A class of alkynyl-substituted quinoline and quinazoline compounds, their preparation and uses

Abstract

The present invention provides a class of alkynyl-substituted quinoline and quinazoline compounds and their preparation and uses. 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 compounds of the present invention have excellent PGK1 kinase inhibitory activity.

Description

Technical Field

[0001] The present invention pertains to the field of medicine, and particularly relates to a class of alkynyl-substituted quinoline and quinazoline compounds, as well as their preparation and uses. Background Art

[0002] The Warburg effect is one of the important characteristics of tumor cells, which represents the transformation of the way tumor cells utilize 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, with a particularly strong demand for energy. Even under normal oxygen concentration, rapidly proliferating tumor cells still prefer to obtain energy through the anaerobic glycolysis pathway. The change in tumor metabolic pathways is 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-bisphosphoglycerate (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. In recent years, research results have shown that PGK1 is closely related to the occurrence and development of tumors. A study in 2016 showed that the severity of liver cancer patients was positively correlated with the expression level of PGK1 protein. After knocking down the pgk1 gene, the glycolytic ability of liver cancer cell lines decreased, energy production decreased, cell proliferation was inhibited, and tumorigenic ability weakened. The results suggest that PGK1 may become a molecular target in the treatment of liver cancer. At the same time, PGK1 is related to the phenomenon of multi-drug resistance in various malignant tumors. For example, PGK1 is a predictor of low survival rate in breast cancer patients and a new prognostic biomarker for resistance to paclitaxel treatment. Moreover, the expression of PGK1 is also significantly up-regulated in various other malignant tumors such as pancreatic cancer, colorectal cancer, neuroblastoma, and glioblastoma. Thus, targeting PGK1 may be an effective strategy for the treatment of malignant tumors.

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

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

[0006] The object of the present invention is to provide a new class of 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] wherein,

[0010] Z is selected from the group consisting of: C(R d ), N;

[0011] R d is selected from the group consisting of: hydrogen, deuterium, substituted or unsubstituted C1-C6 alkyl (preferably C1-C3 alkyl);

[0012] R 1 is selected from the group consisting of: H, -OR a , -SR a , -N(R a ) 2 , unsubstituted or substituted by one or more R s1 of 4-10 membered heterocycloalkyl;

[0013] R a is selected from the group consisting of: H, substituted or unsubstituted C1-C6 alkyl (preferably C1-C3 alkyl);

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

[0015] R 2 and R 3 each independently is selected from the group consisting of: H, halogen, substituted or unsubstituted C1-C6 alkyl, cyano;

[0016] A is a divalent group selected from the group consisting of: -(C(R c )) 2 ) n - and carbonyl (-C(O)-);

[0017] n is 1, 2 or 3;

[0018] R c each independently is selected from the group consisting of: hydrogen, deuterium, substituted or unsubstituted C1-C6 alkyl (preferably C1-C3 alkyl);

[0019] R 4 is selected from the group consisting of: -OR b and -NHR b ;

[0020] R bSelected from the group consisting of: H, unsubstituted or substituted by one or more R s2 substituted C1-C6 alkyl (preferably C1-C3 alkyl), unsubstituted or substituted by one or more R s2 substituted C1-C6 alkyl acyl (preferably C1-C3 alkyl acyl) (i.e., -C(O)-C1-C6 alkyl, preferably -C(O)-C1-C3 alkyl);

[0021] R s2 each independently selected from the group consisting of: halogen, cyano, hydroxy, C1-C3 alkoxy;

[0022] R 5 selected from the group consisting of: hydrogen, substituted or unsubstituted C1-C6 alkyl (preferably C1-C3 alkyl);

[0023] R 6 selected from the group consisting of: unsubstituted or substituted by one or more R S3 substituted C6-C10 aryl, unsubstituted or substituted by one or more R S3 substituted 5-12 membered heteroaryl, unsubstituted or substituted by one or more R S3 substituted C4-C10 cycloalkyl, unsubstituted or substituted by one or more R S3 substituted 5-12 membered heterocycloalkyl;

[0024] R s3 each independently selected from the group consisting of: halogen, substituted or unsubstituted C1-C3 alkyl, substituted or unsubstituted C1-C3 alkyl acyl, cyano, oxo (=O);

[0025] Unless otherwise specified, the substitution means that one or more hydrogens in the group are optionally substituted by substituents selected from the group consisting of: hydroxy, halogen, amino (-NH 2 ), -N(C1-C3 alkyl) 2 , -NH(C1-C3 alkyl), cyano.

[0026] In another preferred embodiment, Z is N or CH.

[0027] In another preferred embodiment, in R 1 the 4-10 membered heterocycloalkyl contains at least one N heteroatom.

[0028] In another preferred embodiment, in R 1 the 4-10 membered heterocycloalkyl is connected to the rest of the compound through the N heteroatom therein.

[0029] In another preferred embodiment, R 1 is H, -N(R a ) 2, a 4- to 10-membered heterocycloalkyl group which is unsubstituted or substituted by one or more R s1 .

[0030] In another preferred embodiment, R 1 is H, -N(R a ), 2 , a 4- to 10-membered heterocycloalkyl group which is unsubstituted or substituted by one or more R s1 ; and wherein the 4- to 10-membered heterocycloalkyl group contains at least one N heterocycloalkane atom and is linked to the remainder of the compound through the N heterocycloalkane atom therein.

[0031] In another preferred embodiment, R 2 is selected from the group consisting of: H, halogen, C1-C6 alkyl (preferably C1-C3 alkyl), cyano.

[0032] In another preferred embodiment, R 2 is halogen.

[0033] In another preferred embodiment, R 2 is Cl.

[0034] In another preferred embodiment, R 3 is H or C1-C6 alkyl (preferably C1-C3 alkyl).

[0035] In another preferred embodiment, R 3 is H.

[0036] In another preferred embodiment, R 2 is selected from the group consisting of: H, halogen, C1-C6 alkyl (preferably C1-C3 alkyl), cyano; and R 3 is H or C1-C6 alkyl (preferably C1-C3 alkyl).

[0037] In another preferred embodiment, R 2 is selected from the group consisting of: H, halogen, C1-C6 alkyl (preferably C1-C3 alkyl), cyano; and R 3 is H.

[0038] In another preferred embodiment, R 2 is halogen (such as Cl); and R 3 is HC1-C6 alkyl (preferably C1-C3 alkyl, more preferably methyl).

[0039] In another preferred embodiment, R 2 is halogen (such as Cl); and R 3 is H.

[0040] In another preferred embodiment, each R c is independently hydrogen or deuterium; preferably H.

[0041] In another preferred example, A is -CH 2 - or -CH 2 CH 2 -; preferably, A is -CH 2 -.

[0042] In another preferred example, A is -CH 2 -.

[0043] In another preferred example, R 4 is -OR b .

[0044] In another preferred example, R b is selected from the group consisting of: H, C1-C6 alkyl (preferably C1-C3 alkyl) which is unsubstituted or substituted by one or more R s2 ; R s2 are each independently selected from the group consisting of: halogen, cyano, hydroxy, C1-C3 alkoxy.

[0045] In another preferred example, R b is selected from the group consisting of: H, C1-C6 alkyl (preferably C1-C3 alkyl) which is unsubstituted or substituted by one or more R s2 ; R s2 are each independently selected from the group consisting of: hydroxy, C1-C3 alkoxy.

[0046] In another preferred example, R b is H.

[0047] In another preferred example, R 4 is -OR b ; and R b is selected from the group consisting of: H, C1-C6 alkyl (preferably, C1-C3 alkyl) which is unsubstituted or substituted by one or more R s2 ; R s2 are each independently selected from the group consisting of hydroxy, C1-C3 alkoxy; preferably, R 4 is -OH.

[0048] In another preferred example, R 5 is selected from the group consisting of: hydrogen, C1-C3 alkyl.

[0049] In another preferred example, R 5 is selected from the group consisting of: hydrogen, methyl, ethyl.

[0050] In another preferred example, R 5 is methyl.

[0051] In another preferred example, R 6 is selected from the group consisting of: unsubstituted or substituted by one or more R S3Substituted phenyl, unsubstituted or substituted by one or more R S3 Substituted 5-6 membered heteroaryl, unsubstituted or substituted by one or more R S3 Substituted C4-C6 cycloalkyl, unsubstituted or substituted by one or more R S3 Substituted 5-7 membered heterocycloalkyl.

[0052] In another preferred embodiment, R 6 In which, the heteroaryl is a sulfur-containing heteroaryl.

[0053] In another preferred embodiment, R 6 In which, the heterocycloalkyl is an oxygen-containing heterocycloalkyl.

[0054] In another preferred embodiment, R 6 Is selected from the group consisting of: unsubstituted or substituted by one or more R S3 Substituted C4-C10 cycloalkyl, unsubstituted or substituted by one or more R S3 Substituted 5-12 membered heterocycloalkyl.

[0055] In another preferred embodiment, R 6 Is unsubstituted or substituted by one or more R S3 Substituted 5-12 membered heterocycloalkyl.

[0056] In another preferred embodiment, R 6 Is unsubstituted or substituted by one or more R S3 Substituted 5-7 membered heterocycloalkyl.

[0057] In another preferred embodiment, R 6 Is unsubstituted or substituted by one or more R S3 Substituted 5-12 membered heterocycloalkyl, and the heterocycloalkyl contains at least one O heteroatom as a ring atom.

[0058] In another preferred embodiment, R 6 Is unsubstituted or substituted by one or more R S3 Substituted 5-7 membered heterocycloalkyl, and the heterocycloalkyl contains at least one O heteroatom as a ring atom.

[0059] In another preferred embodiment, R 6 Is unsubstituted or substituted by one or more R S3 Substituted group as shown below:

[0060]

[0061] Wherein, W 1 、W 2 、and W 3 Are each independently none, -O-, -CO-, -CH 2- or -CH 2 CH 2 -; W 4 is -CO-, -CH 2 - or -CH 2 CH 2 -; provided that W 1 、W 2 、and W 3 at most one of them is O, W 1 、W 2 、W 3 and W 4 at most one of them is -CH 2 CH 2 - and W 1 、W 2 、and W 3 at most one of them is nothing.

[0062] In another preferred example, and W 1 、W 2 、W 3 and W 4 at most 1 or 2 of them are CO.

[0063] In another preferred example, W 1 、W 2 、and W 3 each independently is nothing, O, -CH 2 - or -CH 2 CH 2 -; W 4 is -CH 2 -.

[0064] In another preferred example, the compound of formula I is as shown in formula II

[0065]

[0066] In another preferred example, the compound of formula I is as shown in formula II-1 or II-2

[0067]

[0068] In another preferred example, in the said compound, A, Z, R 1 、R 2 、R 3 、R 4 、R 5 、R 6 、R a 、R b 、R c 、R d 、R s1 、R s2 、RS3 、n, W 1 、W 2 、W 3 and W 4 Each independently represents the corresponding group in the examples or in the specific compounds (such as compounds S1 - S35) described in Table 1.

[0069] In another preferred embodiment, the compound of formula I is a compound selected from Table 1, or a pharmaceutically acceptable salt, enantiomer, diastereomer, optical isomer, racemate, deuterated derivative, solvate or hydrate, metabolite or prodrug thereof.

[0070] In a second aspect of the present invention, there is provided a pharmaceutical composition 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) optionally a pharmaceutically acceptable carrier, excipient or diluent.

[0071] 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).

[0072] In a third aspect of the present invention, there is provided a method for preparing 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, characterized in that

[0073] The preparation method is Preparation Method 1, Preparation Method 2 or Preparation Method 3.

[0074] In another preferred embodiment, Preparation Method 1 includes the steps of:

[0075]

[0076] (vi) In an inert solvent, under Pd catalysis, coupling compound if with an alkyne reagent R 4 -A-C≡CH to obtain compound ig;

[0077] (vii) Removing the protecting group from compound ig, and then performing a condensation reaction with carboxylic acid R 6 COOH to obtain the compound of formula I;

[0078] In each formula, A, Z, R 1 、R 2 、R 3 、R 4 、R5 and R 6 as defined in the first aspect.

[0079] In another preferred example, the first preparation method further comprises the following steps:

[0080]

[0081] (i) In the presence of a chlorinating agent, subject compound ia to a chlorination reaction to obtain compound ib; wherein;

[0082] (ii) In the presence of urea, react compound ib to form compound ic; alternatively, first react compound ib with ethyl chloroformate, then cyclize under basic conditions and remove the ester group to obtain compound ic;

[0083] (iii) In the presence of a chlorinating agent, subject compound ic to a chlorination reaction to obtain compound id;

[0084] (iv) Under basic conditions, subject compound id to a substitution reaction with a nucleophile to obtain compound ie; and

[0085] (v) Under basic conditions, subject compound ie to a substitution reaction with a nucleophile to obtain compound if.

[0086] In another preferred example, the second preparation method comprises the steps of:

[0087]

[0088] (i) Under basic conditions, react compound ie with a nucleophile to carry out a substitution reaction to obtain compound iia;

[0089] (ii) In an inert solvent, under Pd catalysis, couple compound iia with an alkyne reagent R 4 -A-C≡CH to carry out a coupling reaction to obtain the compound of formula I;

[0090] In each formula, A, Z, R 1 , R 2 , R 3 , R 4 , R 5 and R 6 as defined in the first aspect.

[0091] In another preferred example, the third preparation method comprises the steps of:

[0092]

[0093] (i) Remove the protecting group from compound if, and then react with carboxylic acid R6 The condensation reaction is carried out with COOH to obtain compound iiia;

[0094] (ii) In an inert solvent and under Pd catalysis, compound iiia is coupled with an alkyne reagent R 4 -A-C≡CH to obtain the compound of formula I;

[0095] In each formula, A, Z, R 1 、R 2 、R 3 、R 4 、R 5 and R 6 are as defined in the first aspect.

[0096] In the fourth aspect of the present invention, there is provided the use of a compound of formula I as described in the first aspect, or a pharmaceutically acceptable salt, enantiomer, diastereoisomer, optical isomer, racemate, deuterated derivative, solvate or hydrate, metabolite or prodrug thereof, or the pharmaceutical composition as described in the second aspect in the preparation of (i) a PGK1 inhibitor and / or (ii) a drug for treating or preventing a PGK1-related disease.

[0097] In another preferred example, the PGK1-related diseases include: cancer, abnormal cell proliferation, morphological changes, abnormal glucose metabolism, hyperkinesia, tumor growth, diabetes, or a combination thereof.

[0098] In another preferred example, 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.

[0099] In the fifth aspect of the present invention, there is provided a method for treating or preventing a PGK1-related disease, comprising the step of administering to a subject in need a therapeutically effective amount of a compound of formula I as described in the first aspect, or a pharmaceutically acceptable salt, enantiomer, diastereoisomer, optical isomer, racemate, deuterated derivative, solvate or hydrate, metabolite or prodrug thereof, or the pharmaceutical composition as described in the second aspect.

[0100] In another preferred example, the PGK1-related diseases include: cancer, abnormal cell proliferation, morphological changes, abnormal glucose metabolism, hyperkinesia, tumor growth, diabetes, or a combination thereof.

[0101] In another preferred example, 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.

[0102] In another preferred embodiment, the subject is a mammal, preferably a human.

[0103] In a sixth aspect of the present invention, there is provided a method for inhibiting PGK1 activity, comprising the step of contacting PGK1 with a compound of formula I as described in the first aspect, thereby inhibiting the activity of PGK1.

[0104] In another preferred embodiment, the method is non-therapeutic in vitro.

[0105] In a seventh aspect of the present invention, there is provided a method for inhibiting cell proliferation activity, comprising the step of culturing cells in the presence of a compound of formula I as described in the first aspect, thereby inhibiting the proliferation activity of the cells.

[0106] In another preferred embodiment, the method is non-therapeutic in vitro.

[0107] It should be understood that within the scope of the present invention, the above technical features of the present invention and the technical features specifically described below (such as in the examples) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be elaborated one by one here. Detailed Description of the Invention

[0108] The inventors have conducted extensive and in-depth research and found that introducing an alkynyl group at the 7-position of the parent nucleus of existing quinazoline-based PGK1 inhibitors can significantly enhance the inhibitory activity against PGK1, thereby obtaining a novel class of alkynyl-substituted quinoline / quinazoline compounds as PGK1 inhibitors. Based on this, the inventors have completed the present invention.

[0109] Terms

[0110] Unless otherwise defined, the term "alkyl", alone or as part of another substituent, refers to a straight-chain or branched hydrocarbon group having a specified number of carbon atoms (i.e., C1-C6 represents 1-6 carbons). Examples of alkyl include methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, etc.

[0111] 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 ring tops, preferably a fully saturated ring. "Cycloalkyl" can be monocyclic (such as cyclopropyl, cyclobutyl, cyclohexyl, etc.), or can refer to bicyclic and polycyclic hydrocarbon rings (such as fused rings, spiro rings, bridged rings, etc.). The term "heterocycloalkyl", which may also be referred to as "heterocyclic group" herein, refers to a cycloalkyl 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. Heterocycloalkyl can be a monocyclic, bicyclic, or polycyclic system. Generally, heterocyclic groups typically include 5-12 ring atoms (i.e., 5-12 membered heterocycloalkyl), preferably include 5-7 ring atoms (i.e., 5-7 membered heterocyclic group) and contain 1, 2, 3, or 4 heteroatoms. Non-limiting examples of heterocycloalkyl 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, etc. Heterocycloalkyl can be linked to the rest of the molecule via a ring carbon or a heteroatom (such as ring N).

[0112] The term "alkoxy" is used in its conventional sense and refers to those alkyl groups linked to the rest of the molecule via an oxygen atom. In addition, for dialkylamino, the alkyl moieties can be the same or different, or can combine with the nitrogen atom to which each alkyl is attached to form a 3-7 membered ring. Thus, -N(R a ) 2 shown groups represent including piperidinyl, pyrrolidinyl, morpholinyl, azetidinyl, etc.

[0113] Unless otherwise defined, the term "aryl" refers to a polyunsaturated (usually aromatic) hydrocarbon group, which may be monocyclic or polycyclic (up to three rings) fused together or covalently linked. Generally, aryl refers to a group of 6-10 ring atoms in a fully carbon monocyclic or fused polycyclic (i.e., rings sharing adjacent carbon atom pairs) group, and the group has a conjugated π electron system. The aryl ring may be fused to 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, where the nitrogen and sulfur atoms are optionally oxidized and the nitrogen atoms are optionally quaternized. Generally, heteroaryl has 5-10 ring atoms, i.e., 5-10 membered heteroaryl, and preferably has 5-6 ring atoms, i.e., 5 or 6 membered heteroaryl. The heteroaryl can be attached to the rest of the molecule through a heteroatom. Non-limiting examples of aryl include phenyl and naphthyl. The aryl (ring) may be fused to a 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, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, etc. The heteroaryl may be fused to an aryl, heterocycloalkyl or cycloalkyl ring, where the ring attached to the parent structure is the heteroaryl ring.

[0114] As used herein, the term "heteroatom" is intended to include oxygen (O), nitrogen (N) and sulfur (S).

[0115] For the compounds provided herein, the bond from a substituent (usually an R group) to the center of an aromatic ring (such as benzene, pyridine, etc.) will be understood to refer to a bond providing attachment at any available vertex of the aromatic ring. In some embodiments, this description also includes attachments on rings fused to the aromatic ring. For example, a bond drawn to the center of an indole benzene moiety will represent a bond attached to any available vertex of the six-membered or five-membered ring portion of the indole.

[0116] 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 of the hydrogens are replaced by the same or different halogens as defined above.

[0117] Unless otherwise specified, the structural formulas described in the present invention are intended to include all optical and stereoisomeric forms (such as enantiomers, diastereomers, geometric isomers or conformational isomers): for example, the R and S configurations containing an asymmetric center. Therefore, single stereochemical isomers, enantiomers, diastereomers or geometric isomers or conformational isomer mixtures of the compounds of the present invention are within the scope of the present invention.

[0118] As used herein, the terms "comprising", "including" or "containing" mean that various components can be applied together to the mixtures or compositions of the present invention. Thus, the terms "consisting essentially of" and "consisting of" are included in the term "comprising".

[0119] As used herein, the term "pharmaceutically acceptable" ingredient refers to a substance that is suitable for humans and / or animals without excessive adverse side effects (such as toxicity, irritation and allergic reactions), that is, a substance with a reasonable benefit / risk ratio.

[0120] 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 the regression of a disease, and the regression of the disease is manifested as a decrease in the severity of the disease symptoms, an increase in the frequency and duration of the disease-free period, or the prevention of disorders or disabilities caused by the disease. The "therapeutically effective dose" of the drugs of the present invention also includes a "prophylactically 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.

[0121] Compounds containing alkynyl-substituted quinoline and quinazoline

[0122] The present invention relates to a class of alkynyl-substituted quinoline / quinazoline compounds having phosphoglycerate kinase 1 (PGK1) inhibitory activity, and pharmaceutically acceptable salts or pharmaceutically acceptable solvate compounds thereof, a preparation method thereof, and their use in the preparation of drugs for preventing or treating diseases related to abnormal cell proliferation, morphological changes, abnormal glucose metabolism, hyperkinesia, etc. in vivo and related to PGK, especially for treating or preventing tumor growth and metastasis and diabetes.

[0123] As used herein, the term "compounds of the present invention" or "alkynyl-substituted quinoline / quinazoline compounds" refers to the compounds shown in Formula I. This term also includes various crystalline forms, pharmaceutically acceptable salts, hydrates or solvate compounds of the compounds of Formula I.

[0124] Among them, the term "pharmaceutically acceptable salt" refers to a salt formed by a compound of the present invention and an acid or a base, which is suitable for use as a drug. In the present invention, the pharmaceutically acceptable salts are not particularly limited and preferably include: inorganic acid salts, organic acid salts, alkyl sulfonates and aryl sulfonates; the inorganic acid salts include hydrochloride, hydrobromide, nitrate, sulfate, phosphate, etc.; the organic acid salts include formate, acetate, propionate, benzoate, maleate, fumarate, succinate, tartrate, citrate, etc.; the alkyl sulfonates include methyl sulfonate, ethyl sulfonate, etc.; the aryl sulfonates include benzenesulfonate, p-toluenesulfonate, etc.

[0125] The term "solvate" refers to a complex formed by coordination of a compound of the present invention with solvent molecules in a specific ratio. "Hydrate" refers to a complex formed by coordination of a 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.

[0126] In addition, the compounds of the present invention also include prodrugs of the compounds shown in formula I. The term "prodrug" includes those that may be biologically active or inactive per se, and when administered by an appropriate method, they are metabolized or undergo a chemical reaction in the human body to be converted into a class of compounds of formula I, or a salt or solution composed of a compound of formula I. The prodrugs include (but are not limited to) carboxylic acid esters, carbonates, phosphates, nitrates, sulfates, sulfone esters, sulfoxide esters, amino compounds, carbamates, azo compounds, phosphoramides, glucosides, ethers, acetals, etc. of the compounds.

[0127] The object of the present invention is to provide a class of PGK1 kinase inhibitors with novel structures and excellent activities.

[0128] In the first aspect of the present invention, there is provided a compound of formula I shown below, or a pharmaceutically acceptable salt, enantiomer, diastereomer, optical isomer, racemate, deuterated derivative, solvate or hydrate, metabolite or prodrug thereof.

[0129]

[0130] Wherein, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , Z and A are as defined in the first aspect.

[0131] In a specific embodiment,

[0132] R1 Selected from hydrogen, -OR a , -SR a , -NH 2 , a substituted or unsubstituted 4- to 10-membered heterocycloalkyl. Wherein R a is selected from hydrogen, C1-C3 alkyl. The substitution may be selected from halogen, C1-C3 alkyl, cyano. Wherein the C1-C3 alkyl may be further substituted by hydroxy, halogen, amino, cyano; and / or

[0133] R 2 , R 3 is selected from hydrogen, halogen, cyano; and / or

[0134] R 4 is selected from OR b , NHR b . Wherein R b is selected from hydrogen, substituted or unsubstituted C1-C3 alkyl, substituted or unsubstituted C1-C3 alkyl acyl; the substitution may be selected from halogen, cyano, hydroxy, C1-C3 alkoxy;

[0135] R 5 is selected from hydrogen, C1-C3 alkyl; and / or

[0136] R 6 is selected from substituted or unsubstituted 6- to 10-membered aryl, substituted or unsubstituted 5- to 12-membered heteroaryl, substituted or unsubstituted 5- to 12-membered heterocycloalkane; the substitution may be selected from halogen, C1-C3 alkyl, C1-C3 alkyl acyl, cyano. Wherein the C1-C3 alkyl may be further substituted by hydroxy, halogen, amino, cyano; and / or

[0137] Z is selected from a carbon atom, a nitrogen atom; and / or

[0138] A is selected from -(CH 2 ) n -, -CR c R d -, carbonyl. Wherein n is 1-3; R c , R d are each independently selected from hydrogen, deuterium, C1-C3 alkyl.

[0139] In another specific embodiment, in the compound, any one of A, Z, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 is the corresponding group in the specific compound described in the embodiment.

[0140] Preferably, the quinazoline compounds represented by the general formula (I) of the present invention are selected from the compounds in Table 1 below:

[0141] Table 1

[0142]

[0143]

[0144]

[0145] Preparation method

[0146] The present invention also provides a preparation method of the compound of formula I as described in the first aspect of the present invention. The preparation method of 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 compounds of the present invention can also be conveniently prepared by optionally combining various synthesis methods described in this specification or known in the art, and such combinations can be easily carried out by those skilled in the art to which the present invention pertains.

[0147] The compounds of the present invention can be prepared by, for example, Preparation Method 1, 2 or 3.

[0148] Preparation Method 1:

[0149]

[0150] (i) Compound ia undergoes a chlorination reaction under the action of a chlorinating reagent to obtain compound ib;

[0151] (ii) Compound ib forms an intermediate under urea conditions or first reacts with ethyl chloroformate, and then undergoes ring closure and ester group removal under basic conditions to obtain compound ic.

[0152] (iii) Compound ic undergoes a chlorination reaction under a chlorinating reagent to obtain compound id;

[0153] (iv) Compound id undergoes a substitution reaction with a nucleophile under basic conditions to obtain compound ie;

[0154] (v) Compound ie undergoes a substitution reaction with a nucleophile under basic conditions to obtain compound if;

[0155] (vi) Compound if undergoes a Pd-catalyzed coupling reaction with different alkyne reagents in an inert solvent to obtain compound ig;

[0156] (vii) Compound ig removes the protecting group and then undergoes a condensation reaction with different carboxylic acids to obtain compound ih.

[0157] Preparation Method 2:

[0158]

[0159] (i) The compound ie undergoes a substitution reaction with a nucleophile under basic conditions to obtain the compound iia;

[0160] (ii) The compound if undergoes a Pd-catalyzed coupling reaction with different alkyne reagents in an inert solvent to obtain the compound iib.

[0161] Preparation method three:

[0162]

[0163] (i) The protecting group of the compound if is removed and then it undergoes a condensation reaction with different carboxylic acids to obtain the compound iiia;

[0164] (ii) The compound iiia undergoes a Pd-catalyzed coupling reaction with different alkyne reagents in an inert solvent to obtain the compound iiib.

[0165] In each formula, each group is as defined above.

[0166] Generally, 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 - 60 hours, preferably 0.5 - 48 hours.

[0167] Pharmaceutical composition and administration method

[0168] 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 ingredient can be used for the treatment, prevention, and alleviation of diseases related to phosphoglycerate kinase 1 (PGK1). According to the prior art, the compounds of the present invention can be used for the treatment of the following diseases: cancer, abnormal cell proliferation, morphological changes, abnormal glucose metabolism, hyperkinesia, tumor growth, diabetes, or a combination thereof; wherein, 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.

[0169] The present invention also provides the use of the compound of formula I (alkynyl-substituted quinoline / quinazoline compounds) or its isomers or its pharmaceutically acceptable salts, esters, prodrugs or hydrates as a PGK1 inhibitor in the preparation of a drug for preventing and / or treating diseases related to PGK1, and the diseases related to PGK1 include various cancers (such as liver cancer, gastric cancer, colorectal cancer, breast cancer, bladder cancer, pancreatic cancer, and neuroblastoma, etc.).

[0170] 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 thereof, a prodrug thereof, a hydrate or a solvate thereof, and optionally a pharmaceutically acceptable carrier, excipient or diluent, etc.

[0171] 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).

[0172] The pharmaceutical composition of the present invention contains the compound of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient or carrier within the range of a safe and effective amount. The "safe and effective amount" herein refers to: an amount of the compound sufficient to significantly improve the condition without causing serious side effects. Generally, the pharmaceutical composition contains 1 - 2000 mg of the compound of the present invention per dose, more preferably, 10 - 500 mg of the compound of the present invention per dose. Preferably, the "per dose" is a capsule or a tablet.

[0173] "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" herein means that the components in the composition can be mixed with the compound of the present invention and with each other without significantly reducing the efficacy of the compound. Some examples of pharmaceutically acceptable carriers are cellulose and its derivatives (such as sodium carboxymethylcellulose, sodium ethylcellulose, 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 dodecyl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen - free water, etc.

[0174] There is no particular limitation on the administration mode of the compound or pharmaceutical composition of the present invention. Representative administration modes include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

[0175] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is admixed with at least one conventional inert excipient (or carrier) such as sodium citrate or calcium phosphate dibasic, or with the following components: (a) fillers or bulking agents, e.g., starch, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, e.g., hydroxypropylmethyl cellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia; (c) humectants, e.g., glycerol; (d) disintegrants, e.g., agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, e.g., paraffin; (f) absorption accelerators, e.g., quaternary ammonium compounds; (g) wetting agents, e.g., cetyl alcohol and glycerol monostearate; (h) adsorbents, e.g., kaolin; and (i) lubricants, e.g., talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets, and pills, the dosage form may also contain buffering agents.

[0176] Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other materials well known in the art. They may contain opacifying agents, and release of the active compound or compounds in such compositions can be delayed and released in a portion of the digestive tract. Examples of embedding components that can be used are polymeric materials and waxes. If necessary, the active compound can also be in the form of microcapsules with one or more of the above excipients.

[0177] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. Besides the active compound, the liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers, and emulsifying agents, e.g., ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3 - butanediol, dimethylformamide, and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures of these substances.

[0178] Besides these inert diluents, the compositions may also contain adjuvants such as wetting agents, emulsifying agents, and suspending agents, sweetening agents, flavoring agents, and perfumes.

[0179] Besides the active compound, the suspension may contain suspending agents such as ethoxylated isostearyl alcohols, polyoxyethylene sorbitol, and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, and agar, or mixtures of these substances.

[0180] Compositions for parenteral injection may contain a physiologically acceptable sterile aqueous or non-aqueous solution, dispersion, suspension or emulsion, and a sterile powder for reconstitution into a sterile injectable solution or dispersion. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0181] Dosage forms of the compounds of the present invention for topical administration 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, if necessary.

[0182] The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds.

[0183] 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. For a person weighing 60 kg, the daily dosage is usually 1 - 2000 mg, preferably 20 - 500 mg. Of course, the specific dosage should also consider factors such as the route of administration and the health status of the patient, which are within the scope of the skills of a skilled physician.

[0184] The main advantages of the present invention include

[0185] 1. Terazosin (shown below) inhibits PGK1 at high concentrations (2.5 - 25 μM), and the inhibitory IC 50 value of the preferred compound of the present invention for PGK1 is less than 100 nM;

[0186]

[0187] 2. The compounds of the present invention have significant proliferation inhibitory activity against human liver cancer SNU739.

[0188] The present invention will be further illustrated below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods without specific conditions noted in the following examples are generally carried out under conventional conditions or according to the conditions recommended by the manufacturers. Unless otherwise specified, percentages and parts are by weight percentage and weight parts.

[0189] For the following examples, standard operations and purification methods known to those skilled in the art can be used. Unless otherwise specified, starting materials are generally available from commercial sources. Commercially available solvents and reagents are generally used without further purification. Anhydrous solvents are all 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 products is carried out by silica gel (200 - 300 mesh) column chromatography unless otherwise described. The structures of the compounds are determined by nuclear magnetic resonance spectroscopy (NMR). The nuclear magnetic resonance proton chemical shift (δ) is given in parts per million (ppm). The nuclear magnetic resonance proton spectrum is measured on a Varian MercuryAMX300 instrument, using deuterochloroform (CD 3 ), deuterated methanol (CD 3 OD), and deuterated dimethyl sulfoxide (d6-DMSO) as solvents, and tetramethylsilane (TMS) as the internal standard.

[0190] Abbreviations:

[0191] NCS: N-chlorosuccinimide

[0192] TBTU: O-benzotriazol-N,N,N',N'-tetramethyluronium tetrafluoroborate

[0193] DIPEA: N,N-diisopropylethylamine

[0194] DMF: N,N-dimethylformamide

[0195] Et 3 N: triethylamine

[0196] DCC: dicyclohexylcarbodiimide

[0197] TFA: trifluoroacetic acid

[0198] Preparation Example 1: Preparation of Compound S1

[0199]

[0200] Step 1: Weigh Compound 1a (2.28 g, 1 equivalent) into a single-neck flask, dissolve it in DMF, and then add NCS (1.65 g, 1.5 equivalents) under stirring. React at 70 °C for 2 hours. After the reaction is complete, extract with ethyl acetate and water. Wash the organic phase with saturated brine, dry it over anhydrous sodium sulfate, mix the sample and load it onto the column to obtain Compounds 1b, 1c, and 1d respectively. Analytical data of Compound 1b: 1 H NMR (400 MHz, CDCl 3) δ 7.86 (s, 1H), 7.23 (s, 1H), 5.69 (s, 2H), 3.87 (s, 3H); Analytical data of compound 1c: 1 H NMR (400 MHz, CDCl 3 ) δ 8.16 (d, J = 8.9 Hz, 1H), 7.89 (d, J = 8.9 Hz, 1H); Analytical data of compound 1d: 1 H NMR (400 MHz, CDCl 3 ) δ 8.31 (s, 1H).

[0201]

[0202] Step 2: Weigh compound 1b (3.12 g, 1 equivalent) and urea (6.06 g, 10 equivalents) into a sealed tube and react at 200 °C for 5 hours. After the reaction is complete, cool to room temperature, add water to the reaction solution, filter by suction, dry, and obtain compound 1e.

[0203]

[0204] Step 3: Weigh compound 1e (3.06 g, 1 equivalent) into a single-necked flask, add phosphorus oxychloride (8.80 mL, 10 equivalents) and DIPEA (6.30 mL, 4 equivalents), and react at 110 °C for 4 hours. After the reaction is complete, distill off part of 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 over anhydrous sodium sulfate, mix with silica gel and load onto a column to obtain compound 1f. Analytical data of compound 1f: 1 H NMR (400 MHz, d6-DMSO) δ 8.73 (s, 1H), 8.40 (s, 1H).

[0205]

[0206] Step 4: Weigh compound 1f (539 mg, 1 equivalent) into a single-necked flask, add tetrahydrofuran solvent, then add morpholine (0.26 mL, 2 equivalents), and react at room temperature for 1 hour. After the reaction is complete, pour the reaction solution into water, and a large amount of solid precipitates. Filter by suction to obtain compound 1g. Analytical data of compound 1g: 1 H NMR (400 MHz, CDCl 3 ) δ 8.39 (s, 1H), 7.88 (s, 1H), 3.88 (d, J = 5.0 Hz, 8H).

[0207]

[0208] Step 5: Weigh 1 g (375 mg, 1 equivalent) of Compound 1 and 1 h (270 mg, 1.2 equivalents) into a single-necked flask. Add 1,4-dioxane solvent thereto, then add DIPEA (0.50 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 over anhydrous sodium sulfate, mix with silica gel, and load onto a column to obtain Compound 1i. Analytical data of Compound 1i: 1 H NMR(400MHz,CDCl 3 )δ8.09(s,1H),7.68(s,1H),7.46(dd,J=5.0,1.1Hz,1H),7.36(dd,J=3.7,1.0Hz,1H),7.06(dd,J=5.0,3.7Hz,1H),5.01(d,J=13.4Hz,2H),4.63(br,1H),3.90–3.85(m,4H),3.66–3.60(m,4H),3.02(s,3H),2.93(s,2H),1.86–1.76(m,4H).

[0209]

[0210] Step 5: Weigh 1i (60 mg, 1 equivalent), PdCl 2 (PPh 3 ) 2 (7 mg, 0.1 equivalent) and CuI (4 mg, 0.2 equivalent) into a single-necked flask. Add tetrahydrofuran solvent thereto, then add methyl propynyl ether (14 mg, 2 equivalents), Et 3 N(55 μL, 4 equivalents), displace nitrogen, and react by microwave at 80 °C for 1.5 hours. After the reaction is complete, extract with ethyl acetate and water. Wash the organic phase with saturated brine, dry over anhydrous sodium sulfate, mix with silica gel, and load onto a column to obtain Compound S1. Analytical data of Compound S1: 1 H NMR(400MHz,CDCl 3 )δ7.64(s,1H),7.64(s,1H),7.45(dd,J=5.0,1.0Hz,1H),7.35(dd,J=3.6,0.8Hz,1H),7.05(dd,J=5.0,3.7Hz,1H),5.01(d,J=13.2Hz,2H),4.63(br,1H),4.39(s,2H),3.92–3.82(m,4H),3.65–3.57(m,4H),3.48(s,3H),3.02(s,3H),2.92(t,J=12.0Hz,2H),1.86–1.76(m,4H).

[0211] Preparation Example 2: Preparation of Compound S2

[0212] For the synthesis method of Compound S2, refer to Compound S1. Analytical data of Compound S2: 1 H NMR(400MHz,CDCl 3 )δ7.64(s,1H),7.60(s,1H),7.46(dd,J=5.0,1.1Hz,1H),7.35(dd,J=3.7,0.9Hz,1H),7.06(dd,J=5.0,3.7Hz,1H),5.01(d,J=13.3Hz,2H),4.62(br,1H),3.89–3.83(m,6H),3.68–3.55(m,4H),3.02(s,3H),2.96–2.84(m,2H),2.77(t,J=6.2Hz,2H),2.17(t,J=5.7Hz,1H),1.86–1.75(m,4H).

[0213] Preparation Example 3: Preparation of Compound S3

[0214] For the synthesis method of Compound S3, refer to Compound S1. Analytical data of Compound S3: 1 H NMR(400MHz,CDCl 3 )δ7.64(d,J=2.8Hz,2H),7.45(dd,J=5.0,1.0Hz,1H),7.35(dd,J=3.6,0.9Hz,1H),7.05(dd,J=4.9,3.7Hz,1H),5.01(d,J=13.1Hz,2H),4.60(br,1H),4.50(s,2H),3.93–3.84(m,4H),3.81(s,2H),3.79–3.73(m,2H),3.62(dd,J=10.4,5.9Hz,4H),3.02(s,3H),2.92(t,J=15.3Hz,2H),2.19(s,1H),1.86–1.76(m,4H).

[0215] Preparation Example 4: Preparation of Compound S4

[0216] For the synthesis method of Compound S4, refer to Compound S1. Analytical data of Compound S4: 1 H NMR(400MHz,CDCl 3)δ 7.64 (d, J = 2.5 Hz, 2H), 7.45 (d, J = 4.8 Hz, 1H), 7.34 (d, J = 2.8 Hz, 1H), 7.08–7.02 (m, 1H), 5.00 (d, J = 13.1 Hz, 2H), 4.95 (s, 2H), 4.60 (br, 1H), 3.87 (s, 4H), 3.61 (d, J = 3.8 Hz, 4H), 3.01 (s, 3H), 2.91 (t, J = 11.0 Hz, 2H), 2.13 (s, 3H), 1.86–1.76 (m, 4H).

[0217] Preparation Example 5: Preparation of Compound S5

[0218]

[0219] Weigh Compound S4 (57 mg, 1 equivalent) into a single-necked flask, add a mixed solvent of tetrahydrofuran and water (1:1) thereto, and then add LiOH·H 2 O (8 mg, 2 equivalents), and react at room temperature for 2 hours. After the reaction is complete, extract with ethyl acetate and water. Wash the organic phase with saturated brine, dry over anhydrous sodium sulfate, mix with silica gel and load onto a column to obtain Compound S5. Analytical data of Compound S5: 1 H NMR (400 MHz, CDCl 3 )δ 7.62 (d, J = 6.0 Hz, 2H), 7.45 (d, J = 5.0 Hz, 1H), 7.35 (d, J = 3.4 Hz, 1H), 7.07–7.02 (m, 1H), 4.99 (d, J = 13.3 Hz, 2H), 4.62 (br, 1H), 4.53 (s, 2H), 3.92–3.82 (m, 4H), 3.62 (d, J = 4.2 Hz, 4H), 3.01 (s, 3H), 2.97–2.85 (m, 2H), 1.86–7.02 (m, 4H).

[0220] Preparation Example 6: Preparation of Compound S6

[0221]

[0222] Step 1: Weigh Compound 1g (575 mg, 1 equivalent) and 6a (450 mg, 1.5 equivalents) into a single-necked flask, add 1,4-dioxane solvent thereto, and then add DIPEA (0.70 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 over anhydrous sodium sulfate, mix with silica gel and load onto a column to obtain Compound 6b. Analytical data of Compound 6b: 1 H NMR (400 MHz, CDCl 3)δ8.09(s,1H),7.68(s,1H),4.97(d,J=13.4Hz,2H),4.28(s,1H),3.91–3.83(m,4H),3.66–3.58(m,4H),2.89(t,J=12.1Hz,2H),2.71(s,3H),1.79–1.59(m,4H),1.48(s,9H).

[0223]

[0224] Step 2: Weigh compound 6b (528 mg, 1 equiv), PdCl 2 (PPh 3 ) 2 (63 mg, 0.1 equiv), CuI (34 mg, 0.2 equiv) into a single-neck flask, add tetrahydrofuran solvent thereto, then add propargyl acetate (0.18 mL, 2 equiv), DIPEA (0.60 mL, 4 equiv), under nitrogen protection, heat the reaction at 80 °C for 5 h. After the reaction is complete, extract with ethyl acetate and water, wash the organic phase with saturated brine, dry over anhydrous sodium sulfate, mix with silica gel and load onto a column to obtain compound 6c. Analytical data of compound 6c: 1 HNMR(400MHz,CDCl 3 )δ7.64(d,J=3.3Hz,2H),4.98(d,J=10.8Hz,4H),4.30(br,1H),3.93–3.82(m,4H),3.67–3.56(m,4H),2.89(t,J=12.3Hz,2H),2.70(s,3H),2.14(s,3H),1.73(d,J=11.3Hz,2H),1.62(d,J=7.7Hz,2H),1.48(s,9H).

[0225]

[0226] Step 3: Weigh compound 6c (56 mg, 1 equiv) into a single-neck flask, add dichloromethane solvent thereto, then add TFA (74 μL, 10 equiv), react at room temperature for 1.5 h. After the reaction is complete, rotary evaporate the reaction solution. Then add dichloromethane as the solvent, and successively add 2-tetrahydropyran carboxylic acid (20 mg, 1.5 equiv), TBTU (64 mg, 2 equiv), DIPEA (83 μL, 5 equiv), react at room temperature for 3 h. After the reaction is complete, extract with dichloromethane and water, wash the organic phase with saturated brine, dry over anhydrous sodium sulfate, mix with silica gel and load onto a column to obtain compound 6d. Analytical data of compound 6d: 1 H NMR(400MHz,CDCl 3)δ 7.64 (s, 2H), 5.08–4.92 (m, 4H), 4.72 (t, J = 11.6 Hz, 0.7H), 4.15 (d, J = 9.5 Hz, 0.3H), 4.08–4.03 (m, 2H), 3.88 (s, 4H), 3.61 (d, J = 3.5 Hz, 4H), 3.56–3.46 (m, 1H), 3.12–3.00 (m, 1H), 2.98–2.85 (m, 4H), 2.78 (s, 1H), 2.53–2.48 (m, 1H), 2.02–1.71 (m, 6H), 1.54 (d, J = 10.7 Hz, 2H).

[0227]

[0228] Step 4: Weigh compound 6d (54 mg, 1 equivalent) into a single-necked flask, add tetrahydrofuran and water (2:1) as solvents, and then add lithium hydroxide monohydrate (8 mg, 2 equivalents) thereto. React at room temperature for 4 hours. After the reaction is completed, add water and dichloromethane to the reaction solution for extraction. The organic phase is washed with saturated brine, dried over anhydrous sodium sulfate, and then loaded onto a column after mixing to obtain compound S6. Analytical data of compound S6: 1 H NMR (400 MHz, CD 3 OD) δ 7.78 (d, J = 2.0 Hz, 1H), 7.59 (d, J = 2.0 Hz, 1H), 4.99 (d, J = 12.0 Hz, 2H), 4.67–4.58 (m, 0.6H), 4.50 (s, 2H), 4.36–4.33 (m, 0.4H), 4.24–4.21 (m, 0.6H), 4.18–4.10 (m, 0.4H), 4.05–3.98 (m, 1H), 3.91–3.84 (m, 4H), 3.78–3.71 (m, 0.2H), 3.71–3.64 (m, 4H), 3.63–3.51 (m, 0.8H), 3.05–2.95 (m, 2H), 2.94 (s, 1.7H), 2.78 (s, 1.3H), 2.01–1.90 (m, 1H), 1.85–1.50 (m, 9H), 1.38 (t, J = 6.3 Hz, 1H).

[0229] Preparation Example 7: Preparation of Compound S7

[0230] For the synthesis method of compound S7, refer to compound 6d, and replace 2-tetrahydropyran carboxylic acid with (R)-tetrahydrofuran carboxylic acid. Analytical data of compound S7: 1 H NMR (400 MHz, CDCl 3)δ 7.63 (s, 2H), 5.06–4.90 (m, 4H), 4.78–4.66 (m, 1H), 4.65–4.58 (m, 1H), 4.01–3.84 (m, 6H), 3.60 (d, J = 4.2 Hz, 4H), 2.99–2.85 (m, 4H), 2.78 (s, 1H), 2.37 - 2.34 (m, 0.5H), 2.25–2.06 (m, 4.5H), 2.01–1.76 (m, 3H), 1.67 - 1.58 (m, 3H).

[0231] Preparation Example 8: Preparation of Compound S8

[0232] For the synthesis method of Compound S8, refer to Compound 6d, and replace 2 - tetrahydropyran - carboxylic acid with (S) - tetrahydrofuran - carboxylic acid. Analytical data of Compound S8: 1 H NMR (400 MHz, CDCl 3 )δ 7.64 (d, J = 3.5 Hz, 2H), 5.06–4.95 (m, 4H), 4.73 - 4.61 (m, 2H), 4.03–3.84 (m, 6H), 3.61 (s, 4H), 3.00–2.88 (m, 4H), 2.79 (s, 1H), 2.38 (d, J = 10.7 Hz, 0.5H), 2.23 - 2.19 (m, 1H), 2.14 (s, 3H), 2.10–1.89 (m, 3.5H), 1.73–1.66 (m, 3H).

[0233] Preparation Example 9: Preparation of Compound S9

[0234]

[0235] Weigh Compound S7 (28 mg) into a single - necked flask, add methanol, and then add 4N HCl (1 mL). React at room temperature for 3 hours. After the reaction is complete, evaporate the reaction solution to dryness. Then add ethanol to it, and a large amount of solid precipitates. Filter by suction to obtain Compound S9. Analytical data of Compound S9: 1 H NMR (400 MHz, d6 - DMSO) δ 12.83 (s, 1H), 8.24 (s, 1H), 8.08 (s, 1H), 4.87–4.77 (m, 2.5H), 4.66 - 4.57 (m, 1.5H), 4.43 (s, 2H), 4.02 (s, 4H), 3.83–3.70 (m, 6H), 3.21 (br, 2H), 2.85 (s, 1.7H), 2.66 (s, 1.3H), 2.10–1.96 (m, 2H), 1.90 - 1.77 (m, 5H), 1.62 (d, J = 10.1 Hz, 1H).

[0236] Preparation Example 10: Preparation of Compound S10

[0237]

[0238] For the synthesis method of Compound S10, refer to Compound S9. Analytical data of Compound S10: 1 H NMR(400MHz, d6-DMSO) δ 12.94(s, 1H), 8.29(s, 1H), 8.07(s, 1H), 4.90–4.77(m, 2.5H), 4.68–4.51(m, 1.5H), 4.43(s, 2H), 4.01(s, 4H), 3.81–3.71(m, 6H), 3.20(br, 2H), 2.84(s, 1.7H), 2.66(s, 1.3H), 2.13–1.96(m, 2H), 1.91–1.69(m, 5H), 1.61(d, J=12.2Hz, 1H).

[0239] Preparation Example 11: Preparation of Compound S11

[0240] For the synthesis method of Compound S11, refer to Compound S6, and replace 2-tetrahydropyran carboxylic acid with 1,4-dioxane-2-carboxylic acid. Analytical data of Compound S11: 1 H NMR(400MHz, CD 3 OD) δ 7.75(s, 1H), 7.56(s, 1H), 4.96(d, J=11.8Hz, 2H), 4.60–4.50(m, 1H), 4.48(s, 2H), 4.40(dd, J=9.5, 2.7Hz, 0.5H), 4.18–4.10(m, 0.5H), 3.90–3.82(m, 6H), 3.81–3.68(m, 3H), 3.68–3.58(m, 5H), 3.04–2.90(m, 3.7H), 2.75(s, 1.3H), 1.87–1.65(m, 4H).

[0241] Preparation Example 12: Preparation of Compound S12

[0242] For the synthesis route of Compound S12, refer to Compound S6, and replace the raw material 4-N-Boc-4-N-methylaminopiperidine with 4-Boc-aminopiperidine, and 2-tetrahydropyran carboxylic acid with (R)-tetrahydrofuran carboxylic acid. Analytical data of Compound S12: 1 H NMR(400MHz, CDCl 3)δ 7.64 (s, 2H), 6.61 (d, J = 8.4 Hz, 1H), 4.74 (d, J = 10.8 Hz, 2H), 4.56 (s, 2H), 4.34 (dd, J = 8.3, 5.9 Hz, 1H), 4.10–3.99 (m, 1H), 3.94–3.82 (m, 6H), 3.65–3.56 (m, 4H), 3.15–3.03 (m, 2H), 2.34–2.25 (m, 1H), 2.10–1.80 (m, 6H), 1.49–1.36 (m, 2H).

[0243] Preparation Example 13: Preparation of Compound S13

[0244] For the synthesis method of Compound S13, refer to Compound S6, and replace starting material 1b with 1c. Analytical data of Compound S13: 1 HNMR(400 MHz, CD 3 OD) δ 7.64 (dd, J = 8.6, 1.6 Hz, 1H), 7.13 (dd, J = 8.6, 1.6 Hz, 1H), 5.11 (d, J = 11.9 Hz, 2H), 4.85–4.83 (m, 0.5H), 4.73–4.70 (m, 0.5H), 4.65–4.60 (m, 0.5H), 4.49 (s, 2H), 4.21–4.12 (m, 0.5H), 4.02–3.80 (m, 6H), 3.64 (s, 4H), 3.05–2.93 (m, 2H), 2.91 (s, 1.7H), 2.78 (s, 1.3H), 2.32–2.18 (m, 1H), 2.15–1.91 (m, 3H), 1.87–1.67 (m, 4H).

[0245] Preparation Example 14: Preparation of Compound S14

[0246] For the synthesis method of Compound S14, refer to Compound S6, and replace starting material 1b with 1d. Analytical data of Compound S14: 1 HNMR(600 MHz, CD 3OD) δ 7.71 (d, J = 2.6 Hz, 1H), 5.09 (d, J = 11.1 Hz, 2H), 4.86–4.83 (m, 0.5H), 4.72–4.70 (m, 0.5H), 4.65–4.60 (m, 0.5H), 4.55 (s, 2H), 4.16 (br, 0.5H), 3.98–3.93 (m, 1H), 3.90–3.82 (m, 5H), 3.64 (d, J = 4.0 Hz, 4H), 3.02–2.95 (m, 2H), 2.91 (s, 1.7H), 2.77 (s, 1.3H), 2.25–2.20 (m, 1H), 2.14–2.06 (m, 0.5H), 2.03–1.92 (m, 2.5H), 1.88–1.64 (m, 4H).

[0247] Preparation Example 15: Preparation of Compound S15

[0248]

[0249] Step 1: Weigh compound 1f (450 mg) into a single-necked flask, add tetrahydrofuran solvent (4 mL) thereto, and then add aqueous ammonia solution (2 mL). React at room temperature for 3 hours. After the reaction is complete, add water to the reaction solution, and a large amount of white solid precipitates. Filter by suction to obtain compound 15a. Analytical data of compound 15a: 1 H NMR (400 MHz, CDCl 3 ) δ 8.51 (m, 3H), 8.23 (s, 1H).

[0250] For the subsequent synthesis method, refer to the synthesis steps of compound S6. Analytical data of compound S15: 1 H NMR (400 MHz, d6-DMSO) δ 8.20 (s, 1H), 7.59 (br, 2H), 7.34 (s, 1H), 5.47 (t, J = 6.0 Hz, 1H), 4.91 (d, J = 12.3 Hz, 2H), 4.79–4.74 (m, 0.5H), 4.66–4.61 (m, 0.5H), 4.50–4.43 (m, 0.5H), 4.38 (d, J = 6.0 Hz, 2H), 4.13–4.05 (m, 0.5H), 3.81 - 3.71 (m, 2H), 2.91–2.76 (m, 3.7H), 2.64 (s, 1.3H), 2.07–1.96 (m, 2H), 1.90–1.78 (m, 2H), 1.68 - 1.63 (m, 2H), 1.59 - 1.50 (m, 2H).

[0251] Preparation Example 16: Preparation of Compound S16

[0252] For the synthesis method of compound S16, refer to the synthesis steps of compound S15. Analytical data of compound S16: 1 H NMR(400MHz,d6-DMSO)δ8.20(s,1H),7.59(br,2H),7.34(s,1H),5.47(t,J=6.0Hz,1H),4.91(d,J=11.9Hz,2H),4.79–4.74(m,0.5H),4.65-4.62(m,.0.5H),4.48(dd,J=19.7,4.2Hz,0.5H),4.38(d,J=6.0Hz,2H),4.10(dd,J=12.2,5.8Hz,0.5H),3.81–3.71(m,2H),2.89–2.77(m,3.7H),2.64(s,1.3H),2.09–1.96(m,2H),1.90–1.79(m,2H),1.66(t,J=9.6Hz,2H),1.59-1.53(m,2H).

[0253] Preparation Example 17: Preparation of Compound S17

[0254] For the synthesis method of compound S17, refer to the synthesis steps of compound S15. Analytical data of compound S17: 1 H NMR(400MHz,CD 3 OD)δ7.99(d,J=2.0Hz,1H),7.47(s,1H),4.96(d,J=13.2Hz,2H),4.83(t,J=6.8Hz,0.5H),4.78–4.67(m,1.5H),4.65–4.55(m,0.5H),4.16–4.07(m,0.5H),4.00–3.80(m,2H),2.97–2.84(m,3.7H),2.77(s,1.3H),2.24-2.18(m,1H),2.13-1.92(m,3H),1.85–1.62(m,4H),1.53(d,J=6.6Hz,3H).

[0255] Preparation Example 18 Preparation of Compound S18

[0256]

[0257] Step 1: Weigh compound 1f(359mg, 1 equivalent) into a single-necked flask, add methanol as the solvent (8mL), and then add sodium methoxide (162mg, 3 equivalents). React at room temperature for 24 hours. After the reaction is complete, add water to the reaction solution, and a large amount of solid precipitates. Filter by suction to obtain compound 18a. Analytical data of compound 18a: 11H NMR (400 MHz, d6-DMSO) δ 8.52 (d, J = 1.9 Hz, 1H), 8.23 (d, J = 2.5 Hz, 1H), 4.16 (s, 3H).

[0258]

[0259] Step 2: Weigh compound 18a (332 mg, 1 equivalent), 6a (300 mg, 1.5 equivalents) into a single-necked flask, add 1,4-dioxane solvent thereto, then add DIPEA (0.46 mL, 3 equivalents), and react at 100 °C for 5 hours. After the reaction is complete, extract with ethyl acetate and water, wash the organic phase with saturated brine, dry over anhydrous sodium sulfate, mix with silica gel and load onto a column to obtain compound 18b. Analytical data of compound 18b: 1 1H NMR (400 MHz, CDCl 3 ) δ 8.09 (s, 1H), 7.94 (s, 1H), 5.03 (d, J = 13.5 Hz, 2H), 4.33 (br, 1H), 4.09 (s, 3H), 2.95 (t, J = 12.3 Hz, 2H), 2.73 (s, 3H), 1.81–1.65 (m, 4H), 1.50 (s, 9H).

[0260]

[0261] Step 3: Weigh compound 18b (461 mg, 1 equivalent), PdCl 2 (PPh 3 ) 2 (61 mg, 0.1 equivalent), CuI (33 mg, 0.2 equivalent) into a single-necked flask, add tetrahydrofuran solvent thereto, then add propargyl acetate (170 mg, 2 equivalents), DIPEA (0.57 mL, 4 equivalents), protect with nitrogen, and heat and react at 80 °C for 5 hours. After the reaction is complete, extract with ethyl acetate and water, wash the organic phase with saturated brine, dry over anhydrous sodium sulfate, mix with silica gel and load onto a column to obtain compound 18c. Analytical data of compound 18c: 1 1H NMR (400 MHz, d6-DMSO) δ 7.89 (s, 1H), 7.62 (s, 1H), 5.01 (d, J = 13.5 Hz, 2H), 4.97 (s, 2H), 4.30 (br, 1H), 4.07 (s, 3H), 2.92 (t, J = 12.2 Hz, 2H), 2.71 (s, 3H), 1.75 (d, J = 9.6 Hz, 2H), 1.70–1.61 (m, 2H), 1.48 (s, 9H).

[0262]

[0263] Step 4: Weigh compound 18c (320 mg, 1 equivalent) into a single-necked flask, add dichloromethane solvent thereto, and then add TFA (0.48 mL, 10 equivalents). React at room temperature for 3 hours. After the reaction is complete, rotary evaporate the reaction solution. Then add dichloromethane as the solvent, and successively add (R)-tetrahydrofuroic acid (111 mg, 1.5 equivalents), TBTU (411 mg, 2 equivalents), and DIPEA (0.53 mL, 5 equivalents). 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 over anhydrous sodium sulfate, mix with silica gel, and column chromatograph to obtain compound 18d. Analytical data of compound 18d: 1 H NMR(400MHz,CDCl 3 )δ7.91(d,J=4.5Hz,1H),7.64(d,J=4.8Hz,1H),5.07(d,J=14.8Hz,2H),4.98(s,2H),4.79–4.69(m,1H),4.65(dd,J=7.4,5.7Hz,0.5H),4.18(t,J=11.7Hz,0.5H),4.09(d,J=3.2Hz,3H),4.04–3.86(m,2H),3.04–2.94(m,2H),2.92(s,2H),2.81(s,1H),2.43–2.36(m,0.3H),2.29–2.19(m,0.7H),2.16(s,3H),2.13–2.02(m,2H),1.99–1.86(m,2H),1.81–1.61(m,4H).

[0264]

[0265] Step 5: Weigh compound 18d (356 mg, 1 equivalent) into a single-necked flask, add tetrahydrofuran and water (2:1) as the solvent, and then add lithium hydroxide monohydrate (54 mg, 2 equivalents). 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 over anhydrous sodium sulfate, mix with silica gel, and column chromatograph to obtain compound S18. Analytical data of compound S18:

[0266] 1 H NMR(400MHz,CD 3OD) δ 7.85 (d, J = 1.4 Hz, 1H), 7.53 (s, 1H), 5.01 (d, J = 12.9 Hz, 2H), 4.87–4.82 (m, 0.5H), 4.74–4.69 (m, 0.5H), 4.67–4.59 (m, 0.6H), 4.48 (s, 2H), 4.22–4.13 (m, 0.4H), 4.09 (d, J = 2.3 Hz, 3H), 4.00–3.82 (m, 2H), 3.01 (dd, J = 23.8, 10.8 Hz, 2H), 2.91 (s, 1.7H), 2.78 (s, 1.3H), 2.30–2.17 (m, 1H), 2.16–1.93 (m, 3H), 1.93–1.69 (m, 4H).

[0267] Preparation Example 19: Preparation of Compound S19

[0268]

[0269] Weigh Compound S18 (115 mg, 1 equivalent) into a single-necked flask, add 1,2-dichloroethane (8 mL) as the solvent, then add aluminum chloride (167 mg, 5 equivalents). After addition, raise the temperature to 65 °C and react for 3 h. After the reaction is completed, cool the reaction solution to room temperature, extract with dichloromethane and water. Wash the organic phase with saturated brine, dry over anhydrous sodium sulfate, and then load the sample onto a column to obtain Compound S19. Analytical data of Compound S19:

[0270] 1 H NMR (400 MHz, CD 3 OD) δ 7.95 (s, 1H), 7.46 (s, 1H), 4.85–4.80 (m, 0.5H), 4.72–4.69 (m, 0.5H), 4.66–4.42 (m, 4.6H), 4.19 (br, 0.4H), 4.00–3.80 (m, 2H), 3.08 (br, 2H), 2.93 (s, 1.8H), 2.79 (s, 1.2H), 2.29–1.71 (m, 8H).

[0271] Preparation Example 20: Preparation of Compound S20

[0272]

[0273] Step 1: Weigh compound 1f (200 mg, 1 equivalent) into a single-necked flask, add 1,4-dioxane as the solvent (10 mL), then add sodium methyl mercaptide (43 mg, 1.1 equivalents), and react at room temperature for 2 hours. After the reaction is complete, add water to the reaction solution, and a large amount of white solid precipitates. Filter by suction to obtain compound 20a. Analytical data of compound 20a: 1H NMR (400 MHz, CD 3 ) δ 8.48 (s, 1H), 8.11 (s, 1H), 2.74 (s, 3H).

[0274] For the subsequent synthesis method, refer to the synthesis steps of compound S18. Analytical data of compound S20:

[0275] 1 1H NMR (400 MHz, CD 3 OD) δ 7.77 (s, 1H), 7.53 (s, 1H), 5.06 (d, J = 13.0 Hz, 2H), 4.88–4.82 (m, 0.5H), 4.74–4.69 (m, 0.5H), 4.68–4.59 (m, 0.5H), 4.49 (s, 2H), 4.24–4.12 (m, 0.5H), 4.01–3.83 (m, 2H), 3.03 (dd, J = 23.9, 11.1 Hz, 2H), 2.91 (s, 1.7H), 2.78 (s, 1.3H), 2.62 (d, J = 2.3 Hz, 3H), 2.25–2.19 (m, 1H), 2.15–2.07 (m, 0.5H), 2.03–1.93 (m, 2.5H), 1.90–1.69 (m, 4H).

[0276] Preparation Example 21: Preparation of Compound S21

[0277]

[0278] Step 1: Weigh compound 15a (423 mg, 1 equivalent) into a single-necked flask, add tetrahydrofuran as the solvent (10 mL), then add nitroisopentyl ester (0.33 mL, 2 equivalents), and react at 60 °C for 2 days. After the reaction is complete, add water and extract with ethyl acetate. Wash the organic phase with saturated brine, dry over anhydrous sodium sulfate, and then load the sample onto a column to obtain compound 21a. Analytical data of compound 21a: 1 1H NMR (400 MHz, d6-DMSO) δ 9.55 (s, 1H), 8.66 (s, 1H), 8.49 (s, 1H).

[0279]

[0280] Step 2: Weigh compound 21a (180 mg, 1 equivalent), 6a (178 mg, 1.5 equivalents) into a single-necked flask, add 1,4-dioxane solvent thereto, then add DIPEA (0.27 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 over anhydrous sodium sulfate, mix with silica gel and load onto a column to obtain compound 21b. Analytical data of compound 21b: 1 H NMR(400MHz,CDCl 3 )δ8.89(s,1H),8.18(s,1H),7.69(s,1H),5.05(d,J=13.2Hz,2H),4.32(br,1H),2.97(t,J=13.0Hz,2H),2.71(s,3H),1.78(d,J=10.1Hz,2H),1.65(d,J=10.1Hz,2H),1.48(s,9H).

[0281]

[0282] Step 3: Weigh compound 21b (203 mg, 1 equivalent), PdCl 2 (PPh 3 ) 2 (28 mg, 0.1 equivalent), CuI (15 mg, 0.2 equivalent) into a single-necked flask, add tetrahydrofuran solvent thereto, then add propargyl acetate (90 mg, 2 equivalents), DIPEA (0.16 mL, 4 equivalents), and heat and react at 80 °C for 5 hours. After the reaction is complete, extract with ethyl acetate and water, wash the organic phase with saturated brine, dry over anhydrous sodium sulfate, mix with silica gel and load onto a column to obtain compound 21c. Analytical data of compound 21c: 1 H NMR(400MHz,CDCl 3 )δ8.89(s,1H),7.71(s,1H),7.64(s,1H),5.05(d,J=13.7Hz,2H),4.98(s,2H),4.43–4.22(m,1H),2.96(t,J=12.7Hz,2H),2.71(s,3H),2.15(s,3H),1.77(d,J=11.2Hz,2H),1.65(d,J=11.7Hz,2H),1.48(s,9H).

[0283]

[0284] Step 4: Weigh compound 21c (85 mg, 1 equivalent) into a single-necked flask, add dichloromethane solvent thereto, and then add TFA (0.13 mL, 10 equivalents). React at room temperature for 4 hours. After the reaction is complete, rotary evaporate the solvent and TFA. Then add dichloromethane as the solvent, and successively add (R)-tetrahydrofuroic acid (31 mg, 1.5 equivalents), TBTU (115 mg, 2 equivalents), and DIPEA (0.15 mL, 5 equivalents). 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 over anhydrous sodium sulfate, mix with silica gel, and load onto a column to obtain compound 21d.

[0285]

[0286] Step 5: Weigh compound 21d (111 mg, 1 equivalent) into a single-necked flask, add tetrahydrofuran and water (2:1) as the solvent, and then add lithium hydroxide monohydrate (20 mg, 2 equivalents). 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 over anhydrous sodium sulfate, mix with silica gel, and load onto a column to obtain compound S21. Analytical data of compound S21: 1 H NMR(400MHz,CD 3 OD)δ9.01(d,J=3.4Hz,1H),7.82(d,J=2.2Hz,1H),7.64(s,1H),5.07(d,J=12.8Hz,2H),4.88–4.82(m,0.5H),4.74–4.60(m,1H),4.49(s,2H),4.21(br,1H),4.01–3.81(m,2H),3.03(dd,J=23.7,10.6Hz,2H),2.90(s,1.7H),2.77(s,1.3H),2.26–2.17(m,1H),2.14–2.06(m,0.5H),2.03–1.93(m,2.5H),1.92–1.68(m,4H).

[0287] Preparation Example 22: Preparation of Compound S22

[0288] For the synthetic route of compound 22, refer to the synthetic steps of compound 21. Analytical data of compound S22:

[0289] 1 H NMR(400MHz,CD 3OD) δ 9.01 (d, J = 3.6 Hz, 1H), 7.82 (d, J = 2.3 Hz, 1H), 7.63 (s, 1H), 5.07 (d, J = 13.4 Hz, 2H), 4.88–4.81 (m, 0.5H), 4.74–4.60 (m, 1H), 4.49 (s, 2H), 4.23–4.16 (m, 0.5H), 3.99–3.81 (m, 2H), 3.03 (dd, J = 23.6, 10.6 Hz, 2H), 2.90 (s, 1.7H), 2.77 (s, 1.3H), 2.29–2.17 (m, 1H), 2.14–1.91 (m, 3H), 1.88–1.67 (m, 4H).

[0290] Preparation Example 23: Preparation of Compound S23

[0291]

[0292] Step 1: Weigh Compound 1b (1.246 g, 1 equivalent) into a single-necked flask, add dichloromethane as the solvent, then successively add ethyl chloroformate (0.60 mL, 1.1 equivalents) and pyridine (0.35 mL, 1.1 equivalents), and react at room temperature for 2 hours. After the reaction is complete, extract with dichloromethane and water. Wash the organic phase with saturated brine, dry over anhydrous sodium sulfate, and then load the sample onto a column to obtain Compound 23a. Analytical data of Compound 23a: 1 H NMR (400 MHz, d6-DMSO) δ 11.37 (s, 1H), 9.30 (s, 1H), 8.04 (s, 1H), 4.27 (q, J = 7.2 Hz, 2H), 3.95 (s, 3H), 3.51 (s, 2H), 1.31 (t, J = 7.2 Hz, 3H).

[0293]

[0294] Step 2: Weigh Compound 23a (1.14 g, 1 equivalent) into a single-necked flask, add toluene (25 mL) as the solvent, then add a methanol solution of sodium methoxide (5.4 mol / L, 0.50 mL, 1 equivalent). After adding, raise the temperature to 100 °C and react for 5 hours. After the reaction is complete, cool the reaction solution to room temperature, then pour the reaction solution into ice water. Distill the organic phase under reduced pressure, adjust the pH of the remaining aqueous phase to acidic with 4N hydrochloric acid solution, and filter to obtain Compound 23b.

[0295]

[0296] Step 3: Weigh Compound 23b (885 mg, 1 equivalent) into a single-necked flask, add H 2Using O (20 mL) as the solvent, potassium hydroxide (631 mg, 5 equivalents) was added thereto in batches. After the addition, the temperature was raised to 100 °C and the reaction was carried out for 20 hours. After the reaction was complete, the reaction solution was cooled to room temperature, and the pH was adjusted to about 1 with 1N hydrochloric acid solution. Filtration was carried out to obtain compound 23c. Analytical data of compound 23c: 1 HNMR(400MHz, d6-DMSO) δ 11.69 (s, 1H), 11.29 (s, 1H), 7.81 (d, J = 3.4 Hz, 2H), 5.75 (s, 1H).

[0297]

[0298] Step 4: Weigh compound 23c (684 mg, 1 equivalent) into a single-necked flask, add phosphorus oxychloride (1.98 mL, 10 equivalents) thereto, and then slowly add DIPEA (1.40 mL, 5 equivalents). After the addition, the temperature was raised to 110 °C and the reaction was carried out for 3 hours. After the reaction was complete, the reaction solution was cooled to room temperature and poured into ice water. A large amount of solid precipitated. Filtration was carried out, and the filter cake was dried and then passed through a column to obtain compound 23d. Analytical data of compound 23d: 1 H NMR(400MHz, CDCL 3 ) δ 8.62 (s, 1H), 8.25 (s, 1H), 7.54 (s, 1H).

[0299]

[0300] Step 5: Weigh compound 23d (1.576 g, 1 equivalent) into a single-necked flask, add toluene as the solvent (25 mL), and then successively add morpholine (0.77 mL, 2 equivalents), Et 3 N (1.80 mL, 3 equivalents), and heat to 110 °C and react overnight. After the reaction was complete, the reaction solution was cooled to room temperature, extracted with ethyl acetate and water, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and then loaded onto a column after mixing to obtain a mixture of 23e and 23f.

[0301]

[0302] Step 6: Weigh the mixture of 23e and 23f (1.61 g, 1 equivalent) into a sealed tube, add ethanol as the solvent (20 mL), and then successively add compound 6a (1.68 g, 2 equivalents), DIPEA (1.95 mL, 3 equivalents), and heat to 130 °C and react for 1 day. After the reaction was complete, the reaction solution was cooled to room temperature, extracted with dichloromethane and water, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and then loaded onto a column after mixing to obtain a mixture of 23g and 23h.

[0303]

[0304] Step 7: Weigh the mixture of 23g and 23h (270 mg, 1 equivalent) into a single-necked flask, add tetrahydrofuran as the solvent (10 mL), and then successively add PdCl 2 (PPh 3 ) 2 (32 mg, 0.1 equivalent), CuI (18 mg, 0.2 equivalent), propargyl acetate (91 μL, 2 equivalents), DIPEA (0.30 mL, 4 equivalents), displace nitrogen, and heat the reaction at 80 °C for 5 hours. After the reaction is complete, extract with ethyl acetate and water, wash the organic phase with saturated brine, dry over anhydrous sodium sulfate, mix the sample and load it onto a column to obtain a mixture of 23i and 23j.

[0305]

[0306] Step 8: Weigh the mixture of 23i and 23j (170 mg, 1 equivalent) into a single-necked flask, add dichloromethane as the solvent, and then add TFA (0.23 mL, 10 equivalents), and react at room temperature for 4 hours. After the reaction is complete, neutralize the reaction solution with saturated sodium bicarbonate, extract with dichloromethane, wash the organic phase with saturated brine, dry over anhydrous sodium sulfate, mix the sample and load it onto a column to obtain compounds 23l and 23m.

[0307]

[0308] Step 9: Weigh compound 23m (33 mg, 1 equivalent) into a single-necked flask, add dichloromethane as the solvent, and then successively add (R)-tetrahydrofuroic acid (13 mg, 1.5 equivalents), TBTU (46 mg, 2 equivalents), DIPEA (60 μL, 5 equivalents), 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 over anhydrous sodium sulfate, mix the sample and load it onto a column to obtain compound 23n. Analytical data of compound 23n: 1 H NMR (400 MHz, CDCl 3 ) δ 7.85 (s, 1H), 7.79 (d, J = 4.1 Hz, 1H), 6.45 (s, 1H), 5.00 (s, 2H), 4.77–4.59 (m, 4H), 4.03–3.88 (m, 6H), 3.15 (s, 4H), 3.08–2.98 (m, 2H), 2.93 (s, 1.8H), 2.82 (s, 1.2H), 2.18 (s, 3H), 2.14–2.00 (m, 4H), 1.98–1.88 (m, 2H), 1.83–1.70 (m, 3H), 1.68–1.64 (m, 1H).

[0309]

[0310] Step 10: Weigh compound 23n (36 mg, 1 equivalent) into a single-necked flask, add tetrahydrofuran and water (2:1) as solvents, and then add lithium hydroxide monohydrate (5 mg, 2 equivalents) thereto. React at room temperature for 4 hours. After the reaction is completed, add water and dichloromethane to the reaction solution for extraction. The organic phase is washed with saturated brine, dried over anhydrous sodium sulfate, and then loaded onto a column after mixing to obtain compound S23. Analytical data of compound S23:

[0311] 1 H NMR (400 MHz, CD 3 OD) δ 7.79 (d, J = 6.3 Hz, 1H), 7.70 (d, J = 4.5 Hz, 1H), 6.62 (d, J = 3.0 Hz, 1H), 4.84 (t, J = 6.8 Hz, 0.5H), 4.73–4.62 (m, 3H), 4.48 (s, 2H), 4.15 (br, 0.5H), 4.00–3.82 (m, 6H), 3.12 (s, 4H), 3.05–2.97 (m, 2H), 2.90 (s, 1.7H), 2.77 (s, 1.3H), 2.29–2.17 (m, 1H), 2.14–2.06 (m, 0.5H), 2.04–1.91 (m, 2.5H), 1.90–1.66 (m, 4H).

[0312] Preparation Example 24: Preparation of Compound S24

[0313] For the synthetic method of compound S24, refer to compound S23, and replace (S)-tetrahydrofuroic acid with 2-tetrahydropyranic acid. Analytical data of compound S24: 1 H NMR (400 MHz, CDCl 3 ) δ 7.81 (d, J = 6.0 Hz, 1H), 7.76 (d, J = 6.3 Hz, 1H), 6.42 (d, J = 7.3 Hz, 1H), 4.75–4.53 (m, 5H), 4.35 (dd, J = 9.4, 2.9 Hz, 0.3H), 4.28 (dd, J = 9.4, 2.9 Hz, 0.7H), 3.98–3.93 (m, 4H), 3.90 (dd, J = 9.2, 4.6 Hz, 1H), 3.85 (br, 1H), 3.84–3.66 (m, 4H), 3.13 (d, J = 4.1 Hz, 4H), 3.00 (dd, J = 21.4, 12.1 Hz, 2H), 2.91 (s, 2H), 2.78 (s, 1H), 1.90–1.84 (m, 1H), 1.75–1.67 (m, 3H).

[0314] Preparation Example 25: Preparation of Compound S25

[0315]

[0316] Step 1: Weigh compound 6b (176 mg, 1 equivalent) into a single-necked flask, add dichloromethane solvent thereto, and then add TFA (0.22 mL, 10 equivalents). React at room temperature for 5 hours. After the reaction is complete, rotary evaporate the reaction solution. Then add dichloromethane as the solvent, and successively add (R)-(-)-5-oxo-2-tetrahydrofuroic acid (59 mg, 1.5 equivalents), TBTU (193 mg, 2 equivalents), and DIPEA (0.25 mL, 5 equivalents). 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 over anhydrous sodium sulfate, mix with silica gel, and column chromatograph to obtain compound 25a. Analytical data of compound 25a: 1 H NMR (400 MHz, d6-DMSO) δ 8.09 (d, J = 3.6 Hz, 1H), 7.68 (d, J = 3.6 Hz, 1H), 5.30–5.25 (m, 0.3H), 5.18 (dd, J = 8.0, 4.2 Hz, 0.7H), 5.00 (d, J = 12.5 Hz, 2H), 4.73–4.65 (m, 0.6H), 4.03–3.95 (m, 0.4H), 3.91–3.84 (m, 4H), 3.65–3.62 (m, 4H), 3.00–2.89 (m, 4H), 2.83 (s, 1H), 2.77–2.48 (m, 4H), 2.43–2.32 (m, 1H), 1.95–1.66 (m, 4H).

[0317]

[0318] Step 2: Weigh compound 25a (180 mg, 1 equivalent), PdCl 2 (PPh 3 ) 2 (21 mg, 0.1 equivalent), and CuI (11 mg, 0.2 equivalent) into a single-necked flask, add tetrahydrofuran solvent thereto, and then add 2-propyn-1-ol (34 mg, 2 equivalents) and DIPEA (0.20 mL, 4 equivalents). Protect with nitrogen and heat at 80 °C for 5 h. After the reaction is complete, extract with ethyl acetate and water. Wash the organic phase with saturated brine, dry over anhydrous sodium sulfate, mix with silica gel, and column chromatograph to obtain compound S25. Analytical data of compound S25:

[0319] 1 H NMR (400 MHz, CD 3OD) δ 7.73 (s, 1H), 7.54 (s, 1H), 5.63–5.57 (m, 0.3H), 5.44 (t, J = 6.1 Hz, 0.6H), 4.96 (d, J = 11.5 Hz, 2H), 4.67–4.55 (m, 0.7H), 4.48 (s, 2H), 4.02 (br, 0.4H), 3.84 (d, J = 3.6 Hz, 4H), 3.64 (s, 4H), 3.07–2.91 (m, 3.8H), 2.81 (s, 1.2H), 2.64–2.48 (m, 3H), 2.38–2.23 (m, 1H), 1.91–1.66 (m, 4H).

[0320] Preparation Example 26: Preparation of Compound S26

[0321] For the synthesis method of Compound S26, see Compound S25. Analytical data of Compound S26: 1 H NMR (400 MHz, CD 3 OD) δ 7.86 (d, J = 1.4 Hz, 1H), 7.57 (s, 1H), 5.02 (d, J = 12.9 Hz, 2H), 4.89–4.84 (m, 0.5H), 4.75–4.71 (m, 0.5H), 4.67–4.59 (m, 0.6H), 4.25–4.16 (m, 0.4H), 4.03–3.85 (m, 6H), 3.68 (s, 4H), 3.03 (dd, J = 23.8, 10.8 Hz, 2H), 2.94 (s, 1.7H), 2.80 (s, 1.3H), 2.31–2.19 (m, 1H), 2.17–1.65 (m, 7H), 1.48 (s, 6H).

[0322] Preparation Example 27: Preparation of Compound S27

[0323] For the synthesis method of Compound S27, see Compound 6d. Replace propargyl acetate with methyl propiolate. Analytical data of Compound S27:

[0324] 1 H NMR (400 MHz, CD 3OD) δ 7.78 (d, J = 1.8 Hz, 1H), 7.66 (s, 1H), 4.98 (d, J = 13.1 Hz, 2H), 4.86–4.82 (m, 0.5H), 4.73–4.69 (m, 0.5H), 4.66–4.57 (m, 0.5H), 4.21–4.12 (m, 0.5H), 4.00–3.94 (m, 1H), 3.89–3.80 (m, 8H), 3.67 (s, 4H), 2.98 (dd, J = 23.7, 11.0 Hz, 2H), 2.91 (s, 1.7H), 2.77 (s, 1.3H), 2.26–2.18 (m, 1H), 2.14–1.93 (m, 3H), 1.87–1.66 (m, 4H).

[0325] Preparation Example 28: Preparation of Compound S28

[0326]

[0327] Weigh Compound S27 (34 mg, 1 equivalent) into a single-necked flask, add tetrahydrofuran and water (2:1) as solvents, and then add lithium hydroxide monohydrate (6 mg, 2 equivalents). React at room temperature for 4 hours. After the reaction is completed, adjust the pH of the reaction solution to weakly acidic (pH ~ 5) with 1N dilute hydrochloric acid, then add a small amount of water and dichloromethane for extraction. Wash the organic phase with a small amount of saturated brine, dry over anhydrous sodium sulfate, and then load the sample onto a column to obtain Compound S28. Analytical data of Compound S28: 1 H NMR (400 MHz, CD 3 OD) δ 8.10 (s, 1H), 7.88 (t, J = 3.8 Hz, 1H), 4.87–4.82 (m, 0.5H), 4.78–4.59 (m, 3H), 4.30 (br, 0.5H), 4.16–4.08 (m, 4H), 3.99–3.80 (m, 6H), 3.35–3.26 (m, 2H), 2.95 (s, 1.8H), 2.79 (s, 1.2H), 2.29–2.12 (m, 1.5H), 2.04–1.80 (m, 6.5H).

[0328] Preparation Example 29: Preparation of Compound S29

[0329]

[0330] Weigh Compound S27 (25 mg, 1 equivalent) into a single-necked flask, add 3 mL of ammonia methanol solution (7 mol / L), and react at room temperature for 2 hours. After the reaction is completed, evaporate the reaction solution to dryness, then add methanol, and a solid precipitates. Filter by suction to obtain Compound S29. Analytical data of Compound S29:

[0331] 1 1H NMR (400 MHz, CDCl 3 ) δ 7.72 (d, J = 5.1 Hz, 1H), 7.66 (d, J = 4.2 Hz, 1H), 5.99 (s, 1H), 5.87 (s, 1H), 5.00 (t, J = 14.3 Hz, 2H), 4.75–4.68 (m, 1H), 4.63 (dd, J = 7.4, 5.7 Hz, 0.6H), 4.15 (t, J = 11.4 Hz, 0.4H), 4.02–3.91 (m, 1H), 3.90–3.84 (m, 5H), 3.67–3.58 (m, 4H), 3.01–2.87 (m, 4H), 2.79 (s, 1H), 2.42–2.33 (m, 0.5H), 2.25–1.98 (m, 3H), 1.97–1.72 (m, 3H), 1.67–1.58 (m, 1.5H).

[0332] Preparation Example 30: Preparation of Compound S30

[0333]

[0334] Weigh Compound S28 (53 mg, 1 equivalent) into a single-necked flask, add dichloromethane as the solvent, and then successively add dicyclohexylcarbodiimide (DCC, 31 mg, 1.5 equivalents) and DIPEA (66 μL, 2 equivalents), and react at room temperature for 4 hours. After the reaction is completed, extract with dichloromethane and water. Wash the organic phase with saturated brine, dry over anhydrous sodium sulfate, mix the sample and load it onto a column to obtain Compound S30. Analytical data of Compound S30:

[0335] 1 1H NMR (400 MHz, CDCl 3 ) δ 7.80 (d, J = 4.8 Hz, 1H), 7.64 (d, J = 4.8 Hz, 1H), 5.07 (t, J = 12.6 Hz, 2H), 4.70–4.59 (m, 1H), 4.33–4.21 (m, 3.5H), 4.04–3.88 (m, 6.5H), 3.70–3.63 (m, 6H), 3.05–2.90 (m, 4H), 2.81 (s, 1H), 2.44–2.35 (m, 0.5H), 2.27–2.00 (m, 3H), 2.00–1.75 (m, 3H), 1.69–1.60 (m, 1.5H).

[0336] Preparation Example 31: Preparation of Compound S31

[0337]

[0338] Take 2 mL of ultra-dry tetrahydrofuran in a single-necked flask, add lithium aluminum deuteride (8 mg, 2 equivalents), protect with nitrogen, and then add the ultra-dry tetrahydrofuran solution of S27 under an ice bath, and then raise the temperature to room temperature and react for 2 days. After the reaction is completed, quench with saturated ammonium chloride solution, extract with ethyl acetate and water, wash the organic phase with saturated brine, dry over anhydrous sodium sulfate, mix the sample and load the column to obtain compound S31. Analytical data of compound S31: 1 H NMR(400MHz,CD 3 OD)δ7.87(d,J=1.4Hz,1H),7.56(s,1H),5.04(d,J=12.9Hz,2H),4.86–4.82(m,0.5H),4.76–4.72(m,0.5H),4.65–4.57(m,0.6H),4.27–4.18(m,0.4H),4.01–3.82(m,6H),3.65(s,4H),3.10–3.01(m,2H),2.95(s,1.7H),2.78(s,1.3H),2.32–2.19(m,1H),2.18–1.64(m,7H).

[0339] Preparation Example 32: Preparation of Compound S32

[0340] For the synthetic method of compound S32, refer to compound S6, and replace 2-tetrahydropyranoic acid with (R)-1,4-dioxane-2-carboxylic acid. Analytical data of compound S32: 1 H NMR(400MHz,CD 3 OD)δ7.74(s,1H),7.57(s,1H),4.97(d,J=11.8Hz,2H),4.60–4.51(m,1H),4.48(s,2H),4.40(dd,J=9.5,2.7Hz,0.5H),4.18–4.10(m,0.5H),3.90–3.82(m,6H),3.81–3.68(m,3H),3.68–3.58(m,5H),3.04–2.90(m,3.7H),2.75(s,1.3H),1.87–1.65(m,4H).

[0341] Preparation Example 33: Preparation of Compound S33

[0342] For the synthetic method of compound S33, refer to compound S6, and replace 2-tetrahydropyranoic acid with (S)-1,4-dioxane-2-carboxylic acid. Analytical data of compound S33: 1 H NMR(400MHz,CD 3OD) δ 7.77 (s, 1H), 7.55 (s, 1H), 4.96 (d, J = 11.8 Hz, 2H), 4.61–4.50 (m, 1H), 4.48 (s, 2H), 4.40 (dd, J = 9.5, 2.7 Hz, 0.5H), 4.18–4.10 (m, 0.5H), 3.90–3.82 (m, 6H), 3.81–3.68 (m, 3H), 3.68–3.58 (m, 5H), 3.04–2.90 (m, 3.7H), 2.75 (s, 1.3H), 1.87–1.65 (m, 4H).

[0343] Preparation Example 34: Preparation of Compound S34

[0344] For the synthesis method of Compound S34, refer to Compound S23, and replace (R)-tetrahydrofuroic acid with (R)-1,4-dioxane-2-carboxylic acid. Analytical data of Compound S34: 1 H NMR (400 MHz, CDCl 3 ) δ 7.80 (d, J = 6.0 Hz, 1H), 7.75 (d, J = 6.3 Hz, 1H), 6.44 (d, J = 7.3 Hz, 1H), 4.75–4.53 (m, 5H), 4.34 (dd, J = 9.4, 2.9 Hz, 0.3H), 4.28 (dd, J = 9.4, 2.9 Hz, 0.7H), 3.98–3.93 (m, 4H), 3.90 (dd, J = 9.2, 4.6 Hz, 1H), 3.85 (br, 1H), 3.84–3.66 (m, 4H), 3.13 (d, J = 4.1 Hz, 4H), 3.00 (dd, J = 21.4, 12.1 Hz, 2H), 2.91 (s, 2H), 2.78 (s, 1H), 1.90–1.84 (m, 1H), 1.75–1.66 (m, 3H).

[0345] Preparation Example 35: Preparation of Compound S35

[0346] For the synthesis method of Compound S35, refer to Compound S23, and replace (R)-tetrahydrofuroic acid with (S)-1,4-dioxane-2-carboxylic acid. Analytical data of Compound S35: 1 H NMR (400 MHz, CDCl 3)δ 7.83 (d, J = 6.0 Hz, 1H), 7.77 (d, J = 6.3 Hz, 1H), 6.41 (d, J = 7.3 Hz, 1H), 4.75–4.53 (m, 5H), 4.35 (dd, J = 9.4, 2.9 Hz, 0.3H), 4.27 (dd, J = 9.4, 2.9 Hz, 0.7H), 3.99–3.93 (m, 4H), 3.90 (dd, J = 9.2, 4.6 Hz, 1H), 3.85 (br, 1H), 3.84–3.66 (m, 4H), 3.13 (d, J = 4.1 Hz, 4H), 3.00 (dd, J = 21.4, 12.1 Hz, 2H), 2.91 (s, 2H), 2.78 (s, 1H), 1.90–1.84 (m, 1H), 1.75–1.67 (m, 3H).

[0347] II. Examples of Biological Activity Tests

[0348] Example 1. Test for the Inhibitory Activity of the Compound against the PGK1 Enzyme

[0349] Add 20 μL of 50 mM potassium dihydrogen phosphate, pH 7.0, 4 μL of 50 mM GAP, 6 μL of 10 mM βNAD, 4 μL of 10 mM adenosine diphosphate, 10 μL of 100 mM magnesium sulfate, 20 μL of 1 M glycine, 4 μL of 0.25 μg / μL GAPDH, and 32 μL of deionized water to each reaction, for a total of 100 μL of substrate mixture system. Dilute the PGK1 protein to 0.1 ng / μL and add 100 μL of the PGK1 protein dilution to each reaction for mixing. React at 37 °C for 30 min.

[0350] Dilute the ATP detection solution 1:10 with the ATP detection diluent. In a 96-well white plate, add 100 μL of the diluted detection solution to each well, react at room temperature for 5 min (to remove the influence of background ATP), add 100 μL of the enzyme activity reaction solution, react for at least 2 s, and read the fluorescence value in a chemiluminescence detector.

[0351] Set the compound concentration gradient. During the above reaction process, add compounds at different concentrations to the 200 μL 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 a fitting curve using GraphPad Prism, and calculate the IC 50 value.

[0352] Table 2. Inhibitory Activity of the Compound against the PGK1 Enzyme

[0353]

[0354] Example 2. Test of the inhibitory activity of the compound on the proliferation of SNU739 cells

[0355] Experimental procedure:

[0356] 1) Culture the corresponding target cell line, perform cell counting with a hemocytometer and seed the 96-well plate with 1000 cells / well, and wait for the cells to adhere.

[0357] 2) After the cells adhere, set the compound concentration gradient, prepare 10% serum culture medium containing different concentrations of the compound, add it to the 96-well plate, with at least 3 replicates for each concentration.

[0358] 3) After 72 hours, prepare the CCK8 detection solution, dilute it 1:10, add 100 μL to each well, aspirate the original culture medium, add the detection solution, and culture it for 2 hours at 37 °C and 5% CO 2 and detect the OD value at a wavelength of 450 nm using a multimode microplate reader (ELISA reader).

[0359] 4) Compare with the control group, calculate the relative cell number, draw a fitting curve using GraphPad Prism, and calculate the IC 50 value.

[0360] Table 3. Inhibitory activity of the compound on the proliferation of SNU739 cells

[0361]

[0362]

[0363] It can be seen that the present invention provides a novel class of PGK1 inhibitors of alkynyl-substituted quinoline / quinazoline, and this class of compounds has significant advantages compared with existing PGK1 inhibitors.

[0364] III. Comparative example

[0365] The present invention also provides the activities of the compounds with non-alkynyl substitution at the 7th position of the parent nucleus measured by the methods of Example 1 and Example 2 above, and the test results are shown in Table 4

[0366] Table 4

[0367]

[0368] It can be seen that under the condition that the substituents at other positions are the same, the compound with an alkynyl substituent at the 7th position of the parent nucleus of the present invention has significantly improved PGK1 inhibitory enzyme activity and inhibitory activity against the proliferation of SNU739 cells. The compound with an alkynyl substituent at the 7th position of the present invention has an IC50 of PGK1 inhibitory enzyme activity that is only less than 1 / 6, and even less than 1 / 50, compared with the compound with a methoxy group at the 7th position. At the same time, the IC50 of the inhibitory activity against the proliferation of SNU739 cells is also only less than 1 / 5, and even less than 1 / 24.

[0369] All documents mentioned in the present invention are incorporated herein by reference as if each document was individually incorporated by reference. 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 appended claims of the present application.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt thereof; Wherein, Z is N or CH; R 1 selected from the group consisting of: H, -N(R a ) 2 , an unsubstituted or R s1 -substituted 4- to 10-membered heterocycloalkyl; and wherein said 4- to 10-membered heterocycloalkyl contains at least one N heteroatom and is attached to the remainder of the compound through said N heteroatom; R a selected from the group consisting of: H, C1-C6 alkyl; R s1 each independently represents a C1-C3 alkyl group; R 2 is a halogen; R 3 is H; A is -CH 2 - or -CH 2 CH 2 -; R 4 is - OR b ; R b is H; R 5 is a C1-C3 alkyl group; R 6 selected from the group consisting of: unsubstituted or substituted by one or more R S3 substituted phenyl, unsubstituted or substituted by one or more R S3 substituted 5- or 6-membered heteroaryl, unsubstituted or substituted by one or more R S3 substituted C4-C6 cycloalkyl, unsubstituted or substituted by one or more R S3 substituted 5- to 7-membered heterocycloalkyl; wherein the heteroaryl is a sulfur-containing heteroaryl and the heterocycloalkyl is an oxygen-containing heterocycloalkyl; R s3 Each independently is: C1-C3 alkyl or oxo (=O).

2. The compound of formula I according to claim 1, Characterized in that, R 5 is methyl.

3. The compound of formula I according to claim 1, Characterized in that, R a is H.

4. The compound of formula I according to claim 1, Characterized in that, R 1 Selected from the group: H, -NH 2 .

5. The compound of formula I according to claim 1, Characterized in that, R 2 is Cl.

6. The compound of formula I according to claim 1, Characterized in that, A is -CH 2 .

7. The compound of formula I according to claim 1, Characterized in that, R 6 selected from the group consisting of: unsubstituted or substituted by one or more R S3 substituted 5-6-membered heteroaryl, unsubstituted or substituted by one or more R S3 substituted 5-7-membered heterocycloalkyl; wherein the heteroaryl is a sulfur-containing heteroaryl and the heterocycloalkyl is an oxygen-containing heterocycloalkyl.

8. The compound of formula I according to claim 1, Characterized in that, R 6 selected from the group consisting of: unsubstituted or substituted by one or more R S3 -substituted 5- to 7-membered heterocycloalkyl; wherein the heterocycloalkyl is an oxygen-containing heterocycloalkyl.

9. The compound of formula I according to claim 1, Characterized in that, R 6 selected from the group consisting of:

10. The compound of formula I according to claim 1, Characterized in that, The compound of formula I is as shown in formula II wherein, W 1 , W 2 , and W 3 are each independently none, -O-, -CO-, -CH 2 -, or -CH 2 CH 2 -; W 4 is -CO-, -CH 2 -, or -CH 2 CH 2 -; provided that at most one of W 1 , W 2 , and W 3 is O, and at most one of W 1 , W 2 , W 3 , and W 4 is -CH 2 CH 2 - and at most one of W 1 , W 2 , and W 3 is none.

11. A compound of formula I, Characterized in that, The compound of formula I is a compound selected from Table 1 below, Table 1 or a pharmaceutically acceptable salt thereof.

12. A compound of formula I, Characterized in that, The compound of formula I is compound S32, S33, S34 or S35 or a pharmaceutically acceptable salt thereof.

13. A compound of formula I, Characterized in that, The compound of formula I is compound S9 or S10 14. A pharmaceutical composition, Characterized in that, It comprises (i) a therapeutically effective amount of the compound of formula I according to any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof or the compound of formula I according to claim 13, and (ii) optionally a pharmaceutically acceptable carrier.

15. A method for preparing a compound of formula I according to claim 1 or a pharmaceutically acceptable salt thereof, Characterized in that, The preparation method is Preparation Method 1, Preparation Method 2 or Preparation Method 3; Wherein, Preparation Method 1 comprises the steps: (vi) In an inert solvent, in the presence of Pd catalysis, subject compound if to a coupling reaction with an alkyne reagent R 4 -A-C≡CH to obtain compound ig; (vii) Deprotect compound ig, and then carry out a condensation reaction with carboxylic acid R 6 COOH to obtain the compound of formula I; Preparation Method 2 comprises the steps: (i) Under alkaline conditions, subject compound ie to a substitution reaction with a nucleophile to obtain compound iia; (ii) In an inert solvent, under Pd catalysis, compound iia is coupled with an alkyne reagent R 4 -A-C≡CH to obtain a compound of formula I; Preparation Method 3 comprises the steps: (i) Deprotect the compound if and then carry out a condensation reaction with the carboxylic acid R 6 COOH to obtain the compound iiia; (ii) In an inert solvent, under Pd catalysis, compound iiiA is coupled with an alkyne reagent R 4 -A-C≡CH to obtain a compound of formula I; In each case, A, Z, R 1 , R 2 , R 3 , R 4 , R 5 and R 6 are as defined in claim 1.

16. Use of a compound of formula I according to any one of claims 1 - 12 or a pharmaceutically acceptable salt thereof or the compound of formula I according to claim 13 in the preparation of a PGK1 inhibitor.

17. Use of a compound of formula I according to any one of claims 1 - 12 or a pharmaceutically acceptable salt thereof or the compound of formula I according to claim 13 or the pharmaceutical composition according to claim 14 in the preparation of a drug for treating or preventing a disease related to PGK1.

18. The use according to claim 17, Characterized in that, The diseases related to PGK1 are selected from the following group: abnormal cell proliferation, abnormal glucose metabolism, hyperkinesia, or a combination thereof.

19. The use according to claim 17, Characterized in that, The diseases related to PGK1 are selected from the following group: cancer, diabetes, or a combination thereof.

20. The use according to claim 19, Characterized in that, The cancers are selected from the following group: liver cancer, gastric cancer, colorectal cancer, breast cancer, bladder cancer, pancreatic cancer, neuroblastoma, prostate cancer, or a combination thereof.

21. The use according to claim 19, Characterized in that, The cancer is pancreatic ductal adenocarcinoma.

22. An in vitro non - therapeutic and non - diagnostic method for inhibiting PGK1 activity, Characterized in that, comprising the steps of: contacting PGK1 with a compound of formula I or a pharmaceutically acceptable salt thereof as described in any one of claims 1-12 or a compound of formula I as described in claim 13 to inhibit the activity of PGK1.

23. An in vitro non-therapeutic and non-diagnostic method for inhibiting cell proliferation activity, Characterized in that, comprising the steps of: culturing cells in the presence of a compound of formula I or a pharmaceutically acceptable salt thereof as described in any one of claims 1-12 or a compound of formula I as described in claim 13 to inhibit the proliferation activity of the cells.

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