CN116535359A - Indazolyl-containing hydroxamic acid derivative and application thereof - Google Patents

Abstract

The invention discloses a compound represented by general formula (I), its pharmaceutically acceptable salt, hydrate or prodrug, and its preparation for the prevention and/or treatment of tyrosine kinase and/or histone deacetylation The application of the compound in the drug of deacetylase-mediated diseases, the compound can target a variety of targets (especially histone deacetylase 1, histone deacetylase 6, two tyrosine kinases (vascular endothelial cell growth Factor receptor and platelet-derived growth factor receptor)) at the same time play a good inhibitory effect, is expected to be used in the treatment of malignant tumors.

Description

A hydroxamic acid derivative containing indazole and its application

Technical Field

The present invention relates to the field of biomedicine, and in particular to an indazole-containing hydroxamic acid derivative and application thereof.

Background Art

Tyrosine kinases are a class of proteins with tyrosine kinase activity. They can catalyze the transfer of phosphate groups on ATP to tyrosine residues of many important proteins, causing phosphorylation and thus activating downstream signal transduction pathways. Protein tyrosine kinases occupy a very important position in the intracellular signal transduction pathways, regulating a series of physiological and biochemical processes such as cell growth, differentiation, and death. Dysregulation of protein tyrosine kinase function can cause a series of diseases in the body, including tumors. The occurrence, development, and metastasis of many tumors and the formation of tumor angiogenesis are closely related to the abnormal expression of tyrosine kinases. In particular, some tyrosine kinase receptors are abnormally expressed in solid tumor cells, among which vascular endothelial cell growth factor receptor (VEGFR) is highly expressed in many tumor cells and tumor vascular endothelial cells, and platelet-derived growth factor receptor (PDGFR) is abnormally expressed in fibroblasts in the tumor stroma. The autocrine loop formed by its ligand and receptor is directly involved in the occurrence and development of tumor cells. For example, vascular endothelial growth factor receptor (VEGFR) exists in melanoma; platelet-derived growth factor receptor (PDGFR) exists in glioma; stem cell growth factor receptor (KIT) exists in small cell lung cancer, etc. In addition, similar loops also exist in meningiomas, neuroendocrine tumors, ovarian cancer, prostate cancer and pancreatic cancer. This loop is closely related to the occurrence and development of tumors.

In addition, the occurrence, development and metastasis of solid tumors all depend on tumor angiogenesis, which provides the necessary nutrients and oxygen for tumor growth. Tumor angiogenesis is an important process of tumor cell infiltration, migration and proliferation. Among them, the vascular endothelial growth factor receptor family (VEGFR) and the platelet-derived growth factor receptor family (PDGFR) are directly related to the occurrence and development of tumors and the generation of tumor angiogenesis. As the strongest known vascular permeability agent and endothelial cell-specific mitogenic source, vascular endothelial growth factor (VEGF) plays an important role in the proliferation, migration and vascular construction of endothelial cells. Its expression level and the degree of vascularization of tumor tissue show a significant positive correlation. VEGF mainly exerts its biological effects by acting on the high-affinity receptors VEGFR-1 and KDR on endothelial cells to phosphorylate their tyrosine kinases, and the two have different signal transduction pathways. Among them, KDR plays a key role in tumor growth, metastasis and tumor angiogenesis. Platelet-derived growth factor (PDGF) and its receptor (PDGFR) are involved in the pathogenesis of various tumors and play an important role in angiogenesis. Platelet-derived growth factor (PDGF) expresses its cellular biological effects through its receptor (PDGFR). PDGFR maintains the integrity of the vascular wall by regulating the proliferation and migration of pericytes and vascular smooth muscle cells in the vascular wall and promotes the formation of tumor angiogenesis. In addition, it promotes tumor growth by changing the microenvironment within the tumor.

Since abnormal expression of tyrosine kinase is closely related to the occurrence, development and metastasis of tumors and the generation of tumor angiogenesis, the development of drugs targeting tyrosine kinase has become a hot spot in the international research of anti-tumor drugs. In particular, using new blood vessels as a target to inhibit tumor angiogenesis, block the nutrient supply and migration pathways of tumors, and prevent tumor growth and metastasis has become a new strategy for treating tumors. Abnormal expression of KDR or PDGFR receptors plays a key role in the process of tumor angiogenesis, and they have become the most ideal targets for anti-tumor drug treatment. In addition, sorafenib and sunitinib (SU11248, Sunitinib), two anti-tumor drugs approved by the US FDA that mainly inhibit KDR and PDGFR receptor tyrosine kinases, have been fully demonstrated in clinical practice to have high efficacy and few side effects in anti-tumor treatment.

Histone deacetylases (HDACs) are a class of intracellular metalloproteinases that play an important role in the structural modification of chromosomes and the regulation of gene expression. In cancer cells, overexpression of HDACs leads to increased binding of histones to DNA, causing abnormal conformational changes in chromosomes. At the same time, the expression of cell cycle inhibitors is inhibited, the stability and DNA binding ability of the tumor suppressor p53 decrease, and the expression of hypoxia inducible factor-1 (HIF-1) and vascular endothelial growth factor (VEGF) increases. In mammalian cells, the balance between acetylation and deacetylation plays a key role in gene transcription and the functions of different cellular proteins. The acetylation state of histones is regulated by histone acetyltransferases (HATs) and histone deacetylases (HDACs). The dynamic balance between HATs and HDACs controls the structure of chromatin and gene expression, and their dysfunction is one of the important molecular mechanisms for the occurrence and development of tumors. HDACs belong to the deacetylase superfamily. HDACs play an important role in chromatin remodeling, gene repression, cell cycle regulation and differentiation. Abnormal function of histone deacetylase in tumor cells can lead to the inhibition of transcription of many genes and inhibit the expression of tumor suppressor genes. It has been reported that HDAC1mRNA and protein levels are highly expressed in gastric cancer, prostate cancer, colon cancer and liver cancer tissues, and are related to the TNM stage and lymph node metastasis of the tumor. Studies have found that inhibiting the activity of HDACs can effectively inhibit cancer cell proliferation, induce cell cycle arrest and promote cell apoptosis. Therefore, HDACs have become a new target for the design of anticancer drugs, and the development of HDACs inhibitors (HDACi) is regarded as an effective strategy for tumor treatment. Hydroxamic acid small molecule HDACi is a type of HDACi that has received much attention in recent years, showing good antitumor activity both in vivo and in vitro. HDAC has developed relatively maturely as an anticancer target drug. In 2006, the US pharmaceutical company Merck launched Vorinostat for the treatment of cutaneous T-cell lymphoma. This is the first HDAC inhibitor to be launched on the market. Since then, several HDAC inhibitors have been approved for marketing, including romidepsin for the treatment of T-cell lymphoma (launched in 2009, the original manufacturer is Celgene Corporation of the United States), belinostat for the treatment of relapsed or resistant peripheral T-cell lymphoma (launched in 2014, the original manufacturer is Spectrum Biopharmaceuticals), and panobinostat for the treatment of multiple myeloma (launched in 2015, the original manufacturer is Novartis of Switzerland). In 2015, Shenzhen Microchip successfully launched cedabendine, the first HDAC inhibitor for the treatment of lymphoma on the Chinese market, relying on the local Chinese market. More and more HDACi have entered the clinic for the treatment and adjuvant treatment of solid tumors such as colon cancer and lung cancer, as well as malignant tumors of the blood system such as leukemia and lymphoma.

Summary of the invention

One of the purposes of the present invention is to provide a novel indazole-containing hydroxamic acid derivative that can simultaneously exert a good inhibitory effect on multiple targets (especially histone deacetylase 1, histone deacetylase 6, and two tyrosine kinases (vascular endothelial growth factor receptor and platelet-derived growth factor receptor)).

Another object of the present invention is to provide a pharmaceutical composition comprising the novel indazole-containing hydroxamic acid derivative.

Another object of the present invention is to provide the use of the novel indazole-containing hydroxamic acid derivatives or the pharmaceutical composition comprising the novel indazole-containing hydroxamic acid derivatives in the preparation of drugs for treating diseases mediated by tyrosine kinase and/or histone deacetylase.

In order to achieve the above object, a technical solution adopted by the present invention is:

A compound represented by the general formula (I), a pharmaceutically acceptable salt, hydrate or prodrug thereof,

wherein ^Ra and ^Rb are independently selected from hydrogen, _C1-20 alkyl, 3-10 membered heterocycloalkyl, 5-12 membered aromatic group, and 5-12 membered heteroaryl group; wherein the _C1-20 alkyl, 3-10 membered heterocycloalkyl, 5-12 membered aromatic group, and 5-12 membered heteroaryl group are each independently unsubstituted or substituted by one, two, three or more substituents selected from the following substituents: fluorine, chlorine, bromine, iodine, nitro, cyano, mercapto, hydroxyl, amino, amino substituted with _C1-6 alkyl, _C1-10 alkyl, halogenated _C1-6 alkyl, nitro-substituted _C1-6 alkyl, cyano-substituted _C1-6 alkyl, mercapto-substituted _C1-6 alkyl, amino substituted with _C1-6 alkyl, hydroxyl substituted with _C1-6 alkyl, _C1-6 alkoxy, halogenated _C1-6 alkoxy, nitro-substituted _C1-6 alkoxy, cyano-substituted C1-6 alkyl C _1-6 alkoxy, C _1-6 alkoxy substituted with mercapto, C _1-6 alkoxy substituted with amino, C _1-6 alkoxy substituted with hydroxy, C _2-6 alkenyl, C _2-6 alkynyl, -C(=O)R ₁ , -C(=O)NHR ₂ , -C(=O)OR ₃ , -NHC(=O)R ₄ , -OC(=O)R ₅ ;

R ₁ , R ₂ , R ₃ , R ₄ , and R ₅ are independently selected from unsubstituted C _1-20 alkyl groups, or independently selected from C _1-20 alkyl groups substituted by one or more substituents selected from fluorine, chlorine, bromine, iodine, nitro, cyano, mercapto, hydroxyl, and amino;

Each R ^c is the same or different and is independently selected from hydrogen, C _1-20 alkyl, C _1-20 alkoxy, fluorine, chlorine, bromine, iodine, nitro, cyano, mercapto, hydroxyl, amino, amino substituted with C _1-6 alkyl, halogenated C _1-6 alkyl, nitro substituted with C _1-6 alkyl, cyano substituted with C _1-6 alkyl, mercapto substituted with C _1-6 alkyl, amino substituted with C _1-6 alkyl;

R ^d and ^Re are independently selected from hydrogen, C _1-20 alkyl, C _1-6 alkyl substituted by nitro, C _1-6 alkyl substituted by cyano, C _1-6 alkyl substituted by mercapto, and C _1-6 alkyl substituted by amino;

^Rg has a chain structure, the number of atoms constituting the main chain of the chain structure is 3-15 and the number of heteroatoms is no more than 2, and the heteroatoms are selected from oxygen, sulfur, and nitrogen;

x is 1, 2, 3, or 4.

In some embodiments, the C _1-20 alkyl group may be a C _1-10 alkyl group or a C _10-20 alkyl group; including but not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, hexyl, heptyl, and the like.

According to some preferred aspects of the present invention, the number of heteroatoms in the atoms constituting the main chain of the chain structure is 1 or 2, and one of the heteroatoms is adjacent to the indazole ring. The benzene rings are connected.

According to some preferred aspects of the present invention, the chain structure is a saturated straight chain or a saturated branched chain.

According to some preferred and specific aspects of the present invention, the compound has a structure shown in general formula (II):

wherein ^Ra , ^Rb , ^Rc , ^Rd , ^Re and x are respectively the same as ^Ra , ^Rb , ^Rc , ^{Rd, Re and} x in the general formula (I); A and B are independently _CH2 , O, S or -N( _R6 )-; _R6 is hydrogen or _C1-6 alkyl; m and n are independently selected from integers of 0-10, and 3≤m+n≤10.

In some embodiments, m is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10. In some embodiments, n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10.

According to some preferred aspects of the present invention, when any of the following situations exists, B is CH ₂ :

(1) A is CH ₂ ; (2) A is selected from O, S or —N(R ₆ )—, and m is 0; (3) n is 0.

According to some preferred aspects of the present invention, A is selected from CH ₂ , O, S or —NH—, and B is selected from CH ₂ , O, —N(CH ₃ )—, —N(CH ₂ CH ₃ )— or —N(CH ₂ CH ₂ CH ₃ )—.

Furthermore, A is O, B is CH ₂ , and m+n is 3, 4 or 5.

According to some preferred aspects of the present invention, ^Ra and ^Rb are not hydrogen at the same time.

Furthermore, among ^Ra and ^Rb , one is selected from unsubstituted or substituted pyrimidinyl, pyrazolyl, pyridyl, pyrazinyl, imidazolyl, pyridazinyl, thiazolyl, phenyl; and the other is selected from hydrogen, or unsubstituted or substituted _C1-6 alkyl.

In some embodiments, the compound has a structure shown in formula (III):

wherein: R ^c , R ^d , ^Re , x, A, B, m, n are the same as R ^c , R ^d , ^Re , x, A, B, m, n in the general formula (II) respectively; R ^b is selected from hydrogen, or C _1-6 alkyl which is unsubstituted or substituted by 1 to 3 halogens; R ₇ is selected from fluorine, chlorine, bromine, iodine, hydroxyl, mercapto, cyano, amino, methylamino, ethylamino, nitro, C _1-6 alkyl, halogenated C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkoxy, hydroxymethyl, mercaptomethyl, C(═O)CH ₃ , C(═O)CH ₂ CH ₃ , C(═O)NHCH ₃ , C(═O)NHCH ₂ CH ₃ , NHC(═O)CH ₃ , NHC(═O)CH ₂ CH ₃ , NHCH ₃ , N(CH ₃ ) ₂ . NHCH ₂ CH ₃ , t is 0, 1, 2, 3, 4 or 5, and D and E are independently selected from CH or N.

In some embodiments, when t is 4, at least one of D and E is CH, and when t is 5, both D and E are CH.

In some embodiments, the compound has a structure shown in formula (IV):

wherein: R ^b , R ^c , R ^d , ^Re , x, A, B, m, n are respectively the same as R ^b , R ^c , R ^d , ^Re , x, A, B, m, n in the general formula (III); R ₈ and R ₉ are independently selected from hydrogen, fluorine, chlorine, bromine, iodine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, halogenated methyl, halogenated ethyl, halogenated n-propyl, halogenated isopropyl, halogenated n-butyl, halogenated isobutyl, methoxy, ethoxy, halogenated methoxy, halogenated ethoxy.

In some embodiments, R ^b , R ^d , and ^Re are independently selected from hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, or isobutyl.

In some embodiments, each R ^c is the same or different and is independently selected from hydrogen, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, methoxy, or ethoxy.

In some embodiments, R ^b , R ^c , R ^d , and ^Re are all hydrogen.

In some embodiments, ^Rg is selected from:

In some embodiments, the compound has a structure represented by formula (V):

wherein: p is 2, 3, 4, 5, 6, 7, 8, 9 or 10, and R ₈ and R ₉ are independently selected from hydrogen, fluorine, chlorine, bromine, iodine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, halogenated methyl, halogenated ethyl, halogenated n-propyl, halogenated isopropyl, halogenated n-butyl, halogenated isobutyl, methoxy, ethoxy, halogenated methoxy, halogenated ethoxy.

In some embodiments of the present invention, the structural formula of the compound is as shown in any one of formulas (ST-01) to (ST-16):

Another technical solution provided by the present invention is: a pharmaceutical composition, which comprises the above-mentioned compound, its pharmaceutically acceptable salt, hydrate or prodrug, and a pharmaceutically acceptable carrier.

Furthermore, the pharmaceutical composition may also contain a diluent or an excipient.

Furthermore, the pharmaceutical composition is a composition for preventing and/or treating malignant tumors, including but not limited to renal cancer, liver cancer, colon cancer, gastrointestinal stromal tumors, lung cancer, breast cancer, pancreatic cancer, glioma, lymphoma, fibrosarcoma, ovarian cancer, prostate cancer, leukemia, and lymphoma.

Another technical solution provided by the present invention is: use of the above-mentioned compound, its pharmaceutically acceptable salt, hydrate or prodrug, or the above-mentioned pharmaceutical composition in the preparation of drugs for preventing and/or treating diseases mediated by tyrosine kinase and/or histone deacetylase.

Furthermore, the disease mediated by tyrosine kinase and/or histone deacetylase is a malignant tumor, and the malignant tumor includes but is not limited to renal cancer, liver cancer, colon cancer, gastrointestinal stromal tumor, lung cancer, breast cancer, pancreatic cancer, glioma, lymphoma, fibrosarcoma, ovarian cancer, prostate cancer, leukemia, and lymphoma.

According to the present invention, the compound includes not only a single compound form, but also a mixture of multiple compounds whose structures meet the requirements of general formula (I), as well as different isomeric forms of the same compound such as racemates, enantiomers, diastereomers, etc.

The pharmaceutically acceptable salts include, but are not limited to, hydrochloride, hydrobromide, phosphate, sulfate, acetate, trifluoroacetate, maleate, methanesulfonate, benzenesulfonate, benzoate, toluenesulfonate, succinate, fumarate, tartrate, gallate, citrate and the like.

The compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof is usually administered via the oral route in the form of a pharmaceutical preparation comprising an active ingredient or a pharmaceutically acceptable salt or solvate thereof, or a solvate of such a salt, in a pharmaceutically acceptable dosage form. Depending on the disorder to be treated and the patient, the composition can be administered at different doses.

The pharmaceutical formulations of the compounds represented by the general formula (I) described above can be prepared for oral administration, specifically in the form of tablets or capsules, and particularly involve techniques aimed at providing drug release targeted to the colon.

The pharmaceutical formulations of the compounds represented by the general formula (I) described above may conveniently be administered in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy.

Pharmaceutical formulations suitable for oral administration may include one or more physiologically compatible carriers and/or excipients, and may be in the form of solid or liquid. Tablets and capsules may be prepared with adhesives, fillers, lubricants and/or surfactants (such as sodium lauryl sulfate). Liquid compositions may contain conventional additives, such as suspending agents, emulsifiers and/or preservatives. Liquid compositions may be encapsulated in, for example, gelatin to provide a unit dosage form.

Oral formulations are preferred, particularly tablets or capsules, which can be formulated by methods known to those skilled in the art to provide a dose of the active compound in the range of 0.001 mg to 10000 mg, for example, a dose of the active compound in the range of 0.01 mg to 5000 mg, or a dose of the active compound in the range of 0.1 mg to 1000 mg, etc.

In the present invention, the prodrug of the compound represented by general formula (I) refers to a prodrug that can be converted into at least one compound of general formula (I) or its salt by metabolism or chemical reaction (such as enzymatic hydrolysis or hydrolysis) in the subject after being administered by an appropriate method.

Another technical solution provided by the present invention is an intermediate for preparing the above-mentioned compound, its pharmaceutically acceptable salt, hydrate or prodrug, wherein the intermediate has a structure shown in the general formula (VI):

wherein ^Ra , ^Rb , ^Rc , ^Rd , ^Re , ^Rg and x are respectively the same as ^Ra , ^Rb , ^Rc , ^Rd , ^Re , ^Rg and x in the aforementioned general formula; and R is a _C1-6 alkoxy group.

In some embodiments, R may be methoxy, ethoxy, propoxy, or tert-butoxy.

Another technical solution provided by the present invention is a method for preparing the above-mentioned compound, its pharmaceutically acceptable salt, hydrate or prodrug, which comprises the step of reacting the above-mentioned intermediate with hydroxylamine.

In some embodiments, the hydroxylamine is obtained by adding a hydroxylamine inorganic acid salt, and the hydroxylamine inorganic acid salt includes but is not limited to hydroxylamine hydrochloride and/or hydroxylamine phosphate.

In some embodiments, the reaction of the intermediate and hydroxylamine is carried out under alkaline conditions in an alcohol solvent. In some embodiments, the alkaline conditions are formed by adding an alkaline substance, and the alkaline substance includes but is not limited to sodium hydroxide, potassium hydroxide, and the like.

In some embodiments, the alcohol solvent includes but is not limited to methanol and the like.

Due to the application of the above technical solution, the present invention has the following advantages compared with the prior art:

The compound of the present invention can integrate multiple important targets into one, especially can simultaneously have a strong inhibitory effect on histone deacetylase VI (HDAC6, histone deacetylases 6), histone deacetylase 1 (HDAC1) and two tyrosine kinases (vascular endothelial cell growth factor receptor, Vascular endothelial cell growth factor2, VEGFR2) and platelet-derived growth factor receptor (platelet-derived growth factor-β, PDGFR-β), can better avoid the problems caused by different properties and metabolism when multiple drugs are used in combination, can give full play to the synergistic effect to increase biological activity, and has good application prospects.

DETAILED DESCRIPTION

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as understood by one of ordinary skill in the art to which the invention belongs. Although methods and materials similar or equivalent to those described herein can be used for the implementation or testing of the present invention, suitable methods and materials are described below.

The term "C _1-20 alkyl" refers to a straight or branched chain saturated hydrocarbon group of 1 to 20 carbon atoms ("C 1-C 20 -alkyl"). In some embodiments, the alkyl group contains 1 to 10 carbon atoms, or contains 1 to 6 carbon atoms, such as 1, 2, 3, 4, 5 or 6 carbon atoms. Some non-limiting examples include methyl, ethyl, propyl, 2-propyl (isopropyl), n-butyl, isobutyl, sec-butyl, tert-butyl and 2,2-dimethylpropyl.

The term "alkoxy" refers to an alkyl group attached to a parent molecular moiety via an oxygen atom. In some embodiments, the alkoxy group contains 1 to 6 carbon atoms and can be expressed as a C _1-6 alkoxy group. Some non-limiting examples include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, and tert-butoxy.

The term "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br) or iodine (I). Preferably, the term "halogen" refers to fluorine (F), chlorine (Cl) or bromine (Br).

The term "amino-substituted alkyl" refers to an alkyl group in which at least one hydrogen atom of the alkyl group is replaced by an amino group. For example, amino-substituted C _1-6 alkyl refers to an alkyl group in which at least one hydrogen atom of the C _1-6 alkyl group is replaced by an amino group. Preferably, "aminoalkyl" refers to an alkyl group in which 1, 2 or 3 hydrogen atoms of the alkyl group are replaced by an amino group. Non-limiting examples of aminoalkyl groups may be aminomethyl and 1-aminoethyl, etc.

The term "amino-substituted alkoxy" refers to an alkoxy group in which at least one hydrogen atom of the alkoxy group is replaced by an amino group. For example, an amino-substituted C _1-6 alkoxy group refers to a C _1-6 alkoxy group in which at least one hydrogen atom is replaced by an amino group. Preferably, "aminoalkoxy" refers to an alkoxy group in which 1, 2 or 3 hydrogen atoms of the alkoxy group are replaced by an amino group. Non-limiting examples of aminoalkoxy groups are aminomethoxy and 1-aminoethoxy, etc.

The term "nitro-substituted alkyl" refers to an alkyl group in which at least one hydrogen atom of the alkyl group is replaced by a nitro group. For example, a nitro-substituted C _1-6 alkyl group refers to a C _1-6 alkyl group in which at least one hydrogen atom is replaced by a nitro group.

The term "nitro-substituted alkoxy" refers to an alkoxy group in which at least one hydrogen atom of the alkoxy group is replaced by a nitro group. For example, a nitro-substituted C _1-6 alkoxy group refers to a C _1-6 alkoxy group in which at least one hydrogen atom is replaced by a nitro group.

The term "cyano-substituted alkyl" refers to an alkyl group in which at least one hydrogen atom of the alkyl group is replaced by a cyano group. For example, a cyano-substituted C _1-6 alkyl group refers to a C _1-6 alkyl group in which at least one hydrogen atom is replaced by a cyano group.

The term "cyano-substituted alkoxy" refers to an alkoxy group in which at least one hydrogen atom of the alkoxy group is replaced by a cyano group. For example, a cyano-substituted C _1-6 alkoxy group means that at least one hydrogen atom of the C _1-6 alkoxy group is replaced by a cyano group.

The term "thiol-substituted alkyl" refers to an alkyl group in which at least one hydrogen atom of the alkyl group is replaced by a thiol group. For example, a thiol-substituted C _1-6 alkyl group refers to a C _1-6 alkyl group in which at least one hydrogen atom is replaced by a thiol group.

The term "mercapto-substituted alkoxy" refers to an alkoxy group in which at least one hydrogen atom of the alkoxy group is replaced by a mercapto group. For example, a mercapto-substituted C _1-6 alkoxy group means that at least one hydrogen atom of the C _1-6 alkoxy group is replaced by a mercapto group.

The term "hydroxy substituted alkyl" refers to an alkyl group in which at least one hydrogen atom of the alkyl group is replaced by a hydroxy group. For example, a hydroxy substituted C _1-6 alkyl group refers to a C _1-6 alkyl group in which at least one hydrogen atom is replaced by a hydroxy group.

The term "hydroxy substituted alkoxy" refers to an alkoxy group in which at least one hydrogen atom of the alkoxy group is replaced by a hydroxy group. For example, a hydroxy substituted C _1-6 alkoxy group means that at least one hydrogen atom of the C _1-6 alkoxy group is replaced by a hydroxy group.

The term "halogenated alkyl" refers to an alkyl group in which at least one hydrogen atom of the alkyl group is replaced by a halogen, such as fluorine (F), chlorine (Cl), bromine (Br) or iodine (I). For example, a halogenated C _1-6 alkyl group refers to a C _1-6 alkyl group in which at least one hydrogen atom is replaced by a halogen.

The term "halogenated alkoxy" refers to an alkoxy group in which at least one hydrogen atom of the alkoxy group is replaced by a halogen, such as fluorine (F), chlorine (Cl), bromine (Br) or iodine (I). For example, a halogenated C _1-6 alkoxy group means that at least one hydrogen atom of the C _1-6 alkoxy group is replaced by a halogen.

The term "heterocycloalkyl" refers to a saturated monocyclic, bicyclic, or multicyclic ring with more rings condensed. For example, "3-10 membered heterocycloalkyl" refers to a monocyclic, bicyclic, or multicyclic ring with more rings condensed of 3 to 10 ring atoms, wherein 1, 2, 3 or 4 of the ring atoms are heteroatoms selected from N, O and S, and the remaining ring atoms are carbon. Some non-limiting examples of heterocycloalkyl include azetidin-3-yl, azetidin-2-yl, oxetan-3-yl, oxetan-2-yl, 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-piperidinyl, pyrrolidinyl (e.g., pyrrolidin-2-yl), morpholino, morpholin-2-yl, morpholin-3-yl, pyrrolidinyl (e.g., pyrrolidin-3-yl), piperazinyl (e.g., piperazin-1-yl), 3-azabicyclo[3.1.0]hexan-6-yl, or 2,5-diazabicyclo[2.2.1]heptan-2-yl.

The term "aromatic group" refers to an all-carbon monocyclic or fused polycyclic (i.e., rings that share adjacent carbon atom pairs) group with a conjugated π electron system, which is an aromatic cyclic group. When the aromatic group is preceded by a restriction on the number of ring atoms, such as a 5-12-membered aromatic group, it means that the aromatic group has 5-12 ring atoms. Representative examples of aromatic groups include, but are not limited to, phenyl, naphthyl, or similar groups.

The term "heteroaryl" refers to an aromatic heterocyclic group having one to multiple (preferably 1, 2, 3, 4 or 5) heteroatoms, which may be a monocyclic (monocyclic) or polycyclic (bicyclic, tricyclic or polycyclic) group fused together or covalently linked, each heterocyclic group containing heteroatoms may have one or more (e.g., 1, 2, 3, 4) heteroatoms independently selected from the following group: oxygen, sulfur and nitrogen. When there is a number limit before the heteroaryl, it refers to the number of ring atoms of the heteroaryl, for example, a 5-12 membered heteroaryl refers to a heteroaryl having 5-12 ring atoms. Representative examples of heteroaryl include, but are not limited to: pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, thiazolyl, furanyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazolyl, tetrazolyl, 1H-pyrazolo[3,4-d]pyrimidin-6-yl, or similar groups.

The term "hydroxyl" refers to an -OH group.

The term "amino" refers to the _–NH2 group.

The term "cyano" refers to a -CN (nitrile) group.

The term "nitro" refers to the group _-NO2 .

The term "mercapto" refers to the group -SH.

The term "pharmaceutically acceptable salt" refers to those salts that retain the biological effects and properties of free alkali or free acids, which are formed with inorganic acids (such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc., particularly hydrochloric acid) and organic acids (such as acetic acid, trifluoroacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, lactic acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, N-acetylcysteine, etc.). In addition, these salts can be prepared by adding inorganic or organic bases to the free acids. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium salts, etc. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines (including naturally occurring substituted amines), cyclic amines, and basic ion exchange resins (such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, lysine, arginine, N-ethylpiperidine, piperidine, polyimine resins, and the like).

The term "hydrate" refers to an association formed between a solvent molecule being water and a compound of the present invention via one, two, three or more water molecules.

As used herein, the term "treating" includes: (1) inhibiting a state, disorder or condition of at least one clinical or subclinical symptom of a disease (e.g., in the case of maintenance therapy, preventing, alleviating or delaying the development of a disease or its recurrence); and/or (2) relieving a condition (i.e., causing a state, disorder or condition of a disease or at least one clinical or subclinical symptom to subside). The benefit to the patient being treated is statistically significant or at least perceptible to the patient or physician. However, it should be understood that when a drug is administered to a patient to treat a disease, the result may not always be an effective treatment.

When indicating the number of substituents, the term "one or more" refers to the range from one substituent to the highest number of possible substituents, i.e., replacement of one hydrogen by a substituent until all hydrogens are replaced by substituents, for example, "one or more" can mean one, two, three, four, five or six, etc.

The terms "pharmaceutical composition" and "pharmaceutical formulation" (or "formulation") are used interchangeably and refer to a mixture or solution containing a therapeutically effective amount of an active drug ingredient and a pharmaceutically acceptable excipient, which is administered to a mammal, eg, a human in need thereof.

The terms "excipient," "pharmaceutically acceptable carrier," and "therapeutically inert excipient" are used interchangeably and refer to any pharmaceutically acceptable ingredient in a pharmaceutical composition that is not therapeutically active and is non-toxic to the subject to which it is administered, such as a disintegrant, binder, filler, solvent, buffer, tonicity agent, stabilizer, antioxidant, surfactant, carrier, diluent, or lubricant used to formulate the drug product.

The above scheme is further described below in conjunction with specific embodiments; it should be understood that these embodiments are used to illustrate the basic principles, main features and advantages of the present invention, and the present invention is not limited to the scope of the following embodiments; the implementation conditions adopted in the embodiments can be further adjusted according to specific requirements, and the implementation conditions not specified are usually the conditions in conventional experiments.

Unless otherwise specified in the following examples, all raw materials are commercially available or prepared by conventional methods in the art.

Example 1 Synthesis of Compound ST-01

Experimental process:

1) Preparation of Compound 2

At -78°C, compound 1 (11.6 g, 46.03 mmol) was dissolved in 100 ml of anhydrous tetrahydrofuran, and lithium diisopropylamide (30 ml, 59.83 mmol, 2 mol/L tetrahydrofuran solution) was added. After stirring at -78°C for 1 hour, carbon dioxide was bubbled into the reaction and stirring was continued for 1 hour. The reaction mixture was then adjusted to pH 2 with 6M hydrochloric acid, extracted with ethyl acetate, washed with brine, and the organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo to give 14.8 g of crude yellow solid compound 2, which was used directly in the next step. ¹ H NMR (400 MHz, DMSO-d6) δ 13.92 (s, 1H), 7.62 (dd, J = 8.6, 1.5 Hz, 1H), 7.05 (t, J = 8.8 Hz, 1H), 3.85 (s, 3H).

2) Preparation of Compound 3

Compound 2 (16.6 g, 56.07 mmol) and dimethylformamide (0.3 ml) were added to 50 ml of thionyl chloride and heated at 80°C for 3 hours. The reaction mixture was concentrated in vacuo to obtain a crude product of 2-fluoro-6-iodo-3-methoxybenzoyl chloride. 50 ml of anhydrous tetrahydrofuran and 50 ml of aqueous ammonia were added to the crude product at 0°C, and the mixture was stirred at 0°C for 0.5 hours. The reaction mixture was concentrated in vacuo to obtain 10.0 g of brown solid crude compound 3, which was used directly in the next step. ¹ H NMR (400 MHz, DMSO-d6) δ7.99 (s, 1H), 7.73 (s, 1H), 7.57 (dd, J = 8.8, 1.4 Hz, 1H), 6.99 (t, J = 8.8 Hz, 1H), 3.84 (s, 3H).

3) Preparation of Compound 4

Compound 3 (10.7 g, 36.27 mmol) and thionyl chloride (22.0 g, 181.33 mmol) were added to 60 ml of dimethylformamide and heated to 115°C for overnight reaction. The reaction mixture was poured into cooling water and extracted with ethyl acetate. The organic layer was washed with saturated sodium bicarbonate aqueous solution and brine, dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography (petroleum ether-ethyl acetate, 10:1) to obtain 4.4 g of red solid compound 4 with a yield of 43.8%. ¹ H NMR (400 MHz, DMSO-d6) δ7.80 (dd, J=8.8,1.3 Hz, 1H), 7.36 (t, J=8.8 Hz, 1H), 3.89 (s, 3H).

4) Preparation of Compound 5

Compound 4 (1.0 g, 3.61 mmol) was dissolved in 10 ml of dichloromethane at -78°C, and 22 ml of boron tribromide was added. The reaction mixture was placed at room temperature and stirred overnight. The reaction mixture was then poured into cold water and extracted with dimethylformamide. The organic layer was washed with brine, dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography (petroleum ether-ethyl acetate, 7:1) to obtain 850 mg of brown solid compound 5 with a yield of 89.5%. ¹ H NMR (400 MHz, DMSO-d6) δ 10.96 (s, 1H), 7.62 (dd, J = 8.8, 1.2 Hz, 1H), 7.09 (t, J = 8.8 Hz, 1H).

5) Preparation of Compound 7

Compound 5 (1.0 g, 3.80 mmol) was dissolved in 10 ml of acetonitrile, and compound 6 (1.0 g, 4.56 mmol) and potassium carbonate (1.6 g, 11.41 mmol) were added. The reaction mixture was heated at 60°C for 6 hours and then concentrated in vacuo. The residue was purified by silica gel column chromatography (petroleum ether-ethyl acetate, 5:1) to obtain 1.5 g of yellow solid compound 7 with a yield of 97.4%. ¹ H NMR (400MHz, DMSO-d6) δ7.76(dd,J＝8.8,1.3Hz,1H),7.36(t,J＝8.8Hz,1H),4.09(t,J＝6.4Hz,2H),3.58(s,3H),2.30(t,J＝7.2Hz,2H),1.76–1.67(m,2H), 1.58–1.49(m,2H),1.44–1.28(m,4H).LC-MS: (ESI)m/z[M+H] ⁺ =406.0

6) Preparation of Compound 8

Compound 7 (1.0 g, 2.47 mmol) and hydrazine hydrate (927 mg, 18.51 mmol) were added to 10 ml of n-butanol and heated at 110° C. for 1 hour. The reaction mixture was then concentrated in vacuo and purified by silica gel column chromatography (dichloromethane-methanol, 30:1) to obtain 508 mg of yellow solid compound 8 with a yield of 49.3%. ¹ H NMR (400MHz, DMSO-d6) δ11.91(s,1H),7.19(d,J＝7.6Hz,1H),6.50(d,J＝7.6Hz,1H),4.97(s,2H),4.06(t,J＝6.4Hz,2H),3.58(s,3H),2.31(t,J＝7.2Hz,2H ),1.80–1.70(m,2H),1.55(dt,J＝15.2,7.2Hz,2H),1.46(dd,J＝15.2,7.2Hz,2H),1.38–1.29(m,2H).LC-MS: (ESI)m/z[M+H] ⁺ ＝418.0

7) Preparation of Compound 10

Compound 8 (1.1 g, 2.64 mmol) was dissolved in a mixed solution of 12 ml of dimethyl ether and 4 ml of water, and compound 9 (1.2 g, 5.27 mmol), tetrakis(triphenylphosphine)palladium (305 mg, 0.26 mmol) and sodium carbonate (838 mg, 7.91 mmol) were added. The reaction mixture was purged with nitrogen and heated to 80° ^C for overnight reaction. The reaction mixture was then concentrated in vacuo and purified by silica gel column chromatography (dichloromethane-methanol, 15:1) to obtain 868 mg of brown solid compound 10, with a yield of 86.1%. NMR (400MHz, DMSO-d6) δ11.70(s,1H),7.07(d,J＝8.4Hz,2H),6.70(d,J＝7.6Hz,1H),6.65(d,J＝8.4Hz,2H),6.57(d,J＝7.6Hz,1H),5.19(s,2H),4.31(s,2H),4 .09(t,J＝6.4Hz ,2H),3.58(s,3H),2.33(t,J＝7.2Hz,2H),1.82–1.73(m,2H),1.57(dt,J＝14.8,7.2Hz,2H),1.49(dd,J＝15.2,7.6Hz,2H),1.36(dd,J＝14.8,8.0Hz,2H).LC -MS:(ESI)m/z[M+H] ⁺ =383.2

8) Preparation of Compound 12

Compound 10 (860 mg, 2.25 mmol) was dissolved in 8 ml of dimethylformamide at 0°C, and compound 11 (299 mg, 2.25 mmol) was added. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was then concentrated in vacuo and purified by silica gel column chromatography (dichloromethane-methanol, 20:1) to obtain 680 mg of yellow solid compound 12 with a yield of ^58.7 %. NMR (400MHz, DMSO-d6) δ11.81(s,1H),8.75(s,1H),8.63(s,1H),7.55(d,J＝8.4Hz,2H),7.34(d,J＝8.4Hz,2H),7.32(s,1H),7.25(d,J＝8.4Hz,1H),7.16(t,J＝ 7.6Hz,1H),6.80(d,J＝7.2Hz,1H),6.76(d,J＝7.6Hz,1H),6 .67(d,J＝7.6Hz,1H),4.30(s,2H),4.12(t,J＝6.4Hz,2H),3.58(s,3H),2.33(t,J＝7.6Hz,2H),2.29(s,3H),1.84–1.74(m,2H),1.59(dd,J＝15.0,7.6Hz, 2H),1.54–1.46(m,2H),1.40–1.31(m,2H).LC-MS: (ESI)m/z[M+H] ⁺ =516.2

9) Preparation of compound ST-01

4.2 g of hydroxylamine hydrochloride and 2.4 g of sodium hydroxide were added to 25 ml of methanol and stirred at 0°C for half an hour. The reaction mixture was filtered to obtain a filtrate. Compound 12 (1.5 g, 2.91 mmol) and sodium hydroxide (465 mg, 11.64 mmol) were added to 15 ml of the above filtrate and stirred at room temperature for 2 hours. The pH value of the reaction mixture was then adjusted to 7 with 1 mol of hydrochloric acid. The reaction mixture was filtered to obtain a solid, and the solid was washed with methanol and water to obtain 1.0 g of white solid compound ST-01 with a yield of ^66.5 %. NMR (400MHz, DMSO-d6) δ11.83(s,1H),10.38(s,1H),8.80–8.62(m,3H),7.57(d,J＝8.0Hz,2H),7.39–7.31(m,3H),7.27(d,J＝8.0Hz,1H),7.17(t,J＝7.6Hz,1 H),6.80(d,J＝7.2Hz,1H ),6.75(d,J＝7.6Hz,1H),6.68(d,J＝7.6Hz,1H),4.32(s,2H),4.11(t,J＝6.0Hz,2H),2.29(s,3H),1.99(t,J＝7.2Hz,2H),1.83–1.75(m,2H),1.58–1.47( m,4H),1.34(d,J=6.8Hz,2H). ¹³ C NMR (101MHz, DMSO-d6) δ169.7,153.0,148.9,143.9,140.1,139.3,138.5,134.2,133.2,129.7,129.1,127.9,123.1,120.0,119.2,118.5,115.9, 112.3,106.4,68.2,32.7,29.1,28.9,25.6,25.6,21.7.LC-MS: (ESI)m/z[M+H] ⁺ =517.2

Example 2 Synthesis of Compound ST-04

Experimental process:

1) Compound 10 was prepared by the method in Example 1

2) Preparation of Compound 12

Compound 10 (1.0 g, 2.61 mmol) was dissolved in 10 ml of dimethylformamide at 0°C, and then compound 11 (394 mg, 2.61 mmol) was added. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was then concentrated in vacuo and purified by silica gel column chromatography (dichloromethane-methanol, 20:1) to obtain 560 mg of white solid compound 12, ^with a yield of 40.1%. NMR (400MHz, DMSO-d6) δ11.81(s,1H),9.17(s,1H),8.52(s,1H),8.01(d,J＝7.7Hz,1H),7.56(d,J＝8.6Hz,2H),7.36(d,J＝8.5Hz,2H),7.11(dd,J＝11.3,8.3Hz,1 H),6.83–6.78(m,1H),6.76(d,J＝7.8Hz,1H),6.67(d,J＝7 .6Hz,1H),4.29(s,2H),4.12(t,J＝6.4Hz,2H),3.58(s,3H),2.33(t,J＝7.4Hz,2H),2.28(s,3H),1.85–1.75(m,2H),1.59(dd,J＝15.1,7.5Hz,2H),1.50(dd ,J＝15.6,7.5Hz,2H),1.41–1.32(m,2H).LC-MS:(ESI)m/z[M+H] ⁺ ＝534.2

3) Preparation of compound ST-04 and ST-04 hydrochloride

4.2 g of hydroxylamine hydrochloride and 2.4 g of sodium hydroxide were added to 25 ml of methanol and stirred at 0 ° C for half an hour. The reaction mixture was filtered to obtain a filtrate. Compound 12 (760 mg, 1.42 mmol) and sodium hydroxide (227 mg, 5.68 mmol) were added to 10 ml of the above filtrate and stirred at room temperature for 2 hours. The pH value of the reaction mixture was then adjusted to 7 with 1 mol of hydrochloric acid. The reaction mixture was filtered to obtain a solid. The solid was washed with methanol and water to obtain 400 mg of white solid compound ST-04 with a yield of 52.5%. Then 990 mg of compound ST-04 was added to water and heated at 80 ° C for half an hour. The reaction mixture was cooled to room temperature and filtered to obtain a solid. The above solid was added to 30 ml of 2 mol of hydrochloric acid, stirred at room temperature for 1 hour, and freeze-dried to obtain 760 mg of compound ST-04 hydrochloride (containing one molecule of hydrochloric acid).

Compound ST-04: ¹ H NMR (400MHz, DMSO-d6) δ11.81(s,1H),10.34(s,1H),9.21(s,1H),8.65(s,1H),8.54(d,J ＝2.1Hz,1H),8.01(d,J＝6.4Hz,1H),7.56(d,J＝8.5Hz,2H),7.36(d,J＝8.4Hz,2H),7.11(dd,J＝11.3 ,8.4Hz,1H),6. 83–6.78(m,1H),6.76(d,J＝7.8Hz,1H),6.68(d,J＝7.7Hz,1H),4.29(s,2H),4.12(t,J＝6.3Hz,2H ),2.28(s,3H),1.97(t,J＝7.3Hz,2H),1.84–1.73(m,2H),1.59–1.45(m,4H),1.34(dd,J＝14.6,7.6Hz, 2H).

Compound ST-04 hydrochloride:

¹ H NMR (400MHz, DMSO-d6) δ13.26(br,1H),10.42(br,1H),9.79(s,1H),8.78(s,1H),7.98(dd,J＝7.8,1.7Hz ,1H),7.61(d,J＝8.5Hz,2H),7.43(d,J＝8.5Hz,2H),7.09(dd,J＝11.2,8.3Hz,1H ),6.99(d,J＝7.9Hz,1H),6.93(d,J＝7.8Hz,1H),6.85–6.75(m,1H),4.18(t,J＝6.2Hz,2H),2.27(s ,3H),1.98(t,J＝7.3Hz,2H),1.86–1.73(m,2H),1.60–1.44(m,4H),1.36–1.29(m,2H). ¹⁹ F NMR(400MHz,DMSO-d6)δ-134.30. ¹³ C NMR(101MHz,DMSO-d6)δ169.6,152.8,152.0,149.7,144.1,139.7,134.7,133.9,133.9,132.1,129.7,127.6,127.5 , 123.2,122.9,121.6,118.6,115.1,114.9,112.7,108.5,68.6,32.7,28.9,28.8,25.6,25.6,21.2.LC-MS: (ESI)m/z[M+H] ⁺ =535.2.

Example 3 Synthesis of Compound ST-05

Experimental process:

1) Preparation of Compound 2

At 0°C, compound 1 (5 g, 65.71 mmol) was dissolved in 200 ml of anhydrous tetrahydrofuran, and sodium cyanide (2.89 g, 72.28 mmol) was added. After stirring at 0°C for 30 minutes, a solution of tert-butyldiphenylsilyl chloride (18.1 g, 65.71 mmol) dissolved in 50 ml of tetrahydrofuran was added. The reaction mixture was placed at room temperature and stirred overnight. The reaction mixture was then quenched with aqueous ammonium chloride at 0°C and extracted with ethyl acetate. The organic layer was washed with brine, dried over sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column chromatography (petroleum ether-ethyl acetate, 10:1) to obtain 13.6 g of colorless oily compound 2 in a yield of 65.8%. ¹ H NMR (400MHz, DMSO-d6) δ7.63(dd,J＝7.4,1.6Hz,4H),7.50–7.38(m,6H),3.74(t,J＝6.4Hz,2H),3.53(t,J＝6.3Hz,2H),1.70(p,J＝6.3Hz,2H),1.00(s,9H).

2) Preparation of Compound 4

Compound 2 (13.6 g, 43.24 mmol) was dissolved in 170 ml of tert-butanol, and compound 3 (8.31 g, 64.86 mmol) and cesium carbonate (14.09 g, 43.24 mmol) were added. After stirring at room temperature for 6 hours, the reaction mixture was poured into water and extracted with ethyl acetate. The organic layer was washed with brine, dried over sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column chromatography (petroleum ether-ethyl acetate, 10:1) to obtain 11.5 g of colorless oily compound 4 in a yield of 60%. ¹ H NMR (400MHz, DMSO-d6) δ7.63–7.60(m,4H),7.49–7.38(m,6H),3.70(t,J＝6.3Hz,2H),3.53(t,J＝6.2Hz,2H),3.48(t,J＝6.2Hz,2H),2.38(t,J＝6.2Hz,2H) ,1.74(p,J=6.2Hz,2H),1.36(s,9H),0.99(s,9H).

3) Preparation of Compound 5

Compound 4 (0.5 g, 1.13 mmol) was dissolved in 5 ml of anhydrous tetrahydrofuran, and tetrabutylammonium fluoride (2.30 ml, 2.26 mmol, 1 mol tetrahydrofuran solution) was added. After stirring at room temperature for 2 hours, the reaction mixture was poured into water and extracted with ethyl acetate. The organic layer was washed with brine, dried over sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column chromatography (petroleum ether-ethyl acetate, 3:1) to give 180 mg of light yellow oily compound 5 with a yield of 78%. ¹ H NMR (400 MHz, DMSO-d6) δ4.34 (t, J = 5.1 Hz, 1H), 3.53 (t, J = 6.2 Hz, 2H), 3.41 (t, J = 6.8 Hz, 4H), 2.39 (t, J = 6.2 Hz, 2H), 1.68–1.56 (m, 2H), 1.40 (s, 9H).

4) Preparation of Compound 7

At 0°C, compound 6 (1.0 g, 3.80 mmol) was dissolved in 20 ml of anhydrous tetrahydrofuran, and compound 5 (853 mg, 4.18 mmol), triphenylphosphine (1.5 g, 5.7 mmol) and diethyl azodicarboxylate (1.54 g, 7.6 mmol) were added. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was then concentrated in vacuo and purified by silica gel column chromatography (petroleum ether-ethyl acetate, 4:1) to obtain 1.0 g of yellow liquid compound 7 in a yield of 58.5%. ¹ H NMR (400MHz, DMSO-d6) δ7.57(dd,J＝8.8,1.7Hz,1H),6.96(t,J＝8.6Hz,1H),4.13(t,J＝6.3Hz,2H),3.67(t,J＝6.3Hz,2H),3.61(t,J＝5.9Hz,2H),2.47(dd,J＝ 7.5,5.0Hz,2H),2.06(p,J＝6.1Hz,2H),1.43(s,9H).LC-MS:(ESI)m/z[M+Na] ⁺ =472.0

5) Preparation of Compound 8

Compound 7 (180 mg, 0.4 mmol) and hydrazine hydrate (140 mg, 2.8 mmol) were added to 6 ml of n-butanol, and the suspension was heated to 110° C. for 2 hours. The reaction mixture was then concentrated in vacuo and purified by silica gel column chromatography (dichloromethane-methanol, 30:1) to obtain 100 mg of yellow solid compound 8 with a yield of 54.1%. ¹ H NMR (400MHz, CDCl ₃ ) δ7.28(d,J＝8.0Hz,1H),6.42(d,J＝8.0Hz,1H),4.18(t,J＝6.2Hz,2H),3.69(t,J＝6.3Hz,2H),3.63(t,J＝6.0Hz,2H),2.49(t,J＝6.3Hz,2 H), 2.08 (p, J = 6.1Hz, 2H), 1.44 (s, 9H). LC-MS: (ESI) m/z [M+H] ⁺ = 462.2

6) Preparation of Compound 10

Compound 8 (1.4 g, 3.03 mmol) was dissolved in a mixed solution of 12 ml of dimethyl ether and 4 ml of water, and compound 9 (996 mg, 4.54 mmol), tetrakis(triphenylphosphine)palladium (350 mg, 0.3 mmol) and sodium carbonate (963 mg, 9.09 mmol) were added. The reaction mixture was heated to 80 ° C and reacted overnight with nitrogen. The reaction mixture was then concentrated in vacuo and purified by silica gel column chromatography (dichloromethane-methanol, 20:1) to obtain 1.2 g of brown solid compound 10 with a yield of 92.7%. ¹ H NMR (400MHz, DMSO-d6) δ11.71(s,1H),7.07(d,J＝8.4Hz,2H),6.69(d,J＝7.7Hz,1H),6.65(d,J＝8.4Hz,2H),6.57(d,J＝7.7Hz,1H),5.19(s,2H),4.31(s,2 H),4.14(t,J＝6.2Hz,2H),3.61(dt,J＝9.2,6.2Hz,4H),2.43(t,J＝6.1Hz,2H),1.99(p,J＝6.1Hz,2H),1.37(s,9H).LC-MS: (ESI)m/z[M+H] ⁺ ＝427.2

7) Preparation of Compound 12

At 0°C, compound 10 (1.0 g, 2.34 mmol) was dissolved in 10 ml of dimethylformamide, and compound 11 (354 mg, 2.34 mmol) was added. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was then concentrated in vacuo and purified by silica gel column chromatography (dichloromethane-methanol, 20:1) to obtain 760 mg of white solid compound 12, ^with a yield of 56.1%. NMR (400MHz, DMSO-d6) δ11.84(s,1H),9.17(s,1H),8.52(d,J＝2.5Hz,1H),8.01(dd,J＝7.9,1.9Hz,1H),7.56(d,J＝8.6Hz,2H),7.36(d,J＝8.6Hz,2H),7.11(dd,J ＝11.4,8.3Hz,1H),6.84–6.78(m,1H),6.75(d,J＝7.8Hz,1H),6.67(d,J＝7.7Hz,1H),4.30(s,2H),4.17(t ,J＝6.2Hz,2H),3.62(dt,J＝12.2,6.2Hz,4H),2.44(t,J＝6.1Hz,2H),2.28(s,3H),2.01(p,J＝6.1Hz,2H),1.37(s,9H).4.31(s,2H),4.14(t,J＝6.2Hz,2H ),3.61(dt,J＝9.2,6.2Hz,4H),2.43(t,J＝6.1Hz,2H),1.99(p,J＝6.1Hz,2H),1.37(s,9H).LC-MS: (ESI)m/z[M+H] ⁺ ＝578.4

8) Preparation of compound ST-05

8.4 g of hydroxylamine hydrochloride and 4.8 g of sodium hydroxide were added to 50 ml of methanol, and the suspension was stirred at 0°C for half an hour. The reaction mixture was filtered to obtain a filtrate. Compound 12 (2.54 g, 4.40 mmol) and sodium hydroxide (704 mg, 17.6 mmol) were added to 30 ml of the above filtrate and stirred at 60°C for 2 hours. The pH value of the reaction mixture was then adjusted to 7 with 1 mol of hydrochloric acid. The reaction mixture was filtered to obtain a solid. The solid was washed with methanol and water to obtain 724 mg of white solid compound ST-05, with ^a yield of 31%. NMR (400MHz, DMSO-d6) δ11.86(s,1H),10.43(s,1H),9.20(s,1H),8.79(s,1H),8.54(d,J＝2.0Hz,1H),8.03(d,J＝6.4Hz,1H),7.57(d,J＝8.4Hz,2H),7.37(d, J＝8.4Hz,2H),7.12(dd,J＝11.2,8.4Hz ,1H),6.79(dd,J＝16.4,6.4Hz,2H),6.69(d,J＝7.6Hz,1H),4.33(s,2H),4.18(t,J＝6.0Hz,2H),3.62(t,J＝5.2Hz,4H),2.28(s,3H),2.22(t,J＝6.0Hz,2H) ,2.06–1.96(m,2H).LC-MS:(ESI)m/z[M+H] ⁺ =537.2

Example 4 Synthesis of Compound ST-06

Experimental process:

1) Preparation of Compound 3

At 0°C, compound 1 (6.0 g, 22.81 mmol) was dissolved in 60 ml of anhydrous tetrahydrofuran, and compound 2 (5.6 g, 25.09 mmol), triphenylphosphine (8.97 g, 34.21 mmol) and diethyl azodicarboxylate (9.22 g, 45.62 mmol) were added. The reaction mixture was placed at room temperature and stirred for 2 hours. The reaction mixture was then concentrated in vacuo and purified by silica gel column chromatography (petroleum ether-ethyl acetate, 4:1) to obtain 3.5 g of yellow liquid compound 3, with a yield of 36.5%. ¹ H NMR (400MHz, DMSO-d6) δ7.78(d,J＝8.8Hz,1H),7.41(t,J＝8.9Hz,1H),4.25(s,2H),3.54(s,2H),2.84(d,J＝9.5Hz,3H),1.34(d,J＝25.7Hz,9H).LC-MS: (ESI) m/z[M-55] ⁺ =365.0

2) Preparation of Compound 4

Compound 3 (100 mg, 0.21 mmol) was added to 4 ml of 4 mol hydrochloric acid solution dissolved in dioxane, and the suspension was stirred at room temperature for half an hour. The reaction mixture was then concentrated in vacuo to obtain 75 mg of crude yellow solid compound 4, with a yield of 98.4%. LC-MS: (ESI) m/z [M+H] ⁺ = 321.0

3) Preparation of Compound 6

Compound 4 (300 mg, 0.94 mmol) was dissolved in 5 ml of methanol, and compound 5 (149 mg, 0.94 mmol), sodium triacetoxyborohydride (398 mg, 1.88 mmol) and acetic acid (5.83 mg, 0.094 mmol) were added. After stirring at room temperature for half an hour, the reaction mixture was poured into cold water and extracted with dichloromethane. The organic layer was washed with brine, dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography (dichloromethane-methanol, 20:1) to obtain 340 mg of yellow liquid compound 6 with a yield of 78.5%. ¹ H NMR (400MHz, DMSO-d6) δ7.77(dd,J＝8.8,1.3Hz,1H),7.38(t,J＝8.9Hz,1H),4.17(t,J＝5.6Hz,2H),2.70(t,J＝5.6Hz,2H),2.36(t,J＝7.0Hz,2H),2.21(s,3H ),2.17(t,J＝7.3Hz,2H),1.59(p,J＝7.2Hz,2H),1.37(s,9H).LC-MS: (ESI)m/z[M+H]+＝463.2

4) Preparation of Compound 7

Compound 6 (340 mg, 0.74 mmol) and hydrazine hydrate (556 mg, 11.1 mmol) were added to 8 ml of n-butanol, and the suspension was heated at 110°C for 2 hours. The reaction mixture was then concentrated in vacuo and purified by silica gel column chromatography (petroleum ether-ethyl acetate, 1:1) to obtain 280 mg of yellow solid compound 7, with a yield of 80.3%. LC-MS: (ESI) m/z [M+H] + = 475.1

5) Preparation of Compound 9

Compound 7 (500 mg, 1.05 mmol) was dissolved in a mixed solution of 12 ml of dimethyl ether and 4 ml of water, and compound 8 (276 mg, 1.26 mmol), tetrakis(triphenylphosphine)palladium (121 mg, 1.05 mmol) and sodium carbonate (223 mg, 2.1 mmol) were added. The reaction mixture was heated to 80°C and reacted overnight with nitrogen. The reaction mixture was then concentrated in vacuo and purified by silica gel column chromatography (dichloromethane-methanol, 20:1) to obtain 300 mg of yellow solid compound 9 with a yield of 64.8%. ¹ H NMR (400MHz, DMSO-d6) δ11.65(br,1H),7.07(d,J＝8.3Hz,2H),6.74(d,J＝7.7Hz,1H),6.65(d,J＝8.3Hz,2H),6.58(d,J＝7.7Hz,1H),5.18(s,2H),4.31(s,2H ),4.19(t,J＝5.7Hz,2H),2.80(s,2H),2.43(s,2H),2.28(s,3H),2.22(t,J＝7.2Hz,2H),1.70–1.60(m,2H),1.38(s,9H).LC-MS: (ESI)m/z[M+Na] ⁺ ＝462.2

6) Preparation of Compound 11

Compound 9 (1.8 g, 4.1 mmol) was dissolved in 10 ml of dimethylformamide at 0°C, and compound 10 (620 mg, 4.1 mmol) was added. The reaction mixture was placed at room temperature and stirred for 2 hours. The reaction mixture was then concentrated in vacuo and purified by silica gel column chromatography (dichloromethane-methanol, 15:1) to obtain 1.2 g of yellow solid compound 11, with a yield of 49.6%. LC-MS: (ESI) m/z [M+H] ⁺ = 591.3

7) Preparation of compound ST-06

4.2 g of hydroxylamine hydrochloride and 2.4 g of sodium hydroxide were added to 25 ml of methanol and stirred at 0°C for half an hour. The reaction mixture was filtered to obtain a filtrate. Compound 11 (1.2 g, 2.19 mmol) and sodium hydroxide (1.75 g, 4.38 mmol) were added to 10 ml of the above filtrate and heated to 60°C and stirred at constant temperature for 2 hours. The pH value of the reaction mixture was then adjusted to 7 with 1 mol of hydrochloric acid. The reaction mixture was filtered to obtain a solid, which was purified by reverse phase high performance liquid chromatography (0.1% acetic acid-acetonitrile and water). 590 mg of white solid compound ST-06 was obtained with a yield of 49.1%. ¹ H NMR (400MHz, DMSO-d6) δ10.48(br,1H),9.34(s,1H),8.63(s,1H),8.24(s,1H),8.00(dd,J＝7.8,1.4Hz,1H),7.58(d,J＝8.4Hz,2H),7.37(d,J＝8.4Hz,2H),7.11 (dd,J＝11.2,8.4H z,1H),6.81(d,J＝7.7Hz,2H),6.70(d,J＝7.6Hz,1H),4.36–4.24(m,2H),3.03(s,2H),2.64(t,J＝7.0Hz,2H),2.45(s,3H),2.28(s,3H),2.06(t,J＝6.9Hz ,2H),1.87–1.66(m,2H). ¹³ C NMR(101MHz,DMSO-d6)δ169.5,164.4,152.7,152.0,149.6,149.0,143.4,139.1,134.2,134.0,133.3,129.8,128.3,127.6,127.5,123.2,123.2,121.5,112.0,118.4,115.1,114.9,112.3,106.7,65.6,56.8,55.5,42.1,30.4,22.4,21.2. ¹⁹ F NMR(376MHz,DMSO)δ-134.69.LC-MS:(ESI)m/z[M+H] ⁺ ＝550.2

Example 5 Evaluation of the inhibitory effects of compound ST-01, compound ST-04 hydrochloride, compound ST-05, and compound ST-06 on HDAC1, HDAC6, VEGFR2, and PDGFR-β activities, respectively

5.1 Purpose and Background

The purpose of the experiment is to detect the inhibitory ability of small molecule compounds on kinase activity through a kinase-based drug screening system. The experimental principle of the HTRF method is: kinase phosphorylates substrate; Eu-labeled antibody binds to the phosphorylated site of the substrate; streptavidin-XL665 binds to the substrate biotin; when Eu and XL665 are close, Eu as a donor receives excitation from the light source (320nm), emits emission light (615nm) and resonates energy transfer to the adjacent receptor XL665, and the receptor emits emission light (665nm) after being excited; the specific signal is proportional to the phosphorylated substrate. When the inhibitor is added, the phosphorylation level is inhibited, and the emission light of 665nm cannot be detected, and only the emission light of 615nm can be detected. In this way, the inhibitory level of the compound on the kinase activity is evaluated. The experimental principle of the ADP-Glo method is: the kinase and its substrate undergo enzymatic reactions, consuming ATP to produce ADP, and the amount of the product is detected using the ADP-Glo reagent and luminescence method to reflect the activity of the kinase.

5.2 Test methods

5.2.1. Enzyme reaction reagent formula

5.2.1.1 Enzyme reaction formula

Enzyme reaction recipe

name Storage concentration.nM Working concentration.nM HDAC1 2680 5 HDAC6 3040 10 VEGFR2 5900 0.07 PDGFR-β 2272.72 0.227

5.2.1.2 Substrate formulation

HDAC Substrate Formulation

VEGFR2 and PDGFR-β Substrate Formula

5.2.2 Reagents and consumables

Materials and reagents factory Part Number HTRF KinEASE-TK kit Cisbio 62TK0PEC PDGFRβ carna 08-158 VEGFR2 carna 08-191 Sunitinib MCE HY-10255A BIBF1120 MCE HY-50904 MgCl2 Sigma M1028 ATP Promega V915B DMSO Sigma D4540-1L 384-well plate,white,low volume,round-bottom Greiner 784075 384-well plate,black,low volume,round-bottom Corning 4514 96-well polypropylene plate Nunc 249944 HDAC1 Fluorogenic Assay Kit BPS 50061 HDAC6 Fluorogenic Assay Kit BPS 50076

5.2.3 Instruments

Instruments and equipment factory model Multifunctional ELISA reader BMG PHERAstar FSX Microplate low speed centrifuge Xiangyi TDZ5-WS ECHO BECKMAN 655system

5.2.4 Test steps

HDAC detection steps:

1) Dilute the test compound with DMSO to 10 concentrations starting from 10 μM. Take 6 μL of the compound and add it to 54 μL of DMSO to perform a 3-fold dilution.

2) Use Echo to transfer 50 nL of the test substance to a 384-well reaction plate, with two replicate wells for each compound. Centrifuge at 1000 rpm for 1 minute. The final DMSO concentration is 1%.

3) Add 2.5 μL HDAC to the 384 reaction plate, centrifuge at 1000 rpm for 1 minute, and incubate at 25°C for 10 minutes.

4) Add 2.5 μL of substrate to the 384 reaction plate, centrifuge at 1000 rpm for 1 minute, and incubate at 37°C for 30 minutes.

5) Add 5 μL of the developer mixture to the 384 reaction plate, centrifuge at 1000 rpm for 1 minute, and incubate at 25°C for 10 minutes.

6) Read the FI signal using an ELISA reader (em: 340; ex: 450 nm).

VEGFR2 and PDGFR-β detection steps:

1) Dilute the test compound with DMSO to 10 concentrations starting from 10 μM. Take 6 μL of the compound and add it to 54 μL of DMSO to perform a 3-fold dilution.

2) Use Echo to transfer 25 nL of the test substance to a 384-well reaction plate, with two replicate wells for each compound. Centrifuge at 1000 rpm for 1 minute. The final DMSO concentration is 1%.

3) Add 2.5 μL of kinase solution to the 384 reaction plate, centrifuge at 1000 rpm for 1 minute, and incubate at 25°C for 10 minutes.

4) Add 2.5 μL ATP & SubMixture to the 384 reaction plate, centrifuge at 1000 rpm for 1 minute, and incubate at 25°C for 50 minutes.

5) Add 5 μL Sa-XL 665/TK-antibody-Cryptate to the 384 reaction plate, centrifuge at 1000 rpm for 1 minute, and incubate at 25°C for 1 hour.

6) Read the HTRF signal (Ratio 665/620nm) using the BMG high-throughput drug screening multifunctional microplate reader.

5.2.3 Data processing methods

The reading value of the negative control (1% DMSO well) was set as 0% inhibition rate, and the reading value of the positive control (positive drug well) was set as 100% inhibition rate, and the inhibition rate of each test solution was calculated.

10μMReference

1% DMSO

The _IC50 (half maximal inhibitory concentration) of the compound was obtained using the following nonlinear fitting formula:

Y＝Bottom+(Top-Bottom)/(1+10^((LogIC ₅₀ -X)×HillSlope))

X: Log value of compound concentration

Y: Compound inhibition rate (%inhibition)

Z’ factor calculation equation:

Z'＝1-3(SD _min +SD _max )/(AVE _max -AVE _min )

in:

Min is the positive control Ratio value, and Max is the negative control DMSO Ratio value.

SD is standard error, AVE is the mean value of Ratio.

5.2.4 Experimental Results

Positive control: Vorinostat, Vorinostat:

Positive control: Sunitinib, Sunitinib:

Example 6 In vitro efficacy test of compound ST-04 hydrochloride in 20 cell lines

1. Research objectives

use The (CTG) method was used to measure the in vitro efficacy of a test compound and a positive control drug on 20 cell lines.

2. Experimental Design

The IC ₅₀ of one test compound and one positive compound was determined in 20 cell lines. The drug treatment time was 3 days, 9 concentrations were used, and 3 replicates were performed for each concentration.

3. Materials

Cell line:

Note: All cells were cultured at 37°C and 5% _CO2 .

4. Reagents and consumables

96-well plate (Corning, catalog number: 3610)

MEM (Invitrogen, Catalog No.: 11095080)

RPMI 1640 (Invitrogen, Catalog No.: C22400500BT)

FBS (ExCell Bio, Cat. No.: FND500)

CellTiter-Glo (CTG) (Promega, Catalog No.: G7573)

NEAA (GIBCO, item number: 11140050)

Penicillin-Streptomycin (Pen-Strep) (Gibco, Cat. No.: 15070063)

5. Test samples

Drugs to be tested:

Compound Name Weight(mg) Molecular weight purity Powder storage conditions Storage conditions after dissolution Solvents Storage solution concentration ST-04 Hydrochloride 20 570.22 95.96% 2-8℃ 2-8℃ DMSO 20mM

6. Instruments

EnVision 2104 Multilabel Reader: PerkinElmer (Instrument No.: TAREA0011);

Carbon dioxide incubator: SANYO Electric Co., Ltd. (Instrument No.: 02100400059);

Biological safety cabinet, Sujing Antai, BFC-1300IIA2 (instrument number: TBBSC0140);

Inverted microscope: Chongqing Optoelectronics XDS-1B (instrument number: TAMIC0200);

Living cell counter: Vi-Cell XR, Beckman Coulter (instrument number: TACEL0030);

Electronic balance: Mettlertoledo AL104 (instrument number: TBBAL0560);

7. Methods

7.1. 2D inhibitory effect of test compounds on cells

Cell culture

1) Resuscitate cells with culture medium, maintain cell growth to the logarithmic phase, digest with trypsin, and collect cells by centrifugation at 1000 rpm for 5 minutes. After centrifugation, remove the supernatant and add appropriate amount of culture medium to resuspend the cells.

Day 1: Cell plating

2) taking the cell suspension and counting the cells using a cell counter;

3) Plate cells according to the required number of tests, add 90 μL of cell suspension to each well of a 96-well plate, and set up a T0 plate;

4) Cell culture was performed at 37°C, 5% CO ₂ , and 95% humidity;

Day 2: T0 board readings

5) Add 10 μL of medium containing solvent to each well and perform CTG analysis;

6) Thaw the CTG reagent and equilibrate the cell plate to room temperature for 30 minutes;

7) Add 50 μL CTG solution to each well;

8) Shake on an orbital shaker for 2 minutes to lyse the cells;

9) Place the cell plate at room temperature for 10 minutes to stabilize the luminescence signal;

10) Use EnVision to read the luminescence value;

Day 2: Add the drug to be tested

11) Dissolving the test compound in a solvent to form a stock solution and performing gradient dilution, and then diluting it 20 times with a culture medium to obtain a 10-fold solution;

Prepare a 10-fold solution of the positive drug in culture medium;

12) Add 10 μL of 10X drug solution to each well (three replicate wells are set for each cell concentration). The highest concentration of the tested compound is 20 μM, 9 concentrations, 3-fold dilution;

13) Cell culture was performed at 37°C and 5% _CO2 ;

Day 5: Test plate readings (72 hours after drug treatment)

14) Thaw the CTG reagent and equilibrate the cell plate to room temperature for 30 minutes;

15) Add 100 μL CTG solution to each well;

16) Shake on an orbital shaker for 2 minutes to lyse the cells;

17) Place the cell plate at room temperature for 10 minutes to stabilize the luminescence signal;

18) Read the luminescence value using EnVision;

8. Data Analysis

Cell viability was calculated using the formula: (V _sample -V _{Medium control} )/(V _{vehicle control} -V _{Medium control} )×100%. V _sample is the reading of the drug-treated group, and V _{vehicle control} is the average value of the solvent control group. GraphPadPrism 9 software was used to draw a S-shaped dose-survival curve using a nonlinear regression model and calculate the IC ₅₀ value. The data also include the maximum inhibition rate.

9. Experimental Results

9.1. IC ₅₀ and maximum inhibition rate of the test compound on cells

The above experiment proves that the structure of the compound is accurate. Compared with the control sample, the compound of the present invention can simultaneously inhibit histone deacetylase VI (HDAC6, histone deacetylases 6), histone deacetylase 1 (HDAC1) and two tyrosine kinases (vascular endothelial growth factor receptor, Vascular endothelial cell growthfactor2, VEGFR2) and platelet-derived growth factor receptor (platelet-derived growth factor-β, PDGFR-β), indicating that the compound of the present invention integrates multiple targets in one, can be used as a multi-target inhibitor, and has the pharmacological effects of sunitinib and vorinostat at the same time. Therefore, the compound of the present invention can be used to treat diseases mediated by tyrosine kinases and histone deacetylases, such as malignant tumors. Treatable malignant tumors include but are not limited to solid tumors such as renal cancer, liver cancer, colon cancer, gastrointestinal stromal tumors, lung cancer, breast cancer, pancreatic cancer, glioma, lymphoma, fibrosarcoma, ovarian cancer, prostate cancer, and leukemia, lymphoma, and other malignant tumors of the blood system.

This type of compound combines multiple targets in one, which can not only give full play to the synergistic effect to increase biological activity, but also avoid the problems caused by the different properties and metabolism when multiple drugs are used in combination. It has good practicality and good prospects.

In addition, in the in vitro efficacy test of 20 cell lines, the compound ST-04 hydrochloride showed strong activity in 20 tumor cell lines, most of which exceeded the positive control drug, and the rest were also comparable to the positive control drug, showing good prospects for application in the treatment of malignant tumors.

The above embodiments are only for illustrating the technical concept and features of the present invention, and their purpose is to enable people familiar with the technology to understand the content of the present invention and implement it accordingly, and they cannot be used to limit the protection scope of the present invention. Any equivalent changes or modifications made according to the spirit of the present invention should be included in the protection scope of the present invention.

The endpoints and any values of the ranges disclosed in this article are not limited to the precise ranges or values, and these ranges or values should be understood to include values close to these ranges or values. For numerical ranges, the endpoint values of each range, the endpoint values of each range and the individual point values, and the individual point values can be combined with each other to obtain one or more new numerical ranges, which should be regarded as specifically disclosed in this article.

Claims (15)

1. A compound of the general formula (I), a pharmaceutically acceptable salt, hydrate or prodrug thereof,

wherein:

R ^a 、R ^b independently selected from hydrogen, C _1-20 Alkyl, 3-10 membered heterocycloalkyl, 5-12 membered aryl, 5-12 membered heteroaryl; wherein, C is as follows _1-20 Alkyl, 3-10 membered heterocycloalkyl, 5-12 membered aryl, 5-12 membered heteroaryl each independently being unsubstituted or substituted with one, two, three or more substituents selected from: fluorine, chlorine, bromine, iodine, nitro, cyano, mercapto, hydroxy, amino, C _1-6 Alkyl-substituted amino, C _1-10 Alkyl, halogenated C _1-6 Alkyl, nitro substituted C _1-6 Alkyl-and cyano-substituted C _1-6 Alkyl-mercapto-substituted C _1-6 Alkyl, amino substituted C _1-6 Alkyl, hydroxy substituted C _1-6 Alkyl, C _1-6 Alkoxy, halogenated C _1-6 Alkoxy, nitro substituted C _1-6 Alkoxy, cyano-substituted C _1-6 Alkoxy, mercapto-substituted C _1-6 Alkoxy, amino substituted C _1-6 Alkoxy, hydroxy-substituted C _1-6 Alkoxy, C _2-6 Alkenyl, C _2-6 Alkynyl, -C (=o) R ₁ 、-C(＝O)NHR ₂ 、-C(＝O)OR ₃ 、-NHC(＝O)R ₄ 、-OC(＝O)R ₅ ；

R ₁ 、R ₂ 、R ₃ 、R ₄ 、R ₅ Independently selected from unsubstituted C _1-20 Alkyl, or C independently substituted with one or more substituents selected from fluoro, chloro, bromo, iodo, nitro, cyano, mercapto, hydroxy, amino _1-20 An alkyl group;

Each R is ^c Identical or different and are each independently selected from hydrogen, C _1-20 Alkyl, C _1-20 Alkoxy, fluoro, chloro, bromo, iodo, nitro, cyano, mercapto, hydroxy, amino, C _1-6 Alkyl-substituted amino, halogenated C _1-6 Alkyl, nitro substituted C _1-6 Alkyl-and cyano-substituted C _1-6 Alkyl-mercapto-substituted C _1-6 Alkyl, amino substituted C _1-6 An alkyl group;

R ^d 、R ^e independently selected from hydrogen, C _1-20 Alkyl, nitro substituted C _1-6 Alkyl, cyanoSubstituted C _1-6 Alkyl-mercapto-substituted C _1-6 Alkyl, amino substituted C _1-6 An alkyl group;

R ^g having a chain structure in which the number of atoms constituting a main chain is 3 to 15 and in which the number of hetero atoms is not more than 2, the hetero atoms being selected from oxygen, sulfur, nitrogen;

x is 1, 2, 3 or 4.

2. A compound according to claim 1, a pharmaceutically acceptable salt, hydrate or prodrug thereof, wherein the number of heteroatoms in the atoms constituting the backbone of the chain structure is 1 or 2, and wherein one heteroatom is linked to a benzene ring in an indazole ring; and/or, the chain structure is a saturated straight chain or a saturated branched chain.

3. The compound of claim 1, a pharmaceutically acceptable salt, hydrate or prodrug thereof, wherein the compound has the structure of formula (ii):

Wherein:

R ^a 、R ^b 、R ^c 、R ^d 、R ^e x is respectively with R in the general formula (I) ^a 、R ^b 、R ^c 、R ^d 、R ^e X is the same;

A. b is independently CH ₂ O, S or-N (R) ₆ )-，R ₆ Is hydrogen or C _1-6 Alkyl, m, n are independently selected from integers of 0-10, and 3.ltoreq.m+n.ltoreq.10;

b is CH when any of the following conditions exists ₂ ：

(1) A is CH ₂ The method comprises the steps of carrying out a first treatment on the surface of the (2) A is selected from O, S or-N (R) ₆ ) -, and m is 0; (3) n is 0.

4. A compound according to claim 3, a pharmaceutically acceptable salt, hydrate or prodrug thereof, characterized in thatIn which A is selected from CH ₂ O, S or-NH-, B is selected from CH ₂ 、O、-N(CH ₃ )-、-N(CH ₂ CH ₃ ) -or-N (CH) ₂ CH ₂ CH ₃ )-。

5. A compound according to claim 3, wherein a is O and B is CH ₂ M+n is 3, 4 or 5.

6. A compound according to any one of claims 1 to 5, a pharmaceutically acceptable salt, hydrate or prodrug thereof, wherein R ^a 、R ^b Not both hydrogen.

7. The compound of claim 6, pharmaceutically acceptable salts, hydrates or prodrugs thereof, wherein R ^a 、R ^b Wherein one is selected from the group consisting of unsubstituted or substituted pyrimidinyl, pyrazolyl, pyridinyl, pyrazinyl, imidazolyl, pyridazinyl, thiazolyl, phenyl; another is selected from hydrogen, or from C which is unsubstituted or substituted by 1-3 halogens _1-6 An alkyl group.

8. A compound according to any one of claims 3 to 5, a pharmaceutically acceptable salt, hydrate or prodrug thereof, wherein the compound has the structure of formula (iii):

wherein:

R ^c 、R ^d 、R ^e x, A, B, m, n are respectively identical to R in the general formula (II) ^c 、R ^d 、R ^e X, A, B, m, n are identical;

R ^b selected from hydrogen, or from C which is unsubstituted or substituted by 1-3 halogens _1-6 An alkyl group;

R ₇ selected from fluorineChlorine, bromine, iodine, hydroxyl, mercapto, cyano, amino, methylamino, ethylamino, nitro, C _1-6 Alkyl, halogenated C _1-6 Alkyl, C _1-6 Alkoxy, halogenated C _1-6 Alkoxy, hydroxymethyl, mercaptomethyl, C (=o) CH ₃ 、C(＝O)CH ₂ CH ₃ 、C(＝O)NHCH ₃ 、C(＝O)NHCH ₂ CH ₃ 、NHC(＝O)CH ₃ 、NHC(＝O)CH ₂ CH ₃ 、NHCH ₃ 、N(CH ₃ ) ₂ 、NH CH ₂ CH ₃ T is 0, 1, 2, 3, 4 or 5, and D, E are independently selected from CH or N.

9. The compound of claim 8, a pharmaceutically acceptable salt, hydrate or prodrug thereof, wherein the compound has the structure of formula (iv):

wherein: r is R ^b 、R ^c 、R ^d 、R ^e X, A, B, m, n are respectively identical to R in the general formula (II) ^b 、R ^c 、R ^d 、R ^e X, A, B, m, n are identical;

R ₈ 、R ₉ independently selected from the group consisting of hydrogen, fluorine, chlorine, bromine, iodine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, halogenated methyl, halogenated ethyl, halogenated n-propyl, halogenated isopropyl, halogenated n-butyl, halogenated isobutyl, methoxy, ethoxy, halogenated methoxy, halogenated ethoxy.

10. A compound according to any one of claims 1 to 5, a pharmaceutically acceptable salt, hydrate or prodrug thereof, wherein R ^b 、R ^d 、R ^e Independently selected from hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl or isobutyl; and/or the number of the groups of groups,

each R is ^c Identical or notAnd each is independently selected from hydrogen, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, methoxy, or ethoxy; and/or the number of the groups of groups,

R ^b 、R ^c 、R ^d 、R ^e hydrogen at the same time; and/or the number of the groups of groups,

R ^g selected from:

11. the compound of claim 1, a pharmaceutically acceptable salt, hydrate or prodrug thereof, wherein the compound has the structure of formula (v):

wherein: p is 2, 3, 4, 5, 6, 7, 8, 9 or 10; r is R ₈ 、R ₉ Independently selected from the group consisting of hydrogen, fluorine, chlorine, bromine, iodine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, halogenated methyl, halogenated ethyl, halogenated n-propyl, halogenated isopropyl, halogenated n-butyl, halogenated isobutyl, methoxy, ethoxy, halogenated methoxy, halogenated ethoxy.

12. The compound of claim 1, a pharmaceutically acceptable salt, hydrate or prodrug thereof, wherein the compound has the structural formula shown in any one of formulas (ST-01) to (ST-16):

13. A pharmaceutical composition comprising a compound according to any one of claims 1 to 12, a pharmaceutically acceptable salt, hydrate or prodrug thereof, and a pharmaceutically acceptable carrier.

14. The pharmaceutical composition according to claim 13, wherein the pharmaceutical composition is a composition for preventing and/or treating malignant tumors including renal cancer, liver cancer, colon cancer, gastrointestinal stromal tumor, lung cancer, breast cancer, pancreatic cancer, glioma, lymphoma, fibrosarcoma, ovarian cancer, prostate cancer, leukemia, lymphoma.

15. Use of a compound according to any one of claims 1 to 12, a pharmaceutically acceptable salt, hydrate or prodrug thereof, or a pharmaceutical composition according to claim 13, for the manufacture of a medicament for the prevention and/or treatment of a disease mediated by tyrosine kinase and/or histone deacetylase.