Prolyl hydroxylase inhibitor and application thereof

Abstract

The invention discloses a prolyl hydroxylase inhibitor, and belongs to the technical field of medicines. The invention provides a pyridinotriazole compound, the structure of which is shown in the following formula, and the pyridinotriazole compound has higher prolyl hydroxylase inhibitory activity and EPO induction activity, can effectively treat and prevent HIF-related and / or EPO-related diseases, and provides a new thought for anemia, ischemia, hypoxia and other diseases. # imgabs0 #

Description

A prolyl hydroxylase inhibitor and its use Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to a pyridotriazole compound having a prolyl hydroxylase inhibitory effect and a pharmaceutical composition thereof, and further relates to a preparation method and pharmaceutical use thereof. Background Art

[0002] In conditions such as anemia, trauma, tissue necrosis, and defects, tissues or cells are often exposed to hypoxia. Hypoxia triggers the expression of a series of transcriptional inducible factors involved in angiogenesis, iron and glucose metabolism, and cell growth and proliferation. Among these, hypoxia inducible factor (HIF) is a transcription factor activated in somatic cells under hypoxic conditions. It mediates a series of gene regulation responses within biological cells to hypoxia. HIF is a heterodimer consisting of an oxygen-regulated α-subunit (HIFα) and a constitutively expressed β-subunit (HIFβ / ARNT). In oxygenated (normoxic) cells, the HIFα subunit is rapidly degraded through ubiquitination by the von Hippel-Lindau tumor suppressor (pVHL) E3 ligase complex. Under hypoxic conditions, HIFα is not degraded, and active HIFα / β complexes accumulate in the nucleus and activate the expression of various genes, including glycolytic enzymes, glucose transporters, erythropoietin (EPO), and vascular endothelial growth factor (VEGF).

[0003] Erythropoietin (EPO) is a naturally occurring hormone produced with HIFα that stimulates the production of red fat (erythrocytes) that carry oxygen throughout the body. EPO is normally secreted by the kidneys, and endogenous EPO increases under conditions of reduced oxygen (hypoxia). All types of anemia are characterized by a reduced ability of the blood to carry oxygen, and are therefore accompanied by similar signs and symptoms, including pale skin and mucous membranes, weakness, dizziness, fatigue and drowsiness, leading to a decrease in quality of life. Anemia is usually associated with a condition in which there is a lack of blood in red blood cells or in hemoglobin. Common causes of anemia include iron, vitamin B12 and folic acid deficiencies, and are also complicated by chronic diseases, such as inflammatory diseases, including diseases with secondary bone marrow inflammatory suppression. Anemia is also associated with renal dysfunction, and most patients with renal failure who undergo frequent dialysis suffer from chronic anemia.

[0004] Prolyl hydroxylase domain (PHD) is a key regulator of HIF. Under normoxic conditions, PHD hydroxylates two key proline residues, Pro402 and Pro564, in HIFα, increasing its affinity for pVHL and accelerating its degradation. Under hypoxia and other pathological conditions, PHD-catalyzed HIF reactions are blocked, slowing protease degradation and causing HIFα to accumulate within cells, triggering a series of adaptive cellular responses to hypoxia. Inhibiting PHD with PHD inhibitors prolongs HIF action, thereby increasing the expression of genes such as EPO, and effectively treating and preventing HIF- and / or EPO-related conditions, such as anemia, ischemia, and hypoxia.

[0005] For example, U.S. patent application US16757333 discloses a crystalline form of an alkynylpyridine aminoacyl hydroxylase inhibitor and a preparation method thereof. The alkynylpyridine aminoacyl hydroxylase inhibitor has the following structural formula:

[0006] Currently, coenzyme A hydroxylase inhibitor is available on the market, including AstraZeneca's Roxadustat and Japan Tobacco's Enarodustat.

[0007] Due to the limited development and high demand of coagulase inhibitors, there is an urgent need to develop such compounds to treat conditions such as anemia, local ischemia and hypoxia.

[0008] Summary of the Invention

[0009] The present invention aims to provide a pyridotriazole compound and a pharmaceutical composition thereof as a HIF-PHD inhibitor, which can be used to treat various HIF-related or EPO-related diseases, such as anemia.

[0010] In order to achieve the above-mentioned purpose of the invention, the technical solution of the present invention is as follows:

[0011] In one aspect, the present invention provides a compound represented by formula (I) or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs;

[0012] in,

[0013] R is selected from the following atoms or groups:

[0014] L is -CH2- or -CH2O-;

[0015] The value of n is an integer between 0 and 2;

[0016] R 1 、R 2 、R 3 Each is independently selected from substituted or unsubstituted alkyl, cycloalkyl, aryl, and heterocyclic groups.

[0017] Preferably, the aryl group includes an aromatic heterocycle.

[0018] In some embodiments,

[0019] L is -CH2- or -CH2O-;

[0020] The value of n is 0 or 1;

[0021] R 1 is substituted or unsubstituted C 1-4 Alkyl, phenyl, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, 5-6 membered heterocyclic group containing oxygen and / or nitrogen, the substituent is C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 At least one of a haloalkyl group, a halogen group, a cyano group, a phenyl group, a 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, and a 5-6 membered heterocyclic group containing oxygen and / or nitrogen;

[0022] R 2 is substituted or unsubstituted C 1-4 Alkyl, phenyl, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, 5-6 membered heterocyclic group containing oxygen and / or nitrogen, the substituent is C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 At least one of a haloalkyl group, a halogen group, a cyano group, a phenyl group, a 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, and a 5-6 membered heterocyclic group containing oxygen and / or nitrogen;

[0023] R 3 is substituted or unsubstituted C 1-10 Alkyl, phenyl, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, 5-6 membered heterocyclic group containing oxygen and / or nitrogen, the substituent is C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 At least one of a haloalkyl group, a halogen group, a cyano group, a phenyl group, a 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, or a 5-6 membered heterocyclic group containing oxygen and / or nitrogen.

[0024] R 4 At least one selected from hydrogen, substituted or unsubstituted alkyl, cycloalkyl, alkoxy, aryl, and heterocyclic groups.

[0025] In some embodiments,

[0026] L is -CH2- or -CH2O-;

[0027] The value of n is 0 or 1;

[0028] R 1 is a substituted or unsubstituted phenyl group, wherein the substituent is C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 At least one of a haloalkyl group, a halogen group, a cyano group, a phenyl group, a 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, or a 5-6 membered heterocyclic group containing oxygen and / or nitrogen;

[0029] R 2 is a substituted or unsubstituted phenyl group, wherein the substituent is C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 At least one of a haloalkyl group, a halogen group, a cyano group, a phenyl group, a 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, and a 5-6 membered heterocyclic group containing oxygen and / or nitrogen;

[0030] R 3 is substituted or unsubstituted C 1-10 Alkyl, phenyl, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, 5-6 membered heterocyclic group containing oxygen and / or nitrogen, the substituent is C 1-4 Alkyl, C 1-4 At least one of an alkoxy group, a halogen group, a 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, and a 5-6 membered heterocyclic group containing oxygen and / or nitrogen.

[0031] R 4 is at least one of hydrogen, substituted or unsubstituted alkyl, cycloalkyl, alkoxy, aryl, and heterocyclic groups.

[0032] or, R 4 is hydrogen, substituted or unsubstituted C 1-10 Alkyl, C 1-10 Alkoxy, phenyl, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, 5-6 membered heterocyclic group containing oxygen and / or nitrogen, the substituent is C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 At least one of a haloalkyl group, a halogen group, a cyano group, a phenyl group, a 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, or a 5-6 membered heterocyclic group containing oxygen and / or nitrogen.

[0033] or, R 4 is hydrogen, substituted or unsubstituted C 1-10 Alkyl, C 1-10 Alkoxy, phenyl, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, 5-6 membered heterocyclic group containing oxygen and / or nitrogen, the substituent is C 1-4 Alkyl, C 1-4At least one of an alkoxy group, a halogen group, a 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, and a 5-6 membered heterocyclic group containing oxygen and / or nitrogen.

[0034] In some embodiments,

[0035] The value of n is 0 or 1;

[0036] R 1 is a substituted or unsubstituted phenyl group, wherein the substituent is C 1-4 Alkyl, C 1-4 Alkoxy, halogen, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen; preferably, the C 1-4 Alkyl, C 1-4 Alkoxy, oxygen and / or nitrogen-containing 5-6 membered aromatic heterocyclic group or oxygen and / or nitrogen-containing 5-6 membered heterocyclic group is para-substituted relative to the parent core structure, and halogen is ortho-, meta- or para-substituted relative to the parent core structure;

[0037] R 2 is a substituted or unsubstituted phenyl group, wherein the substituent is C 1-4 Alkyl, C 1-4 Alkoxy, halogen or 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen; preferably, the C 1-4 Alkyl, C 1-4 The alkoxy group and the 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or the 5-6 membered heterocyclic group containing oxygen and / or nitrogen are para-substituted relative to the parent core structure, and the halogen group is ortho-, meta- or para-substituted relative to the parent core structure;

[0038] R 3 is substituted or unsubstituted C 1-5 Alkyl, phenyl, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen, the substituent is C 1-4 Alkyl, C 1-4 Alkoxy, halogen, 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen; preferably, the C 1-4 Alkyl, C 1-4 The alkoxy group, the 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen, or the 5-6 membered heterocyclic group containing oxygen and / or nitrogen is para-substituted relative to the parent core structure, and the halogen group is ortho-, meta- or para-substituted relative to the parent core structure.

[0039] R 4 is hydrogen, substituted or unsubstituted C 1-5 Alkyl, C 1-5 Alkoxy, phenyl, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen, the substituent is C 1-4 Alkyl, C1-4 Alkoxy, halogen, 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen; preferably, the C 1-4 Alkyl, C 1-4 The alkoxy group, the 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen, or the 5-6 membered heterocyclic group containing oxygen and / or nitrogen is para-substituted relative to the parent core structure, and the halogen group is ortho-, meta- or para-substituted relative to the parent core structure.

[0040] In some embodiments,

[0041] The value of n is 0 or 1;

[0042] R 1 is a substituted or unsubstituted phenyl group, wherein the substituent is C 1-3 Alkyl, C 1-3 Alkoxy, halogen, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen; preferably, the C 1-3 Alkyl, C 1-3 Alkoxy, oxygen and / or nitrogen-containing 5-6 membered aromatic heterocyclic group or oxygen and / or nitrogen-containing 5-6 membered heterocyclic group is para-substituted relative to the parent core structure, and halogen is ortho-, meta- or para-substituted relative to the parent core structure;

[0043] R 2 is a substituted or unsubstituted phenyl group, wherein the substituent is C 1-3 Alkyl, C 1-3 Alkoxy, halogen, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen; preferably, the C 1-3 Alkyl, C 1-3 Alkoxy, oxygen and / or nitrogen-containing 5-6 membered aromatic heterocyclic group or oxygen and / or nitrogen-containing 5-6 membered heterocyclic group is para-substituted relative to the parent core structure, and halogen is ortho-, meta- or para-substituted relative to the parent core structure;

[0044] R 3 is substituted or unsubstituted C 1-5 Alkyl, phenyl, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen, the substituent is C 1-3 Alkyl, C 1-3 Alkoxy, halogen, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen; preferably, the C 1-3 Alkyl, C 1-3 The alkoxy group, the 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, or the 5-6 membered heterocyclic group containing oxygen and / or nitrogen is para-substituted relative to the parent core structure, and the halogen is ortho-, meta- or para-substituted relative to the parent core structure.

[0045] R4 is hydrogen, substituted or unsubstituted C 1-5 Alkyl, C 1-5 Alkoxy, phenyl, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen, the substituent is C 1-3 Alkyl, C 1-3 Alkoxy, halogen, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen; preferably, the C 1-3 Alkyl, C 1-3 The alkoxy group, the 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, or the 5-6 membered heterocyclic group containing oxygen and / or nitrogen is para-substituted relative to the parent core structure, and the halogen is ortho-, meta- or para-substituted relative to the parent core structure.

[0046] In some embodiments,

[0047] The value of n is 0 or 1;

[0048] R 1 is a substituted or unsubstituted phenyl group, wherein the substituent is C 1-2 Alkyl, C 1-2 Alkoxy, F, Cl, Br, I, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen; preferably, the C 1-2 Alkyl, C 1-2 The alkoxy group, the 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, or the 5-6 membered heterocyclic group containing oxygen and / or nitrogen is para-substituted relative to the parent core structure, and F, Cl, Br, and I are ortho-, meta-, or para-substituted relative to the parent core structure;

[0049] R 2 is a substituted or unsubstituted phenyl group, wherein the substituent is C 1-2 Alkyl, C 1-2 Alkoxy, F, Cl, Br, I, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen; preferably, the C 1-2 Alkyl, C 1-2 The alkoxy group, the 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, or the 5-6 membered heterocyclic group containing oxygen and / or nitrogen is para-substituted relative to the parent core structure, and F, Cl, Br, and I are ortho-, meta-, or para-substituted relative to the parent core structure;

[0050] R 3 is substituted or unsubstituted C 1-4 Alkyl, phenyl, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen, the substituent is C 1-2 Alkyl, C 1-2Alkoxy, F, Cl, Br, I, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen; preferably, the C 1-2 Alkyl, C 1-2 The alkoxy group, the 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, or the 5-6 membered heterocyclic group containing oxygen and / or nitrogen is para-substituted relative to the parent core structure, and the halogen is ortho-, meta- or para-substituted relative to the parent core structure.

[0051] R 4 is hydrogen, substituted or unsubstituted C 1-4 Alkyl, C 1-4 Alkoxy, phenyl, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen, the substituent is C 1-2 Alkyl, C 1-2 Alkoxy, F, Cl, Br, I, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen; preferably, the C 1-2 Alkyl, C 1-2 The alkoxy group, the 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, or the 5-6 membered heterocyclic group containing oxygen and / or nitrogen is para-substituted relative to the parent core structure, and the halogen is ortho-, meta- or para-substituted relative to the parent core structure.

[0052] In some embodiments,

[0053] The value of n is 0 or 1;

[0054] R 1 is a substituted or unsubstituted phenyl group, wherein the substituent is a methyl group, a methoxy group, a F group, a Cl group, a 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, or a 5-6 membered heterocyclic group containing oxygen and / or nitrogen; preferably, the methyl group, the methoxy group, the 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen is para-substituted relative to the parent core structure, and the F group and the Cl group are ortho-, meta-, or para-substituted relative to the parent core structure;

[0055] R 2 is a substituted or unsubstituted phenyl group, wherein the substituent is a methyl group, a methoxy group, a F group, a Cl group, or a 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen; preferably, the methyl group, the methoxy group, and the 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen are para-substituted relative to the parent core structure, and the F group and the Cl group are ortho-, meta-, or para-substituted relative to the parent core structure;

[0056] R 3 is substituted or unsubstituted C 1-3Alkyl, phenyl, 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen, the substituent of the substituted phenyl is methyl, methoxy, F, Cl or 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen; preferably, the methyl, methoxy and 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen are para-substituted relative to the parent core structure, and the halogen is ortho-, meta- or para-substituted relative to the parent core structure.

[0057] R 4 is hydrogen, substituted or unsubstituted C 1-3 Alkyl, C 1-3 Alkoxy, phenyl, 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen, the substituent of the substituted phenyl is methyl, methoxy, F, Cl or 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen; preferably, the methyl, methoxy and 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen are para-substituted relative to the parent core structure, and the halogen is ortho-, meta- or para-substituted relative to the parent core structure.

[0058] In some embodiments,

[0059] The value of n is 0 or 1;

[0060] R 1 is a substituted or unsubstituted phenyl group, wherein the substituent is a methyl group or a 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen; preferably, the methyl group or the 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen is para-substituted relative to the parent core structure;

[0061] R 2 is a substituted or unsubstituted phenyl group, wherein the substituent is a methyl group or a 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen; preferably, the methyl group or the 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen is para-substituted relative to the parent core structure;

[0062] R 3 is substituted or unsubstituted C 1-3 Alkyl, phenyl, 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen, the substituent is methyl, methoxy, F, Cl or 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen; preferably, the methyl, methoxy and 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen are para-substituted relative to the parent core structure, and the halogen is ortho-, meta- or para-substituted relative to the parent core structure.

[0063] R 4 is hydrogen, substituted or unsubstituted C 1-3 Alkyl, C 1-3 Alkoxy, phenyl, 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen, the substituent is methyl, methoxy, F, Cl or 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen; preferably, the methyl, methoxy and 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen are para-substituted relative to the parent core structure, and the halogen is ortho-, meta- or para-substituted relative to the parent core structure.

[0064] In some embodiments,

[0065] The value of n is 0 or 1;

[0066] R 1 is phenyl;

[0067] R 2 is phenyl;

[0068] R 3 is substituted or unsubstituted C 1-3 Alkyl, phenyl, 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen, the substituent is methyl, methoxy, halogen or 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen; preferably, the methyl, methoxy, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen is para-substituted relative to the parent core structure, and the halogen is ortho-, meta- or para-substituted relative to the parent core structure.

[0069] R 4 For hydrogen.

[0070] The term "core structure" refers to the remaining main structure of the compound structure represented by formula (I) excluding the R group.

[0071] In some embodiments, the compound is selected from the following structures:

[0072] In some embodiments, the compound is selected from the following structures:

[0073] In some embodiments, the compound is selected from the following structures:

[0074] In some embodiments, the compound is selected from the following structures:

[0075] In some embodiments, the compound is selected from the following structures:

[0076] In some embodiments, the present invention provides a compound represented by formula (II) or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs;

[0077] in,

[0078] The value of n is an integer between 0 and 3;

[0079] R1 is selected from hydrogen, halogen, substituted or unsubstituted alkyl, cycloalkyl, alkoxy, aryl, and heterocyclic groups which are mono- or poly-substituted on the aromatic ring.

[0080] In some embodiments, the aryl group of the compound of formula (II) or its stereoisomers, geometric isomers, tautomers, N-oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs comprises an aromatic heterocycle.

[0081] In some embodiments, the value of n of the compound of formula (II) or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs is an integer between 0 and 2; R1 is selected from hydrogen, halogen, substituted or unsubstituted alkyl, cycloalkyl, alkoxy, aryl, and heterocyclic groups.

[0082] In some embodiments, the value of n of the compound of formula (II) or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs is 0 or 1; R1 is selected from hydrogen, halogen, substituted or unsubstituted alkyl, cycloalkyl, alkoxy, aryl, heterocyclic group.

[0083] In some embodiments, the value of n in the compound of formula (II) or its stereoisomers, geometric isomers, tautomers, nitrogen oxide hydrates, solvates, pharmaceutically acceptable salts or prodrugs is 0 or 1; R1 is hydrogen, substituted or unsubstituted C 1-10 Alkyl, C 1-10 Alkoxy, phenyl, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, 5-6 membered heterocyclic group containing oxygen and / or nitrogen, the substituent is C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 At least one of a haloalkyl group, a halogen group, a cyano group, a phenyl group, a 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, or a 5-6 membered heterocyclic group containing oxygen and / or nitrogen.

[0084] In some embodiments, the value of n in the compound of formula (II) or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs is 1; and R1 is hydrogen.

[0085] In some embodiments, the compound of formula (II) or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs are selected from the following structures:

[0086] In some embodiments, the preparation method of the compound of formula (II) is characterized by using the following synthetic route:

[0087] In some embodiments, a pharmaceutical composition is provided, comprising a compound represented by formula (II) or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, and one or more pharmaceutically acceptable carriers, diluents, and excipients.

[0088] In some embodiments, the compound represented by formula (II) or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or the above-mentioned pharmaceutical compositions are used in the preparation of drugs for treating diseases mediated by the inhibition of coenzyme A by inhibiting coenzyme A.

[0089] In some embodiments, the disease mediated by the inhibition of coenzyme A is anemia, ischemia, or hypoxia.

[0090] In some embodiments, the disease mediated by inhibition of coenzyme A is renal anemia.

[0091] In the present invention, FG-4592 is roxadustat.

[0092] [(7-Hydroxy-5-(1,2,3,4-tetrahydronaphthalen-2-yl)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid exhibits excellent pharmacological effects in continuous chronic administration, with significant data showing that it can significantly increase the hemoglobin and EPO levels of SD rats, and the overall effect is significantly better than the positive drug Enarodustat.

[0093] Preferably, the purity of the intermediates prepared in each step of the embodiments of the present invention can be above 95%.

[0094] Unless otherwise specified, the term "alkyl" as used herein includes branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms, including all isomers. Common abbreviations for alkyl groups include "Me" or CH3 for methyl, "Et" or CH2CH3 for ethyl, "Pr" or CH2CH2CH3 for propyl, and "Bu" or CH2CH2CH2CH3 for butyl. For example, "C 1-4 "C1-C4 alkyl" refers to a straight or branched chain alkyl group having the specified number of carbon atoms, including all isomers. 1-4 Alkyl includes n-, iso-, sec- and t-butyl, n- and iso-propyl, ethyl and methyl. The term "C 1-10 "Alkyl" and the like have similar meanings.

[0095] The term "alkoxy" represents straight and branched chain alkyl groups having the indicated number of carbon atoms attached through an oxygen bridge.

[0096] The term "halogen" (or "halo") refers to fluorine, chlorine, bromine, and iodine (alternatively referred to as fluoro (F), chloro (Cl), bromo (Br), and iodo (I)).

[0097] The term "aryl" refers to aromatic mono- and polycyclic carbocyclic ring systems wherein the individual carbon rings in the polycyclic ring system are fused or linked to each other by single bonds. Typical aryl groups include phenyl, naphthyl and biphenylene.

[0098] The term "heterocycle" refers to a cyclic structure composed of carbon atoms and non-carbon atoms such as nitrogen, oxygen, and sulfur. Typical heterocyclic groups include pyridine, quinoline, tropane, phenothiazine, benzodiazepine, furan, pyrazolone, and pyrimidine.

[0099] The term "aromatic heterocycle" refers to a 5- or 6-membered monocyclic aromatic ring or a 7- to 12-membered bicyclic ring composed of carbon atoms and one or more heteroatoms selected from N, O and S. Examples of aromatic heterocycles include pyridyl, pyrrolyl, pyrazinyl, pyrimidinyl, pyridazinyl, thienyl (or thiophenyl), thiazolyl, furanyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, diazolyl, thiazolyl, isothiazolyl and thiadiazolyl, benzotriazolyl, indolyl, isoindolyl, indazolyl, indolinyl, isoindolinyl, quinoxalinyl, quinazolinyl, cinnolinyl, chromanyl, isochromanyl, tetrahydroquinolinyl, quinolinyl, tetrahydroisoquinolinyl, isoquinolinyl, 2,3-dihydrobenzofuranyl, 2,3-dihydrobenzo-1,4-dienyl, imidazo(2,1-b)(1,3)thiazole and benzo-1,3-dioxolyl.

[0100] The aryl group in the term "substituted aryl" is as defined above. When the substituent of the substituted aryl group is not specified, the substituent group may be selected from the following groups, including but not limited to: halogen, C1-C 20 Alkyl, CF3, NH2, N(C1-C6 alkyl)2, NO2, oxo, CN, N3, -OH, -O(C1-C6 alkyl), C3-C 10 Cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, (C0-C6 alkyl)S(O) 0-2 -, aryl-S(O) 0-2 -, (C0-C6 alkyl)S(O) 0-2 (C0-C6 alkyl)-, (C0-C6 alkyl)C(O)NH-, H2N-C(NH)-, -O(C1-C6 alkyl)CF3, (C0-C6 alkyl)C(O)-, (C0-C6 alkyl)OC(O)-, (C0-C6 alkyl)2NC(O)-(C0-C6 alkyl)O(C1-C6 alkyl)-, (C0-C6 alkyl)C(O) 1-2(C0-C6 alkyl)-, (C0-C6 alkyl)OC(O)NH-, aryl, aralkyl, heteroaryl, heterocyclylalkyl, halogen-aryl, halogen-aralkyl, halogen-heterocycle, halogen-heterocyclylalkyl, cyano-aryl, cyano-aralkyl, cyano-heterocycle and cyano-heterocyclylalkyl. The term "substituted phenyl" has a similar definition.

[0101] Unless otherwise specified, all ranges listed herein are inclusive. For example, "n is an integer between 0 and 2" means that n can be 0, 1 or 2.

[0102] The term "pharmaceutically acceptable salt" refers to a salt prepared from a pharmaceutically acceptable non-toxic base or acid. When the compound of the present invention is acidic, its corresponding salt can be readily prepared from an inorganic or organic base. Salts derived from such inorganic bases include salts of aluminum, ammonium, calcium, copper (copper and cuprous), iron, ferrous, lithium, magnesium, manganese (manganese and manganous), potassium, sodium, zinc, and the like. Preferred are salts of ammonium, calcium, magnesium, potassium, and sodium. Salts prepared from organic bases include primary, secondary, and tertiary amines derived from natural and synthetic sources. Pharmaceutically acceptable organic non-toxic bases that can form salts include arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucosamine, glucosamine, histidine, hydrabamine, isopropylamine, dicyclohexylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc. When the compound of the present invention is basic, its corresponding salt can be easily prepared from inorganic or organic acids. Such acids include, for example, acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucic acid, nitric acid, pamoic acid, pantothenic acid, phosphoric acid, succinic acid, sulfuric acid, tartaric acid, p-toluenesulfonic acid, and the like.

[0103] The term "solvate" refers to a complex of variable stoichiometry formed by a solute (i.e., a compound of Formula (I)) or a pharmaceutically acceptable salt thereof and a solvent that does not interfere with the biological activity of the solute. Examples of solvents include, but are not limited to, water, ethanol, and acetic acid. When the solvent is water, the solvate is referred to as a hydrate. Hydrates include, but are not limited to, hemihydrates, monohydrates, monosesquihydrates, dihydrates, and trihydrates.

[0104] The term "prodrug" is a functional derivative of the compounds of the invention which is readily convertible in vivo into the required compound.

[0105] On the other hand, the present invention provides a method for preparing the above-mentioned compound, comprising the following four synthetic routes:

[0106] Route 1:

[0107] Route 2:

[0108] Route 3:

[0109] Route 4:

[0110] Among them, R 1 、R 2 、R 3 、R 4 Has the same definition as above.

[0111] In another aspect, the present invention provides a pharmaceutical composition comprising a compound represented by formula (I) or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, and one or more pharmaceutically acceptable carriers, diluents, and excipients.

[0112] In the context of pharmaceutical compositions, the term "composition" includes a product comprising an active ingredient and an inert ingredient (pharmaceutically acceptable excipient) constituting a carrier, as well as any product obtained directly or indirectly by the combination, complexation or aggregation of two or more ingredients, or the decomposition of one or more ingredients, or other types of reactions or interactions of one or more ingredients. Therefore, the pharmaceutical compositions of the present invention include any composition prepared by mixing a compound of formula (I) and / or formula (II), other active ingredients, and a pharmaceutically acceptable excipient.

[0113] The pharmaceutical composition of the present invention comprises a compound represented by formula (I) and / or formula (II) (or a pharmaceutically acceptable salt or solvate thereof) as an active ingredient, a pharmaceutically acceptable carrier, and optionally other therapeutic ingredients or adjuvants. Pharmaceutical compositions include compositions suitable for oral, rectal, topical, and parenteral (including subcutaneous, intramuscular, and intravenous) administration, although the most appropriate route in any particular case depends on the specific subject, the nature and severity of the condition to which the active ingredient is administered. Pharmaceutical compositions can be prepared by any method known in the art of pharmacy.

[0114] The active ingredient can be administered orally in solid dosage forms such as capsules, tablets, lozenges, troches, granules, and powders, or in liquid dosage forms such as elixirs, syrups, emulsions, dispersions, and suspensions. The active ingredient can also be administered parenterally in sterile liquid dosage forms such as dispersions, suspensions, or solutions. Other dosage forms that can be used to administer the active ingredient include ointments, creams, drops, transdermal patches, or powders for topical administration; ophthalmic solutions or suspensions, i.e., eye drops, for administration to the eye; sprays or powder compositions for inhalation or intranasal administration, or creams, ointments, sprays, or suppositories for rectal or vaginal administration. Gelatin capsules contain the active ingredient and a powdered carrier such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, etc. Similar diluents can be used to prepare compressed tablets. Both tablets and capsules can be formulated as sustained-release products to provide sustained release of the drug over several hours. Compressed tablets can be coated with sugar or film to cover any unpleasant taste and protect the tablet from air, or can be enteric-coated for selective disintegration in the gastrointestinal tract. Liquid dosage forms for oral administration can include coloring agents and flavoring agents to increase patient acceptance. Generally speaking, water, suitable oil, saline, dextrose (glucose) aqueous solution and related sugar solutions and glycols such as propylene glycol or polyethylene glycol are suitable carriers of parenteral solutions. The solution for parenteral administration preferably includes a water-soluble salt of the active ingredient, a suitable stabilizer and the buffer substance used as needed. Antioxidants such as sodium bisulfite, sodium sulfite or ascorbic acid, alone or in combination, are suitable stabilizers. Citric acid and its salts and sodium EDTA can also be used. In addition, parenteral solutions can also include preservatives, such as benzalkonium chloride, methylparaben or propylparaben and chlorobutanol. For inhalation administration, the compound of the present invention can be easily delivered in a spray form from a pressurized package or a sprayer. The compound can also be delivered in the form of a powder for preparation, and the powder composition can be inhaled with the help of an insufflation powder inhaler device. The preferred delivery system for inhalation is a metered dose inhalation (MDI) aerosol, which can be prepared as a suspension or solution of the compound of formula (I) and / or formula (II) in a suitable propellant, such as a fluorocarbon or hydrocarbon. For ophthalmic administration, an ophthalmic preparation can be prepared with a solution or suspension of the compound of formula (I) and / or formula (II) in a suitable ophthalmic carrier in an appropriate weight percentage, thereby keeping the compound in contact with the surface of the eye for a sufficient time to allow the compound to penetrate the cornea and internal areas of the eye.

[0115] Useful pharmaceutical dosage forms for administering the compounds of the present invention include, but are not limited to, hard and soft gelatin capsules, tablets, parenteral injection solutions, and oral suspensions.

[0116] When the compounds of the present invention are administered stepwise or in combination with other therapeutic agents, the same dosage forms as described above can be used. When the drugs are administered in a physical combination, the dosage form and route of administration should be selected based on the compatibility of the combined drugs. The compounds of the present invention can be administered as the sole active ingredient or in combination with a second active ingredient, including those known to be useful for increasing erythropoietin levels in patients.

[0117] In another aspect, the present invention provides a compound represented by formula (I) or formula (II) or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or the use of the above-mentioned pharmaceutical compositions in the preparation of a drug for treating diseases mediated by the inhibition of coenzyme A by inhibiting coenzyme A.

[0118] In some embodiments, the disease mediated by the inhibition of coenzyme A is anemia, ischemia, or hypoxia.

[0119] In some embodiments, the anemia is renal anemia.

[0120] The beneficial effects of the present invention are:

[0121] A new compound is proposed, which can be used as a co-aminoacyl hydroxylase inhibitor with high co-aminoacyl hydroxylase inhibitory activity and EPO induction activity, can effectively treat and prevent HIF-related and / or EPO-related diseases, and provide new ideas for diseases such as anemia, local ischemia and hypoxia. BRIEF DESCRIPTION OF THE DRAWINGS

[0122] FIG1 is a microscope photograph of a single crystal sample of chiral isomer A of methyl acetate;

[0123] FIG2 is an asymmetric unit diagram of the crystal structure of chiral isomer A of methyl acetate;

[0124] FIG3 is a unit cell diagram of the crystal structure of chiral isomer A of methyl acetate;

[0125] FIG4 is a stacking diagram of the crystal structure of chiral isomer A of methyl acetate;

[0126] FIG5 is a comparison diagram of the XRPD of a single crystal sample of methyl acetate chiral isomer A and the calculated XRPD;

[0127] FIG6 is a chemical structure diagram of methyl acetate chiral isomer A;

[0128] FIG7 is a microscope photograph of a single crystal sample of methyl acetate chiral isomer B;

[0129] FIG8 is the asymmetric unit of the crystal structure of chiral isomer B of methyl acetate;

[0130] FIG9 is a unit cell diagram of the crystal structure of chiral isomer B of methyl acetate;

[0131] FIG10 is a stacking diagram of the crystal structure of chiral isomer B of methyl acetate;

[0132] FIG11 is a comparison of the XRPD patterns of the actual measured and calculated single crystal samples of methyl acetate chiral isomer B;

[0133] FIG12 is a chemical structure diagram of chiral isomer B of methyl acetate. DETAILED DESCRIPTION

[0134] The following non-limiting examples are provided to enable those skilled in the art to more fully understand the present invention, but are not intended to limit the present invention in any way. The following is merely an illustrative description of the scope of protection claimed in this application. Those skilled in the art may make various changes and modifications to the invention of this application based on the disclosed content, and such changes and modifications should also fall within the scope of protection claimed in this application.

[0135] The present invention is further described below by way of specific examples. Unless otherwise specified, the various chemical reagents used in the examples of the present invention were obtained through conventional commercial channels.

[0136] Example 1

[0137] Preparation of (7-hydroxy-5-(phenoxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid

[0138] Step 1 Preparation of tert-butyl 5-(phenoxy)-7-(benzyloxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylate

[0139] 5-iodo-7-(benzyloxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylic acid tert-butyl ester (1.0 g), phenol (310 mg), 2-picolinic acid (55 mg), cuprous iodide (42 mg), potassium phosphate (942 mg), and dimethyl sulfoxide (5.0 mL) were added sequentially to a reaction flask, and the nitrogen atmosphere was replaced three times. The temperature was raised to 65°C and the reaction was allowed to proceed overnight until the reaction of the raw materials was complete as monitored by TLC. After cooling to room temperature, the mixture was extracted with ethyl acetate and washed with saturated brine. The organic phase was separated, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 450 mg of a white solid.

[0140] MS m / z(ESI)[M+H] + :418.30

[0141] Step 2 Preparation of 5-(phenoxy)-7-(benzyloxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylic acid

[0142] Dissolve tert-butyl 5-(phenoxy)-7-(benzyloxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylate (450 mg) in a 1:1 (volume ratio) mixture of toluene and tetrahydrofuran (10 mL). Add methanesulfonic acid (0.5 mL) and stir, maintaining the reaction temperature between 60-70°C. Stop the reaction until complete reaction is observed by TLC. Cool to room temperature, add a small amount of ethyl acetate, and filter to yield 440 mg of a white solid.

[0143] MS m / z(ESI)[M+H] + :362.22

[0144] Step 3 Preparation of [(7-(benzyloxy)-5-(phenoxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid methyl ester

[0145] 5-(Phenoxy)-7-(benzyloxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylic acid (440 mg), EDCI (280 mg), HoBt (197 mg), N,N-dimethylformamide (5.0 mL), and triethylamine (247 mg) were added to the reaction flask in sequence and stirred at room temperature for 10 minutes. Glycine methyl ester hydrochloride (306 mg) was then added to the system and the temperature was raised to 60°C for 1.5 hours. The reaction was stopped, cooled to room temperature, and quenched by adding saturated aqueous sodium bicarbonate solution. Filtering gave 300 mg of a yellow solid.

[0146] MS m / z(ESI)[M+H] + :433.31

[0147] Step 4 Preparation of [(7-hydroxy-5-(phenoxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid methyl ester

[0148] Methyl [(7-(benzyloxy)-5-(phenoxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetate (300 mg) and palladium on carbon (30 mg) were dissolved in a 1:1 (volume ratio) mixture of methanol and tetrahydrofuran (4.0 mL). The mixture was stirred at room temperature under a hydrogen atmosphere until the reaction was complete as monitored by TLC. The reaction was stopped. The solvent was removed by concentration under reduced pressure, and the product was separated by column chromatography to obtain 170 mg of the product.

[0149] MS m / z(ESI)[M+H] + :343.22

[0150] Step 5 Preparation of [(7-hydroxy-5-(phenoxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid

[0151] Methyl [(7-hydroxy-5-(phenoxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetate (170 mg) was dissolved in methanol (2.0 mL), followed by the addition of 30% aqueous sodium hydroxide solution (1.0 mL). The mixture was stirred at room temperature until the reaction of the raw material was complete as monitored by TLC. The pH was adjusted to between 1 and 2 with dilute hydrochloric acid and filtered to obtain 150 mg of the product.

[0152] 1 H NMR (400MHz, d-DMSO) δ: 14.52 (s, 1H), 12.95 (brs, 1H), 9.71 (s, 1H), 8.59 (s, 1H),7.59–7.53(m,2H),7.44–7.41(m,3H),5.83(s,1H),4.20(d,J=5.6Hz,2H)

[0153] MS m / z(ESI):329.09

[0154] Example 2

[0155] Preparation of (5-(2-chlorophenoxy)-7-hydroxy-[1,2,4]triazolyl[1,5-a]pyridine-8-carbonyl)glycine

[0156] According to the method of Example 1, 428.1 mg of 2-chlorophenol was used to replace phenol to obtain 108.9 mg of product.

[0157] 1 H NMR (400MHz, d-DMSO) δ: 14.58 (s, 1H), 12.97 (brs, 1H), 9.71 (t, J = 5.6Hz, 1H), 8.63 (s, 1H), 7.75 (d, J = 7.6Hz ,1H),7.61(d,J=8.0Hz,1H),7.54(t,J=8.0Hz,1H),7.47(t,J=7.2Hz,1H),5.89(s,1H),4.21(d,J=5.6Hz,2H)

[0158] MS m / z(ESI)[M+H] + :363.17

[0159] Example 3

[0160] Preparation of (5-(3-chlorophenoxy)-7-hydroxy-[1,2,4]triazolyl[1,5-a]pyridine-8-carbonyl)glycine

[0161] According to the method of Example 1, 428.1 mg of 3-chlorophenol was used to replace phenol to obtain 201.6 mg of product.

[0162] 1 H NMR(400MHz,d-DMSO)14.54(s,1H),12.97(brs,1H),9.72(s,1H),8.58(s,1H),7.62(s,1H),7.57 (t,J=8.4Hz,1H),7.47(d,J=8.4Hz,1H),7.41(d,J=8.4Hz,1H),6.10(s,1H),4.21(d,J=5.6Hz,2H)

[0163] MS m / z(ESI)[M+H]+:363.14

[0164] Example 4

[0165] Preparation of (5-(4-chlorophenoxy)-7-hydroxy-[1,2,4]triazolyl[1,5-a]pyridine-8-carbonyl)glycine

[0166] According to the method of Example 1, 428.1 mg of 4-chlorophenol was used to replace phenol to obtain 182.1 mg of product.

[0167] 1 H NMR (400MHz, d-DMSO) δ: 14.54 (s, 1H), 12.96 (s, 1H), 9.72 (t, J = 5.6Hz, 1H), 8.60 (s, 1H),7.62(d,J=8.4Hz,2H),7.48(d,J=8.4Hz,2H),6.02(s,1H),4.21(d,J=5.6Hz,2H)

[0168] MS m / z(ESI)[M+H] + :363.14

[0169] Example 5

[0170] Preparation of (5-(p-tolyloxy)-7-hydroxy-[1,2,4]triazolyl[1,5-a]pyridine-8-carbonyl)glycine

[0171] According to the method of Example 1, 356.9 mg of 4-methylphenol was used to replace phenol to obtain 87.7 mg of product.

[0172] 1H NMR (400MHz, d-DMSO) δ: 14.51 (s, 1H), 12.95 (s, 1H), 9.69 (t, J = 5.6Hz, 1H), 8.59 (s, 1H), 7. 37(d,J=8.4Hz,2H),7.31(d,J=8.4Hz,2H),5.77(s,1H),4.20(d,J=5.6Hz,2H),2.37(s,3H)

[0173] MS m / z(ESI)[M+H] + :343.16

[0174] Example 6

[0175] Preparation of (5-(p-methoxyphenoxy)-7-hydroxy-[1,2,4]triazolyl[1,5-a]pyridine-8-carbonyl)glycine

[0176] According to the method of Example 1, 409.2 mg of 4-methoxyphenol was used to replace phenol to obtain 223.7 mg of the product.

[0177] 1 H NMR (400MHz, d-DMSO) δ: 14.51 (s, 1H), 12.96 (s, 1H), 9.70 (t, J = 5.6Hz, 1H), 8.60 (s, 1H), 7. 39(d,J=8.8Hz,2H),7.11(d,J=8.8Hz,2H),5.72(s,1H),4.21(d,J=5.6Hz,2H),3.82(s,3H)

[0178] MS m / z(ESI)[M+H] + :359.22

[0179] Example 7

[0180] Preparation of (5-(3-morpholinophenoxy)-7-hydroxy-[1,2,4]triazolyl[1,5-a]pyridine-8-carbonyl)glycine

[0181] According to the method of Example 1, 3-morpholinophenol (590 mg) was used to replace phenol to obtain 110.7 mg of the product.

[0182] 1H NMR (400MHz, d-DMSO) δ: 14.51 (s, 1H), 12.95 (brs, 1H), 9.70 (t, J = 5.2Hz, 1H), 8.59 (s, 1H), 7.39 (d, J = 8.0Hz, 2H), 6.99 ( d,J=12.0Hz,2H),6.80(d,J=7.6Hz,2H),5.82(s,1H),4.21(d,J=5.6Hz,2H),3.73(t,J=4.8Hz,4H),3.18(t,J=4.8Hz,4H)

[0183] MS m / z(ESI)[M+H] + :414.23

[0184] Example 8

[0185] Preparation of (5-(4-fluorophenoxy)-7-hydroxy-[1,2,4]triazolyl[1,5-a]pyridine-8-carbonyl)glycine

[0186] According to the method of Example 1, 4-fluorophenol (369.6 mg) was used to replace phenol to obtain 239.3 mg of the product.

[0187] 1 H NMR(400MHz,d-DMSO)14.53(s,1H),12.98(brs,1H),9.71(s,1H),8.60(s,1 H),7.53–7.49(m,2H),7.43–7.38(m,2H),5.86(s,1H),4.20(d,J=5.6Hz,2H)

[0188] MS m / z(ESI)[M+H] + :347.14

[0189] Example 9

[0190] Preparation of (5-(Propoxy)-7-hydroxy-[1,2,4]triazolyl[1,5-a]pyridine-8-carbonyl)glycine

[0191] According to the method of Example 1, phenol was replaced by n-propanol (2 ml) to obtain 20.1 mg of the product.

[0192] 1H NMR(400MHz,d-DMSO)14.51(s,1H),12.92(brs,1H),9.65(t,J=5.6Hz,1H),8.47(s,1H),6.44 (s,1H),4.39(t,J=6.4Hz,2H),4.18(d,J=5.6Hz,2H),1.91–1.82(m,2H),1.03(t,J=7.2Hz,3H)

[0193] MS m / z(ESI)[M+H] + :295.12

[0194] Example 10

[0195] Preparation of [(7-hydroxy-5-(phenylethynyl)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid

[0196] Step 1 Preparation of tert-butyl 7-hydroxy-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylate

[0197] In a reaction flask, tert-butyl 7-(benzyloxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylate (15 g) was dissolved in a mixture of methanol and tetrahydrofuran (25 mL / 50 mL) and stirred at room temperature under a hydrogen atmosphere until the reaction was complete as monitored by TLC. The reaction was stopped. The palladium on carbon was removed by filtration, and the filtrate was then concentrated under reduced pressure to obtain 11.7 g of crude product.

[0198] MS m / z(ESI):[M+H] + :236.24

[0199] Step 2 Preparation of tert-butyl 7-(pivaloyloxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylate

[0200] Tert-butyl 7-hydroxy-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylate (10.7 g) was placed in a reaction flask and tetrahydrofuran (65 mL) was added, followed by triethylamine (8.0 mL) and pivaloyl chloride (9.1 g). The nitrogen was replaced three times and the reaction flask was sealed. The reaction mixture was heated to 60 ° C and stirred overnight. The reaction was stopped until the reaction of the raw materials was complete as monitored by TLC. After the reaction solution was cooled to room temperature, the reaction solution was extracted with ethyl acetate, washed with saturated sodium bicarbonate aqueous solution, saturated ammonium chloride aqueous solution, and saturated brine, and then concentrated under reduced pressure to obtain 12.0 g of a yellow solid.

[0201] MS m / z(ESI):[M+H] + :320.36

[0202] Step 3 Preparation of tert-butyl 5-iodo-7-(pivaloyloxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylate

[0203] Tert-butyl 7-(pivaloyloxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylate (10.0 g) was dissolved in tetrahydrofuran (50 mL) solution, and then elemental iodine (8.75 g) was added, and the reaction temperature was kept below -60 ° C. After 5 minutes, hexamethyldisiloxane lithium (62.7 mL, 1.0 M) was added in batches and stirred until the reaction of the raw materials was complete as monitored by TLC. Subsequently, hydrochloric acid ethyl acetate solution was added to quench the reaction, extracted with ethyl acetate, and washed with sodium sulfite aqueous solution. The organic phase was separated and washed with saturated ammonium chloride aqueous solution, saturated sodium bicarbonate aqueous solution, and saturated brine in sequence. The organic phase was separated and concentrated under reduced pressure, and the crude product was subjected to column chromatography to obtain 5.9 g of the product.

[0204] 1 H NMR (400MHz, CDCl3) δ: 8.36 (d, J = 5.2Hz, 1H), 7.45 (d, J = 5.2Hz, 1H), 2.31 (s, 6H)

[0205] MS m / z(ESI):[M+H] + :446.26

[0206] Step 4 Preparation of tert-butyl 5-(phenylethynyl)-7-hydroxy-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylate

[0207] 5-Iodo-7-(pivaloyloxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylic acid tert-butyl ester (5.9 g), cuprous iodide (70 mg), Pd(dppf)Cl2 (285 mg), tetrahydrofuran (30 mL), triethylamine (6.6 g), and phenylacetylene (2.1 g) were added sequentially to a reaction flask. The atmosphere was replaced with nitrogen three times, and the temperature was raised to 50°C for reaction until the reaction was complete as determined by TLC. Ammonia was then added to quench the reaction. Extraction was performed with ethyl acetate, and the organic phase was separated. The ammonia layer was washed with hydrochloric acid and extracted with ethyl acetate. The organic phases were then combined, concentrated under reduced pressure, and purified by column chromatography to yield 1.9 g of a red solid.

[0208] MS m / z(ESI):[M+H] + :336.36

[0209] Step 5 Preparation of 5-(phenylethynyl)-7-hydroxy-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylic acid

[0210] Dissolve tert-butyl 5-(phenylethynyl)-7-hydroxy-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylate (1.8 g) in a 2:1 volume ratio of toluene and ethyl acetate (24 mL). Add methanesulfonic acid (2.1 g) and stir, maintaining the reaction temperature between 60-70°C. Stop the reaction until the starting material reacts completely, as monitored by TLC. After cooling the reaction solution to room temperature, add ethyl acetate (10 mL), resulting in the precipitation of a large amount of solid, which is filtered to obtain a green solid. The solid is then dissolved in a small amount of N,N-dimethylacetamide, and ethyl acetate is added to the system, resulting in the precipitation of a solid, which is filtered to obtain 1.0 g of crude product.

[0211] MS m / z(ESI):[M+H] + :280.26

[0212] Step 6 Preparation of [(7-hydroxy-5-(phenylethynyl)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid methyl ester

[0213] In reaction flask A, 5-(phenylethynyl)-7-hydroxy-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylic acid (0.8 g) was dissolved in dichloromethane (20 mL). Oxalyl chloride (0.76 g) was added dropwise and stirred at room temperature. In reaction flask B, glycine methyl hydrochloride (0.68 g), dichloromethane (20 mL), and triethylamine (3.0 mL) were added sequentially and stirred at 0°C. TLC monitoring indicated that the reaction of the starting material in reaction flask A was complete. Stirring in reaction flask A was stopped and the material in reaction flask A was added to reaction flask B. Stirring was continued at 0°C until the reaction was complete as measured by TLC. The reaction was quenched with water. The organic phase was extracted with dichloromethane, washed sequentially with saturated aqueous ammonium chloride solution and saturated aqueous sodium bicarbonate solution. The organic phases were separated and combined, concentrated under reduced pressure, and separated by column chromatography to obtain 0.8 g of a yellow solid.

[0214] MS m / z(ESI):[M+H] + :351.33

[0215] Step 7 Preparation of [(7-hydroxy-5-(phenylethynyl)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid

[0216] Methyl [(7-hydroxy-5-phenylethynyl[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetate (0.8 g) was dissolved in ethanol (10 mL), and then an aqueous sodium hydroxide solution (18 mL, 2.0 M) was added. The mixture was stirred at room temperature until the reaction of the raw material was complete as monitored by TLC. The pH was adjusted to 4-5 with dilute hydrochloric acid and filtered to obtain 0.42 g of a light yellow solid.

[0217] 1H NMR (400MHz, d-DMSO) δ: 14.33(s,1H),13.04(s,1H),9.90(s,1H),8.64(s,1 H),7.74–7.72(m,2H),7.61–7.53(m,3H),7.37(s,1H),4.24(d,J=5.6Hz,2H)

[0218] MS m / z(ESI):[M+H] + :337.31

[0219] Example 11

[0220] Preparation of [(7-hydroxy-5-(phenylvinyl)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid

[0221] Step 1 Preparation of tert-butyl 5-(phenylvinyl)-7-(benzyloxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylate

[0222] Dissolve tert-butyl 5-iodo-7-(benzyloxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylate (500 mg), Pd(PPh3)2Cl2 (70.2 mg), sodium carbonate (360 mg), and styrene boronic acid (200 mg) in a 1:1 (volume ratio) mixture of ethylene glycol dimethyl ether and degassed water (30 mL). The atmosphere is replaced with nitrogen three times. Stir at 80°C until the reaction is complete as monitored by TLC, then discontinue heating. After the reaction solution cools to room temperature, extract with ethyl acetate, and wash the organic phase with saturated brine. Separate the organic phase, concentrate under reduced pressure, and purify by column chromatography to obtain 340 mg of a crude product.

[0223] Step 2 Preparation of 5-(phenylvinyl)-7-hydroxy-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylic acid

[0224] Dissolve tert-butyl 5-(phenylvinyl)-7-(benzyloxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylate (290 mg) in dichloromethane (10 mL) and cool to -65°C. Add boron tribromide in dichloromethane (3.4 mL, 1.0 M) and stir until the reaction is complete as monitored by TLC. Stop the reaction, concentrate the solution under reduced pressure, and then add the residue to a mixed solvent of water (10 mL) and methanol (3.0 mL). Stir at room temperature for 1 hour and filter to obtain 250 mg of a solid.

[0225] MS m / z(ESI):[M+H] + :282.04

[0226] Step 3 Preparation of [(7-hydroxy-5-(phenylvinyl)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid methyl ester

[0227] 5-(Vinyl)-7-hydroxy-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylic acid (143 mg), N,N-dimethylformamide (10 mL), DIPEA (329 mg), PyBOP (395 mg), and glycine methyl ester hydrochloride (178 mg) were added to the reaction flask in sequence. The mixture was stirred at room temperature overnight, and the solid was filtered out to obtain 50 mg of a crude product.

[0228] MS m / z(ESI):[M+H] + :353.28

[0229] Step 4 Preparation of [(7-hydroxy-5-(phenylvinyl)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid

[0230] Methyl [(7-hydroxy-5-(phenylvinyl)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetate (50 mg) was dissolved in methanol (2.0 mL), followed by the addition of 30% aqueous sodium hydroxide solution (1.0 mL). The mixture was stirred at room temperature until the reaction of the raw material was complete as monitored by TLC. The pH was adjusted to between 1 and 2 with dilute hydrochloric acid and filtered to obtain 10 mg of the product.

[0231] 1 H NMR (400MHz, d-DMSO) δ: 14.26 (s, 1H), 12.98 (s, 1H), 9.90 (t, J = 16.4Hz, 1H), 8.63 (s, 1 H),8.22(d,J=16.4Hz,1H),7.76–7.71(m,3H),7.50–7.40(m,4H),4.23(d,J=5.6Hz,2H)

[0232] MS m / z(ESI):[M+H] + :339.21

[0233] Example 12

[0234] Preparation of [(7-hydroxy-5-(1,2,3,4-tetrahydronaphthalen-2-yl)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid

[0235] Step 1 Preparation of tert-butyl 5-(3,4-dihydronaphthalen-2-yl)-7-(benzyloxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylate

[0236] Dissolve tert-butyl 5-iodo-7-(benzyloxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylate (5.40 g), Pd(PPh3)2Cl2 (800 mg), potassium carbonate (4.20 g), and 3,4-dihydronaphthyl-2-boronic acid pinacol ester (3.10 g) in a 2:1 volume ratio mixed solvent (30 mL) of ethylene glycol dimethyl ether and degassed water. Replace the nitrogen atmosphere three times. Stir at 80°C until the reaction is complete as monitored by TLC, then stop heating. After the reaction solution cools to room temperature, extract with ethyl acetate, and wash the organic phase with saturated brine. Separate the organic phase, concentrate under reduced pressure, and purify by column chromatography to obtain 3.30 g of crude product.

[0237] Step 2 Preparation of 5-(3,4-dihydronaphthalen-2-yl)-7-(benzyloxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylic acid

[0238] tert-Butyl 5-(3,4-dihydronaphthalen-2-yl)-7-(benzyloxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylate (3.30 g) was dissolved in a mixed solution of toluene and ethyl acetate (28 mL) in a volume ratio of 3:1. Methanesulfonic acid (2.80 g) was then added to the reaction system and stirred until the reaction of the raw materials was complete as monitored by TLC. Ethyl acetate (20.0 mL) was added to the reaction system and stirred for crystallization. The solid was collected by suction filtration. Subsequently, DMF / H2O (20.0 mL / 40.0 mL) was used for slurrying, stirring for crystallization, suction filtration, and solid collection to obtain 2.70 g of a light blue solid.

[0239] Step 3 Preparation of [5-(3,4-dihydronaphthalen-2-yl)-7-(benzyloxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid methyl ester

[0240] Tert-butyl 5-(3,4-dihydronaphthalen-2-yl)-7-(benzyloxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylate (2.70 g), N,N-dimethylformamide (20.0 mL), HOBt (1.10 g), EDCI (1.40 g), glycine methyl ester hydrochloride (0.90 g), and triethylamine (1.50 g) were added to the reaction flask in sequence and stirred at room temperature until the reaction of the raw materials was complete as monitored by TLC. Water (30.0 mL) was added to the reaction system and stirred for crystallization. The solid was collected by suction filtration and dried to obtain an off-white solid (2.2 g).

[0241] Step 4 Preparation of [(7-hydroxy-5-(1,2,3,4-tetrahydronaphthalen-2-yl)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid methyl ester

[0242] [5-(3,4-Dihydronaphthalen-2-yl)-7-(benzyloxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid (1.0 g) was dissolved in tetrahydrofuran solution (8.0 mL), and then palladium carbon (300 mg) was added to replace the hydrogen. The mixture was stirred at room temperature until the reaction of the raw material was complete as monitored by TLC. The mixture was filtered through celite and concentrated under reduced pressure. The mixture was then slurried with isopropyl ether / n-hexane (1.5 mL / 1.5 mL) and filtered to obtain about 500 mg of a similar white solid.

[0243] Step 5 Preparation of [(7-hydroxy-5-(1,2,3,4-tetrahydronaphthalen-2-yl)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid

[0244] Methyl [(7-hydroxy-5-(1,2,3,4-tetrahydronaphthalen-2-yl)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetate (0.20 g) was dissolved in ethanol solution (2.0 mL), followed by addition of aqueous sodium hydroxide solution (2.0 mL, 4.0 M). The mixture was stirred at room temperature until the reaction of the raw material was complete as monitored by TLC. The pH was adjusted to between 6 and 7 with dilute hydrochloric acid and filtered to give 200 mg of an off-white solid.

[0245] 1 H NMR (400MHz, d-DMSO) δ: 14.37(brs,1H),9.93(s,1H),8.56(s,1H),7.13(s,4H),6.78( s,1H),4.12(d,J=4.0Hz,2H),3.81–3.76(m,1H),3.26–2.86(m,4H),2.26–2.05(m,2H),

[0246] MS m / z(ESI):[M+H] + :367.32

[0247] Example 13 Preparation of [(7-hydroxy-5-(1,2,3,4-tetrahydronaphthalen-2-yl)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid chiral isomer A

[0248] Step 1 Preparation of tert-butyl 5-(3,4-dihydronaphthalen-2-yl)-7-(benzyloxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylate

[0249] Dissolve tert-butyl 5-iodo-7-(benzyloxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylate (5.40 g), Pd(PPh3)2Cl2 (800 mg), sodium carbonate (4.20 g), and 3,4-dihydronaphthyl-2-boronic acid pinacol ester (3.10 g) in a 2:1 volume ratio (30 mL) mixed solvent of ethylene glycol dimethyl ether and degassed water. Replace the nitrogen atmosphere three times. Stir at 80°C until the reaction is complete as monitored by TLC, then stop heating. After the reaction solution cools to room temperature, extract with ethyl acetate, and wash the organic phase with saturated brine. Separate the organic phase, concentrate under reduced pressure, and purify by column chromatography to obtain 3.30 g of crude product.

[0250] Step 2 Preparation of 5-(3,4-dihydronaphthalen-2-yl)-7-(benzyloxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylic acid

[0251] tert-Butyl 5-(3,4-dihydronaphthalen-2-yl)-7-(benzyloxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylate (3.30 g) was dissolved in a mixed solution of toluene and ethyl acetate (28 mL) in a volume ratio of 3:1. Methanesulfonic acid (2.80 g) was then added to the reaction system and stirred until the reaction of the raw materials was complete as monitored by TLC. Ethyl acetate (20.0 mL) was added to the reaction system and stirred for crystallization. The solid was collected by suction filtration. Subsequently, DMF / H2O (20.0 mL / 40.0 mL) was used for slurrying, stirring for crystallization, suction filtration, and solid collection to obtain 2.70 g of a light blue solid.

[0252] Step 3 Preparation of [5-(3,4-dihydronaphthalen-2-yl)-7-(benzyloxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid methyl ester

[0253] Tert-butyl 5-(3,4-dihydronaphthalen-2-yl)-7-(benzyloxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carboxylate (2.70 g), N,N-dimethylformamide (20.0 mL), HoBt (1.10 g), EDCI (1.40 g), and glycine methyl hydrochloride (0.90 g) were added to the reaction flask in sequence and stirred at room temperature until the reaction of the raw materials was complete as monitored by TLC. Water (30.0 mL) was added to the reaction system and stirred for crystallization. The solid was collected by suction and dried to obtain an off-white solid (2.2 g).

[0254] Step 4 Preparation of [(7-hydroxy-5-(1,2,3,4-tetrahydronaphthalen-2-yl)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid methyl ester

[0255] [5-(3,4-Dihydronaphthalen-2-yl)-7-(benzyloxy)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid (1.0 g) was dissolved in tetrahydrofuran solution (8.0 mL), and palladium carbon (300 mg) was added. The mixture was stirred at room temperature until the reaction of the raw material was complete as monitored by TLC. The mixture was filtered through celite and concentrated under reduced pressure. The mixture was then slurried with isopropyl ether / n-hexane (1.5 mL / 1.5 mL) and filtered to obtain about 500 mg of a similar white solid.

[0256] Step 5 Preparation of [(7-hydroxy-5-(1,2,3,4-tetrahydronaphthalen-2-yl)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid methyl ester chiral isomer A (methyl acetate chiral isomer A)

[0257] The product synthesized in the previous step was subjected to chiral resolution to obtain 5 g of methyl [(7-hydroxy-5-(1,2,3,4-tetrahydronaphthalen-2-yl)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetate. Resolution conditions: (Chiral column: (S,S) Whelk-O1, 5 x 25 cm, 10 μm; Mobile phase A: CO2, Mobile phase B: ACN:IPA = 1:1; Flow rate: 200 mL / min; Gradient: 50%-50%; B run time: 12 minutes; Wavelength: 220 nm; Peak elution time: 8.50 minutes; Sample solution: Methanol: Dichloromethane = 1:1; Injection volume: 2 mL; Number of injections: 50)) yielding chiral isomer A as a white solid (2.060 g, 41.2% yield).

[0258] Step 6 Preparation of [(7-hydroxy-5-(1,2,3,4-tetrahydronaphthalen-2-yl)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid chiral isomer A

[0259] Methyl [(7-hydroxy-5-(1,2,3,4-tetrahydronaphthalen-2-yl)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetate chiral isomer A (0.20 g) was dissolved in ethanol solution (2.0 mL), followed by addition of aqueous sodium hydroxide solution (1.0 mL, 4.0 M). The mixture was stirred at room temperature until the reaction of the raw material was complete as monitored by TLC. The pH was adjusted to between 1 and 2 with dilute hydrochloric acid and filtered to obtain 200 mg of an off-white solid.

[0260] 1H NMR (400MHz, d-DMSO) δ: 14.37(brs,1H),9.93(s,1H),8.56(s,1H),7.13(s,4H),6.78( s,1H),4.12(d,J=4.0Hz,2H),3.81–3.76(m,1H),3.26–2.86(m,4H),2.26–2.05(m,2H),

[0261] MS m / z(ESI):[M+H] + :367.32

[0262] [α]D 20 =+37.384 (c 1.01, DMF).

[0263] Example 14 Preparation of [(7-hydroxy-5-(1,2,3,4-tetrahydronaphthalen-2-yl)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid chiral isomer B

[0264] Step 1 Preparation of [(7-hydroxy-5-(1,2,3,4-tetrahydronaphthalen-2-yl)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid methyl ester chiral isomer B

[0265] Chiral separation of the synthesized [(7-hydroxy-5-(1,2,3,4-tetrahydronaphthalen-2-yl)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetate was performed, yielding 5g. Separation conditions: (Chiral column: (S,S) Whelk-O1, 5 x 25 cm, 10 μm; Mobile phase A: CO2, Mobile phase B: ACN:IPA = 1:1; Flow rate: 200 mL / min; Gradient: 50%-50%; B over 12 minutes; Wavelength: 220 nm; (OB peak time): 10.09 minutes; Sample solution: Methanol: Dichloromethane = 1:1; Injection volume: 2 mL; Number of injections: 50) yielded Chiral Isomer B (Methyl Acetate Chiral Isomer B): a white solid (1.997 g, 39.94% yield).

[0266] Step 2 Preparation of [(7-hydroxy-5-(1,2,3,4-tetrahydronaphthalen-2-yl)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid chiral isomer B

[0267] Methyl [(7-hydroxy-5-(1,2,3,4-tetrahydronaphthalen-2-yl)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetate chiral isomer B (0.20 g) was dissolved in ethanol solution (2.0 mL), followed by addition of aqueous sodium hydroxide solution (1.0 mL, 4.0 M). The mixture was stirred at room temperature until the reaction of the raw material was complete as monitored by TLC. The pH was adjusted to between 1 and 2 with dilute hydrochloric acid and filtered to give 200 mg of an off-white solid.

[0268] 1 H NMR (400MHz, d-DMSO) δ: 14.37(brs,1H),9.93(s,1H),8.56(s,1H),7.13(s,4H),6.78( s,1H),4.12(d,J=4.0Hz,2H),3.81–3.76(m,1H),3.26–2.86(m,4H),2.26–2.05(m,2H),

[0269] MS m / z(ESI):[M+H] + :367.32

[0270] [α]D 20 =–36.601(c 1.01,DMF).

[0271] Example 15 Biological Test

[0272] 1. PHD2 enzyme activity inhibition assay

[0273] 1.1 Reagents and consumables

[0274] PHD2 enzyme: purchased from Active Motif;

[0275] α-Ketoglutarate sodium salt: purchased from Sigma;

[0276] FITC-HIF1α: purchased from GL;

[0277] Multi-well plate Nunc TM 384: Purchased from Thermo Scientific.

[0278] 1.2 Test methods

[0279] (1) Prepare 1× Assay buffer;

[0280] (2) Preparation of compound concentration gradient: The test compound concentration starts at 10 μM and is diluted 3-fold to 10 concentrations in duplicate. The solution is diluted to 100 times the final concentration in a 384-well plate, and then 100 nL is transferred to the 384-well reaction plate using the non-contact acoustic pipetting system Echo550 for later use. 100 nL of 100% DMSO is added to the negative control well and the positive control well respectively.

[0281] (3) Prepare an enzyme solution with 2x final concentration using 1× Assay buffer;

[0282] (4) Add 5 μL of enzyme solution at 2 times the final concentration to the compound wells and positive control wells respectively;

[0283] (5) Add 5 μL of 1× Assay buffer to the negative control well;

[0284] (6) Centrifuge at 1000 rpm for 30 seconds, shake to mix, and incubate for 15 minutes;

[0285] (7) Prepare a tracer solution with 2x final concentration using 1× Assay buffer and add 5.0 μL of the tracer solution to start the reaction;

[0286] (8) The 384-well plate was centrifuged at 1000 rpm for 30 seconds and mixed by oscillation for 60 minutes. The mP values ​​were read on an Envision Multimode Plate Reader (Perkin Elmer). The data were exported and processed to obtain the inhibition rate of the test compound. Compound numbers 1-12 correspond to the compounds prepared in Examples 1-12, respectively. The results are shown in the following table:

[0287] Table 1

[0288] As can be seen from the above table, the compounds of the present invention have good HIF-prolyl hydroxylase inhibitory activity, and it can be seen that compounds 1-12 have good inhibitory activity.

[0289] 2. In vitro erythropoietin (EPO) induction activity test of the compounds of the present invention

[0290] The in vitro erythropoietin (EPO)-inducing activity of the compounds of Examples 10-12 of the present invention was evaluated using the human hepatocellular carcinoma cell line Hep3B (ATCC). Hep3B cells were cultured in Eagle's Minimum Essential Medium (EMEM) at 37°C in the presence of 10% fetal bovine serum (FBS). The experimental steps were as follows:

[0291] (1) Cell preparation: Cells were seeded in 96-well plates at a density of 2 × 10^4 cells per well and 100 μL per well;

[0292] (2) Preparation of compound concentration gradient: 100 μM, 20 μM, 2 replicates. Prepare a solution with a final concentration of 200 times in a 96-well plate, dilute the compound 200 / 3 times with cell culture medium, and then aspirate 50 μL to administer to the cells. Add 50 μL of culture medium containing DMSO to the negative control well to make the final concentration contain 5‰ DMSO, and add 50 μL of the highest concentration of positive compound to the positive control well, and incubate at 37°C for 24 hours;

[0293] (3) Wash the reaction plate twice with approximately 400 μL of 1X Wash Buffer per well;

[0294] (4) Add 100 μL of diluted standard (including standard blank control) to the appropriate wells;

[0295] (5) Add 50 μL of sample and 50 μL of Sample Diluent to the sample well;

[0296] (6) Add 50 μL of 1X Biotin Conjugated Antibody to all wells and incubate at room temperature for 1 hour;

[0297] (7) Wash the reaction plate six times with approximately 400 μL of 1X Wash Buffer per well;

[0298] (8) Add 100 μL of 1X Streptavidin-HRP to each well. Incubate at room temperature for 15 minutes.

[0299] (9) Wash the reaction plate six times with approximately 400 μL of 1X Wash Buffer per well;

[0300] (10) Add 100 μL of TMB Substrate Solution to each well. Incubate at room temperature for 10 minutes.

[0301] (11) Add 100 μL Stop Solution to each well;

[0302] (12) Read OD450 using EnSight.

[0303] The EPO induction activity of each compound was expressed as half maximal effect concentration (EC50). The experimental results are shown in the following table:

[0304] Table 2

[0305] As can be seen from the above table, the compounds of the present invention have higher EPO induction activity. It can be seen that the compounds obtained in Examples 10-14 have good induction activity.

[0306] Example 16 Pharmacokinetic Evaluation of Compounds

[0307] 1. Purpose of the study

[0308] After intravenous or oral administration of the compounds to SD rats, the concentrations of the relevant compounds in plasma were measured using LC-MS / MS to preliminarily evaluate the pharmacokinetics and absolute bioavailability of the compounds prepared in Examples 13 and 14 in rats.

[0309] 2. Strain: Sprague Dawley rats, sex: male, weight: 200-250 g

[0310] Source: Zhejiang Weitonglihua Experimental Animal Technology Co., Ltd.

[0311] 3. Experimental Operation

[0312] The pharmacokinetic profiles of the compounds following intravenous and oral administration in rodents were tested using standard protocols. Candidate compounds were formulated as clear solutions and administered to rats via single intravenous and oral gavage. The intravenous vehicle consisted of a 10% solution of N,N-dimethylacetamide (DMA) and Solutol HS15 in a specific ratio, while the oral vehicle consisted of a suspension of sodium carboxymethylcellulose (CMC) in a specific ratio. Blood samples were collected from the animals at 5, 15, 30, 1, 2, 4, 6, 8, 10, and 24 hours after administration into K2EDTA tubes and stored on ice until centrifugation.

[0313] Experimental results:

[0314] As can be seen from the above table, the compounds prepared in Examples 13 and 14 of the present invention have good PK properties. It can be seen that the compounds prepared in Examples 13 and 14 have significantly longer half-lives and higher in vivo exposures.

[0315] Example 17 Pharmacological Evaluation of Compounds

[0316] 1. Purpose of the experiment

[0317] The purpose of this experiment is to evaluate the effect of continuous administration of the compounds prepared in Examples 12, 13, and 14 on EPO (erythropoietin) in SD rats.

[0318] 2. Animals

[0319] Species: Rat

[0320] Strain: SD (Sprague Dawley)

[0321] Age and weight: 8 weeks, weight 200-220g

[0322] Gender: Male

[0323] Animal source: Beijing Weitonglihua Experimental Animal Technology Co., Ltd.

[0324] 3. Information on the test compound and positive control drug

[0325] Positive drug: Enarodustat

[0326] Description: Commercialized drugs approved for marketing in Japan

[0327] Specifications and content: 2mg JTZ-951 (Enarodustat), 140 tablets / box

[0328] 4. Experimental plan:

[0329] After one week of adaptive feeding, SD rats were randomly divided into three groups, each with 6 rats: a blank group, a positive drug group (enarodustat), and a compound group according to Example 12. Enarodustat tablets were crushed in a mortar and pestle, and 0.5% methylcellulose was added in a quantitative manner, and magnetic stirring was used to mix. The compound prepared in Example 12 was weighed, and 0.5% methylcellulose was added in a quantitative manner, and magnetic stirring was used to mix. The final concentration of all compounds was 0.3 mg / mL, and the dosing volume was 10 mL / kg. The drug was administered once daily for 14 consecutive days, and blood was collected 4 hours after administration on the 14th day to measure EPO levels.

[0330] 5. Results and Analysis

[0331] The results showed that 14 days after administration, the EPO levels in the serum of rats in the positive drug Enarodustat and compound groups prepared in Examples 12, 13, and 14 increased significantly, and the EPO levels in the plasma of the compounds prepared in Examples 12, 13, and 14 after administration were significantly higher than that of the positive drug Enarodustat.

[0332] Table 3 EPO levels (pg / mL) 14 days after administration

[0333] Note: 2904.6 pg / mL is the upper limit of measurement.

[0334] In summary, the compounds prepared in Examples 12, 13, and 14 can significantly increase the EPO level in SD rats, and the overall effect is significantly better than that of the positive drug Enarodustat, showing excellent pharmacological effects.

[0335] Example 18 Determination of the Chiral Configuration of Methyl Acetate Chiral Isomer A of Example 13

[0336] 1. Compound Preparation

[0337] With reference to the preparation method of [(7-hydroxy-5-(1,2,3,4-tetrahydronaphthalen-2-yl)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid methyl ester chiral isomer A (step five) in Example 13, methyl acetate chiral isomer A was obtained by chiral resolution. This isomer is the compound before the final product in Example 13 (step five product).

[0338] 2. Single crystal cultivation

[0339] Weigh approximately 3-7 mg of methyl acetate chiral isomer A into a vial. Gradually add acetonitrile while stirring at 50°C until the solution is just clear or nearly clear. Filter while hot. Place the filtrate into a clean vial and program the temperature using a Crystal 16 instrument, initially decreasing from 50°C to 30°C at a rate of 0.01°C / min. Then maintain the temperature at 30°C for approximately 2 days.

[0340] Needle-shaped single crystals were obtained from acetonitrile by cooling crystallization, as shown in Figure 1, and used for X-ray single crystal diffraction analysis.

[0341] 3. Instruments and parameters

[0342] Single crystal data of chiral isomer A of methyl acetate were obtained by using a Rigaku XtaLAB Synergy DW diffractometer at 180 K using Cu Kα radiation. The diffraction data were collected and analyzed using the CrysAlisPro program for data reduction and absorption correction, and the structure was solved using the SHELXT program using a dual-space algorithm. All non-hydrogen atoms were located directly from the difference Fourier map, and hydrogen atoms were filled in at the parent atoms. The final structure was refined using the SHELXL program using the F2 full-matrix least-squares method.

[0343] 4. X-ray diffraction analysis

[0344] The single crystal structure of methyl acetate chiral isomer A belongs to the monoclinic system, P21 space group, and the molecular formula of the single crystal is C 20 H 20 N₄O₄. Each asymmetric unit contains one molecule of methyl acetate chiral isomer A, and each unit cell contains two asymmetric units. The absolute configuration of the chiral carbon is "R."

[0345] The refined single crystal structure is shown in Figures 2, 3, and 4. The crystal structure parameters are summarized in Table 4, and the atomic coordinates, anisotropic displacement parameters, torsion angles, hydrogen bonds, bond lengths, and bond angles are detailed in Tables 5 to 11. The XRPD pattern calculated from the single crystal structure is generally consistent with the measured results of the single crystal sample, but with some shift in peak position. This is likely due to the fact that the single crystal data were collected at 180 K, while the experimentally measured XRPD pattern was obtained at room temperature (Figure 5).

[0346] Table 4. Crystal structure parameters of methyl acetate chiral isomer A

[0347] Table 5. Atomic coordinates of chiral isomer A of methyl acetate (×10 4 ) and equivalent isotropic displacement parameters

[0348] Table 6. Anisotropic shift parameters of chiral isomer A of methyl acetate

[0349] Table 7. Bond lengths of chiral isomer A of methyl acetate

[0350] Table 8. Bond angles of chiral isomer A of methyl acetate

[0351] Table 9. Torsion angles of methyl acetate chiral isomer A

[0352] Table 10. Hydrogen bonds of methyl acetate chiral isomer A

[0353] Table 1. Atomic coordinates of chiral isomer A of methyl acetate (×10 4 ) and isotropic displacement parameters

[0354] 5. Analysis methods

[0355] 5.1 Polarized Light Microscopy (PLM)

[0356] PLM analysis was performed using an ECLIPSE LV100POL (Nikon, JPN) polarizing microscope. A small amount of sample was spread onto a glass slide, cedar oil was added to disperse the sample, and a coverslip was applied. The sample was then observed under a microscope using a 10x objective.

[0357] 5.2 X-ray powder diffraction (XRPD)

[0358] The solid sample was tested using an X-ray diffractometer. The sample was spread flat on a zero-background single crystal silicon sample plate, gently pressed flat, and analyzed according to the parameters in Table 12.

[0359] Table 12. XRPD test parameters

[0360] The above single crystal structure analysis shows that the chiral configuration of the methyl acetate chiral isomer A is "R" configuration, as shown in Figure 6. Therefore, it can be seen that in Example 13, when the methyl ester is removed using a sodium hydroxide aqueous solution to obtain the carboxylic acid product, the chiral configuration that has been produced is not changed. Therefore, it can be inferred that the chiral configuration of the product [(7-hydroxy-5-(1,2,3,4-tetrahydronaphthalen-2-yl)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid chiral isomer A in Example 13 is "R" configuration.

[0361] Example 19 Determination of the Chiral Configuration of Methyl Acetate Chiral Isomer B of Example 14

[0362] 1. Compound Preparation

[0363] With reference to the preparation method of [(7-hydroxy-5-(1,2,3,4-tetrahydronaphthalen-2-yl)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid methyl ester chiral isomer B (step 1) in Example 14, methyl acetate chiral isomer B was obtained by chiral resolution. This isomer is the compound before the final product in Example 14.

[0364] 2. Single crystal cultivation

[0365] Weigh an appropriate amount of methyl acetate chiral isomer B into a vial. At room temperature, gradually add dimethyl sulfoxide (DMSO) while stirring until the solution is just clear or nearly clear. Filter the solution and place it into a clean vial. Cover and pierce the vial. Allow the vial to stand at room temperature for slow evaporation.

[0366] Needle-shaped single crystals were obtained from dimethyl sulfoxide by slow volatilization, as shown in FIG7 , and used for X-ray single crystal diffraction analysis.

[0367] 3. Instruments and parameters

[0368] Single crystal data of chiral isomer B of methyl acetate were obtained by using a Rigaku XtaLAB Synergy DW diffractometer at 180 K using Cu Kα radiation. The diffraction data were collected and analyzed using the CrysAlisPro program for data reduction and absorption correction, and the structure was solved using the SHELXT program using a dual-space algorithm. All non-hydrogen atoms were located directly from the difference Fourier map, and hydrogen atoms were filled in at the parent atoms. The final structure was refined using the SHELXL program using the F2 full-matrix least-squares method.

[0369] 4. X-ray diffraction analysis

[0370] The single crystal structure of methyl acetate chiral isomer B belongs to the monoclinic system, P21 space group, and the molecular formula of the single crystal is C 20 H 20 N₄O₄. Each asymmetric unit contains one molecule of methyl acetate chiral isomer B, and each unit cell contains two asymmetric units. The absolute configuration of the chiral carbon is "S."

[0371] The refined single crystal structure is shown in Figures 8, 9, and 10. The crystal structure parameters are summarized in Table 13, and the atomic coordinates, anisotropic displacement parameters, torsion angles, hydrogen bonds, bond lengths, and bond angles are summarized in Tables 14 to 20. The XRPD pattern obtained from the experimental measurement of the single crystal sample is more peaky than the calculated XRPD pattern; this may be due to the near-evaporation of the solvent after the single crystal analysis, resulting in the precipitation of other polymorphs during the solvent evaporation process (Figure 11).

[0372] Table 13. Crystal structure parameters of methyl acetate chiral isomer B

[0373] Table 14. Atomic coordinates of chiral isomer B of methyl acetate (×10 4 ) and equivalent isotropic displacement parameters

[0374] Table 15. Anisotropic shift parameters of methyl acetate chiral isomer B

[0375] Table 16. Bond lengths of methyl acetate chiral isomer B

[0376] Table 17. Bond angles of methyl acetate chiral isomer B

[0377] Table 18. Torsion angles of methyl acetate chiral isomer B

[0378] Table 19. Hydrogen bonds of methyl acetate chiral isomer B

[0379] Table 20. Hydrogen atom coordinates of methyl acetate chiral isomer B (×10 4 ) and isotropic displacement parameters

[0380] 5. Analysis methods

[0381] 5.1 Polarized Light Microscopy (PLM)

[0382] PLM analysis was performed using an ECLIPSE LV100POL (Nikon, JPN) polarizing microscope. A small amount of sample was spread flat on a glass slide, cedar oil was added to disperse the sample, and a coverslip was applied. The sample was then placed under a microscope and observed using an objective lens.

[0383] 5.2 X-ray powder diffraction (XRPD)

[0384] Solid samples were examined using an X-ray diffractometer. The samples were spread flat on a zero-background single-crystal silicon sample plate, gently pressed flat, and analyzed according to the parameters in Table 21.

[0385] Table 21. XRPD test parameters

[0386] The above single crystal structure analysis shows that the chiral configuration of the methyl acetate chiral isomer B is the "S" configuration, as shown in Figure 12. Therefore, it can be seen that in Example 14, when the methyl ester is removed using a sodium hydroxide aqueous solution to obtain the carboxylic acid product, the chiral configuration that has already been produced is not changed. Therefore, it can be inferred that the chiral configuration of the product [(7-hydroxy-5-(1,2,3,4-tetrahydronaphthalen-2-yl)-[1,2,4]triazolo[1,5-a]pyridine-8-carbonyl)amino]acetic acid chiral isomer B in Example 14 is the "S" configuration.

[0387] Example 20 Subacute toxicity test of compounds of Examples 13 and 14

[0388] 1. Experimental Materials

[0389] 1.1 Test sample

[0390] Table 22 Sample information

[0391] 1.2 Prescription preparation

[0392] Weigh appropriate amounts of Example 13 and Example 14 using a balance and place them in appropriate glass bottles. DMSO, Solutol HS-15, and 0.9% saline (v:v:v ratio = 5%:10%:85%) were then added to the bottles in sequence. Vortex and stir the mixture, continuously monitoring the dissolution of the drugs during stirring until they were completely dissolved / suspended. Finally, a dosing solution of Example 13 and Example 14 was prepared at a concentration of 0.67 mg / mL, which was used immediately upon preparation.

[0393] 2. Experimental Animals

[0394] The source and number of experimental animals are shown in Table 23. The animal room was well-ventilated and air-conditioned, maintained at a temperature of 20-25°C and a humidity of 40%-70%. Light and dark illumination were alternating for 12 hours each, and animals had free access to food and water. Sprague-Dawley rats were enrolled in this study after being acclimated for at least three days prior to dosing and in good physical condition as determined by a veterinarian.

[0395] Table 23 Sources and numbers of experimental animals

[0396] 3. Experimental Plan

[0397] 3.1 Trial cycle and dose setting

[0398] Any operation on the experimental animals in this experiment complies with the requirements of the "Experimental Animal Management Regulations" promulgated by the Ministry of Science and Technology of the People's Republic of China. This test sets a solvent control group, an Example 13 compound administration group, and an Example 14 compound administration group. The dosage of the Example 13 compound administration group and the Example 14 compound administration group is (10 mg / kg), once a day, and administered for 14 consecutive days. The blank control group is given an equal amount of blank solvent, and the administration volume is 15 mL / kg. The symptoms and degree of toxic reactions of the animals are observed after each administration. After the last administration, the animals are observed for 7 days. Drinking water is free during the experiment. See Table 24 for details.

[0399] Table 24 Experimental dosage and group design

[0400] After weighing, calculate the theoretical dosage volume for each SD rat using the following formula. The actual dosage and sample collection time for each SD rat should be recorded in detail in the corresponding table.

[0401] 3.2 Experimental observation

[0402] During the experiment, the experimenter and veterinarian must continuously monitor the physical signs and health of the experimental animals. Any abnormal behavior of the animals, such as pain, depression, or decreased activity, must be recorded in the original experimental records. If the abnormal behavior of the experimental animals exceeds the requirements of the relevant IACUC animal welfare documents, the veterinarian may determine whether to terminate the experiment and notify the project leader.

[0403] 3.3 Cage-side observation

[0404] During the experiment, all animals were observed at least twice a day at the cage side and for 2 hours after the end of the drug administration. During the drug administration process, if the animals had serious toxic side effects or died, the observation results should be recorded in the corresponding form. Observation indicators include: animal appearance, behavioral activities, secretions, excretions, diet, death (number of deaths, time of death, pre-mortem reaction), etc. If clinical symptoms appear, the number of observations should be increased to observe in detail the symptoms of the animal's poisoning reaction, the onset time, severity, duration, and whether it is reversible. If an animal is found to be dead or dying, the animal should be dissected and observed in a timely manner in accordance with regulations.

[0405] 3.4 weight

[0406] The animals should be weighed daily before dosing for 14 consecutive days and their weight recorded on the appropriate form. The food intake of the animals should also be monitored during the trial.

[0407] 3.5 Clinical Pathology

[0408] Before the first dose and 24 hours after the last dose, collect approximately 1.8-2.0 mL of whole blood for hematology and biochemistry testing. In addition, collect blood 24 hours after the last dose for coagulation testing. If the animal's condition is abnormal during dosing, the hematology, coagulation, and biochemistry tests will be performed earlier. Fast for at least 12 hours before blood collection.

[0409] Approximately 500 μL of whole blood was placed in an anticoagulant tube containing EDTA-K2 for hematological analysis:

[0410] White blood cell count (WBC)

[0411] Neutrophil differential count (absolute count, NEUT; percentage, %NEUT)

[0412] Lymphocyte differential count (absolute count, LYM; percentage, %LYM)

[0413] Monocyte differential count (absolute count, MONO; percentage, %MONO)

[0414] Eosinophil count (absolute count, EOS; percentage, %EOS)

[0415] Basophil count (absolute count, BASO; percentage, %BASO)

[0416] Red blood cell count (RBC)

[0417] Hemoglobin (HGB)

[0418] Hematocrit (HCT)

[0419] Mean corpuscular volume (MCV)

[0420] Mean corpuscular hemoglobin (MCH)

[0421] Mean corpuscular hemoglobin concentration (MCHC)

[0422] Red blood cell coefficient of variation (RDW-CV)

[0423] Red blood cell distribution width standard deviation (RDW-SD)

[0424] Platelet count (PLT)

[0425] Mean platelet distribution width (MPV)

[0426] Platelet mean volume (PDW)

[0427] Platelet count (PCT)

[0428] Approximately 1.5 mL of whole blood was placed in a separating gel blood collection tube (without anticoagulant) for blood biochemistry analysis:

[0429] Alanine aminotransferase (ALT)

[0430] Aspartate Aminotransferase (AST)

[0431] Alkaline Phosphatase (ALP)

[0432] Albumin (ALB)

[0433] Total cholesterol (Cholesterol, TC)

[0434] Creatinine (CRE)

[0435] Urea (UREA)

[0436] Creatine Phosphokinase (CK)

[0437] Triglyceride (TG)

[0438] Total Bilirubin (TBIL)

[0439] Total Protein (TP)

[0440] Aspartate aminotransferase / alanine aminotransferase (AST / ALT)

[0441] Globulin (GloⅡ)

[0442] Albumin / globulin (A / GⅡ)

[0443] Electrolytes (K+ / Na+ / Cl- / Ca2+)

[0444] 4. Conclusion

[0445] Under the experimental conditions, the compounds of Examples 13 and 14 did not cause obvious gastrointestinal necrosis, black stomach bleeding spots, or flatulence. Their toxicity was lower than that of Enarodustat, and they had better safety.

[0446] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A compound represented by formula (I) or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs: in, R is selected from the following atoms or groups: L is -CH2- or -CH2O-; The value of n is an integer between 0 and 2; R 1 、R 2 、R 3 Each is independently selected from substituted or unsubstituted alkyl, cycloalkyl, aryl, heterocyclic group; R 4 is selected from hydrogen, substituted or unsubstituted alkyl, cycloalkyl, alkoxy, aryl, and heterocyclic groups.

2. The compound according to claim 1 or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, characterized in that: The aryl group includes an aromatic heterocycle.

3. The compound according to claim 1 or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, wherein: L is -CH2- or -CH2O-; The value of n is 0 or 1; R 1 is substituted or unsubstituted C 1-4 Alkyl, phenyl, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, 5-6 membered heterocyclic group containing oxygen and / or nitrogen, the substituent is C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 At least one of a haloalkyl group, a halogen group, a cyano group, a phenyl group, a 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, and a 5-6 membered heterocyclic group containing oxygen and / or nitrogen; R 2 is substituted or unsubstituted C 1-4 Alkyl, phenyl, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, 5-6 membered heterocyclic group containing oxygen and / or nitrogen, the substituent is C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 At least one of a haloalkyl group, a halogen group, a cyano group, a phenyl group, a 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, and a 5-6 membered heterocyclic group containing oxygen and / or nitrogen; R 3 is substituted or unsubstituted C 1-10 Alkyl, phenyl, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, 5-6 membered heterocyclic group containing oxygen and / or nitrogen, the substituent is C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 At least one of a haloalkyl group, a halogen group, a cyano group, a phenyl group, a 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, or a 5-6 membered heterocyclic group containing oxygen and / or nitrogen; R 4 is hydrogen, substituted or unsubstituted C 1-10 Alkyl, C 1-10 Alkoxy, phenyl, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, 5-6 membered heterocyclic group containing oxygen and / or nitrogen, the substituent is C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 At least one of a haloalkyl group, a halogen group, a cyano group, a phenyl group, a 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, or a 5-6 membered heterocyclic group containing oxygen and / or nitrogen.

4. The compound according to claim 2 or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, wherein: L is -CH2- or -CH2O-; The value of n is 0 or 1; R 1 is a substituted or unsubstituted phenyl group, wherein the substituent is C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 At least one of a haloalkyl group, a halogen group, a cyano group, a phenyl group, a 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, or a 5-6 membered heterocyclic group containing oxygen and / or nitrogen; R 2 is a substituted or unsubstituted phenyl group, wherein the substituent is C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 At least one of a haloalkyl group, a halogen group, a cyano group, a phenyl group, a 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, and a 5-6 membered heterocyclic group containing oxygen and / or nitrogen; R 3 is substituted or unsubstituted C 1-10 Alkyl, phenyl, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, 5-6 membered heterocyclic group containing oxygen and / or nitrogen, the substituent is C 1-4 Alkyl, C 1-4 At least one of an alkoxy group, a halogen group, a 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, and a 5-6 membered heterocyclic group containing oxygen and / or nitrogen; R 4 is hydrogen, substituted or unsubstituted C 1-10 Alkyl, C 1-10 Alkoxy, phenyl, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, 5-6 membered heterocyclic group containing oxygen and / or nitrogen, the substituent is C 1-4 Alkyl, C 1-4 At least one of an alkoxy group, a halogen group, a 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, and a 5-6 membered heterocyclic group containing oxygen and / or nitrogen.

5. The compound according to claim 3 or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, wherein: The value of n is 0 or 1; R 1 is a substituted or unsubstituted phenyl group, wherein the substituent is C 1-4 Alkyl, C 1-4 Alkoxy, halogen, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen; preferably, the C 1-4 Alkyl, C 1-4 Alkoxy, oxygen and / or nitrogen-containing 5-6 membered aromatic heterocyclic group or oxygen and / or nitrogen-containing 5-6 membered heterocyclic group is para-substituted relative to the parent core structure, and halogen is ortho-, meta- or para-substituted relative to the parent core structure; R 2 is a substituted or unsubstituted phenyl group, wherein the substituent is C 1-4 Alkyl, C 1-4 Alkoxy, halogen or 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen; preferably, the C 1-4 Alkyl, C 1-4 The alkoxy and the 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or the 5-6 membered heterocyclic group containing oxygen and / or nitrogen are para-substituted relative to the parent core structure, and the halogen is relative to Substitution at the ortho, meta or para position on the parent core structure; R 3 is substituted or unsubstituted C 1-5 Alkyl, phenyl, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen, the substituent is C 1-4 Alkyl, C 1-4 Alkoxy, halogen, 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen; preferably, the C 1-4 Alkyl, C 1-4 The alkoxy group, the 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen, or the 5-6 membered heterocyclic group containing oxygen and / or nitrogen is para-substituted relative to the parent core structure, and the halogen group is ortho-, meta-, or para-substituted relative to the parent core structure; R 4 is hydrogen, substituted or unsubstituted C 1-5 Alkyl, C 1-5 Alkoxy, phenyl, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen, the substituent is C 1-4 Alkyl, C 1-4 Alkoxy, halogen, 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen; preferably, the C 1-4 Alkyl, C 1-4 The alkoxy group, the 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen, or the 5-6 membered heterocyclic group containing oxygen and / or nitrogen is para-substituted relative to the parent core structure, and the halogen group is ortho-, meta-, or para-substituted relative to the parent core structure; Preferably, The value of n is 0 or 1; R 1 is a substituted or unsubstituted phenyl group, wherein the substituent is C 1-3 Alkyl, C 1-3 Alkoxy, halogen, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen; preferably, the C 1-3 Alkyl, C 1-3 Alkoxy, oxygen and / or nitrogen-containing 5-6 membered aromatic heterocyclic group or oxygen and / or nitrogen-containing 5-6 membered heterocyclic group are para-substituted relative to the parent core structure, and halogen is ortho-, meta- or para-substituted; R 2 is a substituted or unsubstituted phenyl group, wherein the substituent is C 1-3 Alkyl, C 1-3 Alkoxy, halogen, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen; preferably, the C 1-3 Alkyl, C 1-3 Alkoxy, oxygen and / or nitrogen-containing 5-6 membered aromatic heterocyclic group or oxygen and / or nitrogen-containing 5-6 membered heterocyclic group is para-substituted relative to the parent core structure, and halogen is ortho-, meta- or para-substituted relative to the parent core structure; R 3 is substituted or unsubstituted C 1-5 Alkyl, phenyl, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen, the substituent is C 1-3 Alkyl, C 1-3 Alkoxy, halogen, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen; preferably, the C 1-3 Alkyl, C 1-3 The alkoxy group, the 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, or the 5-6 membered heterocyclic group containing oxygen and / or nitrogen is para-substituted relative to the parent core structure, and the halogen group is ortho-, meta-, or para-substituted relative to the parent core structure; R 4 is hydrogen, substituted or unsubstituted C 1-5 Alkyl, C 1-5 Alkoxy, phenyl, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen, the substituent is C 1-3 Alkyl, C 1-3 Alkoxy, halogen, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen; preferably, the C 1-3 Alkyl, C 1-3 Alkoxy, oxygen and / or nitrogen-containing 5-6 membered aromatic heterocyclic group or oxygen and / or nitrogen-containing 5-6 membered heterocyclic group are para-substituted relative to the parent core structure, and halogen is relative to The substitution is ortho, meta or para in the parent core structure.

6. The compound according to claim 5 or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, wherein: The value of n is 0 or 1; R 1 is a substituted or unsubstituted phenyl group, wherein the substituent is C 1-2 Alkyl, C 1-2 Alkoxy, F, Cl, Br, I, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen; preferably, the C 1-2 Alkyl, C 1-2 The alkoxy group, the 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, or the 5-6 membered heterocyclic group containing oxygen and / or nitrogen is para-substituted relative to the parent core structure, and F, Cl, Br, and I are ortho-, meta-, or para-substituted relative to the parent core structure; R 2 is a substituted or unsubstituted phenyl group, wherein the substituent is C 1-2 Alkyl, C 1-2 Alkoxy, F, Cl, Br, I, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen; preferably, the C 1-2 Alkyl, C 1-2 The alkoxy group, the 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, or the 5-6 membered heterocyclic group containing oxygen and / or nitrogen is para-substituted relative to the parent core structure, and F, Cl, Br, and I are ortho-, meta-, or para-substituted relative to the parent core structure; R 3 is substituted or unsubstituted C 1-4 Alkyl, phenyl, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen, the substituent is C 1-2 Alkyl, C 1-2 Alkoxy, F, Cl, Br, I, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen; preferably, the C 1-2 Alkyl, C 1-2 The alkoxy group, the 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, or the 5-6 membered heterocyclic group containing oxygen and / or nitrogen is para-substituted relative to the parent core structure, and the halogen group is ortho-, meta-, or para-substituted relative to the parent core structure; R 4 is hydrogen, substituted or unsubstituted C 1-4 Alkyl, C 1-4 Alkoxy, phenyl, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen, the substituent is C 1-2 Alkyl, C 1-2 Alkoxy, F, Cl, Br, I, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen; preferably, the C 1-2 Alkyl, C 1-2 The alkoxy group, the 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, or the 5-6 membered heterocyclic group containing oxygen and / or nitrogen is para-substituted relative to the parent core structure, and the halogen group is ortho-, meta-, or para-substituted relative to the parent core structure; Preferably, The value of n is 0 or 1; R 1 is a substituted or unsubstituted phenyl group, wherein the substituent is a methyl group, a methoxy group, a F group, a Cl group, a 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen, or a 5-6 membered heterocyclic group containing oxygen and / or nitrogen; preferably, the methyl group, the methoxy group, the 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen is para-substituted relative to the parent core structure, and the F group and the Cl group are ortho-, meta-, or para-substituted relative to the parent core structure; R 2 is a substituted or unsubstituted phenyl group, wherein the substituent is a methyl group, a methoxy group, a F group, a Cl group, or a 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen; preferably, the methyl group, the methoxy group, and the 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen are para-substituted relative to the parent core structure, and the F group and the Cl group are ortho-, meta-, or para-substituted relative to the parent core structure; R 3 is substituted or unsubstituted C 1-3 Alkyl, phenyl, 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen, the substituent of the substituted phenyl is methyl, methoxy, F, Cl or 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen; preferably, the methyl, methoxy and 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen are para-substituted relative to the parent core structure, and the halogen is ortho-, meta- or para-substituted relative to the parent core structure; R 4 is hydrogen, substituted or unsubstituted C 1-3 Alkyl, C 1-3 Alkoxy, phenyl, 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen, the substituent of the substituted phenyl is methyl, methoxy, F, Cl or 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen; preferably, the methyl, methoxy and 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen are para-substituted relative to the parent core structure, and the halogen is ortho-, meta- or para-substituted relative to the parent core structure; More preferably, The value of n is 0 or 1; R 1 is a substituted or unsubstituted phenyl group, wherein the substituent is a methyl group or a 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen; preferably, the methyl group or the 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen is para-substituted relative to the parent core structure; R 2 is a substituted or unsubstituted phenyl group, wherein the substituent is a methyl group or a 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen; preferably, the methyl group or the 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen is para-substituted relative to the parent core structure; R 3 is substituted or unsubstituted C 1-3 Alkyl, phenyl, 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen, the substituent is methyl, methoxy, F, Cl or 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen; preferably, the methyl, methoxy and 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen are para-substituted relative to the parent core structure, and the halogen is ortho-, meta- or para-substituted relative to the parent core structure; R 4 is hydrogen, substituted or unsubstituted C 1-3 Alkyl, C 1-3 Alkoxy, phenyl, 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen, the substituent is methyl, methoxy, F, Cl or 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen; preferably, the methyl, methoxy and 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen are para-substituted relative to the parent core structure, and the halogen is ortho-, meta- or para-substituted relative to the parent core structure; More preferably, The value of n is 0 or 1; R 1 is phenyl; R 2 is phenyl; R 3 is substituted or unsubstituted C 1-3 Alkyl, phenyl, 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen, the substituent is methyl, methoxy, halogen or 5-6 membered aromatic heterocycle containing oxygen and / or nitrogen; preferably, the methyl, methoxy, 5-6 membered aromatic heterocyclic group containing oxygen and / or nitrogen or 5-6 membered heterocyclic group containing oxygen and / or nitrogen is para-substituted relative to the parent core structure, and the halogen is ortho-, meta- or para-substituted relative to the parent core structure; R 4 For hydrogen.

7. The compound according to claim 6 or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, wherein: The compound is selected from the following structures: Preferably, the compound is selected from the following structures: Further preferably, the compound is selected from the following structures: More preferably, the compound is selected from the following structures:

8. The method for preparing the compound according to any one of claims 1 to 7, characterized in that: There are four synthetic routes: Route 1: Route 2: Route 3: Route 4: Among them, R 1 、R 2 、R 3 have the same definitions as in the recited claims.

9. A pharmaceutical composition, characterized in that The invention comprises a compound represented by formula (I) or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, and one or more pharmaceutically acceptable carriers, diluents and excipients.

10. Use of a compound of formula (I) or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or the above-mentioned pharmaceutical composition in the preparation of a medicament for treating a disease mediated by inhibition of coenzyme A by inhibiting coenzyme A; preferably, the disease mediated by inhibition of coenzyme A is anemia, local ischemia, hypoxia; more preferably, renal anemia.

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