A class of quinoxaline derivatives and their preparation and use

Abstract

The present invention provides quinoxaline derivatives and their preparation and use. Specifically, the present invention provides a compound of formula I as shown below, or a pharmaceutically acceptable salt, enantiomer, diastereomer, optical isomer, racemate, deuterated derivative, solvate or hydrate, metabolite or prodrug thereof. The compound of the present invention has excellent PGK1 kinase inhibitory activity.

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and specifically relates to a class of quinoxaline derivatives and their preparation and use. Background Art

[0002] The Warburg effect is one of the important characteristics of tumor cells, which represents the change in the way tumor cells use glucose from oxidative phosphorylation to glycolysis. Normal cells obtain ATP energy through the mitochondrial oxidative phosphorylation pathway, while tumor cells are in uncontrolled division and proliferation, and their demand for energy is particularly strong. Even under normal oxygen concentrations, rapidly proliferating tumor cells will still prefer anaerobic glycolysis to obtain energy. Changes in tumor metabolic pathways are considered to be one of the important driving forces for tumor occurrence and development.

[0003] Phosphoglycerate kinase 1 (PGK1) is a key metabolic enzyme in the glycolysis pathway. It catalyzes the conversion of 1,3-diphosphoglycerate (1,3-BPG) into 3-phosphoglycerate (3-PG) and generates the first ATP in the glycolysis pathway, playing an important role in cellular energy metabolism. Research results in recent years have shown that PGK1 is closely related to the occurrence and development of tumors. A 2016 study showed that the severity of liver cancer patients was positively correlated with the expression of PGK1 protein. After knocking down the pgk1 gene, the glycolysis ability of liver cancer cell lines decreased, the production capacity decreased, the proliferation of cells was inhibited, and the tumor formation ability was weakened. The results suggest that PGK1 may become a molecular target in the treatment of liver cancer. At the same time, PGK1 is associated with multidrug resistance in a variety of malignant tumors. For example, PGK1 is a predictor of low survival in breast cancer patients and a new prognostic biomarker for paclitaxel treatment resistance. In addition, the expression of PGK1 is also significantly upregulated in a variety of other malignant tumors, including pancreatic cancer, colorectal cancer, neuroblastoma, and glioma. Therefore, targeting PGK1 may be an effective strategy for the treatment of malignant tumors.

[0004] Although research on targeted PGK1 inhibitors has great potential in the treatment of malignant tumors, research on PGK1 inhibitors is still in its infancy.

[0005] In summary, there is an urgent need to develop a new class of targeted PGK1 inhibitors in this field. Summary of the invention

[0006] The purpose of the present invention is to provide a new class of targeted PGK1 inhibitors.

[0007] In a first aspect of the present invention, there is provided a compound of formula I, or a pharmaceutically acceptable salt, enantiomer, diastereomer, optical isomer, racemate, deuterated derivative, solvate or hydrate, metabolite or prodrug thereof;

[0008]

[0009] in,

[0010] X is selected from the group consisting of N or CH;

[0011] A is selected from the group consisting of: non-existent (chemical bond), substituted or unsubstituted C1-C3 alkylene,

[0012] R a Selected from the group consisting of H, substituted or unsubstituted C1-C3 alkyl;

[0013] R 1 and R 2 Each is independently selected from the following group: H, substituted or unsubstituted C1-C6 alkyl (preferably C1-C3 alkyl), halogen (preferably Cl), cyano, N(R c ) 2 , unsubstituted or substituted with one or more R s1 C6-C10 aromatic ring substituted, unsubstituted or replaced by one or more R s1 A substituted 5-10 membered heteroaromatic ring;

[0014] R s1 Each is independently selected from the following group: halogen, substituted or unsubstituted C1-C3 alkyl, cyano;

[0015] R b Selected from the group consisting of H, unsubstituted or substituted with one or more R s2 Substituted C1-C6 alkyl (preferably C1-C3 alkyl),

[0016] R c Each is independently selected from the following group: H, substituted or unsubstituted C1-C3 alkyl;

[0017] R s2 Each is independently selected from the following group: hydroxy, substituted or unsubstituted C1-C3 alkoxy, halogen, substituted or unsubstituted C1-C3 alkyl, cyano;

[0018] R 3 Selected from the following groups: H, substituted or unsubstituted C1-C6 alkyl (preferably C1-C3 alkyl), N(R d ) 2 ;

[0019] R d Each is independently selected from the following group: H, substituted or unsubstituted C1-C3 alkyl;

[0020] R 4 Selected from the group consisting of: unsubstituted or substituted with one or more R s3 substituted C1-C6 (preferably C1-C3 alkyl), unsubstituted or replaced by one or more R s3 C4-C10 cycloalkyl substituted, unsubstituted or replaced by one or more R s3 substituted 4-10 membered heterocycloalkyl, unsubstituted or substituted with one or more R s3 C6-C10 aryl substituted, unsubstituted or substituted with one or more R s3 substituted 5-10 membered heteroaryl;

[0021] R s3 Each is independently selected from the following group: hydroxy, substituted or unsubstituted C1-C3 alkyl, halogen, substituted or unsubstituted C1-C3 alkylacyl, substituted or unsubstituted C1-C3 alkoxy, substituted or unsubstituted C1-C3 haloalkyl, substituted or unsubstituted C1-C3 hydroxyalkyl;

[0022] Unless otherwise specified, the substitution means that one or more hydrogen atoms in the group are optionally replaced by a substituent selected from the group consisting of hydroxyl, halogen, C1-C3 alkyl, C1-C3 haloalkyl, amino (-NH 2 )、-N(C1-C3 alkyl) 2 , -NH(C1-C3 alkyl), cyano.

[0023] In another preferred embodiment, X is CH.

[0024] In another preferred embodiment, A is

[0025] In another preferred embodiment, R a Selected from the following group: H, C1-C3 alkyl.

[0026] In another preferred embodiment, R a Selected from the group consisting of H, methyl.

[0027] In another preferred embodiment, A is selected from the following groups: absent or Among them, R a Selected from the following group: H, C1-C3 alkyl.

[0028] In another preferred embodiment, A is Among them, R a Selected from the following group: H, C1-C3 alkyl.

[0029] In another preferred embodiment, A is selected from the following groups: absent, -NH-, -N(CH 3 )-.

[0030] In another preferred embodiment, A is selected from the following groups: -NH-, -N(CH 3 )-.

[0031] In another preferred embodiment, R 1 and R 2 Are different groups.

[0032] In another preferred embodiment, R 1 and R 2 At least one of them is a halogen (such as Cl).

[0033] In another preferred embodiment, R 1 It is a halogen.

[0034] In another preferred embodiment, R 1 and R 2 Each independently selected from the group consisting of halogen,

[0035] In another preferred embodiment, R 1 and R 2 One is a halogen and the other is

[0036] In another preferred embodiment, R 1 is halogen and R 2 for

[0037] In another preferred embodiment, R b is unsubstituted or substituted with one or more R s2 Substituted C1-C6 alkyl (preferably C1-C3 alkyl).

[0038] In another preferred embodiment, R s2 Each is independently selected from the group consisting of hydroxy, and substituted or unsubstituted C1-C3 alkoxy.

[0039] In another preferred embodiment, R s2 It is hydroxyl.

[0040] In another preferred embodiment, R b is C1-C6 hydroxyalkyl, preferably C1-C3 hydroxyalkyl; more preferably, R b -CH 2 OH.

[0041] In another preferred embodiment, R 1 and R 2 Each independently selected from the group consisting of halogen, And Rb is unsubstituted or substituted with one or more R s2 Substituted C1-C6 alkyl (preferably C1-C3 alkyl).

[0042] In another preferred embodiment, R 1 and R 2 Each independently selected from the group consisting of halogen, R b is unsubstituted or substituted with one or more R s2 substituted C1-C6 alkyl (preferably C1-C3 alkyl), and R s2 Each is independently selected from the following group: hydroxyl, substituted or unsubstituted C1-C3 alkoxy.

[0043] In another preferred embodiment, R 1 is halogen and R 2 for And R b is unsubstituted or substituted with one or more R s2 Substituted C1-C6 alkyl (preferably C1-C3 alkyl).

[0044] In another preferred embodiment, R 1 is halogen and R 2 for R b is unsubstituted or substituted with one or more R s2 substituted C1-C6 alkyl (preferably C1-C3 alkyl), and R s2 Each is independently selected from the following group: hydroxyl, substituted or unsubstituted C1-C3 alkoxy.

[0045] In another preferred embodiment, R 3 Selected from the following group: H, C1-C3 alkyl, N(R d ) 2 ; Among them, R d Each is independently selected from the following group: H, C1-C3 alkyl.

[0046] In another preferred embodiment, R 3 Selected from the group consisting of H, C1-C3 alkyl, NH(C1-C3 alkyl).

[0047] In another preferred embodiment, R 3 is H or C1-C6 alkyl (preferably, C1-C3 alkyl).

[0048] In another preferred embodiment, R 4 Selected from the group consisting of: unsubstituted or substituted with one or more R s3 substituted C1-C6 alkyl (preferably C1-C3 alkyl), unsubstituted or replaced by one or more R s3C4-C6 cycloalkyl substituted, unsubstituted or replaced by one or more R s3 substituted 5-6 membered heterocycloalkyl, unsubstituted or substituted with one or more R s3 substituted phenyl, unsubstituted or replaced by one or more R s3 substituted 5-6 membered heteroaryl.

[0049] In another preferred embodiment, R 4 Selected from the group consisting of: unsubstituted or substituted with one or more R s3 substituted 5-6 membered heterocycloalkyl, unsubstituted or substituted with one or more R s3 substituted 5-6 membered heteroaryl.

[0050] In another preferred embodiment, R 4 In the embodiment, the heterocycloalkyl group is a saturated oxygen-containing heterocycloalkyl group; preferably, a 5-6-membered heterocycloalkyl group containing 1 or 2 oxygen atoms.

[0051] In another preferred embodiment, R 4 In the above, the heteroaryl group is a sulfur-containing heteroaryl group.

[0052] In another preferred embodiment, R 4 In the above, the heteroaryl group is a five-membered sulfur-containing heteroaryl group.

[0053] In another preferred embodiment, R s3 Each is independently selected from the following group: hydroxy, C1-C3 alkyl, halogen, C1-C3 alkylacyl, C1-C3 alkoxy, C1-C3 haloalkyl, C1-C3 hydroxyalkyl.

[0054] In another preferred embodiment, when R 4 is unsubstituted or substituted with one or more R s3 substituted C4-C10 cycloalkyl (preferably C4-C6 cycloalkyl), unsubstituted or replaced by one or more R s3 When the substituted 4-10 membered heterocycloalkyl (preferably, 5-6 membered heterocycloalkyl), the compound of formula I is as shown in formula I-1 or formula I-2;

[0055]

[0056] In another preferred embodiment, in the compound, A, X, R 1 , R 2 , R 3 , R 4 , R a , R b , R c , R d , R s1 , R s2 and R s3Each is independently a corresponding group in the specific compounds described in the Examples or Table 1 (such as compounds S1-S35).

[0057] In another preferred embodiment, the compound of formula I is a compound selected from the following Table 1:

[0058] Table 1

[0059]

[0060]

[0061]

[0062] or a pharmaceutically acceptable salt, enantiomer, diastereomer, optical isomer, racemate, deuterated derivative, solvate or hydrate, metabolite or prodrug thereof.

[0063] In the second aspect of the present invention, a pharmaceutical composition is provided, comprising (i) a therapeutically effective amount of a compound of formula I as described in the first aspect, or a pharmaceutically acceptable salt, enantiomer, diastereomer, optical isomer, racemate, deuterated derivative, solvate or hydrate, metabolite or prodrug thereof, and (ii) an optional pharmaceutically acceptable carrier, excipient or diluent.

[0064] In another preferred embodiment, the pharmaceutical composition is a pharmaceutical composition for treating tumors, or a pharmaceutical composition for treating diseases related to the activity of energy metabolism enzymes (preferably PGK1 enzyme).

[0065] In the third aspect of the present invention, there is provided a method for preparing the compound of formula I as described in the first aspect, or a pharmaceutically acceptable salt, enantiomer, diastereomer, optical isomer, racemate, deuterated derivative, solvate or hydrate, metabolite or prodrug thereof,

[0066] The preparation method is method one or method two.

[0067] In another preferred embodiment, the method 1 comprises the steps of:

[0068]

[0069] v) removing the protecting group of compound ie and then reacting with R 4 COOH undergoes condensation reaction to obtain compound if;

[0070] vi) optionally removing the protecting group of compound if to obtain a compound of formula I;

[0071] In various types, R 1 'For R 1or R protected by a protecting group 1 , R 2 'For R 2 or R protected by a protecting group 2 ;

[0072] And A, X, R 1 , R 2 , R 3 and R 4 As defined in the first aspect.

[0073] In another preferred embodiment, in method 1, when R 1 or R 2 is a hydroxyl-containing group, and the corresponding R 1 ' or R 2 ' is a hydroxyl group protected (preferably formed into an ester to be protected) R 1 or R 2 When the step vi) is: hydrolyzing the compound if under alkaline conditions to obtain compounds ig-1 and ig-2.

[0074] In another preferred embodiment, the method 2 comprises the steps of:

[0075]

[0076] ii) in an inert solvent, under the catalysis of Pd, compound iia-1 or compound iia-2 or a mixture thereof is reacted with R b -C ≡ CH or R 5 -B(OH) 2 Carrying out a coupling reaction to obtain a compound of formula I;

[0077] Among them, R 5 is unsubstituted or substituted with one or more R s1 The substituted C6-C10 aromatic ring is either unsubstituted or replaced by one or more R s1 substituted 5-10 membered heteroaromatic ring; in the compound of formula I, R 1 and R 2 One of unsubstituted or substituted with one or more R s1 The substituted C6-C10 aromatic ring is either unsubstituted or replaced by one or more R s1 A, X, R 3 and R 4 As defined in the first aspect.

[0078] In another preferred embodiment, in method 2, R 1 and R in compound iia-2 2 For Cl.

[0079] In the fourth aspect of the present invention, there is provided a compound of formula I as described in the first aspect, or a pharmaceutically acceptable salt, enantiomer, diastereomer, optical isomer, racemate, deuterated derivative, solvate or hydrate, metabolite or prodrug thereof, or a pharmaceutical composition as described in the second aspect for use in the preparation of (i) a PGK1 inhibitor and / or (ii) a drug for treating or preventing a PGK1-related disease.

[0080] In another preferred embodiment, the PGK1-related diseases include: cancer, abnormal cell proliferation, morphological changes, abnormal glucose metabolism, hyperkinesia, tumor growth, diabetes, or a combination thereof.

[0081] In another preferred embodiment, the cancer includes: liver cancer, gastric cancer, colorectal cancer, breast cancer, bladder cancer, pancreatic cancer, pancreatic ductal adenocarcinoma, neuroblastoma, prostate cancer, or a combination thereof.

[0082] In the fifth aspect of the present invention, a method for treating or preventing a disease related to PGK1 is provided, comprising the steps of administering to a subject in need thereof a therapeutically effective amount of a compound of formula I as described in the first aspect, or a pharmaceutically acceptable salt, enantiomer, diastereomer, optical isomer, racemate, deuterated derivative, solvate or hydrate, metabolite or prodrug thereof, or a pharmaceutical composition as described in the second aspect.

[0083] In another preferred embodiment, the PGK1-related diseases include: cancer, abnormal cell proliferation, morphological changes, abnormal glucose metabolism, hyperkinesia, tumor growth, diabetes, or a combination thereof.

[0084] In another preferred embodiment, the cancer includes: liver cancer, gastric cancer, colorectal cancer, breast cancer, bladder cancer, pancreatic cancer, pancreatic ductal adenocarcinoma, neuroblastoma, prostate cancer, or a combination thereof.

[0085] In another preferred embodiment, the subject is a mammal, preferably a human.

[0086] In a sixth aspect of the present invention, a method for inhibiting the activity of PGK1 is provided, comprising the step of contacting PGK1 with the compound of formula I as described in the first aspect, thereby inhibiting the activity of PGK1.

[0087] In another preferred embodiment, the method is non-therapeutic in vitro.

[0088] In the seventh aspect of the present invention, a method for inhibiting cell proliferation activity is provided, comprising the steps of: culturing cells in the presence of the compound of formula I as described in the first aspect, thereby inhibiting the proliferation activity of the cells.

[0089] In another preferred embodiment, the method is non-therapeutic in vitro.

[0090] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. DETAILED DESCRIPTION

[0091] After extensive and in-depth research, the inventors found that replacing the core of the existing quinazoline PGK1 inhibitor with quinoxaline to obtain a class of PGK1 inhibitors with a novel core structure still has PGK1 inhibitory activity, and even some preferred compounds have significantly enhanced PGK1 inhibitory activity, thereby obtaining a class of quinoxaline derivatives with quinoxaline as the core that are novel in structure as PGK1 inhibitors. Based on this, the inventors completed the present invention.

[0092] the term

[0093] Unless otherwise defined, the term "alkyl", by itself or as part of another substituent, refers to a straight or branched chain hydrocarbon radical having the specified number of carbon atoms (i.e., C1-C6 means 1-6 carbons). Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, and the like.

[0094] As used herein, the term "cycloalkyl" refers to a hydrocarbon ring having a specified number of ring atoms (e.g., C4-C10 cycloalkyl) and being fully saturated or having no more than one double bond between the ring tops, preferably a fully saturated ring. "Cycloalkyl" can be a monocyclic ring (e.g., cyclopropyl, cyclobutyl, cyclohexyl, etc.), and can also refer to bicyclic and polycyclic hydrocarbon rings (e.g., cyclic, spirocyclic, condensed, bridged, etc.). The term "heterocycloalkyl", which may also be referred to herein as "heterocyclyl", refers to a cycloalkyl group containing one to five heteroatoms selected from N, O and S as ring atoms, wherein the nitrogen and sulfur atoms are optionally oxidized and the nitrogen atom is optionally quaternized. The heterocycloalkyl group may be a monocyclic, bicyclic or polycyclic ring system. In general, the heterocyclyl group usually includes 5-12 ring atoms (i.e., 5-12 membered heterocycloalkyl), preferably, includes 5-7 ring atoms (i.e., 5-7 membered heterocyclyl) and contains 1, 2, 3 or 4 heterocyclic atoms. Non-limiting examples of heterocycloalkyl groups include morpholine ring, piperidine ring, piperazine ring, N-alkyl or acyl substituted piperazine ring, homopiperazine ring, N-alkyl or acyl substituted homopiperazine ring, pyrrole, tetrahydropyrrole, 7H-purine, tetrahydrofuran, tetrahydropyran, and the like. The heterocycloalkyl group may be attached to the rest of the molecule via a ring carbon or a heteroatom (e.g., ring N).

[0095] The term "alkoxy" is used in its conventional sense to refer to those alkyl groups that are attached to the rest of the molecule via an oxygen atom. In addition, for dialkylamino groups, the alkyl moieties may be the same or different and may also be combined with the nitrogen atom to which each alkyl group is attached to form a 3-7 membered ring. Thus, -N(R a ) 2 The groups shown are meant to include piperidinyl, pyrrolidinyl, morpholinyl, azetidinyl, and the like.

[0096] Unless otherwise defined, the term "aryl" represents a polyunsaturated (usually aromatic) hydrocarbon group, which can be a monocyclic or polycyclic (up to three rings) fused together or covalently linked. Generally, aryl refers to a monocyclic or polycyclic (i.e., sharing a ring of adjacent carbon atoms) group having 6-10 ring atoms, and the group has a conjugated π electron system. The aryl ring can be fused with a heterocycloalkyl, heteroaryl or cycloalkyl ring, and non-limiting examples include benzimidazole, benzothiazole, benzoxazole, benzisoxazole, benzopyrazole, quinoline, benzindole, benzodihydrofuran. The term "heteroaryl" refers to a heteroaromatic system containing 1 to 4 heteroatoms selected from N, O and S and 5 to 14 ring atoms, wherein nitrogen and sulfur atoms are optionally oxidized, and nitrogen atoms are optionally quaternized. Generally, heteroaryl has 5-10 ring atoms, i.e., 5-10 yuan heteroaryl, preferably, has 5-6 ring atoms, i.e., 5 or 6 yuan heteroaryl. Heteroaryl can be connected to the rest of the molecule through heteroatoms. Non-limiting examples of aryl include phenyl and naphthyl. The aryl (ring) can be fused with heterocycloalkyl, heteroaryl or cycloalkyl ring, and non-limiting examples include benzimidazole, benzothiazole, benzoxazole, benzisoxazole, benzopyrazole, quinoline, benzindole, benzodihydrofuran, etc. Non-limiting examples of heteroaryl include furyl, thienyl, pyridyl, pyrrolyl, N-alkylpyrrolyl, pyrimidyl, pyrazinyl, imidazolyl, tetrazolyl, etc. The heteroaryl can be fused on aryl, heterocycloalkyl or cycloalkyl ring, and the ring connected with the parent structure is a heteroaryl ring.

[0097] As used herein, the term "heteroatom" is intended to include oxygen (O), nitrogen (N), and sulfur (S).

[0098] For compounds provided herein, a bond from a substituent (typically an R group) to the center of an aromatic ring (e.g., benzene, pyridine, etc.) will be understood to mean a bond providing a connection at any available vertex of the aromatic ring. In some embodiments, this description also includes connections on a ring fused to an aromatic ring. For example, a bond drawn to the center of an indole benzene moiety will represent a bond connected to any available vertex of a six-membered or five-membered ring moiety of indole.

[0099] As used herein, the term "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br) or iodine (I). Similarly, the term "halo" refers to a group in which one or more or all hydrogens are replaced by the same or different halogens as defined above.

[0100] Unless otherwise specified, the structural formula described in the present invention is intended to include all optical isomers and stereoisomeric forms (such as enantiomers, diastereomers, geometric isomers or conformational isomers): for example, R and S configurations containing asymmetric centers. Therefore, single stereochemical isomers, enantiomers, diastereomers or mixtures of geometric isomers or conformational isomers of the compounds of the present invention all fall within the scope of the present invention.

[0101] As used herein, the terms "comprising", "including" or "comprising" indicate that various components can be used together in the mixture or composition of the present invention. Therefore, the terms "consisting essentially of" and "consisting of" are included in the term "comprising".

[0102] As used herein, the term "pharmaceutically acceptable" ingredient refers to a substance that is suitable for use in humans and / or animals without excessive adverse side effects (such as toxicity, irritation, and allergic response), ie, at a reasonable benefit / risk ratio.

[0103] As used herein, the term "therapeutically effective dose" refers to any amount of a drug as described below, which, when used alone or in combination with another therapeutic agent, can promote disease regression, which is manifested as a decrease in the severity of disease symptoms, an increase in the frequency and duration of disease-free periods, or the prevention of disorders or disabilities caused by the disease. The "therapeutically effective dose" of the drug of the present invention also includes a "prophylactic effective dose", which is any amount of a drug as described below, which, when administered alone or in combination with another therapeutic agent to a subject at risk of developing a disease or suffering from a recurrence of the disease, can inhibit the occurrence or recurrence of the disease.

[0104] Quinoxaline derivatives

[0105] The present invention relates to a class of quinoxaline derivatives having phosphoglycerate kinase 1 (PGK1) inhibitory activity and pharmaceutically acceptable salts or pharmaceutically acceptable solvent compounds thereof, a preparation method thereof and use thereof in preparing drugs for preventing or treating diseases related to abnormal cell proliferation, morphological changes, abnormal glucose metabolism, hypermotility and the like associated with PGK in a living body, in particular for treating or preventing tumor growth and metastasis and diabetes.

[0106] As used herein, the term "compound of the present invention" or "quinoxaline derivative" or "quinoxaline compound" refers to a compound of formula I. The term also includes various crystalline forms, pharmaceutically acceptable salts, hydrates or solvates of the compound of formula I.

[0107] The term "pharmaceutically acceptable salt" refers to a salt suitable for use as a drug formed by the compound of the present invention and an acid or base. In the present invention, the pharmaceutically acceptable salt is not particularly limited, and preferably includes: inorganic acid salts, organic acid salts, alkyl sulfonates and aryl sulfonates; the inorganic acid salts include hydrochlorides, hydrobromides, nitrates, sulfates, phosphates, etc.; the organic acid salts include formates, acetates, propionates, benzoates, maleates, fumarates, succinates, tartrates, citrates, etc.; the alkyl sulfonates include methyl sulfonates, ethyl sulfonates, etc.; the aryl sulfonates include benzene sulfonates, p-toluene sulfonates, etc.

[0108] The term "solvate" refers to a complex formed by the coordination of the compound of the present invention with solvent molecules in a specific ratio. "Hydrate" refers to a complex formed by the coordination of the compound of the present invention with water. In the present invention, the pharmaceutically acceptable solvates of the compound represented by the general formula (I) are not particularly limited, and preferably include: solvates of the compound represented by the general formula (I) with water, ethanol, isopropanol, ether, acetone, etc.

[0109] In addition, the compounds of the present invention also include prodrugs of the compounds shown in Formula I. The term "prodrug" includes a compound that can be biologically active or inactive, and when taken by an appropriate method, it undergoes metabolism or chemical reactions in the human body and is converted into a class of compounds of Formula I, or a salt or solution composed of a compound of Formula I. The prodrug includes (but is not limited to) carboxylate, carbonate, phosphate, nitrate, sulfate, sulfone, sulfoxide, amino compound, carbamate, azo compound, phosphoramide, glucoside, ether, acetal and the like forms of the compound.

[0110] The purpose of the present invention is to provide a class of PGK1 kinase inhibitors with novel structure and excellent activity.

[0111] Specifically, the present invention provides a compound of formula I as shown below, or a pharmaceutically acceptable salt, enantiomer, diastereomer, optical isomer, racemate, deuterated derivative, solvate or hydrate, metabolite or prodrug thereof.

[0112]

[0113] Among them, R 1 , R 2 , R 3 , R 4, X and A are as defined in the first aspect.

[0114] In a specific embodiment,

[0115] X is selected from nitrogen atom or CH; and / or

[0116] A is selected from a direct bond or and / or where R a Selected from hydrogen, C1-C3 alkyl; and / or

[0117] R 1 and R 2 are each independently selected from hydrogen, halogen, cyano, substituted or unsubstituted amino, A substituted or unsubstituted 6- to 10-membered aromatic ring, or a substituted or unsubstituted 5- to 10-membered heteroaromatic ring. b Selected from hydrogen, C1-C3 alkyl, The C1-C3 alkyl group may be further substituted by hydroxyl, halogen, cyano. The substitution may be selected from halogen, C1-C3 alkyl, cyano; and / or

[0118] R 3 Selected from hydrogen, amino, C1-C3 alkylamino; and / or

[0119] R 4 Selected from substituted or unsubstituted C1-C3 alkyl, substituted or unsubstituted saturated or unsaturated 4-10-membered cycloalkyl, substituted or unsubstituted 5-10-membered heteroaromatic ring group, substituted or unsubstituted 4-10-membered heterocyclic group. The 5-10-membered heteroaromatic ring group and the 4-10-membered heterocyclic group contain 1-3 heteroatoms selected from O, N, and S. The substituted group is further substituted by one or more substituents selected from the following group: hydroxyl, C1-C3 alkyl, halogen, C1-C3 alkyl acyl. The C1-C3 alkyl group can be further substituted by hydroxyl, halogen, amino, and cyano.

[0120] In another preferred embodiment, in the compound, X, A, R 1 , R 2 , R 3 , R 4 Any one of them is the corresponding group in the specific compound described in the embodiment.

[0121] Preferably, the quinoxaline compound represented by the general formula (I) of the present invention is selected from the compounds in Table 1 below:

[0122] Table 1

[0123]

[0124]

[0125] Preparation method

[0126] The present invention also provides a method for preparing the compound of formula I as described in the first aspect of the present invention. The method for preparing the compound of formula I of the present invention is described in more detail below. However, these specific methods do not constitute any limitation to the present invention. The compound of the present invention can also be conveniently prepared by optionally combining various synthetic methods described in this specification or known in the art, and such a combination can be easily performed by a technician in the field to which the present invention belongs.

[0127] The compound of the present invention can be prepared by, for example, method 1 or 2.

[0128] In another preferred embodiment, the method 1 and the method 2 are as defined in the second aspect.

[0129] In another preferred embodiment, method 1 comprises the following steps:

[0130]

[0131] i) Compound iiia-1 or iiia-2 or a mixture thereof and a basic nucleophile (such as ) undergoes a substitution reaction to obtain compound iiib-1 or iiib-2 or a mixture thereof;

[0132] ii) Compound iiib-1 or iiib-2 or a mixture thereof is reacted with an alkyne reagent (such as R b -C≡CH) by Pd-catalyzed coupling reaction and optionally separation to obtain compound iiic-1 and / or iiic-2;

[0133] iii) removing the protecting group from compound iiic-1 and / or iiic-2 and then reacting with different carboxylic acids (such as R 4 COOH) to obtain compound iiid-1 and / or iiid-2 through a condensation reaction;

[0134] iv) optionally hydrolyzing compound iiid-1 and / or iiid-2 under alkaline conditions to obtain compound iiie-1 and iiie-2 (i.e., compound of formula I, wherein R 1 or R 2 One is Cl and the other is );

[0135] Wherein, unless otherwise specified, each group is as defined in the first aspect. In another preferred embodiment, when in method 1, R 3 When H is H, the method 1 is method 1 (1), and comprises the following steps:

[0136]

[0137] i) Compound ia undergoes a ring-closing reaction with ethyl glyoxylate to obtain compound ib;

[0138] ii) Compound ib is treated with a chlorinating agent to obtain compound ic-1 or ic-2 or a mixture thereof;

[0139] iii) Compound ic-1 or ic-2 or a mixture thereof is reacted with a basic nucleophile (such as ) undergoes a substitution reaction to obtain compound id-1 or id-2 or a mixture thereof;

[0140] iv) reacting compound id-1 or id-2 or a mixture thereof with an alkyne reagent (such as R b -C≡CH or R protected by a protecting group b -C≡CH) by Pd-catalyzed coupling reaction and optionally separation to obtain compound ie-1 and / or ie-2;

[0141] v) Compound ie-1 and / or ie-2 is deprotected and then reacted with a different carboxylic acid (such as R 4 COOH) through condensation reaction to obtain compounds if-1 and / or if-2 respectively;

[0142] vi) optionally hydrolyzing compound if-1 and / or if-2 under alkaline conditions to obtain compound ig-1 and / or ig-2 (i.e., a compound of formula I, wherein R 1 or R 2 One is Cl and the other is );

[0143] Unless otherwise specified, each group is as defined in the first aspect.

[0144] Preparation method 2:

[0145]

[0146] i) removing the protecting group of compound id-1 or id-2 or a mixture thereof and then reacting with different carboxylic acids (such as R 4 COOH) by condensation reaction and optionally separation to obtain a mixture of iia-1 and / or iia-2;

[0147] ii) a mixture of iia-1 and / or iia-2 is reacted with different boric acids (such as R 5 -B(OH) 2 , where R 5 as defined above) or a different alkyne reagent (such as R b -C≡CH) through a Pd-catalyzed coupling reaction to obtain compound iib-1 and / or iib-2 (i.e., a compound of formula I, wherein R1 and R 2 One is Cl and the other is unsubstituted or substituted with one or more R s1 The substituted C6-C10 aromatic ring is either unsubstituted or replaced by one or more R s1 The substituted 5-10 membered heteroaromatic ring (ie, R 5 )).

[0148] Usually, in the preparation process, each reaction is usually carried out in an inert solvent at room temperature to reflux temperature. The reaction time is usually 0.1 hour to 60 hours, preferably 0.5 to 48 hours.

[0149] Pharmaceutical compositions and methods of administration

[0150] Since the compounds of the present invention have excellent inhibitory activity against phosphoglycerate kinase 1 (PGK1), the compounds of the present invention and their various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and pharmaceutical compositions containing the compounds of the present invention as the main active ingredients can be used to treat, prevent and alleviate diseases related to phosphoglycerate kinase 1 (PGK1). According to the prior art, the compounds of the present invention can be used to treat the following diseases: cancer, abnormal cell proliferation, morphological changes, abnormal glucose metabolism, hypermotility, tumor growth, diabetes, or a combination thereof; wherein the cancers include: liver cancer, gastric cancer, colorectal cancer, breast cancer, bladder cancer, pancreatic cancer, pancreatic ductal adenocarcinoma, neuroblastoma, prostate cancer, or a combination thereof.

[0151] The present invention also provides the use of a compound of formula I (quinoxaline derivative) or its isomer or a pharmaceutically acceptable salt, ester, prodrug or hydrate thereof as a PGK1 inhibitor in the preparation of a drug for preventing and / or treating diseases associated with PGK1, wherein the diseases associated with PGK1 include various cancers (liver cancer, gastric cancer, colorectal cancer, breast cancer, bladder cancer, pancreatic cancer and neuroblastoma, etc.).

[0152] The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of the compound as described in the first aspect of the present invention, or a pharmaceutically acceptable salt, prodrug, hydrate or solvate thereof, and optionally a pharmaceutically acceptable carrier, excipient or diluent.

[0153] In another preferred embodiment, the pharmaceutical composition is a pharmaceutical composition for treating tumors, or a pharmaceutical composition for treating diseases related to the activity of energy metabolism enzymes (preferably PGK1 enzyme).

[0154] The pharmaceutical composition of the present invention comprises a safe and effective amount of the compound of the present invention or a pharmacologically acceptable salt thereof and a pharmacologically acceptable excipient or carrier. Wherein "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Usually, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, and more preferably, contains 10-500 mg of the compound of the present invention per dose. Preferably, the "one dose" is a capsule or tablet.

[0155] "Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as ), wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0156] There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administrations include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

[0157] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0158] Solid dosage forms such as tablets, pills, capsules, pills and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active compound or compounds in such compositions can be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into microencapsulated form with one or more of the above-mentioned excipients.

[0159] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.

[0160] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0161] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanol and agar, or mixtures of these substances, and the like.

[0162] Compositions for parenteral injection may include physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0163] Dosage forms for topical administration of the compounds of the invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.

[0164] The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds.

[0165] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage during administration is a pharmaceutically effective dosage, and for a person weighing 60 kg, the daily dosage is usually 1 to 2000 mg, preferably 20 to 500 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the health status of the patient, which are all within the skill of a skilled physician.

[0166] The main advantages of the present invention include:

[0167] 1. The compounds of the present invention (especially the preferred compounds) have strong inhibitory activity against PGK1 and show strong proliferation inhibitory activity on human liver cancer SNU739 or HepG2 cells.

[0168] The details are as follows:

[0169] 1.1 Terazosin (as shown below) inhibits PGK1 at high concentrations (2.5-25 μM). The inhibitory IC values ​​of the preferred compounds of the present invention on PGK1 are 50 Values ​​less than 100 nM;

[0170] 1.2. The preferred compounds of the present invention have significant proliferation inhibitory activity against human liver cancer SNU739 or HepG2.

[0171] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended only to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods for the unrecorded specific conditions in the following examples are usually based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are weight percentages and weight parts.

[0172] 1. Preparation Example

[0173] For the following examples, standard operations and purification methods known to those skilled in the art can be used. Unless otherwise specified, raw materials are generally available from commercial sources. Commercially available solvents and reagents are generally used without further purification, anhydrous solvents are treated by standard methods, and other reagents are commercially available analytical grade. Unless otherwise stated, all temperatures are expressed in ° C (degrees Celsius), and room temperature or ambient temperature refers to 20-25 ° C. The purification of the products uses silica gel (200-30 mesh) column chromatography unless otherwise stated. The structure of the compound is determined by nuclear magnetic resonance spectroscopy (NMR). The nuclear magnetic resonance hydrogen spectrum shift (δ) is given in parts per million (ppm). The nuclear magnetic resonance hydrogen spectrum was measured using a Varian Mercury AMX300 instrument, and deuterated chloroform (CDCl 3 ), deuterated methanol (CD 3 OD) and deuterated dimethyl sulfoxide (d6-DMSO) were used as solvents, and tetramethylsilane (TMS) was used as the internal standard.

[0174] abbreviation

[0175] DIPEA: N,N-diisopropylethylamine

[0176] TBTU: O-Benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate

[0177] TFA: trifluoroacetic acid

[0178] DAST: Diethylaminosulfur trifluoride

[0179] Pd 2 (dba) 3 :Tris(dibenzylideneacetone)dipalladium

[0180] Xantphos: 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene

[0181] DMF: N,N-dimethylformamide

[0182] Preparation Example 1 Preparation of Compound S1

[0183]

[0184] Step 1: Weigh compound 1a (243 mg, 1 equivalent) into a single-necked bottle, add ethanol as solvent, and then add ethyl glyoxylate (50% toluene solution, 222 mg, 1.2 equivalents), and heat to 80°C for 2 hours. After the reaction is completed, cool to room temperature, a large amount of solid precipitates, and filter to obtain a mixture of 1b and 1c.

[0185]

[0186] Step 2: Weigh the mixture of 1b and 1c (1.385 g, 1 equivalent) into a single-mouth bottle, add phosphorus oxychloride (4.2 mL, 10 equivalents) and DIPEA (3.0 mL, 4 equivalents), and react at 110°C for 4 hours. After the reaction is complete, partially distill off the phosphorus oxychloride under reduced pressure, then pour the reaction solution into ice water, neutralize with saturated sodium bicarbonate solution, extract with ethyl acetate, wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, mix and load on a column to obtain a mixture of 1d and 1e.

[0187]

[0188] Step 3: Weigh a mixture of 1d and 1e (486 mg, 1 equivalent) and 1f (482 mg, 1.5 equivalents) in a single-mouth bottle, add 1,4-dioxane solvent, then add DIPEA (0.74 mL, 3 equivalents), and react at 100°C for 3 hours. After the reaction is complete, extract with ethyl acetate and water, wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, mix and load on the column to obtain a mixture of 1g and 1h.

[0189]

[0190] Step 4: Weigh a mixture of 1g and 1h (168 mg, 1 equivalent), PdCl 2 (PPh3 ) 2 (23 mg, 0.1 equivalent) and CuI (13 mg, 0.2 equivalent) were placed in a single-mouth bottle, tetrahydrofuran solvent was added, and then propargyl acetate (66 μL, 2 equivalents) and DIPEA (0.60 mL, 4 equivalents) were added, and the reaction was heated at 80°C for 5 hours under nitrogen protection. After the reaction was complete, it was extracted with ethyl acetate and water, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and then mixed and loaded onto the column to obtain compounds 1i and 1j. Analytical data of compound 1i: 1 H NMR (400 MHz, CDCl 3 )δ8.57(s,1H),8.00(s,1H),7.67(s,1H),4.99(s,2H),4.71(d,J=13.4Hz,2H),4.33(br,1H),3.06(t,J=12.4Hz,2H),2.72(s,3H),2.16(s,3H),1.84(d,J=12.7Hz,2H),1.73(t,J=13.6Hz,2H),1.48(s,9H). Analytical data of compound 1j: 1 H NMR (400 MHz, CDCl 3 )δ8.56(s,1H),7.88(s,1H),7.82(s,1H),4.99(s,2H),4.67(d,J=13.4Hz,2H),4.40–4.21(m,1H),3.05(t, J=12.7Hz,2H),2.72(s,3H),2.16(s,3H),1.83(d,J=9.8Hz,2H),1.78–1.68(t,J=7.7Hz,2H),1.48(s,9H).

[0191]

[0192] Step 5: Weigh compound 1j (78 mg, 1 equivalent) in a single-mouth bottle, add dichloromethane solvent, then add TFA (0.12 mL, 10 equivalents), and react at room temperature for 3 hours. After the reaction is complete, spin dry the reaction solution. Then add dichloromethane as a solvent, add (R)-tetrahydrofurancarboxylic acid (29 mg, 1.5 equivalents), TBTU (106 mg, 2 equivalents), DIPEA (0.14 mL 5 equivalents) in sequence, and react at room temperature for 3 hours. After the reaction is complete, extract with dichloromethane and water, wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, mix the sample and load it on the column to obtain compound 1k. Analytical data of compound 1k: 1 H NMR (400 MHz, CDCl 3)δ8.53(s,1H),7.85(s,1H),7.78(s,1H),4.96(s,2H),4.78–4.69(m,1H),4.68–4.56(m,3H),4.00–3.79(m,2H),3.0 7(t,J=12.0Hz,2H),2.89(s,2H),2.77(s,1H),2.13–2.17(m,1H),2.13(s,3H),2.09–1.84(m,5H),1.80–1.66(m,3H).

[0193]

[0194] Step 6: Weigh compound 1k (73 mg, 1 equivalent) into a single-mouth bottle, add tetrahydrofuran and water (2:1) as solvent, and then add lithium hydroxide monohydrate (13 mg, 2 equivalents) to it, and react at room temperature for 4 hours. After the reaction is completed, add water and dichloromethane to the reaction solution for extraction, wash the organic phase with saturated salt water, dry it with anhydrous sodium sulfate, mix the sample and load it on the column to obtain compound S1. Analytical data of compound S1:

[0195] 1 H NMR (400MHz, CD 3 OD)δ8.70(d,J=3.0Hz,1H),7.82(s,1H),7.72(s,1H),4.84(d,J=7.5Hz,0.4H),4.82–4.69(m,2.6H),4.68–4.60(m,0.6H),4.49(s,2H) ,4.27–4.19(m,0.4H),4.02–3.82(m,2H),3.18–3.04(m,2H),2.92(s,1.8H),2.78(s,1.2H),2.27–2.07(m,1.5H),2.03–1.73(m,6.5H).

[0196] Preparation Example 2 Preparation of Compound S2

[0197]

[0198] Step 1: Weigh compound 1i (63 mg, 1 equivalent) in a single-mouth bottle, add dichloromethane solvent, then add TFA (99 μL, 10 equivalents), and react at room temperature for 3 hours. After the reaction is complete, spin dry the reaction solution. Then add dichloromethane as a solvent, add (R)-tetrahydrofurancarboxylic acid (23 mg, 1.5 equivalents), TBTU (86 mg, 2 equivalents), DIPEA (0.11 mL 5 equivalents) in sequence, and react at room temperature for 3 hours. After the reaction is complete, extract with dichloromethane and water, wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, mix the sample and load it on the column to obtain compound 2a. Analytical data of compound 2a: 1 H NMR (400 MHz, CDCl 3 )δ8.99(s,1H),8.42(s,1H),8.09(s,1H),5.42(s,2H),5.21–5.06(m,4H),4.36(d,J=36.2Hz,2H),3.54(t, J=14.8Hz,2H),3.34(s,2H),3.22(s,1H),2.63(br,1H),2.58(s,3H),2.37–2.31(m,4H),2.24–2.14(m,4H).

[0199]

[0200] Step 2: Weigh compound 2a (53 mg, 1 equivalent) into a single-mouth bottle, add tetrahydrofuran and water (2:1) as solvent, then add lithium hydroxide monohydrate (9 mg, 2 equivalents) into it, and react at room temperature for 4 hours. After the reaction is completed, add water and dichloromethane to the reaction solution for extraction, wash the organic phase with saturated brine, dry it with anhydrous sodium sulfate, mix the sample and load it on the column to obtain compound S2. Analytical data of compound S2: 1 H NMR(400MHz,d6-DMSO)δ8.89(s,1H),7.93(s,1H),7.67(s,1H),5.42(t,J=6.0Hz,1 H),4.81–4.74(m,2.5H),4.63(dd,J=7.3,5.9Hz,0.5H),4.59–4.52(m,0.5H),4.38 (d,J=6.0Hz,2H),4.22–4.16(m,0.5H),3.81–3.70(m,2H),3.17–3.04(m,2H),2.83 (s,1.7H),2.64(s,1.3H),2.12–1.94(m,2H),1.90–1.66(m,5H),1.62–1.59(m,1H).

[0201] Preparation Example 3 Preparation of Compound S3

[0202] The synthesis method of compound S3 refers to compound S1, except that (R)-tetrahydrofurancarboxylic acid is replaced with thiophene-2-carboxylic acid. The analytical data of compound S3 are as follows: 1 H NMR (400MHz, CD 3 OD)δ8.72(s,1H),7.83(s,1H),7.74(s,1H),7.65(d,J=5.0Hz,1H),7.47(d,J=3.5Hz,1H),7.17–7 .10(m,1H),4.80(d,J=13.3Hz,2H),4.60(br,1H),4.49(s,2H),3.06(br,5H),2.04–1.90(m,4H).

[0203] Preparation Example 4 Preparation of Compound S4

[0204] The synthesis method of compound S4 refers to compound S1, except that (R)-tetrahydrofurancarboxylic acid is replaced with 4,5-dichlorothiophene-2-carboxylic acid. The analytical data of compound S4 are as follows: 1 H NMR (400 MHz, CDCl 3 )δ8.57(s,1H),7.90(s,1H),7.80(s,1H),7.16(s,1H),4.73–4.70(m,3H),4.60(s,2H) ,3.13(d,J=13.1Hz,2H),3.06(s,3H),1.92(d,J=10.0Hz,3H),1.84(d,J=10.3Hz,1H).

[0205] Preparation Example 5 Preparation of Compound S5

[0206] The synthesis method of compound S5 refers to compound S1, except that (R)-tetrahydrofurancarboxylic acid is replaced with 4-acetylthiophene-2-carboxylic acid (the synthesis method refers to patent WO2001096286). The analytical data of compound S5 are: 1 H NMR(400MHz,d6-DMSO)δ8.91(s,1H),8.65(d,J=1.1Hz,1H),7.96(s,1H),7.75(br,1H),7.70(s,1H),5.47(br ,1H),4.78(d,J=13.1Hz,2H),4.54(br,1H),4.40(s,2H),3.04(d,J=22.0Hz,5H),2.52(s,3H),1.82(br,4H).

[0207] Preparation Example 6 Preparation of Compound S6

[0208] The synthesis method of compound S6 refers to compound S1, except that (R)-tetrahydrofurancarboxylic acid is replaced with 4-(hydroxymethyl)thiophene-2-carboxylic acid. The analytical data of compound S6 are as follows: 1 H NMR(400MHz,d6-DMSO)δ8.92(s,1H),7.96(s,1H),7.70(s,1H),7.50(s,1H),7.40(s,1H),5.48(t,J=6.0Hz,1H),5.20(t,J=5.7Hz,1H),4 .77(d,J=13.0Hz,2H),4.54(s,1H),4.47(d,J=5.6Hz,2H),4.40(d,J=6.0Hz,2H),3.07(t,J=12.0Hz,2H),2.97(s,3H),1.89–1.73(m,4H).

[0209] Preparation Example 7 Preparation of Compound S7

[0210]

[0211] The synthesis method of compound 7a is shown in compound 1k;

[0212] Step 1: Weigh compound 7a (82 mg, 1 equivalent) in a single-mouth bottle, add dichloromethane solvent, add DAST (31 μL, 1.5 equivalent) under ice bath, and react at room temperature for 0.5 hours. After the reaction is complete, neutralize the reaction solution with sodium bicarbonate, then extract with dichloromethane and water, wash the organic phase with saturated salt water, dry with anhydrous sodium sulfate, mix the sample and load it on the column to obtain compound 7b. Analytical data of compound 7b: 1 H NMR (400 MHz, CDCl 3 )δ8.58(s,1H),7.89(s,1H),7.83(s,1H),7.47(s,1H),7.37(s,1H),5.35(d,J=47.8Hz,2H ),4.99(s,2H),4.71(d,J=13.1Hz,3H),3.16–3.00(m,5H),2.16(s,3H),1.96–1.82(m,4H).

[0213]

[0214] Step 6: Weigh compound 7b (30 mg, 1 equivalent) into a single-mouth bottle, add tetrahydrofuran and water (2:1) as solvent, then add lithium hydroxide monohydrate (5 mg, 2 equivalents) into the bottle, and react at room temperature for 3 hours. After the reaction is completed, water and dichloromethane are added to the reaction solution for extraction, the organic phase is washed with saturated brine, dried with anhydrous sodium sulfate, and then mixed with the sample and loaded onto the column to obtain compound S7. Analytical data of compound S7: 1H NMR (400 MHz, CDCl 3 )δ8.57(s,1H),7.90(s,1H),7.80(s,1H),7.48(s,1H),7.38(s,1H),5.35(d,J=47.9Hz,2H),4.73–4.69(m,3H), 4.60(s,2H),3.12(d,J=12.0Hz,2H),3.06(s,3H),2.08(br,1H),1.94(d,J=10.8Hz,2H),1.86(d,J=10.2Hz,1H).

[0215] Preparation Example 8 Preparation of Compound S8

[0216] The synthesis method of compound S8 refers to compound S1, except that (R)-tetrahydrofurancarboxylic acid is replaced with 4-difluoromethylthiophene-2-carboxylic acid (the synthesis method is described in the literature European Journal of Medicinal Chemistry, 2018, 159, 23-34). The analytical data of compound S8 are as follows: 1 H NMR (400 MHz, CDCl 3 )δ8.57(s,1H),7.89(s,1H),7.80(s,1H),7.67(s,1H),7.43(s,1H),6.67(t,J=56.2Hz,1H),4.71(d,J=13.3Hz,3H) ,4.60(d,J=4.8Hz,2H),3.12(d,J=12.4Hz,2H),3.06(s,3H),2.01(t,J=6.4Hz,1H),1.90(dd,J=33.1,11.6Hz,4H).

[0217] Preparation Example 9 Preparation of Compound S9

[0218]

[0219] Weigh compound S5 (48 mg, 1 equivalent) into a single-mouth bottle, add ethanol as solvent, then add sodium borohydride (11 mg, 3 equivalents) in an ice bath, warm to room temperature and react for 6 hours. After the reaction is completed, add water and ethyl acetate to the reaction solution for extraction, wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, mix the sample and load it on the column to obtain compound S9. Analytical data of compound S9: 1 H NMR (400 MHz, CDCl 3)δ8.57(s,1H),7.89(s,1H),7.79(s,1H),7.38(s,1H),7.33(s,1H),5.00–4.91(m,1H),4.71(d,J=13.2Hz,3H),4.60(d,J=6.2Hz,2H ), 3.11 (d, J = 13.3Hz, 2H), 3.05 (s, 3H), 2.04 (t, J = 6.3Hz, 1H), 1.95 (d, J = 14.8Hz, 3H), 1.86 (d, J = 10.9Hz, 2H), 1.53 (d, J = 6.4Hz, 3H).

[0220] Preparation Example 10 Preparation of Compound S10

[0221]

[0222] Weigh compound S5 (135 mg, 1 equivalent) into a single-mouth bottle, add tetrahydrofuran as solvent, protect with nitrogen, then add methylmagnesium bromide (3M tetrahydrofuran solution, 0.38 mL, 3 equivalents) under ice bath, warm to room temperature and react for 8 hours. After the reaction is completed, add saturated ammonium chloride solution to the reaction solution to quench, then add water and ethyl acetate to extract, wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, mix and load on column to obtain compound S10. Analytical data of compound S10: 1 H NMR(600MHz,d6-DMSO)δ8.91(s,1H),7.96(s,1H),7.70(s,1H),7.45(d,J=4.4Hz,2H),5.46(t,J=6.0Hz,1H),5.11(s,1H), 4.78(d,J=13.3Hz,2H),4.51(br,1H),4.40(d,J=6.0Hz,2H),3.05(br,2H),2.97(br,3H),1.87–1.80(m,4H),1.43(s,6H).

[0223] Preparation Example 11 Preparation of Compound S11

[0224] The synthesis method of compound S11 refers to compound S1, except that (R)-tetrahydrofurancarboxylic acid is replaced with 1,4-dioxane-2-carboxylic acid. The analytical data of compound S11 are as follows:

[0225] 1 H NMR (400MHz, CD 3OD)δ8.72(d,J=5.0Hz,1H),7.83(d,J=2.5Hz,1H),7.74(d,J=2.7Hz,1H),4.78(d,J=13 .1Hz,2H),4.65–4.57(m,0.6H),4.53(dd,J=9.4,2.7Hz,0.4H),4.49(s,2H),4.41(dd,J =9.4,2.7Hz,0.6H),4.24–4.17(m,0.4H),3.90–3.71(m,4H),3.70–3.59(m,2H),3.19– 3.05(m,2H),2.95(s,1.7H),2.76(s,1.3H),1.98–1.78(m,3H),1.74(d,J=12.7Hz,1H).

[0226] Preparation Example 12 Preparation of Compound S12

[0227]

[0228] Weigh compound S7 (24 mg, 1 equivalent) into a single-mouth bottle, add dichloromethane solvent, add DAST (10 μL, 1.5 equivalents) under ice bath, and react at room temperature for 0.5 hours. After the reaction is complete, neutralize the reaction solution with sodium bicarbonate, then extract with dichloromethane and water, wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, mix the sample and load it on the column to obtain compound S12. Analytical data of compound S12: 1 H NMR (400 MHz, CDCl 3 )δ8.59(s,1H),7.91(s,1H),7.84(s,1H),7.48(s,1H),7.37(s,1H),5.41(s,1H),5.33(s,1H),5.2 9(s,1H),5.22(s,1H),4.73–4.70(m,3H),3.13(d,J=15.2Hz,2H),3.06(s,3H),1.96–1.85(m,4H).

[0229] Preparation Example 13 Preparation of Compound S13

[0230] The synthetic route of compound S13 is similar to compound S1, except that the raw material 4-N-tert-butyloxycarbonyl-4-N-methylaminopiperidine is replaced by 4-tert-butyloxycarbonylaminopiperidine, and (R)-tetrahydrofurancarboxylic acid is replaced by 4-(hydroxymethyl)thiophene-2-carboxylic acid. Analytical data of compound S13:

[0231] 1H NMR(500MHz,d6-DMSO)δ8.91(s,1H),8.30(d,J=7.8Hz,1H),7.96(s,1H),7.72 (d,J=10.2Hz,2H),7.47(s,1H),5.45(t,J=6.0Hz,1H),5.17(t,J=5.5Hz,1H), 4.60(d,J=13.4Hz,2H),4.43(d,J=5.5Hz,2H),4.40(d,J=6.0Hz,2H),4.15–4. 06(m,1H),3.18(t,J=12.0Hz,2H),1.93(d,J=10.3Hz,2H),1.63–1.53(m,2H).

[0232] Preparation Example 14 Preparation of Compound S14

[0233] The synthetic route of compound S14 is similar to that of compound S1, except that the raw material 4-N-tert-butyloxycarbonyl-4-N-methylaminopiperidine is replaced by 4-tert-butyloxycarbonylaminopiperidine. Analytical data of compound S14:

[0234] 1 H NMR (500 MHz, CDCl 3 )δ8.55(s,1H),7.89(s,1H),7.79(s,1H),6.63(d,J=8.1Hz,1H),4.59(s,2H),4.49(d,J=13.8Hz,2H),4.34(dd,J=8.3,6.0Hz,1H),4 .15–4.05(m,1H),3.95–3.84(m,2H),3.34–3.08(m,2H),2.35–2.27(m,1H),2.12–2.02(m,3H),1.95–1.83(m,2H),1.57–1.48(m,3H).

[0235] Preparation Example 15 Preparation of Compound S15

[0236]

[0237] Step 1: Weigh a mixture of 1d and 1e (650 mg, 1 equivalent) and tert-butyl piperazine-1-carboxylate (559 mg, 1.5 equivalents) in a single-mouth bottle, add 1,4-dioxane solvent, then add DIPEA (0.99 mL, 3 equivalents), and react at 100°C for 3 hours. After the reaction is complete, extract with ethyl acetate and water, wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, mix and load on a column to obtain a mixture of 15a and 15b.

[0238]

[0239] Step 2: Weigh the mixture of 15a and 15b (768 mg, 1 equivalent), PdCl 2 (PPh 3 ) 2 (131 mg, 0.1 equivalent) and CuI (71 mg, 0.2 equivalent) were placed in a single-mouth bottle, tetrahydrofuran solvent was added, and then propargyl acetate (0.37 mL, 2 equivalents) and DIPEA (1.24 mL, 4 equivalents) were added, and the reaction was heated at 80°C for 5 hours under nitrogen protection. After the reaction was complete, it was extracted with ethyl acetate and water, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and then mixed and loaded onto a column to obtain compounds 15c and 15d. Analytical data of compound 15c: 1 H NMR (400 MHz, CDCl 3 )δ8.54(s,1H),8.01(s,1H),7.68(s,1H),4.99(s,2H),3.85–3.76(m,4H),3.64–3.55(m,4H),2.15(s,3H),1.49(s,9H); Analytical data of compound 15d: 1 H NMR (400 MHz, CDCl 3 )δ8.53(s,1H),7.90(s,1H),7.83(s,1H),4.99(s,2H),3.78–3.72(m,4H),3.63–3.55(m,4H),2.15(s,3H),1.49(s,9H).

[0240]

[0241] Step 3: Weigh compound 15d (60 mg, 1 equivalent) in a single-mouth bottle, add dichloromethane solvent, then add TFA (0.10 mL, 10 equivalents), and react at room temperature for 3 hours. After the reaction is complete, spin dry the reaction solution. Then add dichloromethane as a solvent, add (R)-tetrahydrofurancarboxylic acid (23 mg, 1.5 equivalents), TBTU (87 mg, 2 equivalents), DIPEA (0.11 mL, 5 equivalents) in sequence, and react at room temperature for 3 hours. After the reaction is complete, extract with dichloromethane and water, wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, mix the sample and load it on the column to obtain compound 15e. Analytical data of compound 15e: 1 H NMR (400 MHz, CDCl 3)δ8.55(s,1H),7.91(s,1H),7.84(s,1H),4.99(s,2H),4.69–4.62(m,1H),3.99–3.83(m, 6H),3.80–3.62(m,4H),2.80(s,3H),2.44–2.34(m,1H),2.16(s,3H),2.10–1.99(m,3H).

[0242]

[0243] Step 4: Weigh compound 15e (48 mg, 1 equivalent) into a single-mouth bottle, add tetrahydrofuran and water (2:1) as solvent, then add lithium hydroxide monohydrate (9 mg, 2 equivalents) into the bottle, and react at room temperature for 4 hours. After the reaction is completed, water and dichloromethane are added to the reaction solution for extraction, the organic phase is washed with saturated brine, dried with anhydrous sodium sulfate, and then mixed with the sample and loaded onto the column to obtain compound S15. Analytical data of compound S15: 1 H NMR (400MHz, CD 3 OD)δ8.69(s,1H),7.83(s,1H),7.73(s,1H),4.79(dd,J=7.8,6.0Hz,1H) ,4.49(s,2H),4.01–3.63(m,10H),2.30–2.06(m,2H),2.04–1.91(m,2H).

[0244] Preparation Example 16 Preparation of Compound S16

[0245]

[0246] Step 1: Weigh compound 15c (60 mg, 1 equivalent) in a single-mouth bottle, add dichloromethane solvent, then add TFA (0.10 mL, 10 equivalents), and react at room temperature for 3 hours. After the reaction is complete, spin dry the reaction solution. Then add dichloromethane as a solvent, add (R)-tetrahydrofurancarboxylic acid (23 mg, 1.5 equivalents), TBTU (87 mg, 2 equivalents), DIPEA (0.11 mL, 5 equivalents) in sequence, and react at room temperature for 3 hours. After the reaction is complete, extract with dichloromethane and water, wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, mix and load on the column to obtain compound 16a. Analytical data of compound 16a: 1 H NMR (400 MHz, CDCl 3)δ8.56(s,1H),8.03(s,1H),7.70(s,1H),4.99(s,2H),4.68–4.61(m,1H) ,3.97–3.67(m,10H),2.46–2.34(m,1H),2.15(s,3H),2.11–1.90(m,3H).

[0247]

[0248] Step 2: Weigh compound 16a (57 mg, 1 equivalent) into a single-mouth bottle, add tetrahydrofuran and water (2:1) as solvent, then add lithium hydroxide monohydrate (11 mg, 2 equivalents) into the bottle, and react at room temperature for 4 hours. After the reaction is completed, water and dichloromethane are added to the reaction solution for extraction, the organic phase is washed with saturated brine, dried with anhydrous sodium sulfate, and then mixed with the sample and loaded onto the column to obtain compound S16. Analytical data of compound S16:

[0249] 1 H NMR (400MHz, CD 3 OD)δ8.70(s,1H),7.92(s,1H),7.65(s,1H),4.79(dd,J=7.8,6.0Hz,1H),4.48(s,2H),3.96(dd, J=14.6,7.0Hz,3H),3.91–3.78(m,5H),3.75–3.65(m,2H),2.26–2.08(m,2H),2.01–1.92(m,2H).

[0250] Preparation Example 17 Preparation of Compound S17

[0251] The synthesis method of compound S17 refers to compound S15, except that (R)-tetrahydrofurancarboxylic acid is replaced with 4-(hydroxymethyl)thiophene-2-carboxylic acid. The analytical data of compound S17 are as follows: 1 H NMR (400MHz, CD 3 OD)δ8.71(s,1H),7.86(s,1H),7.77(s,1H),7.52(s,1H),7.44(s,1H),4.62(s,2H),4.49(s,2H),3.94(s,8H).

[0252] Preparation Example 18 Preparation of Compound S18

[0253] The synthesis method of compound S18 refers to compound S16, except that (R)-tetrahydrofurancarboxylic acid is replaced with 4-(hydroxymethyl)thiophene-2-carboxylic acid. The analytical data of compound S18 are as follows: 1H NMR(400MHz,d6-DMSO)δ8.87(s,1H),7.98(s,1H),7.72(s,1H),7.54(s,1H),7.42(d,J=1.0Hz,1H) ,5.44(br,1H),5.23(br,1H),4.48(s,2H),4.39(s,2H),3.94(dd,J=6.6,3.5Hz,4H),3.84(s,4H).

[0254] Preparation Example 19 Preparation of Compound S19

[0255] The synthesis method of compound S19 refers to compound S15, except that (R)-tetrahydrofurancarboxylic acid is replaced with (R)-2-trifluoromethyl-2-hydroxypropionic acid. The analytical data of compound S19 are as follows: 1 H NMR (400MHz, CD 3 OD)δ8.71(s,1H),7.86(s,1H),7.78(s,1H),4.49(s,2H),4.24(br,2H),3.87(br,4H),3.84–3.70(m,2H),1.65(s,3H).

[0256] Preparation Example 20 Preparation of Compound S20

[0257]

[0258] For the synthesis method of compound 20a, please refer to patent WO2012045196A.

[0259] Step 1: Weigh compound 20a (359 mg, 1 equivalent) in a microwave tube, add tetrahydrofuran as solvent, then add ethylamine aqueous solution (12 mol / L, 0.17 mL, 2 equivalents), and microwave for 20 minutes. After the reaction, extract with ethyl acetate and water, wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, mix and load on a column to obtain a mixture of 20b and 20c.

[0260]

[0261] Step 2: Weigh a mixture of 20b and 20c (405 mg, 1 equivalent) and 1f (471 mg, 1.5 equivalents) in a single-mouth bottle, add 1,4-dioxane solvent, then add DIPEA (0.55 mL, 3 equivalents), and react at 100°C for 16 hours. After the reaction is complete, extract with ethyl acetate and water, wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, mix and load on a column to obtain a mixture of 20d and 20e.

[0262]

[0263] Step 5: Weigh a mixture of 20d and 20e (582 mg, 1 eq), PdCl 2 (PPh 3 ) 2 (74 mg, 0.1 equivalent) and CuI (42 mg, 0.2 equivalent) were placed in a single-mouth bottle, tetrahydrofuran solvent was added, propargyl acetate (0.21 mL, 2 equivalents) and DIPEA (0.70 mL, 4 equivalents) were added, and the mixture was heated at 80°C for 5 hours under nitrogen protection. After the reaction was complete, the mixture was extracted with ethyl acetate and water, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and then mixed and loaded onto a column to obtain compounds 20f and 20g. Analytical data of compound 20f: 1 H NMR (400 MHz, CDCl 3 )δ7.84(s,1H),7.64(s,1H),5.36(t,J=5.5Hz,1H),4.97(s,2H),4.17(br,1H),3.67(d,J=11.6Hz,2H),3.62–3.53(m,2H),2.91(t,J=11.1Hz,2H),2.80(s,3H),2.14(s,3H),1.93–1.80(m,4H),1.48(s,9H),1.32(t,J=7.2Hz,3H).Analytical data of compound 20g: 1 H NMR (400 MHz, CDCl 3 )δ7.79(s,1H),7.70(s,1H),4.98(s,2H),4.15(br,1H),3.74(d,J=13.8Hz,2H),3.60–3.52(m,2H),2. 91(t,J=11.8Hz,2H),2.80(s,3H),2.15(s,3H),1.92–1.79(m,4H),1.48(s,9H),1.32(t,J=7.1Hz,3H).

[0264]

[0265] Step 6: Weigh compound 20f (65 mg, 1 equivalent) in a single-mouth bottle, add dichloromethane solvent, then add TFA (94 μL, 10 equivalents), and react at room temperature for 3 hours. After the reaction is complete, spin dry the reaction solution. Then add dichloromethane as a solvent, add (R)-tetrahydrofurancarboxylic acid (22 mg, 1.5 equivalents), TBTU (81 mg, 2 equivalents), DIPEA (0.10 mL 5 equivalents) in sequence, and react at room temperature for 3 hours. After the reaction is complete, extract with dichloromethane and water, wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, mix the sample and load it on the column to obtain compound 20h. Analytical data of compound 20h:1 H NMR (400 MHz, CDCl 3 )δ7.83(d,J=3.6Hz,1H),7.65(d,J=5.9Hz,1H),5.39–5.32(m,1H),4.97(s,2H),4.68–4.59(m,1.7H),4.12(br,0.3H),4.01–3.85(m,2 H),3.70(d,J=13.2Hz,2H),3.63–3.54(m,2H),3.03–2.92(m,4H),2.89(s,1H),2.14(s,3H),2.09–1.76(m,8H),1.32(t,J=3.5Hz,3H).

[0266]

[0267] Step 7: Weigh compound 20h (64 mg, 1 equivalent) into a single-mouth bottle, add tetrahydrofuran and water (2:1) as solvent, and then add lithium hydroxide monohydrate (11 mg, 2 equivalents) to it, and react at room temperature for 4 hours. After the reaction is completed, water and dichloromethane are added to the reaction solution for extraction, the organic phase is washed with saturated brine, dried with anhydrous sodium sulfate, and then mixed with the sample and loaded onto the column to obtain compound S20. Analytical data of compound S20: 1 H NMR (400MHz, CD 3 OD)δ7.72(s,1H),7.56(d,J=1.8Hz,1H),4.83–4.79(m,0.5H),4.75–4.72(m,0.5H),4.58–4 .51(m,0.5H),4.46(s,2H),4.12–4.03(m,0.5H),4.12–4.03(m,1H),3.91–3.83(m,1H),3.76 (d,J=12.0Hz,2H),3.57(qd,J=7.1,2.5Hz,2H),3.01(s,1.7H),2.95–2.92(m,2H),2.89(s,1 .3H),2.29–2.16(m,2H),2.13–1.92(m,4H),1.84–1.69(m,2H),1.29(td,J=7.1,2.5Hz,3H).

[0268] Preparation Example 21 Preparation of Compound S21

[0269]

[0270] Step 1: Weigh compound 20g (65mg, 1 equivalent) in a single-mouth bottle, add dichloromethane solvent, then add TFA (94μL, 10 equivalents), and react at room temperature for 3 hours. After the reaction is complete, spin dry the reaction solution. Then add dichloromethane as solvent, add (R)-tetrahydrofurancarboxylic acid (22mg, 1.5 equivalents), TBTU (81mg, 2 equivalents), DIPEA (0.10mL 5 equivalents) in sequence, and react at room temperature for 3 hours. After the reaction is complete, extract with dichloromethane and water, wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, mix the sample and load it on the column to obtain compound 21a. Analytical data of compound 21a: 1 H NMR (400 MHz, CDCl 3 )δ7.79(d,J=6.2Hz,1H),7.70(s,1H),5.35(d,J=7.2Hz,1H),5.17(br,1H),4.98(s,2H),4.65(t,J=6.5Hz,1H),4.02–3.7 1(m,4H),3.61–3.51(m,2H),3.03–2.93(m,3.7H),2.89(s,1.3H),2.15(s,3H),2.04–1.73(m,8H),1.31(t,J=3.8HZ,3H).

[0271]

[0272] Step 2: Weigh compound 21a (52 mg, 1 equivalent) into a single-mouth bottle, add tetrahydrofuran and water (2:1) as solvent, then add lithium hydroxide monohydrate (8 mg, 2 equivalents) into it, and react at room temperature for 4 hours. After the reaction is completed, add water and dichloromethane to the reaction solution for extraction, wash the organic phase with saturated brine, dry it with anhydrous sodium sulfate, mix the sample and load it on the column to obtain compound S21. Analytical data of compound S21: 1 H NMR (400MHz, CD 3 OD)δ7.67(d,J=1.7Hz,1H),7.62(s,1H),4.83–4.79(m,0.5H),4.75–4.72(m,0.5 H),4.58–4.51(m,0.5H),4.47(s,2H),4.12–4.03(m,0.5H),3.99–3.92(m,1H),3. 89–3.82(m,3H),3.55(qd,J=6.7,2.4Hz,2H),3.00(s,1.7H),2.94(t,J=12.4Hz, 2H),2.88(s,1.3H),2.29–1.93(m,6H),1.85–1.69(m,2H),1.29(t,J=6.7Hz,3H).

[0273] Preparation Example 22 Preparation of Compound S22

[0274] The synthesis method of compound S22 refers to compound S20, except that (R)-tetrahydrofurancarboxylic acid is replaced with 4-(hydroxymethyl)thiophene-2-carboxylic acid. The analytical data of compound S22 are as follows: 1 H NMR (400MHz, CD 3 OD)δ7.72(s,1H),7.57(s,1H),7.49(s,1H),7.43(s,1H),4.60(s,2H),4.46(br,3H),3.78(d,J=11.4Hz,2H),3.5 7(q,J=7.1Hz,2H),3.15(br,3H),2.90(br,2H),2.26–2.17(m,2H),1.86(d,J=11.4Hz,2H),1.29(t,J=7.1Hz,3H).

[0275] Preparation Example 23 Preparation of Compound S23

[0276] The synthesis method of compound S23 refers to compound S21, except that (R)-tetrahydrofurancarboxylic acid is replaced with 4-(hydroxymethyl)thiophene-2-carboxylic acid. The analytical data of compound S23 are as follows: 1 H NMR (400MHz, CD 3 OD)δ7.67(s,1H),7.63(s,1H),7.49(s,1H),7.43(s,1H),4.60(s,2H),4.47(br,3H),3.85(d,J=12.8Hz,2H),3.5 6(q,J=7.2Hz,2H),3.14(br,3H),2.92(br,2H),2.27–2.16(m,2H),1.86(d,J=10.9Hz,2H),1.30(t,J=7.2Hz,3H).

[0277] Preparation Example 24 Preparation of Compound S24

[0278]

[0279] Step 1: Weigh the mixture 20b and 20c (405 mg, 1 equivalent) and tert-butyl piperazine-1-carboxylate (410 mg, 1.5 equivalents) in a single-mouth bottle, add 1,4-dioxane solvent, then add DIPEA (0.55 mL, 3 equivalents), and react at 100°C for 16 hours. After the reaction is complete, extract with ethyl acetate and water, wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, mix and load on the column to obtain a mixture of 24a and 24b.

[0280]

[0281] Step 2: Weigh the mixture of 24a and 24b (493 mg, 1 eq), PdCl 2 (PPh 3 ) 2 (67 mg, 0.1 equivalent) and CuI (38 mg, 0.2 equivalent) were placed in a single-mouth bottle, tetrahydrofuran solvent was added, propargyl acetate (0.19 mL, 2 equivalents) and DIPEA (0.63 mL, 4 equivalents) were added, and the mixture was heated at 80°C for 5 hours under nitrogen protection. After the reaction was complete, the mixture was extracted with ethyl acetate and water, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and then mixed and loaded onto a column to obtain compounds 24c and 24d. Analytical data of compound 24c: 1 H NMR (400 MHz, CDCl 3 )δ7.83(s,1H),7.66(s,1H),5.36(t,J=4.9Hz,1H),4.98(s,2H),3.63–3.54(m,6H),3.24–3.14(m,4H),2.14(s,3H),1.49(s,9H),1.31(t,J=7.2Hz,3H); Analytical data of compound 24d: 1 H NMR (400 MHz, CDCl 3 )δ7.80(s,1H),7.70(s,1H),4.98(s,2H),3.65–3.59(m,4H),3.58–3.52(m ,2H),3.27–3.21(m,4H),2.15(s,3H),1.49(s,9H),1.31(t,J=7.2Hz,3H).

[0282]

[0283] Step 3: Weigh compound 24c (64 mg, 1 equivalent) in a single-mouth bottle, add dichloromethane solvent, then add TFA (97 μL, 10 equivalents), and react at room temperature for 3 hours. After the reaction is complete, spin dry the reaction solution. Then add dichloromethane as a solvent, add (R)-tetrahydrofurancarboxylic acid (23 mg, 1.5 equivalents), TBTU (84 mg, 2 equivalents), DIPEA (0.11 mL 5 equivalents) in sequence, and react at room temperature for 3 hours. After the reaction is complete, extract with dichloromethane and water, wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, mix the sample and load it on the column to obtain compound 24e. Analytical data of compound 24e: 1 H NMR (400 MHz, CDCl 3)δ7.84(s,1H),7.67(s,1H),5.37–5.34(m,1H),4.98(s,2H),4.66(dd,J=7.4,5.3Hz,1H),3.98–3.73(m,6H),3.58(dt ,J=12.7,6.4Hz,2H),3.30(t,J=5.0Hz,2H),3.27–3.21(m,2H),2.15(s,3H),2.12–1.98(m,4H),1.32(t,J=7.2Hz,3H).

[0284]

[0285] Step 4: Weigh compound 24e (62 mg, 1 equivalent) into a single-mouth bottle, add tetrahydrofuran and water (2:1) as solvent, then add lithium hydroxide monohydrate (11 mg, 2 equivalents) into the bottle, and react at room temperature for 4 hours. After the reaction is completed, water and dichloromethane are added to the reaction solution for extraction, the organic phase is washed with saturated brine, dried with anhydrous sodium sulfate, and then mixed with the sample and loaded onto the column to obtain compound S24. Analytical data of compound S24: 1 H NMR (400MHz, CD 3 OD)δ7.73(d,J=3.7Hz,1H),7.58(d,J=3.1Hz,1H),4.78(dd,J=7.7,6.0Hz,1H),4.46(s,2H),3.96(dd,J=14.7,6.8Hz,1H),3.87(dd,J=14.6,6.6 Hz,3H),3.82–3.75(m,2H),3.57(q,J=7.1Hz,2H),3.30–3.20(m,4H),2. 24–2.17(m,1H),2.13–2.05(m,1H),2.00–1.92(m,2H),(t,J=7.1Hz,3H).

[0286] Preparation Example 25 Preparation of Compound S25

[0287]

[0288] Step 1: Weigh compound 24d (60 mg, 1 equivalent) in a single-mouth bottle, add dichloromethane solvent, then add TFA (91 μL, 10 equivalents), and react at room temperature for 3 hours. After the reaction is complete, spin dry the reaction solution. Then add dichloromethane as a solvent, add (R)-tetrahydrofurancarboxylic acid (21 mg, 1.5 equivalents), TBTU (79 mg, 2 equivalents), DIPEA (0.10 mL 5 equivalents) in sequence, and react at room temperature for 3 hours. After the reaction is complete, extract with dichloromethane and water, wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, mix and load on the column to obtain compound 25a. Analytical data of compound 25a: 1 H NMR (400 MHz, CDCl 3 )δ7.81(s,1H),7.70(s,1H),5.19(t,J=5.6Hz,1H),4.98(s,2H),4.66(dd,J=7.1,5.6Hz,1H),3.98–3.78(m,6H), 3.57(dt,J=14.3,7.0Hz,2H),3.34(br,2H),3.28(br,2H),2.15(s,3H),2.11–1.91(m,4H),1.32(t,J=7.2Hz,3H).

[0289]

[0290] Step 2: Weigh compound 25a (52 mg, 1 equivalent) into a single-mouth bottle, add tetrahydrofuran and water (2:1) as solvent, then add lithium hydroxide monohydrate (9 mg, 2 equivalents) into the bottle, and react at room temperature for 4 hours. After the reaction is completed, water and dichloromethane are added to the reaction solution for extraction, the organic phase is washed with saturated brine, dried with anhydrous sodium sulfate, and then mixed with the sample and loaded onto the column to obtain compound S25. Analytical data of compound S25:

[0291] 1 H NMR (400MHz, CD 3 OD)δ7.68(s,1H),7.63(d,J=2.4Hz,1H),4.78(dd,J=7.6,6.1Hz,1H),4.47(s,2H),3.95(dd,J=14.6,6.9Hz,1H),3.87(dd,J=14.4,6.6Hz,3 H),3.81–3.74(m,2H),3.56(q,J=7.1Hz,2H),3.35(br,4H),2.26–2.17(m,1H),2.12–2.05(m,1H),2.00–1.92(m,2H),1.29(t,J=7.1Hz,3H).

[0292] Preparation Example 26 Preparation of Compound S26

[0293]

[0294] Step 1: Weigh the mixture of 1g and 1h (519mg, 1 equivalent) in a single-mouth bottle, add dichloromethane solvent, then add trifluoroacetic acid (TFA, 0.74mL, 10 equivalents), and react at room temperature for 3 hours. After the reaction is complete, spin dry the reaction solution. Then add dichloromethane as solvent, add (R)-tetrahydrofurancarboxylic acid (174mg, 1.5 equivalents), TBTU (642mg, 2 equivalents), DIPEA (0.83mL, 5 equivalents) in sequence, and react at room temperature for 3 hours. After the reaction is complete, extract with dichloromethane and water, wash the organic phase with saturated brine, dry with anhydrous sodium sulfate, mix and load on the column to obtain a mixture of 26a and 26b.

[0295]

[0296] Step 2: Weigh the mixture of 26a and 26b (460 mg, 1 eq), tert-butyl carbamate (138 mg, 1.3 eq), Pd 2 (dba) 3 (83 mg, 0.1 equivalent), Xantphos (110 mg, 0.2 equivalent), cesium carbonate (598 mg, 2 equivalent) were placed in a single-mouth bottle, 1,4-dioxane was added as solvent, nitrogen was protected, and the reaction was heated at 100°C for 6 hours. After the reaction was complete, it was extracted with ethyl acetate and water, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and then mixed and loaded onto a column to obtain compounds 26c and 26d. Analytical data of compound 26c: 1 H NMR (400 MHz, CDCl 3 )δ8.63(d,J=5.9Hz,1H),8.59(d,J=4.3Hz,1H),7.72(d,J=5.2Hz,1H),7.08(s,1H),4.81–4.75(m,0.6H),4.73–4.62(m,3H),4.27–4.20(m,0.4H),4.05–3.86(m,2H),3.16–3.03(m,2H),2.93(s,2H),2.82(s,1H),2.29–2.18(m,1H),2.15–2.03(m,2H),1.99–1.89(m,2H),1.84–1.77(m,3H),1.58(s,9H). Analytical data of compound 26d: 1 H NMR (400 MHz, CDCl 3)δ8.47(d,J=7.5Hz,1H),8.45(d,J=4.2Hz,1H),7.85(d,J=5.1Hz,1H),7.22(d ,J=5.3Hz,1H),4.80–4.59(m,3.5H),4.27–4.17(m,0.5H),4.03–3.85(m,2H),3 .12–3.01(m,2H),2.89(s,2H),2.78(s,1H),2.44–2.37(m,0.5H),2.26–2.16(m ,1H),2.13–2.00(m,2.5H),1.97–1.87(m,3H),1.81–1.75(m,1H),1.56(s,9H).

[0297]

[0298] Step 3: Weigh compound 26d (52 mg, 1 equivalent) into a single-mouth bottle, add dichloromethane solvent, then add trifluoroacetic acid (TFA, 79 μL, 10 equivalents), and react at room temperature for 5 hours. After the reaction is complete, spin dry the reaction solution. Neutralize with saturated sodium bicarbonate solution, extract with dichloromethane and water, wash the organic phase with saturated salt water, dry with anhydrous sodium sulfate, mix the sample and load on the column to obtain compound S26. Analytical data of compound S26: 1 H NMR (400MHz, CD 3 OD)δ8.35(d,J=2.8Hz,1H),7.65(d,J=2.3Hz,1H),6.89(s,1H),4.86–4.81(m,0.5H),4.73–4.59(m,3H),4.22–4.13(m, 0.5H),4.00–3.82(m,2H),3.05(q,J=13.0Hz,2H),2.91(s,1.7H),2.78(s,1.3H),2.27–2.17(m,1H),2.14–1.68(m,7H).

[0299] Preparation Example 27 Preparation of Compound S27

[0300]

[0301] Weigh compound 26c (147 mg, 1 equivalent) in a single-mouth bottle, add dichloromethane solvent, then add TFA (0.22 mL, 10 equivalents), and react at room temperature for 5 hours. After the reaction is complete, spin dry the reaction solution. Neutralize with saturated sodium bicarbonate solution, extract with dichloromethane and water, wash the organic phase with saturated salt water, dry with anhydrous sodium sulfate, mix the sample and load it on the column to obtain compound S27. Analytical data of compound S27: 1 H NMR (400MHz, CD 3OD)δ8.55(s,1H),7.59(s,1H),7.15(s,1H),4.86–4.81(m,0.5H),4.71(dd,J=6.9 ,4.2Hz,0.5H),4.59(d,J=11.3Hz,2H),4.15(t,J=10.6Hz,0.5H),3.99–3.82(m,2H ),3.03(dd,J=23.2,10.3Hz,2H),2.91(s,1.7H),2.78(s,1.3H),2.29–2.17(m,1H) ,2.14–2.06(m,0.5H),2.02–1.89(m,3.5H),1.87–1.76(m,2H),1.74–1.66(m,1H).

[0302] Preparation Example 28 Preparation of Compound S28

[0303]

[0304] The mixture of 26a and 26b (200 mg, 1 eq.), 1H-pyrazole-3-boronic acid (78 mg, 1.5 eq.), Pd(PPh 3 ) 4 (92 mg, 0.2 equivalents), sodium carbonate (130 mg, 3 equivalents) were placed in a single-mouth bottle, 1,4-dioxane (10 mL) and water (0.6 mL) were added as solvents, nitrogen was protected, and the reaction was heated at 100°C for 3 hours. After the reaction was complete, it was extracted with ethyl acetate and water, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and then mixed and loaded onto a column to obtain compound S28 and compound S29. Analytical data of compound S28: 1 H NMR (400MHz, CD 3 OD)δ8.70(s,1H),7.88(br,2H),7.76(br,1H),6.78(s,1H),4.86–4.81(m,0.5H),4.79–4.60(m,3H),4.23–4.17(m,0.5H ),3.99–3.80(m,2H),3.08(t,J=10.9Hz,2H),2.90(s,1.7H),2.77(s,1.3H),2.26–2.06(m,1.5H),2.01–1.69(m,6.5H).

[0305] Preparation Example 29 Preparation of Compound S29

[0306] The synthesis method of compound S29 refers to compound S28, and the analytical data of compound S29 is: 1 H NMR (400MHz, CD 3OD)δ8.72(s,1H),8.04(br,1H),7.72(s,2H),6.73(s,1H),4.85–4.61(m,3.5H),4.22(br,0.5H),4.00–3.8 2(m,2H),3.10(t,J=12.5Hz,2H),2.92(s,1.7H),2.78(s,1.3H),2.25–2.07(m,1.5H),2.02–1.74(m,6.5H).

[0307] Preparation Example 30 Preparation of Compound S30

[0308] The synthesis method of compound S30 refers to compound S28, and the analytical data of compound S30 is: 1 H NMR (400MHz, CD 3 OD)δ8.66(t,J=7.8Hz,1H),8.07(br,2H),7.85(t,J=6.9Hz,1H),7.74(t,J=7.0 Hz,1H),4.85(t,J=6.8Hz,0.5H),4.80–4.69(m,2.5H),4.64(dd,J=11.9,7.9Hz, 0.5H),4.21(br,0.5H),4.01–3.81(m,2H),3.08(t,J=12.8Hz,2H),2.91(d,J=2. 7Hz,1.7H),2.78(d,J=1.7Hz,1.3H),2.29–2.07(m,1.5H),2.04–1.71(m,6.5H).

[0309] Preparation Example 31: Preparation of Compound S31

[0310] The synthesis method of compound S31 refers to compound S28, and the analytical data of compound S31 are: 1 H NMR (400MHz, CD 3 OD)δ8.73(d,J=3.0Hz,1H),8.03(br,2H),7.92(d,J=2.4Hz,1H),7.74(d,J=2.0Hz,1H),4.85–4.60(m,4H),4.27–4.20 (m,0.5H),3.98–3.86(m,2H),3.18–3.07(m,2H),2.93(s,1.7H),2.79(s,1.3H),2.28–2.12(m,2H),2.06–1.82(m,6H).

[0311] Preparation Example 32 Preparation of Compound S32

[0312] The synthesis method of compound S32 refers to compound S28, and the analytical data of compound S32 is: 1 H NMR (400MHz, CD 3 OD)δ8.71(d,J=2.5Hz,1H),7.88(d,J=2.7Hz,1H),7.84(d,J=3.0Hz,1H),6.52 (s,1H),4.87–4.83(m,1H),4.81–4.69(m,3H),4.69–4.62(m,0.5H),4.27–4.1 7(m,0.5H),3.99–3.83(m,2H),3.16–3.03(m,2H),2.91(s,1.7H),2.78(s,1.3 H),2.37(s,3H),2.28–2.17(m,1H),2.14–2.06(m,0.5H),2.02–1.75(m,6.5H).

[0313] Preparation Example 33 Preparation of Compound S33

[0314] The synthesis method of compound S33 refers to compound S28, and the analytical data of compound S33 are: 1 H NMR (400MHz, CD 3 OD)δ8.70(s,1H),7.99(s,1H),7.69(s,1H),6.48(s,1H),4.87–4.59(m,3.5H),4.21(br,0.5H),4.03–3.80(m,2H ),3.09(t,J=11.1Hz,2H),2.91(s,1.7H),2.78(s,1.3H),2.36(s,3H),2.27–2.08(m,1.5H),2.02–1.73(m,6.5H).

[0315] Preparation Example 34 Preparation of Compound S34 The synthesis method of compound S34 refers to compound S1, except that (R)-tetrahydrofurancarboxylic acid is replaced by (2R)-1,4-dioxane-2-carboxylic acid. The analytical data of compound S34 are as follows: 1 H NMR (400MHz, CD 3OD)δ8.69(d,J=5.5Hz,1H),7.80(d,J=2.6Hz,1H),7.71(d,J=3.0Hz,1H),4.76(d,J=13 .3Hz,2H),4.64–4.56(m,0.6H),4.49(s,2H),4.53(dd,J=9.4,2.8Hz,0.4H),4.40(dd,J =9.4,2.8Hz,0.6H),4.23–4.16(m,0.4H),3.90–3.71(m,4H),3.70–3.59(m,2H),3.18– 3.03(m,2H),2.95(s,1.7H),2.76(s,1.3H),1.99–1.78(m,3H),1.73(d,J=10.3Hz,1H).

[0316] Preparation Example 35 Preparation of Compound S35 The synthesis method of compound S35 refers to compound S1, except that (R)-tetrahydrofurancarboxylic acid is replaced by (2S)-1,4-dioxane-2-carboxylic acid. The analytical data of compound S35 are as follows: 1 H NMR (400MHz, CD 3 OD)δ8.70(d,J=5.5Hz,1H),7.83(d,J=2.6Hz,1H),7.71(d,J=3.0Hz,1H),4.76(d,J=13 .3Hz,2H),4.64–4.56(m,0.6H),4.49(s,2H),4.53(dd,J=9.4,2.8Hz,0.4H),4.40(dd,J =9.4,2.8Hz,0.6H),4.23–4.16(m,0.4H),3.90–3.71(m,4H),3.70–3.59(m,2H),3.18– 3.03(m,2H),2.95(s,1.7H),2.76(s,1.3H),1.99–1.78(m,3H),1.72(d,J=10.3Hz,1H).

[0317] 2. Biological Activity Test Example

[0318] Example 1. Test of the PGK1 enzyme inhibitory activity of compounds

[0319] To each reaction, add 20 μL 50 mM potassium dihydrogen phosphate, pH 7.0, 4 μL 50 mM GAP, 6 μL 10 mM βNAD, 4 μL 10 mM adenosine diphosphate, 10 μL 100 mM magnesium sulfate, 20 μL 1 M Glycine, 4 μL 0.25 μg / μL GAPDH, and 32 μL deionized water, for a total of 100 μL substrate mixture. Dilute PGK1 protein to 0.1 ng / μL, and add 100 μL PGK1 protein dilution to each reaction. Incubate at 37°C for 30 min.

[0320] Dilute the ATP detection solution with ATP detection diluent at a ratio of 1:10. In a 96-well white plate, add 100uL of the diluted detection solution to each well, react at room temperature for 5min (to remove the influence of background ATP), add 100uL of enzyme activity reaction solution, react for at least 2s, and read the fluorescence value in a chemiluminescence detector.

[0321] Set the compound concentration gradient. In the above reaction process, add different concentrations of compounds into the 200uL reaction system. Set at least 3 replicate wells. After reading the fluorescence value, compare it with the control group, calculate the relative enzyme activity, draw the fitting curve with GraphPad Prism, and calculate the IC 50 value.

[0322] Table 2. Inhibitory activity of compounds on PGK1 enzyme

[0323]

[0324] Example 2. Test of the inhibitory activity of compounds on SNU739 cell proliferation

[0325] Experimental steps:

[0326] 1) Cultivate the corresponding target cell line, count the cells using a hemocytometer, and plate 96-well plates, 1000 cells / well, and wait for the cells to adhere.

[0327] 2) After the cells adhered, a compound concentration gradient was set, and 10% serum culture medium containing different concentrations of the compound was prepared and added to a 96-well plate, with at least 3 replicate wells for each concentration.

[0328] 3) After 72 hours, prepare CCK8 detection solution, dilute 1:10, 100uL per well, remove the original culture medium, add detection solution, and incubate at 37°C and 5% CO 2 The samples were cultured for 2 hours and the OD value was measured at a wavelength of 450 nm using a multi-mode microplate reader (enzyme reader).

[0329] 4) Compare with the control group, calculate the relative cell number, draw a fitting curve using GraphPad Prism, and calculate the IC50 value.

[0330] Table 3. Inhibitory activity of compounds on proliferation of human hepatoma cell line SNU739 or HepG2

[0331]

[0332]

[0333] ND: Not tested, no data.

[0334] It can be seen that the present invention provides a new type of quinoxaline derivatives as PGK1 inhibitors, some of which have strong inhibitory activity on PGK1 and show strong proliferation inhibition activity on human liver cancer SNU739 or HepG2 cells, which have significant advantages over existing PGK1 inhibitors.

[0335] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt, enantiomer, diastereomer, racemate or deuterated derivative thereof; in, X is selected from the group consisting of N or CH; A is selected from the following group: does not exist, R a Selected from the group consisting of H, substituted or unsubstituted C1-C3 alkyl; R1 is Cl and R2 is R b For an R s2 a substituted methyl group; R s2 is hydroxyl group; R3 is selected from the group consisting of H, NH (C1-C3 alkyl); R4 is selected from the group consisting of: unsubstituted or substituted with one or more R s3 substituted 5-6 membered heterocycloalkyl, unsubstituted or substituted with one or more R s3 substituted 5-6 membered heteroaryl; wherein the heterocycloalkyl is a saturated oxygen-containing heterocycloalkyl; and the heteroaryl is a sulfur-containing heteroaryl; R s3 Each is independently selected from the following group: hydroxy, substituted or unsubstituted C1-C3 alkyl, halogen, substituted or unsubstituted C1-C3 alkylacyl, substituted or unsubstituted C1-C3 alkoxy, substituted or unsubstituted C1-C3 haloalkyl, substituted or unsubstituted C1-C3 hydroxyalkyl; The substitution means that one or more hydrogen atoms in the group are optionally replaced by a substituent selected from the group consisting of halogen, C1-C3 alkyl, and C1-C3 haloalkyl.

2. The compound of formula I or a pharmaceutically acceptable salt, enantiomer, diastereomer, racemate or deuterated derivative thereof according to claim 1, characterized in that: X is CH; and / or, A is Among them, R a Selected from the following group: H, C1-C3 alkyl.

3. The compound of formula I or a pharmaceutically acceptable salt, enantiomer, diastereomer, racemate or deuterated derivative thereof according to claim 1, characterized in that: R3 is H.

4. The compound of formula I or a pharmaceutically acceptable salt, enantiomer, diastereomer, racemate or deuterated derivative thereof according to claim 1, characterized in that: The compound of formula I is shown in formula I-1 or formula I-2; 5. A compound or a pharmaceutically acceptable salt, enantiomer, diastereomer, racemate or deuterated derivative thereof, characterized in that: The compound is selected from the following Table 1, Table 1 6. A pharmaceutical composition, characterized in that The invention comprises (i) a therapeutically effective amount of a compound of formula I as claimed in claim 1 or a pharmaceutically acceptable salt, enantiomer, diastereomer, racemate or deuterated derivative thereof, and (ii) an optional pharmaceutically acceptable carrier.

7. A method for preparing the compound of formula I or a pharmaceutically acceptable salt, enantiomer, diastereomer, racemate or deuterated derivative thereof as claimed in claim 1, characterized in that: The preparation method is method one or method two; in, The method 1 comprises the steps of: v) removing the protecting group from compound ie, and then condensing with R4COOH to obtain compound if; vi) optionally removing the protecting group of compound if to obtain a compound of formula I; In each formula, R1' is R1 or R1 protected by a protecting group, and R2' is R2 or R2 protected by a protecting group; A, X, R1, R2, R3 and R4 are as defined in claim 1; The second method comprises the steps of: ii) in an inert solvent, under the catalysis of Pd, compound iia-1 is reacted with R b -C≡CH to carry out coupling reaction, thereby obtaining a compound of formula I; in, In the compound of formula I, A, X, R1, R2, R3, R4, R b and R s1 As defined in claim 1.

8. Use of a compound of formula I as claimed in claim 1 or a pharmaceutically acceptable salt, enantiomer, diastereomer, racemate or deuterated derivative thereof, or a pharmaceutical composition as claimed in claim 6 in the preparation of (i) a PGK1 inhibitor and / or (ii) a medicament for treating or preventing a disease associated with PGK1.

9. The use according to claim 8, characterized in that The PGK1-related disease is selected from: abnormal cell proliferation, abnormal glucose metabolism, hyperkinesia, or a combination thereof.

10. The use according to claim 8, characterized in that The PGK1-related disease is selected from: cancer, hyperkinesia, diabetes, or a combination thereof.

11. The use according to claim 10, characterized in that The cancer is selected from the group consisting of liver cancer, gastric cancer, colorectal cancer, breast cancer, bladder cancer, pancreatic cancer, pancreatic ductal adenocarcinoma, neuroblastoma, prostate cancer, or a combination thereof.

12. An in vitro non-therapeutic method for inhibiting PGK1 activity, comprising the step of contacting PGK1 with the compound of formula I as claimed in claim 1 or a pharmaceutically acceptable salt, enantiomer, diastereomer, racemate or deuterated derivative thereof, thereby inhibiting the activity of PGK1.

13. An in vitro non-therapeutic method for inhibiting cell proliferation activity, comprising the steps of: culturing cells in the presence of a compound of formula I as claimed in claim 1 or a pharmaceutically acceptable salt, enantiomer, diastereomer, racemate or deuterated derivative thereof, thereby inhibiting cell proliferation activity; and, The cells are PGK1-mediated liver cancer cells SNU739 or HepG2.

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