0.88 mmol).

[1018] 1 H NMR (400MHz, DMSO) δ9.57(s,1H),8.74(s,1H),8.70(d,J=2.2Hz,1H),8.64(d,J=2.6Hz,1H),8.50(q,J=4.5Hz,1H),8.21–8.09(m,2H),8.0 2(d,J=8.2Hz,1H),7.98–7.90(m,2H),7.90–7.79(m,2H),7.33(s,1H),6.60(t,J=2.7Hz,1H),4.62(d,J=5.9Hz,2H),2.81(d,J=4.5Hz,3H).

[1019] LRMS m / z is calculated as C 23 H 21 N6O3[M+H] + :429.17, Actual value: 429.10, [M+2H] 2+ / 2 = 215.1

[1020] Example 122 Synthesis of N-((6-(4-carbamoylphenyl)pyridin-3-yl)methyl)-4-hydroxy-6-(1H-pyrazol-1-yl)nicotinamide (146)

[1021]

[1022] The title compound (8 mg, 0.019 mmol, 38%) was obtained by conventional method D:139 (22 mg, 0.05 mmol) and lithium chloride (21.59 mg, 0.51 mmol).

[1023] 1H NMR (400MHz, DMSO) δ9.72 (s, 1H), 8.75–8.66 (m, 2H), 8.62 (d, J = 2.6Hz, 1H), 8.18–8.10 (m, 2H), 8.06–7.94 (m,4H),7.92–7.78(m,2H),7.40(s,1H),7.30(s,1H),6.58(dd,J=2.7,1.7Hz,1H),4.61(d,J=5.9Hz,2H).

[1024] LRMS m / z is calculated as C 22 H 19 N6O3[M+H] + :415.15, Actual value: 415.20, [M+2H] 2+ / 2 = 208.40

[1025] Example 123 – Synthesis of 4-hydroxy-N-((6-(4-morpholinophenyl)pyridin-3-yl)methyl)-6-(1H-pyrazol-1-yl)nicotinamide (147)

[1026]

[1027] The title compound (6 mg, 0.0160 mmol, 81%) was obtained by conventional method D:140 (35 mg, 0.11 mmol) and lithium chloride (44.68 mg, 1.06 mmol).

[1028] 1 H NMR (400MHz, DMSO) δ9.65(s,1H),8.72(s,1H),8.63(dd,J=2.6,0.7Hz,1H),8.58(dd,J=2.3,0.9Hz,1H),8.00–7.92(m,2H),7.88–7.80(m,2H),7.77(dd ,J=8.2,2.3Hz,1H),7.29(s,1H),7.06–6.98(m,2H),6.59(dd,J=2.6,1.7Hz ,1H),4.56(d,J=5.9Hz,2H),3.75(dd,J=5.8,3.9Hz,4H),3.22–3.15(m,4H).

[1029] LRMS m / z is calculated as C 25 H 25 N6O3[M+H] + :457.20, Actual value: 457.40, [M+2H] 2+ / 2 = 229.2

[1030] Example 123 Synthesis of N-((6-(benzo[d]thiazolyl)pyridin-3-yl)methyl)-4-hydroxy-6-(1H-pyrazol-1-yl)nicotinamide (148)

[1031]

[1032] The title compound (14 mg, 0.032 mmol, 65%) was obtained by conventional method D:141 (22 mg, 0.05 mmol) and lithium chloride (21 mg, 0.5 mmol).

[1033] 1 H NMR(400MHz,DMSO)δ9.45(s,1H),8.84–8.59(m,3H),8.35–8.19(m,2H),8.17–8.07(m,2 H),7.95–7.82(m,2H),7.34(s,1H),6.60(dd,J=3.8,3.0Hz,1H),4.62(d,J=6.2Hz,2H).

[1034] LRMS m / z is calculated as C 22 H 17 N6O2S[M+H] + :429.11, Actual value: 429.10, [M+2H] 2+ / 2 = 215.2

[1035] Synthesis of methyl 6-chloro-4-methoxypyridazine-3-carboxylic acid ester (149) as described in Example 120

[1036]

[1037] Methyl 4,6-dichloropyridazine-3-carboxylic acid (1000 mg, 4.93 mmol) was dissolved in THF (10 mL). The resulting mixture was cooled to 0 °C, and sodium methoxide (319 mg, 5.91 mmol) was slowly added. The reaction mixture was heated to room temperature and stirred for 6 hours. The solvent was then removed under vacuum, and the resulting residue was dissolved in a mixture of EtOAc (50 mL) and H2O (50 mL). The organic and aqueous layers were separated, and the aqueous layer was washed at least twice with EtOAc (50 mL). The organic layers were combined, dried (Na2SO4), and purified by rapid column chromatography (0%–100% EtOAc, 100%–0% cyclohexane) to give the title compound (103 mg, 0.509 mmol, 10%).

[1038] LRMS m / z calculation yields C7H8ClN2O3[M+H] + 203.02, Actual value: 203.1

[1039] Synthesis of methyl 4-methoxy-6-(1H-pyrazol-1-yl)pyridazine-3-carboxylate (150) from Example 125

[1040]

[1041] According to the general method C: 149 (103 mg, 0.29 mmol), pyrazole (30 mg, 0.44 mmol), Pd t BuXPhos G3 (23 mg, 0.03 mmol) and cesium carbonate (284 mg, 0.88 mmol) yielded the title compound (37 mg, 0.16 mmol, 55%).

[1042] LRMS m / z is calculated as C 10 H 11 N4O3[M+H] + 235.08, Actual value: 235.2

[1043] Reference Example 126– Synthesis of 4-methoxy-6-(1H-pyrazol-1-yl)pyridazine-3-carboxylic acid (151)

[1044]

[1045] 150 (37 mg, 0.16 mmol) was dissolved in a mixture of methanol (5 mL) and H₂O (0.5 mL), and then lithium hydroxide (33.21 mg, 0.79 mmol) was added. The resulting mixture was stirred at room temperature for 16 hours. 1 M HCl (5 mL) was added to the reaction mixture. The precipitate was then collected and washed with Et₂O to give the title compound (22 mg, 0.099 mmol, 62%).

[1046] LRMS m / z calculation yields C9H9N4O4[M+H] + 221.07, Actual Received: 221.1

[1047] Example 127 – Synthesis of 4-methoxy-6-(1H-pyrazol-1-yl)-N-((4-(trifluoromethyl)cyclohexyl)methyl)pyridazine-3-carboxamide (152)

[1048]

[1049] The title compound (34 mg, 0.088 mmol, 89%) was obtained by conventional method B: 151 (22 mg, 0.10 mmol), HATU (76 mg, 0.20 mmol), (4-trifluoromethyl)cyclohexyl)methylamine (15 μL, 0.1 mmol) and DIPEA (51 μL, 0.3 mmol).

[1050] LRMS m / z is calculated as C 17 H 21 F3N5O2[M+H] + 384.16, Actual value: 384.2

[1051] Example 128 – Synthesis of 4-hydroxy-6-(1H-pyrazol-1-yl)-N-((4-(trifluoromethyl)cyclohexyl)methyl)pyridazine-3-carboxamide (153)

[1052]

[1053] The title compound (8 mg, 0.0216 mmol, 11%) was obtained by conventional method D:152 (34 mg, 0.08 mmol) and lithium chloride (78 mg, 1.86 mmol).

[1054] 1 H NMR (400MHz, DMSO) δ9.92(s,1H),8.79–8.74(m,1H),7.94(d,J=1.7Hz,1H),7.46(s,1H),6.68(dd,J=2.7,1.7Hz,1H) ,3.22(t,J=6.5Hz,2H),2.32–2.14(m,1H),1.93–1.79(m,4H),1.63(ddt,J=11.7,8.3,4.4Hz,1H),1.32–0.99(m,4H).

[1055] LRMS m / z is calculated as C 16 H 19 N5O2[M+H] + 370.15, Actual value: 370.2

[1056] Example 129 – Synthesis of methyl 1-(3-hydroxy-5-(((4-(trifluoromethyl)cyclohexyl)methyl)carbamoyl)pyridin-2-yl)-1H-pyrazole-4-carboxylic acid (154)

[1057]

[1058] 128 (10 mg, 0.02 mmol) was added to a 10 mL round-bottom flask, rinsed with N2, and then MeOH (3 mL) was added. A 10% sodium methoxide methanol solution (500 μL) was added to the reaction mixture, and the resulting mixture was stirred at room temperature for 16 hours. Formic acid was added to neutralize the reaction mixture, and then diatomaceous earth was added. The solvent was removed under vacuum, and the crude mixture was purified by rapid column chromatography (0–10% MeOH in CH2Cl2 solution) to give the title compound (4.5 mg, 0.010 mmol, 52%).

[1059] 1 H NMR (400MHz, DMSO) δ8.93(d,J=0.7Hz,1H),8.71(t,J=6.0Hz,1H),8.47(d,J=1.9Hz,1H),8.28(d,J=0.7H z,1H),7.90(d,J=1.9Hz,1H),3.82(s,3H),3.15(t,J=6.5Hz,2H),1.95–1.78(m,5H),1.30–0.96(m,5H).

[1060] LRMS m / z is calculated as C 19 H 22 F3N4O4[M+H] + :427.16, Actual yield: 427.2 Reference Example 130 – Synthesis of 1-aminocarbamoyl-1H-pyrazole-4-carboxylic acid ethyl ester (155)

[1061]

[1062] A solution of 4M HCl in dioxane (20 mL) was added to a solution of ethyl 1H-pyrazole-4-carboxylate (6.66 g, 47.62 mmol), cyanamide (2 g, 47.62 mmol), and dioxane (30 mL). The reaction mixture was heated to 100 °C and maintained for 3 hours. The reaction was cooled to room temperature, and Et₂O (30 mL) was added. The precipitate was collected to give the title compound (3 g, 16.4 mmol, 35%).

[1063] LRMS m / z calculation is C7H 11 N4O2[M+H] + :183.09, Actual value: 183.1

[1064] Synthesis of methyl 1-(5-cyano-4-hydroxypyrimidin-2-yl)-1H-pyrazole-4-carboxylic acid (156) as described in Example 131

[1065]

[1066] Ethyl (E)-2-cyano-3-ethoxyacrylate (3.0 g, 17.75 mmol) was added to a solution of 155 μL (3.23 g, 17.75 mmol), K₂CO₃ (4.9 g, 35.50 mmol), and methanol (100 mL). The resulting mixture was stirred at room temperature for 1 hour. H₂O (50 mL) was added, and a white precipitate was filtered off to give the title compound (3.52 g, 13.59 mmol, 76%).

[1067] LRMS m / z is calculated as C 10 H8N5O3[M+H] + 246.06, Actual value: 246.1

[1068] Reference Example 132 – Synthesis of 1-(5-cyano-4-hydroxypyrimidin-2-yl)-1H-pyrazole-4-carboxylic acid (157)

[1069]

[1070] A solution of 156 (3.0 g, 13.1 mmol) in MeOH (15 mL) and H₂O (10 mL) was prepared, followed by the addition of lithium hydroxide monohydrate (904 mg, 39.3 mmol). The reaction mixture was stirred for 4 hours. The reaction mixture was neutralized with 1 N HCl, and methanol was removed under vacuum. The precipitate was collected to give the title compound (2.49 g, 10.8 mmol, 83%).

[1071] LRMS m / z calculation yields C9H4N5O3[MH] - 230.03, Actual value: 230.4

[1072] Example 133 Synthesis of N-([1,1'-biphenyl]-4-ylmethyl)-1-(5-cyano-4-hydroxypyrimidin-2-yl)-1H-pyrazole-4-carboxamide (158)

[1073]

[1074] The title compound (8 mg, 0.020 mmol, 57%) was obtained by conventional method B: 157 (10 mg, 0.04 mmol), HATU (33 mg, 0.09 mmol), 4-phenylbenzylamine (11.88 mg, 0.06 mmol) and DIPEA (22 μL, 0.13 mmol).

[1075] 1H NMR (400MHz, DMSO) δ9.15 (s, 1H), 8.99 (t, J = 6.0Hz, 1H), 8.59 (s, 1H), 8.23 ​​(s ,1H),7.64(dd,J=7.9,4.7Hz,4H),7.51–7.31(m,5H),4.48(d,J=6.0Hz,2H).

[1076] LRMS m / z is calculated as C 22 H 17 N6O2[M+H] + 397.14, Actual value: 397.2

[1077] Example 134 – Synthesis of 1-(5-cyano-4-hydroxypyrimidin-2-yl)-N-((4-(trifluoromethyl)cyclohexyl)methyl)-1H-pyrazole-4-carboxamide (159)

[1078]

[1079] The title compound (9 mg, 0.022 mmol, 14%) was obtained by general method B: 157 (37 mg, 0.16 mmol), C-4 trifluoromethyl-cyclohexyl-methylamine (28 μL, 0.19 mmol), T3P (190 μL, 0.32 mmol), and DIPEA (83 μL, 0.48 mmol).

[1080] 1 H NMR (400MHz, DMSO) δ9.05 (s, 1H), 8.27 (d, J = 6.7Hz, 2H), 7.98 (s, 1H), 3.07 (t, J = 6.0Hz, 2H), 2.2 9–2.12(m,1H),1.85(t,J=16.4Hz,4H),1.64–1.42(m,1H),1.27–1.13(m,2H),1.05–0.91(m,2H).

[1081] LRMS m / z is calculated as C 17 H 18 F3N6O2[M+H] + :395.14, Actual value: 395.2

[1082] Example 135: Synthesis of 1-(5-cyano-4-hydroxypyrimidin-2-yl)-N-((6-phenylpyridin-3-yl)methyl)-1H-pyrazole-4-carboxamide (160)

[1083]

[1084] The title compound (65 mg, 0.163 mmol, 38%) was obtained by conventional method B: 157 (100 mg, 0.43 mmol), (6-phenylpyridin-3-yl)methylamine (119 mg, 0.65 mmol), T3P (515 μL, 0.87 mmol), and DIPEA (223 μL, 0.130 mmol).

[1085] 1 H NMR (400MHz, DMSO) δ9.28–8.99(m,2H),8.87–8.60(m,2H),8.43–8.19(m,1H),8.09–8.03(m,2H),7 .95(d,J=8.1Hz,1H),7.82(ddd,J=16.8,8.3,2.4Hz,1H),7.54–7.39(m,3H),4.46(d,J=5.5Hz,2H).

[1086] LRMS m / z is calculated as C 21 H 16 N7O2[M+H] + :398.14, Actual amount received: 398.1

[1087] Example 136 – Synthesis of methyl 4-(5-((1-(5-cyano-4-hydroxypyrimidin-2-yl)-1H-pyrazol-4-carboxamido)-methyl)-pyridin-2-yl)-benzoate (161)

[1088]

[1089] The title compound (11 mg, 0.0241 mmol, 6%) was obtained by conventional method B: 157 (100 mg, 0.43 mmol), methyl 4-(5-(aminomethyl)pyridin-2-yl)benzoate (157 mg, 0.65 mmol), T3P (515 μL, 0.87 mmol), and DIPEA (223 μL, 0.130 mmol).

[1090] 1 H NMR (400MHz, DMSO) δ9.05 (s, 1H), 8.27 (d, J = 6.7Hz, 2H), 7.98 (s, 1H), 3.07 (t, J = 6.0Hz, 2H), 2.2 9–2.12(m,1H),1.85(t,J=16.4Hz,4H),1.64–1.42(m,1H),1.27–1.13(m,2H),1.05–0.91(m,2H).

[1091] LRMS m / z is calculated as C 23 H18 N7O4[M+H] + :456.14, actual value: 456.2.

[1092] Example 137 – Synthesis of 4-(5-((1-(5-cyano-4-hydroxypyrimidin-2-yl)-1H-pyrazol-4-carboxamido)methyl)pyridin-2-yl)benzoic acid (162)

[1093]

[1094] The title compound (2 mg, 0.0045 mmol, 45%) was obtained by DMF solution (1 mL) of D:161 (5 mg, 0.01 mmol) and LiCl (4.51 mg, 0.11 mmol) according to the general method.

[1095] 1 H NMR (400MHz, DMSO) δ9.13 (s, 1H), 9.04 (t, J = 5.5Hz, 1H), 8.68 (d, J = 2.2Hz, 1H), 8.57 (s, 1H), 8.2 3–8.16(m,3H),8.04(dd,J=8.3,4.0Hz,3H),7.86(dd,J=8.3,2.3Hz,1H),4.52(d,J=5.5Hz,2H).

[1096] LRMS m / z is calculated as C 22 H 14 N7O4[MH] - :440.11, Actual value: 439.3, [M-2H] 2- / 2 = 220.0

[1097] Example 138 – Synthesis of 6-(4-(([1,1'-biphenyl]-4-ylmethyl)carbamoyl)-1H-pyrazol-1-yl)-5-methoxynicotinic acid ethyl ester (163a)

[1098]

[1099] A dioxane solution (5 mL) of ethyl 6-chloro-5-methoxynicotinic acid (30 mg, 0.14 mmol), 92 (30 mg, 0.108 mmol), RockPhos G3 (10 mg, 0.014 mmol), and Cs2CO3 (77 mg, 0.238 mmol) was reacted for 7 days to yield 163a (5 mg, 0.01 mmol, 9%), which was a transparent oil.

[1100] 1H NMR(400MHz,THF-d8)δ8.71(s,1H,),8.65(d,J=1.0Hz,1H),8.07–8.05(m,1H),7.91(t,J=6.0Hz,1H),7.75(d,J= 1.0Hz, 1H), 7.46–7.24 (m, 9H), 4.58 (d, J = 6.0Hz, 2H), 4.40 (q, J = 7.0Hz, 2H), 3.98 (s, 3H), 1.42 (t, J = 7.0Hz, 3H).

[1101] HRMS (ESI-TOF) is calculated as C 26 H 25 O4N4[M+H] + :457.1870, actual value: 457.1867.

[1102] Example 139 – Synthesis of 6-(4-(([1,1'-biphenyl]-4-ylmethyl)carbamoyl)-1H-pyrazol-1-yl)-5-methoxynicotinic acid (164a)

[1103]

[1104] 163a (5 mg, 0.0109 mmol) was dissolved in a mixture of THF and water (1.5 mL (10:1)). Lithium hydroxide monohydrate (1.0 mg, 0.021 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature for 16 hours. The reaction was confirmed to be complete by TLC, and HCLaq (5 mL, 1 M) was added to the reaction mixture. The mixture was extracted with EtOAc (3 × 10 mL), washed with brine, dried over anhydrous Na2SO4, and purified by rapid column chromatography (CH2Cl2, MeOH 0-5%, 1% formic acid) for 20 column volumes to give 164a (3.5 mg, 0.0082 mmol, 76%) as a clear oil.

[1105] 1H NMR(400MHz,DMSO-d6)δ8.69(s,1H),8.47(s,1H),8.12(s,1H),7.99(s,1H) ,7.70–7.61(m,4H),7.50–7.32(m,5H),4.50(d,J=6.0Hz,2H),3.89(s,3H).

[1106] HRMS (ESI-TOF) is calculated as C 24 H 21 O4N4[M+H] + :429.1557, actual value: 429.1557.

[1107] Synthesis of 2,5-dimethylpyrazolo[1,5-a]pyrimidine-7-amine (163) from Example 140

[1108]

[1109] 3-Aminocrotonitrile (10 g, 0.12 mol) and 3-methyl-1H-pyrazole-5-amine (11.83 g, 0.12 mol) were mixed and heated to 140 °C for 2 hours. The resulting mixture was allowed to cool. The crude mixture was then recrystallized from EtOH to give the title compound (6.68 g, 0.041 mol, 35%).

[1110] LRMS m / z calculation is C8H 11 N4[M+H] + :163.1, Actual value: 163.10

[1111] Synthesis of 2-(((2,5-dimethylpyrazolo[1,5-a]pyrimidin-7-yl)amino)methylene)diethyl malonate (164) as described in Example 141

[1112]

[1113] 163 (1.50 g, 0.01 mmol) was dissolved in toluene (10 mL), and then diethyl 2-(ethoxymethylene)malonate (2.24 mL, 0.01 mmol) was added. The resulting mixture was heated to 120 °C and maintained for 48 hours. The reaction mixture was cooled to room temperature. Diatomaceous earth was added to the reaction mixture, and then the solvent was removed under vacuum. The crude compound was purified by rapid chromatography (0–10% methanol) to give the title compound (2.26 g, 0.0068 mmol, 74%).

[1114] LRMS m / z is calculated as C 16 H 21 N4O4[M+H] + 333.16, Actual value: 333.2

[1115] Synthesis of 2–6-hydroxy-2,5-dimethylpyrazolo[1,5-a]pyrido[3,2-e]pyrimidine-7-carboxylic acid ethyl ester (165) from Example 14

[1116]

[1117] 164 (6.15 g, 18.52 mmol) was dissolved in Eaton reagent (10 mL). The resulting mixture was heated to 70 °C and held for 4 hours, with the process monitored by LCMS. After the starting material was consumed, the reaction mixture was cooled to 0 °C and slowly poured into a saturated NaHCO3 solution to quench the Eaton reagent. The resulting mixture was then extracted with a mixture of CHCl3 and IPA (3 × 100 mL (3:1)). The organic fractions were combined, dried (Na2SO4), and the solvent was removed under vacuum. The crude compound was purified by rapid column chromatography (CHCl3:MeOH (0–20%)) to give the title compound (4.67 g, 16.32 mmol, 88%).

[1118] LRMS m / z is calculated as C 14 H 15 N4O3[M+H] + :287.11, Actual amount received: 287.20

[1119] Reference Example 143 – Synthesis of 6-chloro-2,5-dimethylpyrazolo[1,5-a]pyrido[3,2-e]pyrimidine-7-carboxylic acid ethyl ester (166)

[1120]

[1121] 165 (4.67 g, 16.32 mmol) was dissolved in phosphorus oxychloride (25 mL) and refluxed at 100 °C for 3 hours. The reaction mixture was cooled to 0 °C and slowly quenched with saturated NaHCO3 solution. The resulting mixture was extracted with a CHCl3:IPA mixture (3:1 (3 × 100 mL)). The organic fractions were combined, dried (Na2SO4), and the solvent was removed under vacuum. The crude mixture was then proceeded to the next step without further purification (4.54 g, 14.93 mmol, 91%).

[1122] LRMS m / z is calculated as C 14 H 14 ClN4O4[M+H] + :305.08, Actual yield: 305.2 Reference Example 144 – Synthesis of methyl 6-methoxy-2,5-dimethylpyrazolo[1,5-a]pyrido[3,2-e]pyrimidine-7-carboxylic acid ester (167)

[1123]

[1124] 166 (156 mg, 0.51 mmol) was dissolved in MeOH (10 mL), and then sodium methoxide solution (5 mL, 0.5 M) was added. The resulting mixture was stirred at room temperature for 4 hours. Diatomaceous earth was added to the reaction mixture, and the solvent was removed under vacuum. The crude mixture was then purified by rapid column chromatography (CHCl3 solution of MeOH (0–10%)) to give the title compound (38 mg, 0.13 mmol, 26%).

[1125] LRMS m / z is calculated as C 14 H 15 N4O3[M+H] + 287.11, Actual value: 287.2

[1126] Example 145–Synthesis of 6-methoxy-2,5-dimethyl-N-((4-(trifluoromethyl)cyclohexyl)methyl)pyrazolo[1,5-a]pyridolo[3,2-e]pyrimidine-7-carboxamide (168)

[1127]

[1128] The title compound (9 mg, 0.02 mmol, 20%) was obtained by general method A: 167 (30 mg, 0.1 mmol), 4-trifluoromethyl-cyclohexylamine (19 μL, 0.16 mmol) and DABAL (26.85 mg, 0.1 mmol).

[1129] Solvent system used for purification: 0%-15% MeOH in CH3Cl solution.

[1130] LRMS m / z is calculated as C 21 H 25 F3N5O2[M+H] + :436.20,found:436.2.

[1131] Example 146 Synthesis of N-([1,1'-biphenyl]-4-ylmethyl)-6-hydroxypyrazolo[1,5-a]pyrido[3,2-e]pyrimidine-7-carboxamide (169)

[1132]

[1133] The title compound (1.5 mg, 0.0035 mmol, 18%) was obtained by general method with a DMSO solution (1 mL) of D:168 (9 mg, 0.02 mmol) and LiCl (8.6 mg, 0.21 mmol).

[1134] Solvent system used for purification: reversed phase (0%-100% ACN (0.1% formic acid) in H2O solution (0.1% formic acid)).

[1135] 1 H NMR (400MHz, DMSO) δ8.91(s,1H),6.42(s,1H),3.23(d,J=5.1Hz,2H),2.66(s,3H),2.41(s,3H),2.07(s,1H ),1.83–1.75(m,2H),1.65(d,J=12.7Hz,2H),1.48(d,J=9.6Hz,1H),1.25–1.08(m,2H),0.95–0.81(m,2H).

[1136] LRMS m / z is calculated as C 20 H 23 F3N5O2[M+H] + :422.18, Actual measurement + 422.2

[1137] Example 147 – Synthesis of N-((6-chloropyridin-3-yl)methyl)-6-methoxy-2,5-dimethylpyrazolo[1,5-a]pyridolo[3,2-e]pyrimidine-7-carboxamide (170)

[1138]

[1139] The title compound (364 mg, 0.91 mmol, 29%) was obtained by general method A: 167 (900 mg, 3.15 mmol), (6-chloropyridin-3-yl)methylamine (532 mg, 3.78 mmol) and DABAL (805 mg, 3.15 mmol).

[1140] Solvent system used for purification: 0%-15% MeOH in CH3Cl solution.

[1141] LRMS m / z is calculated as C 19 H 18 ClN6O2[M+H] + :397.12, Actual amount received: 397.10

[1142] Example 148 – Synthesis of N-((6-(4-carbamoylphenyl)pyridin-3-yl)methyl)-6-hydroxy-2,5-dimethylpyrazolo[1,5-a]pyridolo[3,2-e]pyrimidine-7-carboxamide (171)

[1143]

[1144] The title compound (2 mg, 0.0042 mmol, 11%) was obtained by conventional method C:170 (15 mg, 0.04 mmol), (4-(carbamoyl)phenyl)boronic acid (9.75 mg, 0.06 mmol), palladium aminophosphine (2.77 mg, 0.0039 mmol) and cesium carbonate (38.19 mg, 0.12 mmol).

[1145] 1 H NMR (400MHz, DMSO) δ8.77(s,1H),8.69–8.66(m,1H),8.31(s,1H),8.14(t,J=4.2Hz,3H),7.99(dd,J=17.9,8.4H z, 3H), 7.86 (dd, J = 8.1, 2.5Hz, 1H), 6.53 (s, 2H), 6.20 (s, 1H), 4.59 (d, J = 6.0Hz, 2H), 2.87 (s, 3H), 2.36 (s, 3H).

[1146] LRMS m / z is calculated as C 25 H 22 N7O3[M+H] + :468.18, Actual value: 468.1, [M+2H] 2+ / 2 = 234.70

[1147] Example 149 – Synthesis of 6-hydroxy-2,5-dimethyl-N-((6-(pyrimidin-5-yl)pyridin-3-yl)methyl)pyrazolo[1,5-a]pyridolo[3,2-e]pyrimidin-7-carboxamide (172)

[1148]

[1149] The title compound (2.5 mg, 0.0058 mmol, 15%) was obtained by conventional method C:170 (15 mg, 0.04 mmol), 5-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)pyrimidine (12.1 mg, 0.06 mmol), palladium aminophosphine (1.39 mg, 0.0019 mmol) and cesium carbonate (38.19 mg, 0.12 mmol).

[1150] 1H NMR (400MHz, DMSO) δ10.69(s,1H),9.42(s,2H),9.24(s,1H),8.74(d,J=2.2Hz,1H),8.59(s,1H),8.14–8.09(m ,1H),7.93(dd,J=8.2,2.3Hz,1H),6.52(s,1H),6.42(s,1H),4.63(d,J=6.0Hz,2H),2.89(s,3H),2.45(s,3H).

[1151] LRMS m / z is calculated as C 22 H 19 N8O2[M+H] + :427.16, Actual value: 427.1, [M+2H] 2+ / 2 = 214.20

[1152] Example 150 – Synthesis of 6-hydroxy-2,5-dimethyl-N-((6-(4-morpholinophenyl)pyridin-3-yl)methyl)pyrazolo[1,5-a]pyridolo[3,2-e]pyrimidine-7-carboxamide (173)

[1153]

[1154] The title compound (5 mg, 0.009 mmol, 25%) was obtained by conventional method C:170 (15 mg, 0.04 mmol), 4-morpholine-phenylboronic acid (12.1 mg, 0.06 mmol), palladium aminophosphine (1.39 mg, 0.0019 mmol) and cesium carbonate (38.19 mg, 0.12 mmol).

[1155] 1 H NMR (400MHz, DMSO) δ10.50(s,1H),8.59–8.48(m,2H),7.98–7.93(m,2H),7.83(dd,J=8.3,0.9Hz,1H),7.76(dd,J=8.2,2.3Hz,1H),7 .05–6.99(m,2H),6.53(s,1H),6.45(s,1H),4.57(d,J=5.9Hz,2H),3.77–3.71(m,4H),3.23–3.14(m,4H),2.88(s,3H),2.46(s,3H).

[1156] LRMS m / z is calculated as C 28 H 28 N7O3[M+H] + :510.22, Actual measurement [M+2H] 2+ / 2 = 255.70

[1157] Example 151 – Synthesis of 6-hydroxy-2,5-dimethyl-N-((6-(4-(methylcarbamoyl)phenyl)pyridin-3-yl)methyl)pyrazolo[1,5-a]pyridolo[3,2-e]pyrimidine-7-carboxamide (174)

[1158]

[1159] The title compound (1 mg, 0.0021 mmol, 5%) was obtained by conventional method C:170 (15 mg, 0.04 mmol), 4-N-methylcarbonyl-phenylboronic acid (10.52 mg, 0.06 mmol), palladium aminophosphine (2.77 mg, 0.0039 mmol) and cesium carbonate (38.19 mg, 0.12 mmol).

[1160] 1 H NMR (400MHz, DMSO) δ8.67(d,J=6.2Hz,2H),8.51(d,J=5.0Hz,1H),8.18–8.10(m,2H),8.02(d,J=8.2Hz,1H),7.93(d,J=8.4H z,2H),7.88–7.78(m,1H),6.52(s,1H),6.34(s,1H),4.61(d,J=5.8Hz,2H),2.88(s,3H),2.80(d,J=2.3Hz,3H),2.41(s,3H).

[1161] LRMS m / z is calculated as C 26 H 24 N7O3[M+H] + :482.19, Actual value: [M+2H] 2+ / 2 = 241.50

[1162] Example 152 – Synthesis of 6-hydroxy-2,5-dimethyl-N-((6-phenylpyridin-3-yl)methyl)pyrazolo[1,5-a]pyridolo[3,2-e]pyrimidine-7-carboxamide (175)

[1163]

[1164] The title compound (9 mg, 0.0212 mmol, 22%) was obtained by conventional method C:170 (50 mg, 0.13 mmol), phenylboronic acid (46.21 mg, 0.38 mmol), palladium aminophosphine (8.94 mg, 0.01 mmol) and cesium carbonate (123 mg, 0.38 mmol).

[1165] 1H NMR (400MHz, DMSO) δ9.42(s,1H),8.95(s,1H),8.49(s,1H),8.09–8.00(m,2H),7.89(d,J=8.2Hz ,1H),7.69–7.62(m,1H),7.51–7.36(m,3H),6.47(s,1H),4.64(s,2H),2.84(s,3H),2.43(s,3H).

[1166] LRMS m / z is calculated as C 24 H 21 N6O2[M+H] + :425.17, Actual value: 425.2, [M+2H] 2+ / 2 = 213.2

[1167] Example 153 – Synthesis of 6-hydroxy-2,5-dimethyl-N-((6-(3-morpholinophenyl)pyridin-3-yl)methyl)pyrazolo[1,5-a]pyridolo[3,2-e]pyrimidine-7-carboxamide (176)

[1168]

[1169] The title compound (14 mg, 0.275 mmol, 28%) was obtained by conventional method C:170 (40 mg, 0.10 mmol), 3-morpholino-phenylboronic acid (63 mg, 0.30 mmol), palladium aminophosphine (7.15 mg, 0.01 mmol) and cesium carbonate (99 mg, 0.30 mmol).

[1170] 1 H NMR (400MHz, DMSO) δ9.52(s,1H),8.95(s,1H),8.46(d,J=2.2Hz,1H),7.88(d,J=8.2Hz,1H),7.68–7.56(m,2H),7.46(d,J=7.6Hz,1H),7.31(t,J=7. 9Hz,1H),7.00(dd,J=8.3,2.5Hz,1H),6.46(s,1H),4.67–4.60(m,2H),3.7 5(t,J=4.7Hz,4H),3.16(dd,J=6.6,3.2Hz,4H),2.84(s,3H),2.43(s,3H).

[1171] LRMS m / z is calculated as C 28 H 28 N7O3[M+H] + :510.22, Actual value: 510.30, [M+2H] 2+ / 2 = 255.80

[1172] Example 154 – Synthesis of N-((6-(3-carbamoylphenyl)pyridin-3-yl)methyl)-6-hydroxy-2,5-dimethylpyrazolo[1,5-a]pyridolo[3,2-e]pyrimidine-7-carboxamide (177)

[1173]

[1174] The title compound (11 mg, 0.0235 mmol, 24%) was obtained by conventional method C:170 (40 mg, 0.10 mmol), 3-acylamino-phenylboronic acid (50 mg, 0.30 mmol), palladium aminophosphine (7.15 mg, 0.01 mmol) and cesium carbonate (99 mg, 0.30 mmol).

[1175] 1 H NMR (400MHz, DMSO) δ9.41(s,1H),8.95(s,1H),8.56–8.49(m,2H),8.21–8.06(m,2H),7.97(d,J=8.2Hz,1H),7.91(d,J=7.7Hz,1H ), 7.71 (dd, J = 8.3, 2.3Hz, 1H), 7.55 (t, J = 7.7Hz, 1H), 7.43 (s, 1H), 6.47 (s, 1H), 4.66 (d, J = 4.1Hz, 2H), 2.85 (s, 3H), 2.43 (s, 3H).

[1176] LRMS m / z is calculated as C 25 H 22 N7O3[M+H] + :468.18, Actual value: 468.2, [M+2H] 2+ / 2 = 234.80

[1177] Example 155 Synthesis of N-((6-(benzo[d]thiazo-5-yl)pyridin-3-yl)methyl)-6-hydroxy-2,5-dimethylpyrazolo[1,5-a]pyrido[3,2-e]pyrimidine-7-carboxamide (178)

[1178]

[1179] The title compound (5 mg, 0.0103 mmol, 8%) was obtained by conventional method C:170 (50 mg, 0.13 mmol), 5-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)benzo[d]thiazole (98.86 mg, 0.38 mmol), palladium aminophosphine (8.94 mg, 0.01 mmol) and cesium carbonate (123 mg, 0.38 mmol).

[1180] 1 H NMR (400MHz, DMSO) δ9.44(s,1H),8.96(s,1H),8.73(d,J=1.6Hz,1H),8.54(d,J=2.3Hz,1H),8.27–8.19(m,2H),8 .08(d,J=8.2Hz,1H),7.71(dd,J=8.3,2.3Hz,1H),6.46(s,1H),4.65(d,J=4.6Hz,2H),2.85(s,3H),2.43(s,3H).

[1181] LRMS m / z is calculated as C 25 H 20 N7O2S[M+H] + :482.14, measured 482.20, [M+2H] 2+ / 2 = 241.70

[1182] Example 156 – Synthesis of 6-hydroxy-2,5-dimethyl-N-((6-(3-(methylcarbamoyl)phenyl)pyridin-3-yl)methyl)pyrazolo[1,5-a]pyridolo[3,2-e]pyrimidine-7-carboxamide (179)

[1183]

[1184] The title compound (11 mg, 0.0228 mmol, 32%) was obtained by conventional method C:170 (29 mg, 0.07 mmol), 3-(methylcarbamoyl)phenyl)boronic acid (39.33 mg, 0.22 mmol), palladium aminophosphine (5.18 mg, 0.01 mmol) and cesium carbonate (71 mg, 0.22 mmol).

[1185] 1H NMR (400MHz, DMSO) δ9.96(s,1H),8.96(s,1H),8.60–8.44(m,3H),8.16(d,J=7.9Hz,1H),7.95(d,J=8.2Hz,1H),7.86(d,J=7.7Hz,1 H), 7.70 (dd, J = 8.2, 2.3Hz, 1H), 7.54 (t, J = 7.7Hz, 1H), 6.45 (s, 1H), 4.64 (s, 2H), 2.84 (s, 3H), 2.80 (d, J = 4.4Hz, 3H), 2.42 (s, 3H).

[1186] LRMS m / z is calculated as C 26 H 23 N7O3[M+H] + :482.19, Actual value: 482.2, [M+2H] 2+ / 2 = 241.60

[1187] Example 157 – Synthesis of (S)-6-hydroxy-N-(1-(4-methoxyphenyl)ethyl)-2,5-dimethylpyrazolo[1,5-a]pyridolo[3,2-e]pyrimidine-7-carboxamide (180)

[1188]

[1189] The title compound (7 mg, 0.0179 mmol, 11%) was obtained by general method A: 167 (50 mg, 0.17 mmol), (S)-1-(4-methoxyphenyl)ethyl-1-amine (31 μL, 0.21 mmol) and DABAL-Me3 (44.76 mg, 0.17 mmol).

[1190] Solvent system used for purification: 0%-15% MeOH in CH3Cl solution.

[1191] 1 H NMR (400MHz, DMSO) δ10.40(d,J=7.9Hz,1H),8.51(s,1H),7.35–7.28(m,2H),6.91(dd,J=8.5,1.9Hz ,2H),6.47(s,1H),5.16–5.05(m,1H),3.74(s,3H),2.89(s,3H),2.46(s,3H),1.47(d,J=6.9Hz,3H).

[1192] LRMS m / z is calculated as C 21 H 22 N5O3[M+H] +:392.17, Actual amount received: 392.20

[1193] Active Examples

[1194] General experimental methods

[1195] Preparation of tPHD2 (residues 181-426)

[1196] In summary, cDNA encoding the tPHD2 catalytic domain (residues 181-426) was cloned into the pET28a(+) / pET24a(+) vector (Novagen) to generate tPHD2 (residues 181-426) protein with or without an N-terminal His 6-tag. The tPHD2 (residues 181-426) encoding construct was transformed into the *E. coli* BL21 DE3 cell line; protein production was induced with 0.5 mM isopropyl-bD-thiogalactosidase (3-5 h at 28°C). Cells were harvested and lysed by sonication in 20 mM Tris-HCl (pH 7.0) and 0.3 M NaCl. Soluble proteins (approximately 5% of the total soluble extract) were purified by Ni-ion affinity chromatography immobilized on pentadentate tricarboxymethyl ethylenediamine resin, followed by thrombin cleavage of the His6 tag (or cation exchange chromatography as an alternative), and finally purification by gel filtration chromatography. Proteins were exchanged into 50 mM Tris-HCl buffer (pH 7.5) and concentrated to 40 mg / mL. The purity of the protein was >95% as determined by SDS-PAGE and electrospray ionization mass spectrometry.

[1197] PHD2 hydroxylation test

[1198] PHD2 RF-MS RapidFire chromatography-mass spectrometry (RF-MS) analysis monitored the turnover of the C-terminal oxygenase-dependent domain (CODD) peptide substrate DLDLEMLAPYIPMDDDFQL-CONH2 and the appearance of the hydroxylated peptide product (Pro564) in endpoint-type analysis (typical enzyme incubation time 15 min). Tris(hydroxymethyl)aminomethane was obtained from Fisher. Ferrous ammonium sulfate (FAS), 2-oxoglutaric acid (2OG), and L-ascorbic acid were obtained from Sigma Aldrich; these solutions were prepared fresh daily. All inhibition assays were performed on 384-well polypropylene plates (Greiner Bio-One). PHD2 assays were performed in test buffer (50 mM Tris.Cl pH 7.5, 50 mM NaCl). Preparation for IC was done using an ECHO 550 acoustic dispenser (Labcyte). 50 The compound titrant to be determined (3 times and 11-point IC50) 50The DMSO was dried and dispensed into 384-well polypropylene test plates. The final test concentration of DMSO was kept constant at 0.5%. A 300 nM concentration of PHD2 protein was prepared in assay buffer, and 25 μl was dispensed into each 384-well plate. The PHD2 solution was allowed to equilibrate with the inhibitor at room temperature for 15 minutes; then the enzyme reaction was initiated by dispensing 25 μl of substrate (20 μM FAS, 200 μM M ascorbic acid, 10 μM CODD peptide, and 20 μM 2OG) into the assay buffer. The enzyme reaction was allowed to proceed at room temperature for 20 minutes, and then terminated by adding 10% formic acid (5 μl). The test plates were then transferred to a RapidFire RF360 sampling robot (Agilent) connected to an Agilent 6530 Precision Mass Quadrupole Time-of-Flight (Q-TOF) mass spectrometer. The test samples were aspirated under vacuum and loaded onto a C4 solid-phase extraction (SPE) column. After loading, the C4 SPE was washed with 0.1% formic acid aqueous solution to remove non-volatile buffer salts, and then the peptide was eluted from the SPE to the mass spectrometer with 85% acetonitrile and 15% water containing 0.1% formic acid. The charge state of the peptide was monitored in positive ion mode. Ion chromatogram data of the +2 charge state were extracted and peak area data were integrated using RapidFire Integrator software (Agilent Technologies). The percentage of CODD peptide substrate converted to +16 hydroxylated peptide was calculated using the following equation: % conversion = 100 × hydroxylated / (hydroxylated + non-hydroxylated peptide). IC50 was determined from the nonlinear regression plot using GraphPad Prism. 50 value.

[1199] Hep3B cell culture and immunoblotting

[1200] These were conducted according to TLYeh et al., Chem Sci, 2017, 8, 7651-7668.

[1201] Cell culture using HEK293T cells:

[1202] HEK293T cells were grown in a 37°C incubator at 5% CO2 in Dulbecco modified Eagle's medium (DMEM, high glucose, pyruvate, glutamine-free, Gibco) supplemented with 10% FBS (Sigma Aldrich F7524-500ML) and 1% GlutaMAX (Gibco). Cells reached 90% confluence.

[1203] Inhibitor addition and culture:

[1204] Cells were loaded at 1.2 × 10⁻⁶ 6Cells were seeded at a density in T75 flasks. Cells were exposed to an inhibitor at a final concentration of 1% DMSO and incubated at 37°C for 3–18 hours.

[1205] Protein extraction and analysis using HEK293T cells:

[1206] Wash cells twice with ice-cold PBS (Sigma, D8537). 1x RIPA buffer [Sigma, R0278] and protease inhibitor [Complete] TM Mini, EDTA-free protease inhibitor-free mixture, Roche] was used for protein extraction. Adherent cells were scraped off with a spatula and transferred to ice-cold microcentrifuge tubes. The cell suspension was frozen at -20°C or incubated on ice for 45 minutes, followed by three rounds of 10-second pulses of sonication at 5-second intervals. After centrifugation (16,000 g, 15 minutes, 4°C), the cell supernatant was collected in new microcentrifuge tubes. Protein was extracted using the BCA protein assay kit (ThermoScientific). TM Pierce TM The BCA protein assay kit is used to determine protein concentration.

[1207] SDS-PAGE analysis was performed using HEK293T cells.

[1208] For polyacrylamide gel electrophoresis, all samples were loaded into pre-cast NuPAGE 4-12% Bis-Tris protein gels (Life Tech). The gels were run at 180V for 45 minutes in 1x Tris / glycine / SDS run buffer (20x NuPAGEMESSDS run buffer, Life Tech) (Mini Gel Tank, Life Technologies). Protein sizes were compared using 5 μl of Page Ruler pre-contained Protein Ladder markers (Thermo Scientific).

[1209] Western blot analysis using HEK293T cells

[1210] The gel containing the decomposed proteins was then transferred to a nitrocellulose membrane (Amersham Protran Premium 0.2NC 300mm, GE Healthcare) using 1x transfer buffer (20x NuPAGE transfer buffer, Invitrogen). Mini Protean Tetra Cell, Bio-Rad was used for the transfer. The transfer was performed at 100 volts for 1 hour. The membrane was blocked for 30 minutes with 5% milk powder in 1x PBS-T, and then incubated overnight at 4°C with primary antibody (1:1000 dilution) prepared with 1% milk powder in 1x PBS-T buffer. The membrane was washed three times with 1x PBS-T for 10 minutes each time, and then incubated a second time for 1 hour at room temperature with horseradish peroxidase (HRP) conjugated secondary antibody (1:5000 dilution) prepared with 1% milk powder in 1x PBS-T. The blot was then washed three times with 1x PBS-T for 10 minutes each time, and then incubated for 1 hour at room temperature with GE Healthcare Amersham TM ECL TM Washing with Prime Western blot assay kit (RPN2236) and protein levels were measured using Bio-Rad Universal Hoodiii via densitometry. Values ​​were normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and corresponding controls, such as 1% (v / v) dimethyl sulfoxide (DMSO) and 20 μM roxadustat. The primary antibody used was purified mouse anti-human HIF-1α [catalog number 610959]. The secondary antibody used was rabbit anti-mouse IgG (D3V2A) mAb (HRP conjugate).

[1211] Blood cell count – reticulocytes and hemoglobin

[1212] A group of seven C57BL / 6 mice was administered 15 mg / kg or 30 mg / kg of a mediator (1% methylcellulose), compound 68 (Example 46), or 30 mg / kg of dapoxetine (2) via intraperitoneal (IP) injection twice daily. Blood samples were collected before treatment, on day 4, or on day 8 of treatment. Blood samples were analyzed using a Celltac Alpha MEK-6500K. Statistically significant increases in red blood cell count (RBC), hemoglobin (HGB), and hematocrit (HCT) were observed in the compound-treated mice.

[1213] Example 158 – Structure-activity relationship (SAR) study of PHD suppression

[1214] SAR studies on the PHD2 inhibition of the compounds of the present invention were conducted. Compounds were screened for PHD2 using an RF-MS hydroxylation assay. The results are shown in Table 1, indicating that the compounds of the present invention have high potency as PHD2 inhibitors.

[1215] Table 1

[1216]

[1217] Example 159 – 2OG oxygenase selectivity 68 (Example 46)

[1218] The selectivity of 68 (Example 46) for the following purified human 2OG-dependent oxygenases was determined using a reported experimental procedure (TLYeh et al., Chem Sci., 2017, 8, 7651-7668): FIH (a factor that inhibits HIF) (IC50 of 68) was used to detect the selectivity of 68 (Example 46) for the following purified human 2OG-dependent oxygenases: FIH (a factor that inhibits HIF) (IC50 of 68). 50 =>100μM), KDM4A: Lysine-specific demethylase 4A (IC50 68) 50 =>100μM), KDM5B: Lysine-specific demethylase 5B (IC50 68) 50 =>100μM) and KDM6B: lysine-specific demethylase 6B (IC50, 68%) 50 =>100μM). These results indicate that 68 is selective for PHD.

[1219] Example 160 – Immunoblotting of Hep3B cells treated with the compounds of the present invention

[1220] Hep3B cells were treated with compounds 117, 119, 122, and 123 of the present invention (Examples 93, 95, 98, and 99) at 100 μM (A) and 20 μM (B) PHD inhibitors for 3 hours. Western blot analysis showed the protein levels of HIF1-α and β-actin at the 3rd hour. Cell culture and immunoblotting protocols were performed as described in TLYeh et al., Chem Sci, 2017, 8, 7651-7668. Figure 1 As shown, the results of this embodiment demonstrate that the compound of the present invention is stable for HIF-1α.

[1221] Example 161 – Immunoblotting of HEK293 cells treated with lower concentrations of the compounds of the present invention

[1222] HEK293 T cells were treated with the 68 method of the present invention (Example 46). Cells were treated at 0.5, 1, 5, 10, 20, 50, and 100 μM for 18 hours. Immunoblots show the protein levels of HIF1-α and GAPDH detected at specific hourly time points after treatment. Established methods were used for cell culture and immunoblotting. Figure 2 As shown in the figure, the results of this embodiment demonstrate that the compound of the present invention stabilizes HIF-1α cells.

[1223] Example 162 – In vivo efficacy study

[1224] A group of seven C57BL / 6 mice was administered intraperitoneally (IP) twice daily with 15 mg / kg or 30 mg / kg of a mediator (1% methylcellulose), compound 68 (Example 46), or 30 mg / kg of dapoxetine (2). Figure 3 Samples were taken before treatment, on day 4 of treatment, or on day 8 of treatment. Blood samples were analyzed using a Celltac Alpha MEK-6500K. Statistically significant increases in red blood cell count (RBC), hemoglobin (HGB), and hematocrit (HCT) were observed in mice treated with the compound. 68 showed similar levels to 2. 15 mg / kg of 68 produced the same levels of RBC, HCT, and HGB as 30 mg / kg of 2. Indeed, the similarity between different doses of 68 suggests that the dose can be reduced. No side effects were observed in mice with the administered compound.

[1225] Example 163 – Selectivity of PHD

[1226] The lack of selectivity of enzyme inhibitors can lead to unpredictable and undesirable off-target effects. Currently available PHD inhibitors, including roxadustat, dapoxetine, molistat, decistat, and vadastat, show only limited target selectivity for PHD. For example, one or more of these inhibitors have been observed to inhibit collagen prolyl hydroxylases (CPHs), 1-oxoglutarate-containing iron-dependent oxygenase domain (OFGOD1), and 6-jumonji domain (JMJD6).

[1227] Compound 68 (Example 46) was evaluated to observe its inhibitory activity against sites that are typically inhibited by existing PHD inhibitors.

[1228] Production, purification and SPE-MS IC of recombinant PHD2 50 Measurement

[1229] such as Yeh et al 1 The production, purification, and IC50 determination of recombinant PHD2 were reported.

[1230] Production, purification and SPE-MS IC of recombinant FIH 50 Measurement

[1231] such as Yeh et al 1 The report describes the production, purification, and determination of the IC50 of recombinant FIH.

[1232] Production and purification of recombinant JMJD6

[1233] Such as Cockman et al and Islam et al 2,3 The recombinant JMJD6 was reported to be produced and purified as a full-length protein in Escherichia coli.

[1234] JMJD6 IC 50 Measurement

[1235] All reagents were from Sigma Aldrich and were of the highest grade available. Ferrous ammonium sulfate (FAS) was freshly prepared by dissolving in 20 mM HCl to 400 mM and then diluting to 1 mM in deionized water. 2-O-glutaric acid (2OG, 10 mM) and L-ascorbic acid (LAA, 50 mM) were freshly prepared by dissolving in deionized water. The inhibition of catalytic activity of recombinant human JMJD6 was assessed using the N-terminal peptide (RSKKRKKSKSRS) of RNA-binding motif protein 39 (RBM39 residues 31-42) and monitoring the appearance of the hydroxylated peptide product in 50 mM Tris.Cl at pH 7.5. Titrations of 68°C were performed using an ECHO 550 acoustic dispenser (Labcyte) for IC50 assays. 50 Measurement (3 times and 11 points IC) 50 The DMSO was dried and dispensed into 384-well polypropylene test plates. The final test concentration of DMSO was kept constant at 0.5% (v / v). Full-length JMJD6 was prepared at a concentration of 1.0 mM in 50 mM Tris.Cl (pH 7.5), and 25 μl was dispensed into 384-well plates. JMJD6 was pre-incubated with the compound dilution buffer for 15 min. The substrate (20 μM ferrous sulfate, 200 μM L-ascorbic acid, 10 μM RBM39) was then dispensed into the test plates. 31-42The reaction was initiated by dispensing 20 μM 2-oxoglutaric acid into each 384-well plate. The reaction was allowed to proceed for 30 minutes and then terminated by dispensing 10% formic acid (5 μl). Peptide analysis was performed by liquid chromatography-mass spectrometry (LCMS) using an Agilent 1290 Infinity II LC system equipped with an Agilent 1290 multisampler and an Agilent 1290 high-speed pump connected to an Agilent 6550 Precision Quadrupole Time-of-Flight (QTOF) mass spectrometer. 10 mL of the analytical mixture was injected into a ZORBAX RRHDEclipse Plus C18 column (Agilent). Solvent A consisted of LCMS-grade water containing 0.1% (v / v) formic acid, and solvent B consisted of acetonitrile containing 0.1% (v / v) formic acid. Peptides were separated using a stepwise gradient (0 min - 95% solvent A, 1 min - 80% solvent A, 3 min - 45% solvent A, 4 min - 45% solvent A, 5 min - 0% solvent A, 6 min - 0% solvent A, 7 min - 95% solvent A). The column was then reequilibrated with 95% (v / v / ) solvent A for 1 min, with all flow rates at 0.2 mL / min. The mass spectrometer was operated in positive ion mode (dry gas temperature (280 °C), dry gas flow rate (13 L / min), nebulizer pressure (40 psig), sheath gas temperature (350 °C), sheath gas flow rate (12 L / min), capillary voltage (4000 V), nozzle voltage (1000 V)). All acquired data were analyzed using Agilent MassHunter Qualitative Analysis (version B.07.00) software.

[1236] JMJD6 Solid Phase Extraction Mass Spectrometry (SPE-MS) Experiment

[1237] Use protein 4 (BRD4) containing the bromine domain. 511–550 ) 2The inhibitory activity of dapoxetine, roxadustat, and molistat against JMJD6 was assessed by monitoring the hydroxylation of the peptide product in 50 mM Tris.HCl pH 7.5 buffer using 40-mer peptide substrates. Titration of the compounds was prepared using an ECHO 550 acoustic dispenser (Labcyte). Eleven-point and three-fold dilutions of each compound were prepared and dried before being dispensed into 384-well polypropylene plates. A 1.0 mM full-length JMJD6 solution was prepared, and 25 μL of this solution was dispensed into each well of the plate using a multichannel dispenser (Thermo) equipped with a low-volume dispensing cassette. The compound dilutions were pre-incubated with JMJD6 for 15 min. Titration was performed using 50 mM Tris.HCl pH 7.5 (200 mM L-ascorbate, 20 mM ferrous ammonium sulfate, 20 mM 2-oxoglutarate, and 10 mM JMJD6 substrate BRD4). 511–550 2 Dispense 25 μl of the substrate mixture to initiate the enzyme reaction. Allow the reaction to proceed at room temperature for 15 minutes and stop by dispensing 10% (v / v) formic acid (5 μl). The final concentration of DMSO is 0.5% (v / v). Transfer the test plate to a RapidFire RF365 high-throughput sampling robot (Agilent) connected to an Agilent 6550 quadrupole time-of-flight (Q-TOF) mass spectrometer. The sample is aspirated under vacuum and loaded onto a C4 solid-phase extraction (SPE) column. Wash the C4 SPE column with 0.1% (v / v) aqueous formic acid at a flow rate of 1.5 mL / min for 5.5 s to remove nonvolatile buffer salts. Then elute the peptide from the SPE to the mass spectrometer with 80% (v / v) acetonitrile and 20% (v / v) water containing 0.1% (v / v) formic acid at a flow rate of 1.6 mL / min. The mass spectrometer was operated in positive ion mode, with a dry gas temperature of 280°C, a dry gas flow rate of 13 L / min, a nebulizer pressure of 40 psig, a sheath gas temperature of 350°C, a sheath gas flow rate of 12 L / min, a capillary voltage of 4000 V, and a nozzle voltage of 1000 V. Peak area data for the +8 charge state were integrated using RapidFire Integrator software (Agilent). BRD4 was calculated using the following formula. 511-550 Percentage converted to hydroxylated products:

[1238] % conversion rate = 100 × hydroxylated / (hydroxylated + non-hydroxylated peptides): IC 50 The data was determined from the nonlinear regression plot using GraphPadprism 6.0.

[1239] KDM4A, KDM5B and KDM6B liquid chromatography-mass spectrometry (LCMS) analysis

[1240] The inhibitory activity of 68 was assessed by monitoring the demethylation of the corresponding peptide substrates of KDM4A, KDM5B, and KDM6B. The peptide substrate of KDM4A is a 15-mer histone-H3 derivative (ARTAQTARK(me3)STGGI), which was developed by Hutchinson et al. 4 The reported results were synthesized by GL Biochem (Shanghai) Co., Ltd. (Shanghai, China). The peptide substrate for KDM5B is a 21-mer histone H3 peptide (ARTK(me3)QTARKSTGGKAPRKQLA), synthesized by the Peptide Protein Institute (Hampshire, UK). The peptide substrate for KDM6B is a 17-mer histone H3 peptide (LATKAARK(me3)SAPATGGVK), synthesized by GL Biochem (Shanghai) Co., Ltd. (Shanghai, China). Recombinant KDM4A, residues M1–L359, was produced and purified in *E. coli*, as described by Ng et al. 5 As reported, recombinant KDM5B, residues M1–R822, was expressed and purified in a baculovirus expression system, as described by Johansson et al. 6 As previously mentioned, KDM6B, residues D1141–E1590, is expressed and purified in *E. coli*, as described by Rose et al. 7 As previously stated.

[1241] The KDM4A reaction was performed under optimized buffer conditions (50 mM MES, pH 7.0). KDM4A (0.15 mM) was pre-incubated at 68°C (100 mM) for 15 min, and the enzyme reaction was initiated by adding substrate (100 mM L-ascorbate, 10 mM ferrous ammonium sulfate, 10 mM 2-oxoglutarate, and 10 mM peptide substrate). The enzyme reaction was allowed to proceed for 50 min and terminated by adding formic acid to a final concentration of 1% (v / v). A control reaction was also set up with 0.5% (v / v) DMSO, and a reaction was also set up with a known KDM4A inhibitor (50 mM 2,4-pyridinedicarboxylic acid). 8 ( ) is a comparison.

[1242] The KDM5B enzyme reaction was performed under optimized buffer conditions (50 mM MES pH 7.0, 50 mM NaCl, 1 mM TPCEP). KDM5A (0.15 mM) was pre-incubated at 68°C (100 mM) for 15 min, and the enzyme reaction was initiated by adding substrate (100 mM L-ascorbate, 10 mM ferrous ammonium sulfate, 10 mM 2-oxoglutarate, and 5 mM peptide). The enzyme reaction was allowed to proceed for 30 min and terminated by adding formic acid to a final concentration of 1% (v / v). Control reactions included a 0.5% DMSO control and a reaction with a known KDM5B inhibitor (10 mM KDOAM25).9 ) reaction.

[1243] The KDM6B reaction was performed under optimized buffer conditions (50 mM MES, pH 7.0). KDM6B (0.15 mM) was pre-incubated at 68°C (100 mM) for 15 min, and the enzyme reaction was initiated by adding substrate (100 mM L-ascorbate, 10 mM ferrous ammonium sulfate, 10 mM 2-oxoglutarate, and 5 mM peptide). The enzyme reaction was allowed to proceed for 30 min and terminated by adding formic acid to a final concentration of 1% (v / v). Control reactions included a 0.5% DMSO control and a reaction with a known KDM6B inhibitor (10 mM GSKJ1). 10 ) reaction.

[1244] The enzyme reaction was transferred to a 96-well polypropylene plate, and peptide analysis was performed by LCMS using an Agilent 1290 Infinity II liquid chromatography system equipped with an Agilent 1290 multisampler and an Agilent 1290 high-speed pump connected to an Agilent 6550 Precision Quadrupole Time-of-Flight (QTOF) mass spectrometer. 4 ml of the enzyme reaction solution was injected and loaded onto a ZORBAXRRHD Eclipse Plus C18 column (Agilent Technologies, CA, US). Solvent A consisted of LCMS-grade water containing 0.1% (v / v) formic acid, and solvent B consisted of acetonitrile containing 0.1% (v / v) formic acid. Peptides were separated using a stepwise gradient (0 min - 95% solvent A, 1.0 min - 80% solvent A, 3.0 min - 45% solvent A, 4.0 min - 45% solvent A, 5.0 min - 0% solvent A, 6.0 min - 0% solvent A, 7.0 min - 95% solvent A). The column was then reequilibrated with 95% solvent A for 3 minutes, with all flow rates at 0.2 mL / min. The mass spectrometer was operated in positive ion mode with the following settings: dry gas temperature 280 °C, dry gas flow rate 13 L / min, nebulizer pressure 40 psig, sheath gas temperature 350 °C, sheath gas flow rate 12 L / min, capillary voltage 4000 V, and nozzle voltage 1000 V. All acquired data were analyzed using Agilent MassHunter Qualitative Analysis (version B.07.00) software.

[1245] OGFOD1 solid phase extraction mass spectrometry (SPE-MS) assay

[1246] The OGFOD1 gene was cloned into the pET-28a vector and expressed and purified in *E. coli* strain BL21(DE3) to obtain a full-length enzyme (Met1–Glu542) with an N-terminal 6-His tag. The synthetic peptide substrate RPS 23 (Ala47–Lys76AKGIVLEKVGVEAKQPNSAIRKAVRVQLIK-NH2) was synthesized by GL Biochem (Shanghai, China) with a purity >95%. Ferrous ammonium sulfate (FAS), 2-oxoglutaric acid (2-OG), and L-ascorbic acid (LAA) were obtained from Sigma Aldrich. Ferrous ammonium sulfate was freshly prepared by dissolving 50–100 mg of ferrous ammonium sulfate to a concentration of 400 mM in 20 mM HCl, followed by further dilution to 1 mM in deionized water. 2-OG (10 mM) and L-AA (50 mM) were both freshly prepared in deionized water.

[1247] IC 50 The assays were performed using polypropylene plates in 384-well plate format (Greiner Bio One, catalog number 781096). Compounds were prepared as 20 mM DMSO stock solutions, and all compounds were partitioned using an ECHO 550 acoustic dispenser (Labcyte, Sunnyvale, CA). The positive control compound (2,4-PDCA, 100 mM) was partitioned into column 1 (250 nmol), and DMSO into column 13 (250 nmol). All test compounds were serially diluted (throughout 11 IC50 intervals). 50The OGFOD1 was diluted approximately 3-fold in series, and 250 nmol of each dilution was dispensed in duplicate into polypropylene plates. OGFOD1 was diluted to 0.3 mM in test buffer (50 mM Tris.Cl, pH 7.5) and dispensed (25 mL) into 384-well compound plates using a multichannel combination reagent dispenser (Thermo Scientific, code 5840300) with a small plastic tip dispensing box (Thermo Scientific, code 24073290). The compound was pre-incubated with OGFOD1 for 15 minutes, and the enzyme reaction was initiated by dispensing 25 mL of substrate solution (200 mL M AA, 20 mM FAS, 20 mM 2-OG, 10 mM RPS23 (47-76) peptide) into test buffer. The final concentration of DMSO used in the experiment was 0.5%. The reaction was allowed to proceed for 20 minutes, then stopped by adding 10% (v / v) formic acid (5 ml), and the plate was transferred to a RapidFire RF 365 coupled with an Agilent 6550 Accurate-Mass Quadrupole Time-of-Flight (QTOF) mass spectrometer. The sample was aspirated under vacuum and loaded onto a C4 solid-phase extraction (SPE) column. The SPE column was washed for 5.5 seconds with LCMS-grade water containing 0.1% (v / v) formic acid at a flow rate of 1.5 ml / min to remove non-volatile buffer components. After washing with water, the peptides were eluted from the C4 SPE column in an organic elution step (80% (v / v) acetonitrile, 20% (v / v) LCMS-grade water containing 0.1% formic acid) at a flow rate of 1.6 ml / min for 5.5 seconds. The mass spectrometer was operated in positive ion mode with the following settings: dry gas temperature (280 °C), dry gas flow rate (13 L / min), nebulizer pressure (40 psig), sheath gas temperature (350 °C), sheath gas flow rate (12 L / min), capillary voltage (4000 V), and nozzle voltage (1000 V). Ion data for the substrate and the hydroxylated (+16) peptide product were extracted, and peak area data were integrated using RapidFire Integrator software (version 4.3.017235, Agilent). The percentage of peptide substrate converted to the hydroxylated product was calculated in Excel, and IC50 curves were generated using GraphPad Prism version 7.0.

[1248] in conclusion

[1249] As in this application Figure 4 As shown, compound 68 (Example 46) exhibited very high IC50 values ​​for each off-target site tested. 50This indicates that the compound exhibits good selectivity and specificity in inhibiting PHD. Therefore, compared to existing PHD inhibitors, the compound of the present invention can be expected to have reduced side effects.

[1250] References

[1251] 1Yeh, TLet al. Molecular and cellular mechanisms of HIF prolylhydroxylase inhibitors in clinical trials. Chem Sci 8, 7651-7668 (2017). https: / / doi.org / 10.1039 / c7sc02103h

[1252] 2Cockman, ME et al. Widespread hydroxylation of unstructured lysine-rich protein domains by JMJD6. Proc Natl Acad Sci USA 119, e2201483119 (2022). https: / / doi.org / 10.1073 / pnas.2201483119

[1253] 3Islam, MS et al. Biochemical and structural investigations clarify the substrate selectivity of the 2-oxoglutarate oxygenase JMJD6.J Biol Chem294, 11637-11652 (2019). https: / / doi.org / 10.1074 / jbc.RA119.008693

[1254] 4Hutchinson, SE et al. Enabling Lead Discovery for Histone LysineDemethylases by High-Throughput RapidFire Mass Spectrometry. Journal of Biomolecular Screening 17, 39-48 (2011). https: / / doi.org / 10.1177 / 1087057111416660

[1255] 5Ng,S.S.et al.Crystal structures of histone demethylase JMJD2A revealbasis for substrate specificity.Nature 448,87-91(2007). https: / / doi.org / 10.1038 / nature05971

[1256] 6Johansson,C.et al.Structural analysis of human KDM5B guides histonedemethylase inhibitor development.Nature Chemical Biology 12,539-545(2016). https: / / doi.org / 10.1038 / nchembio.2087

[1257] 7Rose,N.R.et al.Plant Growth Regulator Daminozide Is a SelectiveInhibitor of Human KDM2 / 7Histone Demethylases.Journal of Medicinal Chemistry55,6639-6643(2012). https: / / doi.org / 10.1021 / jm300677j

[1258] 8Rose,N.R.et al.Inhibitor scaffolds for 2-oxoglutarate-dependenthistone lysine demethylases.J Med Chem 51,7053-7056(2008). https: / / doi.org / 10.1021 / jm800936s

[1259] 9Tumber,A.et al.Potent and Selective KDM5 Inhibitor Stops CellularDemethylation of H3K4me3 at Transcription Start Sites and Proliferation ofMM1SMyeloma Cells.Cell Chemical Biology 24,371-380(2017). https: / / doi.org / https: / / doi.org / 10.1016 / j.chembiol.2017.02.006

[1260] 10Kruidenier,L.et al.A selective jumonji H3K27 demethylase inhibitormodulates the proinflammatory macrophage response.Nature 488,404-408(2012). https: / / doi.org / 10.1038 / nature11262

[1261] Other aspects and embodiments of the invention are defined in the following numbered clauses.

[1262] 1. A compound that is a substituted azine of formula (I) or a pharmaceutically acceptable salt thereof.

[1263]

[1264] in

[1265] X is CR 6 Or N;

[1266] R 0 It is H or unsubstituted or substituted C 1-6 alkyl;

[1267] R 1 It is H, unsubstituted or substituted C 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN, –N(R) t )C(O)N(R u )R v –C(O)OR w or –C(O)N(R) x )R 7 ;

[1268] R 2 It is H, –OR q C, whether substituted or not 1-6 Alkyl; and R 3 It is H, –OR 8 C, whether substituted or not 1-6 Alkyl; or R 2 It is –N= and R 3 =C(R) y ) – and R 2 and R 3 Together forming formula – N = C(R) y )– groups;

[1269] R 4 It is H, unsubstituted or substituted C 1-6 Alkyl, –OR 9 Or –C(O)OR10 ;

[1270] R 5 It is H, unsubstituted or substituted C 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN, –N(R) t )C(O)N(R u )R v –C(O)OR w or –C(O)N(R) x )R 7 ;

[1271] R 6 It is H or unsubstituted or substituted C 1-6 alkyl;

[1272] R 7 It is –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar, –Ary–Ar, –CH(R 11 –Cyc, –CH2C≡CCH3, –Cyc, or –Ar, where Ar is an unsubstituted or substituted aryl or an unsubstituted or substituted heteroaryl, Ary is an unsubstituted or substituted arylene or an unsubstituted or substituted heteroaryl, and Cyc is an unsubstituted or substituted C 3-10 cycloalkyl, R 11 It is H, –C(O)OR z C, whether substituted or not 1-4 alkyl;

[1273] R 8 R 9 and R 10 Each is independently selected from H and unsubstituted or substituted C. 1-6 Alkyl groups; and

[1274] R t R u R v R w R x R y and R z Each is independently selected from H, unsubstituted or substituted C. 1-6 Alkyl groups and unsubstituted or substituted phenyl groups;

[1275] R q It is H, unsubstituted or substituted C 1-6 Alkyl groups, or unsubstituted or substituted phenyl groups;

[1276] The condition is R 1 and R 5 One of them is –C(O)N(R)x )R 7 And R 1 and R 5 The other one is H, unsubstituted or substituted C. 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN, –N(R) t )C(O)N(R u )R v Or –C(O)OR w .

[1277] 2. The compound according to Clause 1, wherein:

[1278] R 0 Is it H or unsubstituted C? 1-6 alkyl;

[1279] R 1 It is H, –CN, –C(O)OR w or –C(O)N(R) x )R 7 ;

[1280] R 2 Is it H or unsubstituted C? 1-6 Alkyl; and R 3 Is it H or –OR 8 Or R 2 It is –N= and R 3 =C(R) y ) – and R 2 and R 3 Together forming formula – N = C(R) y )– groups;

[1281] R 4 It is H, –OR 9 Or –C(O)OR 10 ;

[1282] R 5 It is H, –CN, –C(O)OR w or –C(O)N(R) x )R 7 ;

[1283] R 6 It is H;

[1284] R 7 It is –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar, –Ary–Ar or –CH(R 11)–Cyc, where Ar is an unsubstituted or substituted aryl or an unsubstituted or substituted heteroaryl, Ary is an unsubstituted arylene or an unsubstituted heteroaryl, and Cyc is an unsubstituted or substituted C 3-10 cycloalkyl, R 11 It is H, –C(O)OR z or unreplaced C 1-4 alkyl;

[1285] R 8 R 9 and R 10 Each is independently selected from H and unsubstituted or substituted C. 1-6 Alkyl; and

[1286] R x It is H, R z It is H, R y Is it H or unsubstituted C? 1-6 Alkyl, R w It is H, unsubstituted C 1-6 Alkyl or phenyl or –OC(O)R ww Replacement C 1-6 Alkyl, wherein R ww It is phenyl, unsubstituted C 1-6 Alkyl, –N(R) a (R) b ), –C(O)R c –OR d Or amino acids, of which R a R b R c and R d Each is independently selected from H, unsubstituted or substituted C. 1-6 Alkyl groups and amino acids;

[1287] The condition is R 1 and R 5 One of them is –C(O)N(R) x )R 7 And R 1 and R 5 Another one is H, –CN, or –C(O)ORw.

[1288] 3. The compound according to Clause 1 or Clause 2, wherein:

[1289] R 0 It is hydrogen or methyl;

[1290] R 1 It is H, –CN, –C(O)OR w or –C(O)N(R) x )R 7 ;

[1291] R 2 It is H or methyl; and R 3 Is it H or –OR 8 Or R 2 It is –N= and R 3 =C(R) y ) – and R 2 and R 3 Together forming formula – N = C(R) y )– groups;

[1292] R 4 It is H, –OR 9 Or –C(O)OR 10 ;

[1293] R 5 It is H, –CN, –C(O)OR w or –C(O)N(R) x )R 7 ;

[1294] R 6 It is H;

[1295] R 7 It is –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar, –Ary–Ar or –CH(R 11 –Cyc; where Ar is an unsubstituted phenyl, unsubstituted pyrimidinyl, unsubstituted benzothiazole, or a phenyl substituted with –C(O)OH, –C(O)OMe, –C(O)Oet, –C(O)NH2, –C(O)N(H)Me, –Ome, or N-morpholino; Ary is an unsubstituted phenylene or unsubstituted pyridylene; Cyc is an unsubstituted cyclohexyl or a cyclohexyl substituted with –CF3 or –OCF3; and R 11 It is H, –C(O)OR z or methyl;

[1296] R 8 R 9 and R 10 Each is independently selected from H and unsubstituted C. 1-6 Alkyl, or phenyl or –OC(O)R 99 Replacement C 1-6 Alkyl, wherein R 99 It is phenyl, unsubstituted C 1-6 Alkyl, –N(R) a (R) b ), –C(O)R c –OR d Or amino acids, of which R aR b R c and R d Each is independently selected from H, unsubstituted or substituted C. 1-6 Alkyl groups and amino acids; and

[1297] R x It is H;

[1298] R z It is H;

[1299] R w It is H, unsubstituted C 1-6 Alkyl groups or phenyl groups or -OC(O)R ww Replacement C 1-6 Alkyl, wherein R ww It is a phenyl or unsubstituted C 1-6 Alkyl; and

[1300] R y It is H or methyl;

[1301] The condition is R 1 and R 5 One of them is –C(O)N(R) x )R 7 And R 1 and R 5 Another one is H, –CN, or –C(O)ORw.

[1302] 4. A compound according to any one of clauses 1 to 3, wherein:

[1303] (1)(a)R 5 It is –C(O)N(R) x )R 7 , and (b)R 3 Yes – OR 8 or R 4 Yes – OR 9 ;or

[1304] (2)(a)R 1 It is –C(O)N(Rx)R7, and (b)R 4 Yes – OR 9 Or –C(O)OR 10 , or R 5 It is –C(O)OR w .

[1305] 5. The compound according to any one of the preceding clauses, wherein said substituted azazine has formula (Ia).

[1306]

[1307] in

[1308] X is CR 6 Or N;

[1309] R 0 It is H or unsubstituted or substituted C 1-6 alkyl;

[1310] R 1 It is H, unsubstituted or substituted C 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN or –C(O)OR w ;

[1311] R 2 It is H, –OR q C, whether substituted or not 1-6 alkyl;

[1312] R 3 It is H or unsubstituted or substituted C 1-6 alkyl;

[1313] R 6 It is H or unsubstituted or substituted C 1-6 alkyl;

[1314] R 7 It is –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar, –Ary–Ar, –CH(R 11 –Cyc, –CH2C≡CCH3, –Cyc, or –Ar, where Ar is an unsubstituted or substituted aryl or an unsubstituted or substituted heteroaryl, Ary is an unsubstituted or substituted arylene or an unsubstituted or substituted heteroaryl, and Cyc is an unsubstituted or substituted C 3-10 cycloalkyl, R 11 It is H, –C(O)OR z C, whether substituted or not 1-4 alkyl;

[1315] R 9 It is H or unsubstituted or substituted C 1-6 Alkyl; and

[1316] R w R x and R z Each is independently selected from H, unsubstituted or substituted C. 1-4 Alkyl groups and unsubstituted or substituted phenyl groups; and

[1317] R q It is H, unsubstituted or substituted C 1-6 Alkyl, or unsubstituted or substituted phenyl.

[1318] 6. The compound according to clause 5, wherein R 9 It's H.

[1319] 7. The compound according to any one of the preceding clauses, wherein said substituted acridine has any one of the following structures:

[1320]

[1321]

[1322]

[1323]

[1324] 8. The compound according to any one of clauses 1 to 4, wherein said substituted azazine has the formula (Ib).

[1325]

[1326] in

[1327] R 0 It is H or unsubstituted or substituted C 1-6 alkyl;

[1328] R 1 It is H, unsubstituted or substituted C 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN or –C(O)OR w ;

[1329] R 2 It is H, –OR q C, whether substituted or not 1-6 alkyl;

[1330] R 4 It is H or unsubstituted or substituted C 1-6 alkyl;

[1331] R 6 It is H or unsubstituted or substituted C 1-6 alkyl;

[1332] R 7 It is –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar, –Ary–Ar, –CH(R 11 –Cyc, –Cyc, or –Ar, where Ar is an unsubstituted or substituted aryl or an unsubstituted or substituted heteroaryl, Ary is an unsubstituted or substituted arylene or an unsubstituted or substituted heteroaryl, and Cyc is an unsubstituted or substituted C3-10 cycloalkyl, R 11 It is H, –C(O)OR z C, whether substituted or not 1-4 alkyl;

[1333] R 8 It is H or unsubstituted or substituted C 1-6 alkyl;

[1334] R w R x and R z Each is independently selected from H, unsubstituted or substituted C. 1-4 Alkyl groups and unsubstituted or substituted phenyl groups; and

[1335] R q It is H, unsubstituted or substituted C 1-6 Alkyl, or unsubstituted or substituted phenyl.

[1336] 9. The compound according to clause 8, wherein R 8 It is H.

[1337] 10. The compound according to Clause 8 or Clause 9, wherein the substituted azazine has any of the following structures.

[1338]

[1339] 11. The compound according to any one of clauses 1 to 4, wherein said substituted azazine has the formula (Ic).

[1340]

[1341] in

[1342] R 0 It is H or unsubstituted or substituted C 1-6 alkyl;

[1343] R 2 It is H, –OR q C, whether substituted or not 1-6 alkyl;

[1344] R 3 It is H, –OR 8 C, whether substituted or not 1-6 alkyl;

[1345] R 4 It is H, unsubstituted or s...

Claims

1. A compound that is a substituted azine of formula (I) or a pharmaceutically acceptable salt thereof. in X is CR 6 Or N; R 0 It is H or unsubstituted or substituted C 1-6 alkyl; R 1 It is H, unsubstituted or substituted C 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN, –N(R) t )C(O)N(R u )R v –C(O)OR w or –C(O)N(R) x )R 7 ; R 2 It is H, –OR q C, whether substituted or not 1-6 Alkyl; and R 3 It is H, –OR 8 C, whether substituted or not 1-6 Alkyl; or R 2 It is –N= and R 3 =C(R) y ) – and R 2 and R 3 Together forming formula – N = C(R) y )– groups; R 4 It is H, unsubstituted or substituted C 1-6 Alkyl, –OR 9 Or –C(O)OR 10 ; R 5 It is H, unsubstituted or substituted C 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN, –N(R) t )C(O)N(R u )R v –C(O)OR w or –C(O)N(R) x )R 7 ; R 6 It is H or unsubstituted or substituted C 1-6 alkyl; R 7 It is –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar, –Ary–Ar, –CH(R 11 –Cyc, –CH2C≡CCH3, –Cyc, or –Ar, where Ar is an unsubstituted or substituted aryl or an unsubstituted or substituted heteroaryl, Ary is an unsubstituted or substituted arylene or an unsubstituted or substituted heteroaryl, and Cyc is an unsubstituted or substituted C 3-10 Cycloalkyl, and R 11 It is H, –C(O)OR z C, whether substituted or not 1-4 Alkyl; R 8 R 9 and R 10 Each is independently selected from H and unsubstituted or substituted C. 1-6 Alkyl groups; and R t R u R v R w R x R y and R z Each is independently selected from H, unsubstituted or substituted C. 1-6 Alkyl groups and unsubstituted or substituted phenyl groups; R q It is H, unsubstituted or substituted C. 1-6 Alkyl groups, or unsubstituted or substituted phenyl groups; The condition is R 1 and R 5 One of them is –C(O)N(R) x )R 7 And R 1 and R 5 The other one is H, unsubstituted or substituted C. 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN, –N(R) t )C(O)N(R u )R v Or –C(O)OR w .

2. The compound according to claim 1, wherein: R 0 Is it H or unsubstituted C? 1-6 alkyl; R 1 is H, –CN, –C(O)OR w or –C(O)N(R x )R 7 ; R 2 Is it H or unsubstituted C? 1-6 Alkyl; and R 3 Is it H or –OR 8 Or R 2 It is –N= and R 3 =C(R) y ) – and R 2 and R 3 Together forming formula – N = C(R) y )– groups; R 4 is H, –OR 9 or –C(O)OR 10 ; R 5 is H, –CN, –C(O)OR w or –C(O)N(R x )R 7 ; R 6 It is H; R 7 It is –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar, –Ary–Ar or –CH(R 11 )–Cyc, where Ar is an unsubstituted or substituted aryl or an unsubstituted or substituted heteroaryl, Ary is an unsubstituted arylene or an unsubstituted heteroaryl, and Cyc is an unsubstituted or substituted C 3-10 Cycloalkyl, and R 11 It is H, –C(O)OR z or unreplaced C 1-4 alkyl; R 8 R 9 and R 10 Each is independently selected from H and unsubstituted or substituted C. 1-6 Alkyl; and R x It is H, R z It is H, R y Is it H or unsubstituted C? 1-6 Alkyl, R w It is H, unsubstituted C 1-6 Alkyl or phenyl or –OC(O)R ww Replacement C 1-6 Alkyl, wherein R ww It is phenyl, unsubstituted C 1-6 Alkyl, –N(R) a (R) b –C(O)R c –OR d Or amino acids, of which R a R b R c and R d Each is independently selected from H, unsubstituted or substituted C. 1-6 Alkyl groups and amino acids; The condition is R 1 and R 5 One of them is –C(O)N(R) x )R 7 And R 1 and R 5 The other one is H, –CN, or –C(O)OR. w .

3. The compound according to claim 1 or claim 2, wherein: R 0 It is hydrogen or methyl; R 1 is H, –CN, –C(O)OR w or –C(O)N(R x )R 7 ; R 2 It is H or methyl; and R 3 Is it H or –OR 8 Or R 2 It is –N= and R 3 =C(R) y )–, and R 2 and R 3 Together forming formula – N = C(R) y )– groups; R 4 is H, –OR 9 or –C(O)OR 10 ; R 5 is H, –CN, –C(O)OR w or –C(O)N(R x )R 7 ; R 6 It is H; R 7 It is –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar, –Ary–Ar or –CH(R 11 –Cyc; where Ar is an unsubstituted phenyl, unsubstituted pyrimidinyl, unsubstituted benzothiazole, or a phenyl substituted with –C(O)OH, –C(O)Ome, –C(O)Oet, –C(O)NH2, –C(O)N(H)Me, –Ome, or N-morpholino; Ary is an unsubstituted phenylene or unsubstituted pyridylene; Cyc is an unsubstituted cyclohexyl or a cyclohexyl substituted with –CF3 or –OCF3; and R 11 It is H, –C(O)OR z or methyl; R 8 R 9 and R 10 Each is independently selected from H and unsubstituted C. 1-6 Alkyl groups and those with phenyl groups or –OC(O)R 99 Replacement C 1-6 Alkyl, wherein R 99 It is phenyl, unsubstituted C 1-6 Alkyl, –N(R) a (R) b –C(O)R c –OR d Or amino acids, of which R a R b R c and R d Each is independently selected from H, unsubstituted or substituted C. 1-6 Alkyl groups and amino acids; and R x It is H; R z It is H; R w It is H, unsubstituted C 1-6 Alkyl groups or phenyl groups or -OC(O)R ww Replacement C 1-6 Alkyl, wherein R ww It is a phenyl or unsubstituted C 1-6 Alkyl; and R y It is H or methyl; The condition is R 1 and R 5 One of them is –C(O)N(R) x )R 7 And R 1 and R 5 The other one is H, –CN, or –C(O)OR. w .

4. The compound according to any one of claims 1 to 3, wherein: (1)(a) R 5 is –C(O)N(R x )R 7 , and (b) R 3 is –OR 8 or R 4 is –OR 9 ; or (2)(a)R 1 It is –C(O)N(Rx)R7, and (b)R 4 Yes – OR 9 Or –C(O)OR 10 , or R 5 It is –C(O)OR w .

5. The compound according to any one of the preceding claims, wherein the substituted azazine has the formula (Ia). in X is CR 6 Or N; R 0 It is H or unsubstituted or substituted C 1-6 alkyl; R 1 It is H, unsubstituted or substituted C 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN or –C(O)OR w ; R 2 It is H, –OR q C, whether substituted or not 1-6 alkyl; R 3 It is H or unsubstituted or substituted C 1-6 alkyl; R 6 It is H or unsubstituted or substituted C 1-6 alkyl; R 7 It is –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar, –Ary–Ar, –CH(R 11 –Cyc, –CH2C≡CCH3, –Cyc, or –Ar, where Ar is an unsubstituted or substituted aryl or an unsubstituted or substituted heteroaryl, Ary is an unsubstituted or substituted arylene or an unsubstituted or substituted heteroaryl, and Cyc is an unsubstituted or substituted C 3-10 cycloalkyl, R 11 It is H, –C(O)OR z C, whether substituted or not 1-4 alkyl; R 9 It is H or unsubstituted or substituted C 1-6 Alkyl; and R w R x and R z Each is independently selected from H, unsubstituted or substituted C. 1-4 Alkyl groups and unsubstituted or substituted phenyl groups; and R q It is H, unsubstituted or substituted C 1-6 Alkyl, or unsubstituted or substituted phenyl.

6. The compound according to claim 5, wherein R 9 It is H.

7. The compound according to any one of the preceding claims, wherein the substituted azazine has any one of the following structures:

8. The compound according to any one of claims 1 to 4, wherein the substituted azazine has the formula (Ib). in R 0 It is H or unsubstituted or substituted C 1-6 alkyl; R 1 It is H, unsubstituted or substituted C 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN or –C(O)OR w ; R 2 It is H, –OR q C, whether substituted or not 1-6 alkyl; R 4 It is H or unsubstituted or substituted C 1-6 alkyl; R 6 It is H or unsubstituted or substituted C 1-6 alkyl; R 7 It is –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar, –Ary–Ar, –CH(R 11 –Cyc, –Cyc, or –Ar, where Ar is an unsubstituted or substituted aryl or an unsubstituted or substituted heteroaryl, Ary is an unsubstituted or substituted arylene or an unsubstituted or substituted heteroaryl, and Cyc is an unsubstituted or substituted C 3-10 Cycloalkyl, and R 11 It is H, –C(O)OR z C, whether substituted or not 1-4 alkyl; R 8 It is H or unsubstituted or substituted C 1-6 alkyl; R w R x and R z Each is independently selected from H, unsubstituted or substituted C. 1-4 Alkyl groups and unsubstituted or substituted phenyl groups; and R q It is H, unsubstituted or substituted C 1-6 Alkyl, or unsubstituted or substituted phenyl.

9. The compound according to claim 8, wherein R 8 It is H.

10. The compound according to claim 8 or 9, wherein the substituted azazine has any one of the following structures.

11. The compound according to any one of claims 1 to 4, wherein the substituted azazine has the formula (Ic). in R 0 It is H or unsubstituted or substituted C 1-6 alkyl; R 2 It is H, –OR q C, whether substituted or not 1-6 alkyl; R 3 It is H, –OR 8 C, whether substituted or not 1-6 alkyl; R 4 It is H, unsubstituted or substituted C 1-6 Alkyl, –OR 9 Or –C(O)OR 10 ; R 5 It is H, unsubstituted or substituted C 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN or –C(O)OR w ; R 6 It is H or unsubstituted or substituted C 1-6 alkyl; R 7 It is –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar, –Ary–Ar, –CH(R 11 –Cyc, –Cyc, or –Ar, where Ar is an unsubstituted or substituted aryl or an unsubstituted or substituted heteroaryl, Ary is an unsubstituted or substituted arylene or an unsubstituted or substituted heteroaryl, and Cyc is an unsubstituted or substituted C 3-10 Cycloalkyl, and R 11 It is H, –C(O)OR z C, whether substituted or not 1-4 alkyl; R 8 R 9 and R 10 Each is independently selected from H and unsubstituted or substituted C. 1-6 alkyl; R w R x and R z Each is independently selected from H, unsubstituted or substituted C. 1-4 Alkyl groups and unsubstituted or substituted phenyl groups; and R q It is H, unsubstituted or substituted C 1-6 Alkyl, or unsubstituted or substituted phenyl.

12. The compound according to claim 11, wherein the substituted azazine has any one of the following structures:

13. The compound according to any one of claims 1 to 4, wherein the substituted azazine has the formula (Id). in R 0 It is H or unsubstituted or substituted C 1-6 alkyl; R 1 It is H, unsubstituted or substituted C 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN or –C(O)OR w ; R 6 It is H or unsubstituted or substituted C 1-6 alkyl; R 7 It is –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar, –Ary–Ar, –CH(R 11 –Cyc, –Cyc, or –Ar, where Ar is an unsubstituted or substituted aryl or an unsubstituted or substituted heteroaryl, Ary is an unsubstituted or substituted arylene or an unsubstituted or substituted heteroaryl, and Cyc is an unsubstituted or substituted C 3-10 cycloalkyl, R 11 It is H, –C(O)OR z C, whether substituted or not 1-4 alkyl; R 9 It is H or unsubstituted or substituted C 1-6 Alkyl; and R w R x R y and R z Each is independently selected from H, unsubstituted or substituted C. 1-4 Alkyl groups and unsubstituted or substituted phenyl groups.

14. The compound according to claim 13, wherein the substituted azazine has any one of the following structures:

15. The compound according to any one of claims 1 to 3, wherein the substituted azazine has any one of the following structures.

16. The compound according to claim 5, wherein R 9 Is it unsubstituted or substituted C? 1-6 Alkyl groups, optionally substituted azazines of formula (Ia) having any of the following structures 17. The compound according to claim 8, wherein R 8 Is it unsubstituted or substituted C? 1-6 Alkyl groups, optionally substituted azazines of formula (Ib) having any of the following structures 18. The compound according to claim 11, wherein R 4 Yes – OR 9 Or –C(O)OR 10 , and / or R 5 It is –C(O)OR w , where R 9 R 10 and R w Each is an unsubstituted or substituted C. 1-6 Alkyl; optionally, wherein the substituted azazine of formula (Ic) has any of the following structures.

19. The compound according to claim 13, wherein R 9 Is it unsubstituted or substituted C? 1-6 Alkyl; optionally, the substituted azazine of formula (Id) has the following structure 20. A compound that is a substituted pyrimidine of formula (IV) or a pharmaceutically acceptable salt thereof. in R 0 It is H or unsubstituted or substituted C 1-6 alkyl; R 2 It is H, –OR q C, whether substituted or not 1-6 alkyl; R 4 Yes – OR 9 , where R 9 Selected from H and unsubstituted or substituted C 1-6 alkyl; R 5 It is H, unsubstituted or substituted C 1-6 Alkyl, unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, –CN or –C(O)OR w ; R 6 It is H or unsubstituted or substituted C 1-6 alkyl; R 7 It is –CH(R) 11 –Ar、–CH(R) 11 )–Ary–Ar, –Ary–Ar, –CH(R 11 –Cyc, –Cyc, or –Ar, where Ar is an unsubstituted or substituted aryl or an unsubstituted or substituted heteroaryl, Ary is an unsubstituted or substituted arylene or an unsubstituted or substituted heteroaryl, and Cyc is an unsubstituted or substituted C 3-10 cycloalkyl, R 11 It is H, –C(O)OR z C, whether substituted or not 1-4 alkyl; R x It is H, unsubstituted or substituted C 1-4 Alkyl groups, or unsubstituted or substituted phenyl groups; R w and R z Each is independently selected from H, unsubstituted or substituted C. 1-4 Alkyl groups and unsubstituted or substituted phenyl groups; and R q It is H, unsubstituted or substituted C. 1-6 Alkyl, or unsubstituted or substituted phenyl.

21. The compound according to claim 20, wherein R 4 It is OH, and preferably R is wherein 4 It is OH and R 5 It's CN.

22. The compound according to claim 20 or 21, wherein the substituted pyrimidine has any of the following structures:

23. A pharmaceutical composition comprising a compound as defined in any one of the preceding claims and a pharmaceutically acceptable carrier or diluent; Optionally, the pharmaceutical composition further comprises one or more additional active agents selected from the following: ACE inhibitors, angiotensin II receptor agonists, beta-blockers, calcium channel blockers, PDE inhibitors, mineralocorticoid receptor antagonists, diuretics, aspirin, iron supplements, vitamin B12 and folic acid supplements, statins, digitalis (digoxin) derivatives, tumor chemotherapeutic agents, and antibiotics.

24. A compound as defined in any one of claims 1 to 22 or a pharmaceutical composition as defined in claim 23, for use in treating a human or animal body by means of a therapy.

25. The compound as defined in any one of claims 1 to 22 or the pharmaceutical composition as defined in claim 23, for the treatment of anemia, ischemia, inflammation, Parkinson's disease, Alzheimer's disease, non-fatty liver disease, irritable bowel disease, cardiovascular disease, heart failure, chronic kidney disease, renal insufficiency or sickle cell anemia, or for the repair of skeletal muscle injury, increasing red blood cell count (RBC), increasing hemoglobin (HGB) production, increasing hematocrit (HCT) production, increasing erythropoietin (EPO) production, wound healing, angiogenesis, vascular regeneration, stem cell activation or cardiac protection after myocardial infarction; Optionally, the anemia described is renal anemia, such as anemia associated with chronic kidney disease or anemia in dialysis patients; chemotherapy-induced anemia; age-related anemia; or anemia caused by cancers such as leukemia, multiple myeloma, and pyogenic myeloma. Optionally, the ischemia is ischemia in circulatory or cardiovascular disease, myocardial infarction, ischemia during surgery, organ ischemia, ischemic disease, diabetic limb ischemia, or sickle cell anemia.

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