WO2016107541A1 - Pyrrole amide compound, preparation method therefor, and use thereof - Google Patents

Abstract

Disclosed are a pyrrole amide compound as represented in formula I, or pharmaceutically acceptable salts, polymorphs, hydrates, or solvate compounds. R₁ is chosen from hydrogen, hydroxyl, the cyano group, halogen, carboxyl, etc. R₂ and R₃ are chosen from hydrogen, hydroxyl, the cyano group, halogen, carboxyl, and alkyl of C₁ to C₆, etc., R₂ and R₃ being same or different. R₄ are chosen from hydroxyl, the sulfhydryl group, amino-substituted phenyl, or the epoxy ketone group. The new compound as represented in formula I of the present invention expresses good deacetylase inhibitory activity and has the potential for preventing and/or treating diseases caused by histone deacetylase activity abnormality. Additionally, the new compound of the present invention has good inhibitory activity against different liver cancer cells, and has good prospect for clinical application.

Description

Pyrrolamide compound and preparation method and use thereof Technical field

The present invention relates to a pyrrolamide compound, a preparation method and use thereof, and particularly to a pyrrolamide compound having histone deacetylase inhibitory activity, a preparation method thereof and use thereof.

Background technique

Inactivation of genes that control cell growth in the body is a hallmark of tumorigenesis. The epigenetic mechanisms that cause gene inactivation mainly include DNA methylation, histone acetylation, and modification of other components in the chromatin high-level structure. These modifications alter the chromatin configuration, leading to changes in gene transcriptional regulation, and dysregulation of gene transcription. Cell proliferation is abnormal, resulting in tumor production.

More than 40 years ago, Allfrey et al. recognized that histone acetylation is closely related to eukaryotic gene transcriptional regulation (Allfrey VG, Faulkner R, Mirsky AE. Acetylation and methylation of histones and their possible role in the regulation of RNA synthesis [ J]. Proc Natl Acad Sci USA, 1964, 51: 786-794). Histone acetylation plays a central role in the transcriptional regulation of eukaryotic cells. The acetylation of histones occurs on the ε-amino group of the N-terminal evolutionarily conserved lysine residue. The modification on H3 and H4 is more common than H2A and H2B. The more important acetylation site is H3. Lys ⁹ and Lys ¹⁴ , and Lys ⁵ , Lys ⁸ , Lys ¹² and Lys ¹⁶ on H4. The acetylation of HAT causes the amino group of the N-terminal lysine of the histone to be acetylated, and the positive charge on the amino group is eliminated. The negative charge carried by the DNA molecule itself facilitates the unfolding of the DNA conformation, and the structure of the nucleosome becomes slack. Conducive to the contact of transcription factors and co-transcriptional activators with DNA molecules, histone acetylation can activate the transcriptional expression of specific genes. In contrast, histone deacetylation is not conducive to the expression of specific genes (eg, Rb, p21, p27). The acetylation and deacetylation of histones becomes a switch for specific gene expression (Thiagalingam S, Cheng KH, Lee HJ, et al. Histone deacetylases: unique players in shaping the epigenetic histone code [J]. Ann NY Acad Sci, 2003, 983:84-100).

Histone acetylation is regulated by a pair of functionally antagonistic protease histone acetyltransferases (HATs) and histone deacetylases (HDACs). In normal cells, this pair of enzymes is in a state of dynamic equilibrium. In general, increased levels of histone acetylation are associated with increased transcriptional activity, while low levels of acetylation are associated with inhibition of gene expression (Forsberg EC, Bresnick EH. Histone acetylation beyond promoters: long-range acetylation patterns in the chromatin world [ J]. Bioessays, 2001, 23(9): 820-830). Studies have found that HDAC is overexpressed and recruited by transcription factors, leading to abnormal inhibition of specific genes, leading to tumors and other diseases; while inhibition of HDAC activity will cause growth inhibition and apoptosis of many cancer cells (Somech R, Izraeli S, J Simon A. Histone deacetylase inhibitors-a new tool to treat cancer [J]. Cancer Treat Rev, 2004, 30(5): 461-472). Therefore, HDAC has become the latest and most popular target in the field of anti-tumor drug research and development.

HDAC inhibitors inhibit HDAC enzyme activity by inhibiting HDAC, blocking gene expression inhibition due to HDAC recruitment dysfunction, and altering staining by altering the degree of histone acetylation Quality structure, which regulates gene expression to treat cancer. It is effective in treating hematological tumors and solid tumors by inducing growth arrest, differentiation or apoptosis of tumor cells. HDAC inhibitors are tumor-specific and have cytotoxic effects on both proliferating and resting variant cells, whereas normal cells are more than 10 times more tolerant and do not cause normal cell growth arrest and apoptosis. Moreover, the clinical dose of HDAC inhibitors is much lower than the maximum tolerated dose of the human body, and the toxicity to the body is low. The development and utilization of HDAC inhibitors has become a new hot spot in cancer therapy.

At present, HDAC inhibitors that have been researched and developed can be divided into five categories: (1) hydroxamic acid compounds, functional groups are hydroxamic acid, and representatives are TSA, SAHA (Curtin ML, Garland RB, Heyman HR, et A1.Succinimide hydroxamic acids as potent inhibitors of histone deacetylase [J]. Bioorg Med Chem Lett, 2002, 12(20): 2919-2923), LAQ824 (Atadja P, Hsu M, Kwon P, et a1. Moleculer and cellular basis For the anti-proliferative effects of the HDAC inhibitor LAQ824.Novartis Found Symp, 2004, 259: 249-266); (2) containing or not containing 2-amino-8-oxo-9,10-epoxydecanoyl Group of tetrapeptides, such as FK-228; (3) benzamides, the representative MS-275 has entered clinical research; (4) short-chain fatty acids, such as butyric acid and phenylbutyric acid; (5) other Classes, such HDAC inhibitors do not have the structural features of a general HDAC, but all contain some or all of the structural subunits that inhibit HDAC activity requirements.

For example, Chinese Patent No. 103420917A discloses a benzamide compound having a fused ring structure, as shown in Formula A, for histone deacetylase inhibitory activity and in the treatment of malignant tumors and diseases associated with differentiation and proliferation. Application; Chinese Patent CN 103288728A discloses a naphthalenecarboxamide derivative, as shown in Formula B, which is effective for treating some diseases caused by dysregulation of protein kinases; Chinese Patent CN 103539695A discloses a substituted diphenyl ether histone a deacetylase inhibitor, as shown in Formula C; Chinese Patent CN 103467359A discloses a cinnamamide-containing histone deacetylase inhibitor containing hydrazine, as shown in Formula D; Chinese Patent CN 102659630A discloses a different Hydroxamic acid compounds, as shown in formula E.

Chinese patent CN 102786458 A discloses a pyrrolecarboxamide derivative, as shown in Formula F, for use as an anti-malignant drug, particularly for the preparation of a medicament for the treatment of breast cancer, lung cancer and gastric cancer.

R ₁ , R ₂ , R ₃ , R ₄ are: a C1-C6 straight or branched alkyl group, a C3-C6 cycloalkyl group;

R ₅ and R _{6 are} simultaneously or respectively: hydrogen, C1-C6 alkyl; hydroxy, halogen, C1-C4 alkoxy, nitrate-substituted C1-C6 alkyl.

At present, the SAHA developed by Merck is a listed histone deacetylase inhibitor, which is limited to the treatment of cutaneous T-cell lymphoma, and is not effective for many other cancers. Other HDAC inhibitors developed have certain problems in anticancer activity, toxic side effects, and subtype selectivity. Therefore, the development of a novel compound having histone deacetylase inhibitory activity has important social and economic significance.

Summary of the invention

The object of the invention is a pyrrolamide compound.

The pyrrole amide compound of the formula I, or a pharmaceutically acceptable salt, crystal form, hydrate or solvate thereof, of the invention:

among them,

R _{1 is} selected from the group consisting of hydrogen, hydroxy, cyano, halogen, carboxyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₂ -C ₆ amide, C ₁ -C ₆ amino An alkyl group, a C ₂ -C ₆ aminoacyl group, a C ₃ -C ₆ heterocyclic group, a C ₃ -C ₆ heterocycloalkenyl group, a phenoxy group, a phenyl group or a substituted phenyl group;

R ₂ and R ₃ are respectively or simultaneously selected from the group consisting of hydrogen, hydroxy, cyano, halogen, carboxyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ aminoalkyl, C ₂ -C ₆ amide group, C ₂ -C ₆ aminoacyl group, C ₃ -C ₆ heterocyclic group, C ₃ -C ₆ heterocycloalkenyl group, phenoxy group, phenyl group, substituted phenyl group , piperazinyl or substituted piperazinyl;

R _{4 is} selected from a hydroxy, thiol, amino substituted phenyl or epoxy ketone group.

further,

R _{1 is} selected from the group consisting of hydrogen, hydroxy, cyano, fluoro, chloro, bromo, carboxy, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentylene, hexyl Alkyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentyloxy, hexyloxy, aminomethyl, Aminoethyl, aminopropyl, aminobutylalkyl, aminopentyl, aminohexane, carboxamido, acetylamino, n-propionylamino, isopropylamide, n-butyryl, isobutyryl, Tert-butylamide, pentanoamide, hexane amide, methyl, acetyl, n-alanyl, isopropylyl, n-butyryl, isobutyryl, tert-butyryl, pentylamine Acyl group, hexyl group, nitrogen heterocyclic group of C ₃ , nitrogen heterocyclic group of C ₄ , nitrogen heterocyclic group of C ₅ , nitrogen heterocyclic group of C ₆ , azacycloheteroyl group of C ₃ , C ₄ Azacycloheteroenyl, C ₅ azacycloalkenyl, C ₆ azacycloalkenyl, phenoxy, phenyl or substituted phenyl;

R ₂ and R ₃ are respectively or simultaneously selected from the group consisting of hydrogen, hydroxyl, cyano, fluorine, chlorine, bromine, carboxyl, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl , pentyl, hexane, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentaneoxy, hexaneoxy Base, aminomethyl, aminoethyl, aminopropyl, aminobutylalkyl, aminopentyl, aminohexane, carboxamide, acetylamino, n-propionyl, isopropylamide, n-butyryl , isobutyryl amide, tert-butyryl amide, pentane amide, hexane amide, carbamoyl, acetyl, n-alanyl, isopropyl, n-butyryl, isobutyryl, uncle Butyryl group, valyl group, hexyl group, nitrogen heterocyclic group of C ₃ , nitrogen heterocyclic group of C ₄ , nitrogen heterocyclic group of C ₅ , nitrogen heterocyclic group of C ₆ , nitrogen heterocyclic ring of C ₃ Alkenyl, C ₄ azacycloalkenyl, C ₅ azacycloalkenyl, C ₆ azacycloalkenyl, phenoxy, phenyl, substituted phenyl, piperazinyl, methylpiperazine Base, ethyl piperazinyl, propyl piperazinyl , butyryl piperazinyl, pentyl piperazinyl or hexyl piperazinyl;

R _{4 is} selected from hydroxy, decyl or amino substituted phenyl.

Further, the compound of formula I is:

The invention also provides a preparation method of the pyrrole amide compound represented by the above formula I,

When R ₄ is a hydroxyl group, its synthetic route is:

Wherein, Boc represents tert-butoxycarbonyl; TFA represents trifluoroacetic acid; Fmoc-Cl represents fluorenylmethoxycarbonyl chloride; and HATU represents 2-(7-azobenzotriazole)-N,N,N',N' -tetramethylurea hexafluorophosphate; DIEA stands for N,N-diisopropylethylamine; DCM stands for dichloromethane;

R _{1 is} selected from the group consisting of hydrogen, hydroxy, cyano, halogen, carboxyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₂ -C ₆ amide, C ₁ -C ₆ amino An alkyl group, a C ₂ -C ₆ aminoacyl group, a C ₃ -C ₆ heterocyclic group, a C ₃ -C ₆ heterocycloalkenyl group, a phenoxy group, a phenyl group or a substituted phenyl group;

R ₂ and R ₃ are respectively or simultaneously selected from the group consisting of hydrogen, hydroxy, cyano, halogen, carboxyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ aminoalkyl, C ₂ -C ₆ amide group, C ₂ -C ₆ aminoacyl group, C ₃ -C ₆ heterocyclic group, C ₃ -C ₆ heterocycloalkenyl group, phenoxy group, phenyl group, substituted phenyl group , piperazinyl or substituted piperazinyl;

R ₅ is a halogen;

Includes the following steps:

a, compound IM-1a, lithium hydroxide and ether solvent / water mixed solvent, stirred at 20 ° C ~ 30 ° C for 1 h ~ 6h, remove the organic solvent, diluted with water, adjust the pH = 3 ~ 6, precipitated solid, Filtration to obtain a solid; washing the solid with water and drying to give N-tert-butoxycarbonyl-2,5-dihydro-1H-pyrrole-3-carboxylic acid;

The molar ratio of the compound IM-1a to lithium hydroxide is 1:1 to 10; the mass to volume ratio of the compound IM-1a to the mixed solvent is 1:7 to 20 g/ml; in the mixed solvent, the ether The volume ratio of solvent to water is 1 to 2:1;

b, 0 ° C ~ 5 ° C, the N-tert-butoxycarbonyl-2,5-dihydro-1H-pyrrole-3-carboxylic acid obtained in step a is dissolved in a halogen hydrocarbon solvent, added trifluoroacetic acid, at 20 After stirring at °C ~ 30 ° C for 2h ~ 12h, the reaction liquid is obtained; the reaction liquid is concentrated to obtain a yellow oil, which is the compound IM-2a;

a mass to volume ratio of the N-tert-butoxycarbonyl-2,5-dihydro-1H-pyrrole-3-carboxylic acid to a halocarbon solvent: 1:5 to 20 g/ml; the N-tert-butoxycarbonyl-2 , the mass to volume ratio of 5-dihydro-1H-pyrrole-3-carboxylic acid to trifluoroacetic acid is 1:2 to 10 g/ml;

c. The compound IM-2a obtained in step b, a mixed solvent of sodium carbonate, decanoyl chloride and ether solvent/water is stirred at 20 ° C to 30 ° C for 12 h to 16 h, diluted with water to adjust pH = 1 to 3 , ester solvent extraction, the organic phase is combined, the organic phase is dried, filtered, concentrated to give the compound IM-3a;

The molar ratio of the compound IM-2a, sodium carbonate and decanoyl chloride is 1:1 to 5: 0.9 to 1.5; the compound The mass to volume ratio of the IM-2a to the mixed solvent is 1:10 to 25 g / ml; in the mixed solvent, the volume ratio of the ether solvent to water is 1 to 2:1;

d, compound IM-3a, O-(tetrahydro-2H-pyran-2-yl)hydroxylamine, 2-(7-azobenzotriazole)-N,N,N' obtained in step c, N'-tetramethylurea hexafluorophosphate, N,N-diisopropylethylamine and halogen hydrocarbon solvent, stirred at 25 ° C ~ 30 ° C for 12h ~ 16h, diluted with water, ester solvent extraction, combined The organic phase is dried, filtered and concentrated to obtain a crude product; the crude product is purified by column chromatography to give compound IM-4a;

The compound IM-3a, O-(tetrahydro-2H-pyran-2-yl)hydroxylamine, 2-(7-azobenzotriazole)-N,N,N',N'-four The molar ratio of methyl urea hexafluorophosphate to N,N-diisopropylethylamine is 1:1 to 2:1 to 2:2 to 4; the mass volume of the compound IM-3a and the halogen hydrocarbon solvent The ratio is 1:9-20g/ml;

e, the compound IM-4a obtained in the step d, the piperidine and the nitrogen-containing solvent are stirred at 25 ° C to 30 ° C for 4 h to 6 h, diluted with water, extracted with an ester solvent, and the organic phase is combined to dry the organic phase. Filtration and concentration to obtain compound IM-5a;

The mass ratio of the compound IM-4a to piperidine is 1:1 to 4 g / ml; the mass ratio of the compound IM-4a to the nitrogen-containing solvent is 1: 5 to 20 g / ml;

f, the compound IM-5a, the triethylamine, the compound IM-6a, and the halocarbon solvent in the step e are stirred at 25 ° C to 30 ° C for 1 h to 10 h, and then the solvent is removed to obtain a crude product; the crude product is purified by column chromatography. , obtaining compound TM-1a;

The molar ratio of the compound IM-5a, triethylamine, and compound IM-6a is 1:1 to 5:1 to 2; the mass to volume ratio of the compound IM-5a to the halocarbon solvent is 1:50 to 100 g. /ml;

g, 0 ° C ~ 5 ° C, the compound TM-1a obtained in step f is dissolved in a halocarbon solvent, trifluoroacetic acid, stirred at 25 ° C ~ 30 ° C for 1 h ~ 12h, the solvent is removed, to obtain a crude product; The crude product is purified by preparative high performance liquid chromatography to give the compound of formula I;

The mass to volume ratio of the compound TM-1a to the halocarbon solvent is 1:50 to 100 g/ml; and the mass ratio of the compound TM-1a to trifluoroacetic acid is 1:10 to 50 g/ml.

further,

a, compound IM-1a, lithium hydroxide and ether solvent / water mixed solvent, stirred at 25 ° C for 2h, remove the organic solvent, diluted with water, adjust the pH = 5, precipitate a solid, filtered to give a solid; Washing with water and drying to obtain N-tert-butoxycarbonyl-2,5-dihydro-1H-pyrrole-3-carboxylic acid;

The molar ratio of the compound IM-1a to lithium hydroxide is 1:4.5 to 5; the mass to volume ratio of the compound IM-1a to the mixed solvent is 1:10 to 12 g/ml; in the mixed solvent, the ether The volume ratio of solvent to water is 2:1;

b, at 0 ° C, the N-tert-butoxycarbonyl-2,5-dihydro-1H-pyrrole-3-carboxylic acid obtained in step a is dissolved in a halogen hydrocarbon solvent, trifluoroacetic acid is added, and the reaction is stirred at 25 ° C After 2h, the reaction liquid is obtained; the reaction liquid is concentrated to obtain a yellow oil, which is the compound IM-2a;

a mass to volume ratio of the N-tert-butoxycarbonyl-2,5-dihydro-1H-pyrrole-3-carboxylic acid to a halocarbon solvent: 1:10 g/ml; the N-tert-butoxycarbonyl-2,5 - a mass to volume ratio of dihydro-1H-pyrrole-3-carboxylic acid to trifluoroacetic acid of 1:4 to 5 g/ml;

c. A mixed solvent of the compound IM-2a, sodium carbonate, decyloxycarbonyl chloride and ether solvent/water obtained in the step b, After stirring at 25 ° C for 12 h to 16 h, dilute with water, adjust pH = 1, extract the ester solvent, combine the organic phase, dry the organic phase, filter and concentrate to obtain compound IM-3a;

The molar ratio of the compound IM-2a, sodium carbonate and decyloxycarbonyl chloride is 1:3:1; the mass to volume ratio of the compound IM-2a to the mixed solvent is 1:20 g/ml; in the mixed solvent, The volume ratio of the ether solvent to water is 5:3;

d, compound IM-3a, O-(tetrahydro-2H-pyran-2-yl)hydroxylamine, 2-(7-azobenzotriazole)-N,N,N' obtained in step c, N'-tetramethyluronium hexafluorophosphate, N,N-diisopropylethylamine and a halogenated hydrocarbon solvent are stirred at 25 ° C for 12 h to 16 h, diluted with water, extracted with an ester solvent, and combined with an organic phase. Drying the organic phase, filtering and concentrating to obtain a crude product; the crude product is purified by column chromatography to give compound IM-4a;

The compound IM-3a, O-(tetrahydro-2H-pyran-2-yl)hydroxylamine, 2-(7-azobenzotriazole)-N,N,N',N'-four The molar ratio of methyl urea hexafluorophosphate to N,N-diisopropylethylamine is 1:1.1:1.2:3; the mass to volume ratio of the compound IM-3a to the halocarbon solvent is 1:9~ 10g/ml;

e, the compound IM-4a obtained in step d, piperidine and a nitrogen-containing solvent are stirred at 25 ° C for 4 h to 6 h, diluted with water, extracted with an ester solvent, and the organic phase is combined, and the organic phase is dried, filtered and concentrated. , obtaining compound IM-5a;

The mass ratio of the compound IM-4a to piperidine is 1:2g / ml; the mass ratio of the compound IM-4a to the nitrogen-containing solvent is 1:10g / ml;

f, the compound IM-5a, the triethylamine, the compound IM-6a, and the halocarbon solvent in the step e are stirred at 25 ° C for 2 h, and then the solvent is removed to obtain a crude product; the crude product is purified by column chromatography to give the compound TM- 1a;

The molar ratio of the compound IM-5a, triethylamine, and compound IM-6a is 1:1.4:1 to 1.2; the mass to volume ratio of the compound IM-5a to the halocarbon solvent is 1:80 g/ml;

g, at 0 ° C, the compound TM-1a obtained in the step f is dissolved in a halogen hydrocarbon solvent, trifluoroacetic acid is added, and the reaction is stirred at 25 ° C for 2 h, then the solvent is removed to obtain a crude product; the crude product is purified by preparative high performance liquid chromatography. Obtaining a compound of formula I;

The mass-to-volume ratio of the compound TM-1a to the halocarbon solvent is 1:60 to 65 g/ml; and the mass ratio of the compound TM-1a to trifluoroacetic acid is 1:25 g/ml.

further,

R _{1 is} selected from the group consisting of hydrogen, hydroxy, cyano, fluoro, chloro, bromo, carboxy, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentylene, hexyl Alkyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentyloxy, hexyloxy, aminomethyl, Aminoethyl, aminopropyl, aminobutylalkyl, aminopentyl, aminohexane, carboxamido, acetylamino, n-propionylamino, isopropylamide, n-butyryl, isobutyryl, Tert-butylamide, pentanoamide, hexane amide, methyl, acetyl, n-alanyl, isopropylyl, n-butyryl, isobutyryl, tert-butyryl, pentylamine Acyl group, hexyl group, nitrogen heterocyclic group of C ₃ , nitrogen heterocyclic group of C ₄ , nitrogen heterocyclic group of C ₅ , nitrogen heterocyclic group of C ₆ , azacycloheteroyl group of C ₃ , C ₄ Azacycloheteroenyl, C ₅ azacycloalkenyl, C ₆ azacycloalkenyl, phenoxy, phenyl or substituted phenyl;

R ₂ and R ₃ are respectively or simultaneously selected from the group consisting of hydrogen, hydroxyl, cyano, fluorine, chlorine, bromine, carboxyl, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl , pentyl, hexane, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentaneoxy, hexaneoxy Base, aminomethyl, aminoethyl, aminopropyl, aminobutylalkyl, aminopentyl, aminohexane, carboxamide, acetylamino, n-propionyl, isopropylamide, n-butyryl , isobutyryl amide, tert-butyryl amide, pentane amide, hexane amide, carbamoyl, acetyl, n-alanyl, isopropyl, n-butyryl, isobutyryl, uncle Butyryl group, valyl group, hexyl group, nitrogen heterocyclic group of C ₃ , nitrogen heterocyclic group of C ₄ , nitrogen heterocyclic group of C ₅ , nitrogen heterocyclic group of C ₆ , nitrogen heterocyclic ring of C ₃ Alkenyl, C ₄ azacycloalkenyl, C ₅ azacycloalkenyl, C ₆ azacycloalkenyl, phenoxy, phenyl, substituted phenyl, piperazinyl, methylpiperazine Base, ethyl piperazinyl, propyl piperazine , D piperazinyl, piperazinyl amyl or hexyl alkyl piperazinyl group;

R _{5 is} selected from the group consisting of fluorine, chlorine, bromine or iodine.

Further, in step f, the compound IM-6a is:

Further, in the steps a to g, the ether solvent is selected from any one or two or more selected from the group consisting of tetrahydrofuran, diethyl ether, tert-butyl methyl ether, diisopropyl ether and dibutyl ether; and the halocarbon solvent is selected from two. Any one or two or more of methyl chloride, ethyl chloride, dichloroethane, chloroform, and carbon tetrachloride; and the ester solvent is selected from any one of ethyl acetate and ethyl formate or Two or more kinds; the nitrogen-containing solvent is selected from any one or more selected from the group consisting of N,N-dimethylformamide, N,N-dimethylacetamide, acetonitrile, and pyridine.

The present invention also provides another preparation method of the pyrrole amide compound represented by the above formula I,

When R ₄ is a hydroxyl group, its synthetic route is:

among them,

R _{1 is} selected from the group consisting of hydrogen, hydroxy, cyano, halogen, carboxyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₂ -C ₆ amide, C ₁ -C ₆ amino An alkyl group, a C ₂ -C ₆ aminoacyl group, a C ₃ -C ₆ heterocyclic group, a C ₃ -C ₆ heterocycloalkenyl group, a phenoxy group, a phenyl group or a substituted phenyl group;

R ₂ and R ₃ are respectively or simultaneously selected from the group consisting of hydrogen, hydroxy, cyano, halogen, carboxyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ aminoalkyl, C ₂ -C ₆ amide group, C ₂ -C ₆ aminoacyl group, C ₃ -C ₆ heterocyclic group, C ₃ -C ₆ heterocycloalkenyl group, phenoxy group, phenyl group, substituted phenyl group , piperazinyl or substituted piperazinyl;

R ₅ and R ₆ are each a halogen;

Includes the following steps:

i, compound IM-5b, compound IM-6b, sodium bicarbonate and ether solvent / water mixed solvent, stirring reaction at 25 ° C ~ 30 ° C for 1h ~ 10h, remove the solvent, add water, ester solvent extraction, combined organic The phase is washed with saturated brine, the organic phase is dried, filtered, and concentrated to give a crude product. The crude product is purified by column chromatography to give compound IM-7b;

The molar ratio of the compound IM-5b, the compound IM-6b, and the sodium hydrogencarbonate is 1:1 to 2:1 to 5; the mass to volume ratio of the mixed solvent of the compound IM-5b and the ether solvent/water is 1 : 20～100g/ml; in the mixed solvent, the volume ratio of the ether solvent to water is 1 to 5:1;

Ii, a mixed solvent of the compound IM-7b, the compound IM-8b, sodium carbonate, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride and an ether solvent/water obtained in the step i After the reaction is stirred at 50 ° C to 100 ° C for 1 h to 10 h under an inert gas atmosphere, the solvent is removed, water is added, and the organic phase is extracted with a halogenated solvent, and the organic phase is dried, filtered, and concentrated. Crude; crude product is purified by column chromatography to give compound TM-1b;

The molar ratio of the compound IM-7b, the compound IM-8b, and the sodium carbonate is 1:1 to 2:1 to 5; the compound IM-7b and [1,1'-bis(diphenylphosphino)2 The mass ratio of the ferromagnetic] palladium dichloride is 1:0.05 to 0.2; the mass ratio of the mixed solvent of the compound IM-7b to the ether solvent/water is 1:20 to 100 g/ml; in the mixed solvent , the volume ratio of the ether solvent to water is from 1 to 10:1;

Iii, the compound TM-1b of the step II is dissolved in an alcohol solvent, hydrochloric acid is added, and the reaction is stirred at 25 ° C to 30 ° C for 1 h to 10 h, then separated and purified to obtain a compound of the formula I;

The mass to volume ratio of the compound TM-1b to the alcohol solvent is 1:18 to 100 g/ml; the compound The mass-to-volume ratio of TM-1b to hydrochloric acid is 1:3 to 20 g/ml; and the concentration of the hydrochloric acid is in the range of 0.5 N to 2 N.

further,

i, compound IM-5b, compound IM-6b, sodium hydrogencarbonate and ether solvent / water mixed solvent, stirred at 25 ° C for 2h, remove the solvent, add water, extract the ester solvent, combine the organic phase, with saturated salt Washing with water, drying the organic phase, filtering and concentrating to obtain a crude product; the crude product is purified by column chromatography to give compound IM-7b;

The molar ratio of the compound IM-5b, the compound IM-6b, and the sodium hydrogencarbonate is 1:1:2 to 3; the mass ratio of the compound IM-5b to the ether solvent/water mixed solvent is 1:20. ～40g / ml; in the mixed solvent, the volume ratio of the ether solvent to water is 1 to 2:1;

Ii, a mixed solvent of the compound IM-7b, the compound IM-8b, sodium carbonate, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride and an ether solvent/water obtained in the step i After stirring at 80 ° C for 2 h under inert gas atmosphere, the solvent is removed, water is added, and the organic solvent is extracted with a halogenated solvent. The organic phase is washed with saturated brine, and the organic phase is dried, filtered and concentrated to give a crude product. Purification by chromatography to give the compound TM-1b;

The molar ratio of the compound IM-7b, the compound IM-8b, and the sodium carbonate is 1:1:2 to 3; the compound IM-7b and [1,1'-bis(diphenylphosphino)ferrocene The mass ratio of palladium dichloride to 1:0.08 to 0.12; the mass ratio of the mixed solvent of the compound IM-7b to the ether solvent/water is 1:20 to 40 g/ml; in the mixed solvent, the ether The volume ratio of solvent to water is 4 to 6:1;

Iii, the compound TM-1b of the step II is dissolved in an alcohol solvent, hydrochloric acid is added, and the reaction is stirred at 25 ° C for 2 h, then separated and purified to obtain a compound of the formula I;

The mass to volume ratio of the compound TM-1b to the alcohol solvent is 1:18 to 40 g/ml; the mass ratio of the compound TM-1b to hydrochloric acid is 1:3 to 10 g/ml; The concentration ranges from 0.5N to 2N.

further,

R _{1 is} selected from the group consisting of hydrogen, hydroxy, cyano, fluoro, chloro, bromo, carboxy, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentylene, hexyl Alkyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentyloxy, hexyloxy, aminomethyl, Aminoethyl, aminopropyl, aminobutylalkyl, aminopentyl, aminohexane, carboxamido, acetylamino, n-propionylamino, isopropylamide, n-butyryl, isobutyryl, Tert-butylamide, pentanoamide, hexane amide, methyl, acetyl, n-alanyl, isopropylyl, n-butyryl, isobutyryl, tert-butyryl, pentylamine Acyl group, hexyl group, nitrogen heterocyclic group of C ₃ , nitrogen heterocyclic group of C ₄ , nitrogen heterocyclic group of C ₅ , nitrogen heterocyclic group of C ₆ , azacycloheteroyl group of C ₃ , C ₄ Azacycloheteroenyl, C ₅ azacycloalkenyl, C ₆ azacycloalkenyl, phenoxy, phenyl or substituted phenyl;

R ₂ and R ₃ are respectively or simultaneously selected from the group consisting of hydrogen, hydroxyl, cyano, fluorine, chlorine, bromine, carboxyl, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl , pentyl, hexane, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentaneoxy, hexaneoxy Base, aminomethyl, aminoethyl, aminopropyl, aminobutylalkyl, aminopentyl, aminohexane, carboxamide, acetylamino, n-propionyl, isopropylamide, n-butyryl , isobutyryl amide, tert-butyryl amide, pentane amide, hexane amide, carbamoyl, acetyl, n-alanyl, isopropyl, n-butyryl, isobutyryl, uncle Butyryl, pentyl, hexyl, C ₃ nitrogen heterocyclyl, C ₄ nitrogen heterocyclyl, C ₅ nitrogen heterocyclyl, C ₆ nitrogen heterocyclyl, C ₃ nitrogen heterocycle Alkenyl, C ₄ azacycloalkenyl, C ₅ azacycloalkenyl, C ₆ azacycloalkenyl, phenoxy, phenyl, substituted phenyl, piperazinyl, methylpiperazine Base, ethyl piperazinyl, propyl piperazine , D piperazinyl, piperazinyl amyl or hexyl alkyl piperazinyl group;

R ₅ and R _{6 are} selected from fluorine, chlorine, bromine or iodine.

further,

In step i, the compound IM-6b is:

In step ii, the compound IM-8b is:

Further, in the steps i to iii, the ether solvent is selected from any one or more selected from the group consisting of tetrahydrofuran, dioxane, diethyl ether, tert-butyl methyl ether, diisopropyl ether and dibutyl ether; The solvent is selected from any one or more selected from the group consisting of dichloromethane, ethyl chloride, dichloroethane, chloroform, and carbon tetrachloride; and the ester solvent is selected from ethyl acetate and ethyl formate. Any one or two or more kinds; the alcohol solvent is selected from any one or two or more selected from the group consisting of methanol, ethanol, n-propanol, and isopropanol.

The present invention also provides another preparation method of the pyrrole amide compound represented by the above formula I,

When R ₄ is an amino-substituted phenyl group, its synthetic route is:

Wherein, Boc represents a tert-butoxycarbonyl group; TFA represents trifluoroacetic acid; and HATU represents 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate; DIEA stands for N,N-diisopropylethylamine; LiOH stands for lithium hydroxide;

R _{1 is} selected from the group consisting of hydrogen, hydroxy, cyano, halogen, carboxyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₂ -C ₆ amide, C ₁ -C ₆ amino An alkyl group, a C ₂ -C ₆ aminoacyl group, a C ₃ -C ₆ heterocyclic group, a C ₃ -C ₆ heterocycloalkenyl group, a phenoxy group, a phenyl group or a substituted phenyl group;

R ₂ and R ₃ are respectively or simultaneously selected from the group consisting of hydrogen, hydroxy, cyano, halogen, carboxyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ aminoalkyl, C ₂ -C ₆ amide group, C ₂ -C ₆ aminoacyl group, C ₃ -C ₆ heterocyclic group, C ₃ -C ₆ heterocycloalkenyl group, phenoxy group, phenyl group, substituted phenyl group , piperazinyl or substituted piperazinyl;

R _{5 is} selected from halogen;

Includes the following steps:

1. At 0 ° C to 5 ° C, the compound IM-1c is dissolved in a halogen hydrocarbon solvent, trifluoroacetic acid is added, and the reaction is stirred at 20 ° C to 30 ° C for 2 h to 12 h to obtain a reaction liquid; the reaction liquid is concentrated to obtain a yellow oil, which is the compound IM-2c;

The mass ratio of the compound IM-1c to the halocarbon solvent is 1:5-20 g/ml; the mass ratio of the compound IM-1c to trifluoroacetic acid is 1:2-10 g/ml;

2. The compound IM-2c, the compound IM-3c, the triethylamine and the halocarbon solvent obtained in the step 1 are stirred at 25 ° C to 30 ° C for 1 h to 10 h, and then concentrated to obtain a crude product; the crude product is purified by column chromatography. Obtaining compound IM-4c;

The molar ratio of the compound IM-2c, the compound IM-3c, and the triethylamine is 1:1 to 2:1 to 5; the compound The mass to volume ratio of IM-2c to the halocarbon solvent is 1:50 to 100 g/ml;

3. The compound IM-4c obtained in the second step, a mixed solvent of lithium hydroxide and an ether solvent/water, and stirred at 20 ° C to 30 ° C for 2 h to 16 h to obtain a reaction liquid; separated and purified to obtain a compound IM-5c. ;

The molar ratio of the compound IM-4c to lithium hydroxide is 1:1 to 10; the mass ratio of the compound IM-4c to the mixed solvent is 1:55 to 60 g/ml; in the mixed solvent, the ether The volume ratio of solvent to water is 1 to 5:1;

4. Compound IM-5c, 1,2-diaminobenzene, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate obtained in step 3. The ester, N,N-diisopropylethylamine and a halogenated hydrocarbon solvent are stirred at 25 ° C to 30 ° C for 12 h to 16 h to obtain a reaction liquid; isolated and purified to obtain a compound of the formula I;

The compound IM-5c, 1,2-diaminobenzene, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluron hexafluorophosphate, N, The molar ratio of N-diisopropylethylamine is 1:0.8 to 2:0.8 to 2:1.5 to 4; and the mass ratio of the compound IM-5c to the halogen hydrocarbon solvent is 1:40 to 100 g/ml.

further,

1. At 0 ° C, the compound IM-1c is dissolved in a halogen hydrocarbon solvent, trifluoroacetic acid is added, and the reaction mixture is stirred at 25 ° C for 2 h to obtain a reaction liquid; the reaction liquid is concentrated to obtain a yellow oil, which is a compound IM. -2c;

The mass to volume ratio of the compound IM-1c to the halocarbon solvent is 1:20 g/ml; the mass to volume ratio of the compound IM-1c to trifluoroacetic acid is 1:8 g/ml;

2. The compound IM-2c, the compound IM-3c, the triethylamine and the halogen hydrocarbon solvent obtained in the step 1 are stirred and reacted at 25 ° C for 2 hours, and then concentrated to obtain a crude product; the crude product is purified by column chromatography to give the compound IM-4c. ;

The molar ratio of the compound IM-2c, the compound IM-3c, and the triethylamine is 1:1.1 to 1.2:2.5 to 3; the mass to volume ratio of the compound IM-2c to the halocarbon solvent is 1:65 to 70 g. /ml;

3, the compound IM-4c obtained in step 2, lithium hydroxide and ether solvent / water mixed solvent, stirring reaction at 25 ° C for 2h ~ 16h, the reaction liquid is obtained; isolated, purified, to obtain the compound IM-5c;

The molar ratio of the compound IM-4c to lithium hydroxide is 1:4.5; the mass to volume ratio of the compound IM-4c to the mixed solvent is 1:55-60 g/ml; in the mixed solvent, the ether solvent and The volume ratio of water is 3:1;

4. Compound IM-5c, 1,2-diaminobenzene, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate obtained in step 3. The ester, N,N-diisopropylethylamine and a halogenated hydrocarbon solvent are stirred at 25 ° C for 12 h to 16 h to obtain a reaction liquid; isolated and purified to obtain a compound of the formula I;

The compound IM-5c, 1,2-diaminobenzene, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluron hexafluorophosphate, N, The molar ratio of N-diisopropylethylamine is 1:0.8 to 1.2:0.8 to 1.2:1.5 to 2; the mass to volume ratio of the compound IM-5c to the halocarbon solvent is 1:40 to 45 g/ml.

further,

R _{1 is} selected from the group consisting of hydrogen, hydroxy, cyano, fluoro, chloro, bromo, carboxy, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentylene, hexyl Alkyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentyloxy, hexyloxy, aminomethyl, Aminoethyl, aminopropyl, aminobutylalkyl, aminopentyl, aminohexane, carboxamido, acetylamino, n-propionylamino, isopropylamide, n-butyryl, isobutyryl, Tert-butylamide, pentanoamide, hexane amide, methyl, acetyl, n-alanyl, isopropylyl, n-butyryl, isobutyryl, tert-butyryl, pentylamine Acyl group, hexyl group, nitrogen heterocyclic group of C ₃ , nitrogen heterocyclic group of C ₄ , nitrogen heterocyclic group of C ₅ , nitrogen heterocyclic group of C ₆ , azacycloheteroyl group of C ₃ , C ₄ Azacycloheteroenyl, C ₅ azacycloalkenyl, C ₆ azacycloalkenyl, phenoxy, phenyl or substituted phenyl;

R ₂ and R ₃ are respectively or simultaneously selected from the group consisting of hydrogen, hydroxyl, cyano, fluorine, chlorine, bromine, carboxyl, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl , pentyl, hexane, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentaneoxy, hexaneoxy Base, aminomethyl, aminoethyl, aminopropyl, aminobutylalkyl, aminopentyl, aminohexane, carboxamide, acetylamino, n-propionyl, isopropylamide, n-butyryl , isobutyryl amide, tert-butyryl amide, pentane amide, hexane amide, carbamoyl, acetyl, n-alanyl, isopropyl, n-butyryl, isobutyryl, uncle Butyryl group, valyl group, hexyl group, nitrogen heterocyclic group of C ₃ , nitrogen heterocyclic group of C ₄ , nitrogen heterocyclic group of C ₅ , nitrogen heterocyclic group of C ₆ , nitrogen heterocyclic ring of C ₃ Alkenyl, C ₄ azacycloalkenyl, C ₅ azacycloalkenyl, C ₆ azacycloalkenyl, phenoxy, phenyl, substituted phenyl, piperazinyl, methylpiperazine Base, ethyl piperazinyl, propyl piperazinyl , butyryl piperazinyl, pentyl piperazinyl or hexyl piperazinyl;

R _{5 is} selected from the group consisting of fluorine, chlorine, bromine or iodine.

Further, in step 2, the compound IM-3c is:

Further, in the steps 1 to 4, the halogen hydrocarbon solvent is selected from the group consisting of dichloromethane, ethyl chloride, dichloroethane, and trichlorochloride. Any one or two or more of methane and carbon tetrachloride; and the ether solvent is selected from any one or two of tetrahydrofuran, dioxane, diethyl ether, tert-butyl methyl ether, diisopropyl ether, and dibutyl ether. More than one species.

The present invention also provides the use of the above-mentioned pyrrole amide compound or a pharmaceutically acceptable salt, crystal form, hydrate or solvate thereof for the preparation of a histone deacetylase inhibitor.

The histone deacetylase inhibitor drug is a drug for preventing and/or treating a disease caused by abnormal histone deacetylase activity.

Further, the disease is any one or more of a cell proliferative disease, an autoimmune disease, an inflammation, a neurodegenerative disease, or a viral disease.

Further, the disease is cancer.

The present invention also provides a pharmaceutical composition for inhibiting histone deacetylase activity, which comprises the above-mentioned pyrrole amide compound or a pharmaceutically acceptable salt, crystal form, hydrate or solvate thereof as an active ingredient. A preparation prepared by adding pharmaceutically acceptable excipients or auxiliary ingredients.

Further, the preparation includes an orally administered preparation, a sublingual preparation, a buccal preparation, a transdermal absorption preparation or an injection preparation.

The novel compound of the formula I provided by the present invention exhibits good deacetylase inhibitory activity and has the potential to prevent and/or treat diseases caused by abnormal histone deacetylase activity; The compound has good inhibitory activity against different liver cancer cells and has a prospect of clinical application. The present invention provides the following compounds 1 to 11 having good deacetylase inhibitory activity, as shown in Table 1:

Table 1. Compounds 1 to 11 prepared by the present invention

Histone deacetylase plays an important role in gene transcription and regulation, signal transduction, growth and development, differentiation and apoptosis, metabolic diseases and tumors. If the histone deacetylase activity is abnormal, it will trigger a series of abnormalities in histone deacetylase activity. These include cell proliferative diseases, autoimmune diseases, inflammation, neurodegenerative diseases, viral diseases (for example, a review of diseases applicable to HDAC6 inhibitors in World Patent WO2011011186).

The compounds and derivatives provided in the present invention may be named according to the IUPAC (International Union of Pure and Applied Chemistry) or CAS (Chemical Abstracts Service, Columbus, OH) nomenclature system.

Definitions of terms of use in connection with the present invention: Unless otherwise stated, the initial definitions provided herein by the group or term apply to the group or term of the entire specification; for terms not specifically defined herein, it should be based on the disclosure and context. Given the meanings that those skilled in the art can give to them.

"Substitution" means that a hydrogen atom in a molecule is replaced by a different atom or molecule.

The minimum and maximum values of the carbon atom content in the hydrocarbon group are represented by a prefix, for example, the prefix (Ca to b) alkyl group indicates any alkyl group having "a" to "b" carbon atoms. Thus, for example, C1-4 alkyl refers to an alkyl group containing from 1 to 4 carbon atoms.

The term "pharmaceutically acceptable" means that a carrier, carrier, diluent, adjuvant, and/or salt formed is generally chemically or physically compatible with the other ingredients that constitute a pharmaceutical dosage form, and is physiologically Compatible with the receptor.

The terms "salt" and "pharmaceutically acceptable salt" refer to the above-mentioned compounds or stereoisomers thereof, acid and/or basic salts formed with inorganic and/or organic acids and bases, and also includes zwitterionic salts (within Salts) also include quaternary ammonium salts such as alkylammonium salts. These salts can be obtained directly in the final isolation and purification of the compounds. It can also be obtained by mixing the above compound, or a stereoisomer thereof, with a certain amount of an acid or a base as appropriate (for example, an equivalent amount). These salts may be precipitated in a solution and collected by filtration, or recovered after evaporation of the solvent, or may be obtained by lyophilization after reaction in an aqueous medium. The salt in the present invention may be a hydrochloride, a sulfate, a citrate, a besylate, a hydrobromide, a hydrofluoride, a phosphate, an acetate, a propionate or a dibutyl compound. An acid salt, an oxalate salt, a malate salt, a succinate salt, a fumarate salt, a maleate salt, a tartrate salt or a trifluoroacetate salt.

In certain embodiments of the invention, the invention includes isotopically labeled compounds, which are the same as the compounds listed herein, but wherein one or more of the atoms are replaced by another atom, the atomic The atomic mass or mass number is different from the atomic mass or mass number that is common in nature. Isotopes which may be introduced into the compounds of formula (I) include hydrogen, carbon, nitrogen, oxygen, sulfur, i.e., ² H, ³ H, ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁷ O, ¹⁸ O, ³⁵ S. Compounds of formula (I) and stereoisomers thereof containing such isotopes and/or other atomic isotopes, as well as pharmaceutically acceptable salts of such compounds, stereoisomers, are intended to be encompassed within the scope of the invention.

The key intermediates and compounds in the present invention are isolated and purified in a manner common to separation and purification methods in organic chemistry and examples of such methods include filtration, extraction, drying, spin drying, and various types of chromatography. Alternatively, the intermediate can be subjected to the next reaction without purification.

In certain embodiments, one or more compounds of the invention may be used in combination with one another. Alternatively, the compounds of the invention may be used in combination with any other active agent for the preparation of a medicament or pharmaceutical composition that modulates cellular function or treats a disease. If a group of compounds is used, the compounds can be administered to the subject simultaneously, separately or sequentially.

The mode of administration of the compound or pharmaceutical composition of the present invention is not particularly limited, and representative modes of administration include, but are not limited to, oral, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or mixed with: (a) a filler or compatibilizer, for example, Starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and gum arabic; (c) humectants, For example, glycerin; (d) a disintegrant such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) a slow solvent such as paraffin; (f) Absorbing accelerators, for example, quaternary amine compounds; (g) wetting agents, such as cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, hard Calcium citrate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or a mixture thereof. In capsules, tablets and pills, the dosage form may also contain a buffer.

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 known in the art. They may contain opacifying agents and the release of the active compound or compound in such compositions may be released in a portion of the digestive tract in a delayed manner. Examples of embedding components that can be employed are polymeric and waxy materials. If necessary, the active compound may also be in microencapsulated form with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or elixirs. In addition to the active compound, the liquid dosage form may contain inert diluents conventionally employed in the art, such as water or other solvents, solubilizers and emulsifiers, for example, 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 a mixture of these substances.

In addition to these inert diluents, the compositions may contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents and perfumes.

In addition to the active compound, the suspension may contain suspending agents, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar or mixtures of these and the like.

Compositions for parenteral injection may comprise a physiologically acceptable sterile aqueous or nonaqueous solution, dispersion, suspension or emulsion, and a sterile powder for reconstitution into a sterile injectable solution or dispersion. Suitable aqueous and nonaqueous vehicles, diluents, solvents or vehicles include water, ethanol, polyols, and suitable mixtures thereof.

Dosage forms for the compounds of the invention for topical administration include ointments, powders, patches, propellants and inhalants. The active ingredient is admixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or, if necessary, propellants.

The pharmaceutically acceptable excipient of the present invention means a substance which is contained in a dosage form in addition to the active ingredient.

The pharmaceutically acceptable auxiliary component of the present invention has certain physiological activity, but the addition of the component does not change the dominant position of the above pharmaceutical composition in the course of disease treatment, but only plays an auxiliary effect, and the auxiliary effects are only It is the utilization of the known activity of the component, and is an auxiliary treatment method conventionally used in the medical field. It is still within the scope of the present invention to use the above auxiliary ingredients in combination with the pharmaceutical composition of the present invention.

The compound of the present invention has the functions of inducing differentiation, immunoregulation, hindering cell cycle, promoting apoptosis, and good HDAC subtype selectivity, and aims to have better curative effect on various cancers while overcoming the current Side effects of HDAC inhibitors such as anemia, ischemic stroke, deep vein thrombosis, thrombocytopenia, and vomiting.

The compound of the present invention has HDAC inhibitory activity and can be used for treating diseases associated with abnormal HDAC activity, and particularly has excellent effects on liver cancer.

It is apparent that various other modifications, substitutions and changes can be made in the form of the above-described embodiments of the present invention.

The above content of the present invention will be further described in detail below by way of specific embodiments in the form of embodiments. However, the scope of the above-mentioned subject matter of the present invention should not be construed as being limited to the following examples. Any technique implemented based on the above description of the present invention is within the scope of the present invention.

detailed description

The raw materials and equipment used in the specific embodiments of the present invention are known products and are obtained by purchasing commercially available products.

In the present invention, Boc represents a tert-butoxycarbonyl group; TFA represents trifluoroacetic acid; Fmoc-Cl represents an anthraceneoxycarbonyl chloride; and HATU represents 2-(7-azobenzotriazole)-N,N,N',N. '-Tetramethylurea hexafluorophosphate; DIEA stands for N,N-diisopropylethylamine; DCM stands for dichloromethane.

Example 1. N-Hydroxy-1-(4'-methoxy-[1,1'-biphenyl]-4-sulfonyl) 2,5-dihydro-1H-pyrrole-3-carboxamide Preparation of 1,2,5-dihydro-1H-pyrrole-3-carboxylic acid

Ethyl N-tert-butoxycarbonyl-2,5-dihydro-1H-pyrrole-3-carboxylate (8.5 g, 36 mmol; manufacturer: 韶远科技 (Shanghai) Co., Ltd.) was dissolved in 60 mL of tetrahydrofuran and 30 mL of water. After mixing the solution, lithium hydroxide (4.2 g, 176 mmol) was added, and the reaction was stirred at 25 ° C for 2 hours, and the organic solvent was removed in vacuo to give a residue. The residue was diluted with water and adjusted to pH = 5 with 1 N hydrochloric acid. The solid was precipitated and filtered to give a solid. EtOAcjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj

MS (ESI) m / z214 ( M + 1) +.

In an ice bath, N-tert-butoxycarbonyl-2,5-dihydro-1H-pyrrole-3-carboxylic acid (7 g, 36 mmol) was dissolved in 70 mL of dichloromethane, then 30 mL of trifluoroacetic acid was added dropwise and stirred. The mixture was slowly stirred to 25 ° C and the reaction was stirred for 2 h to obtain a reaction mixture. The reaction mixture was concentrated to give a yellow oil, 2,5-dihydro-1H-pyrrole-3-carboxylic acid (4.0 g, 99% yield) .

MS (ESI) m / z114 ( M + 1) +.

2. Preparation of N-methoxycarbonyl-2,5-dihydro-1H-pyrrole-3-carboxylic acid

2,5-Dihydro-1H-pyrrole-3-carboxylic acid (4.0 g, 35.4 mmol) was dissolved in 50 mL of tetrahydrofuran and 30 mL of water, then sodium carbonate (11.2 g, 106 mmol) and oxalyl chloride (9.2 g). , 35.4mmol; manufacturer: Alfa Aesar (China) Chemical Co., Ltd.), stirring at 25 ° C overnight; after the end of the reaction, add 200mL of water to dilute, adjust the pH with 2N hydrochloric acid, extract ethyl acetate, and combine organic The organic phase was dried <RTI ID=0.0></RTI> to <RTI ID=0.0></RTI> to <RTI ID=0.0></RTI> <RTIgt;

MS (ESI) m / z336 ( M + 1) +.

3. Preparation of N-methoxycarbonyl-2,5-dihydro-1H-pyrrole-3-(tetrahydropyran-2-oxo)-carboxamide

N-Methoxycarbonyl-2,5-dihydro-1H-pyrrole-3-carboxylic acid (11.0 g, 32.8 mmol) was dissolved in 100 mL of dichloromethane, and then O-(tetrahydro-2H-pyran- 2-yl)hydroxylamine (4.2 g, 36 mmol), HATU (15 g, 39.4 mmol), DIEA (12.8 g, 98.4 mmol; manufacturer: Belling Technology Co., Ltd.), stirring at 25 ° C overnight to obtain a reaction solution; The reaction mixture was diluted with 50 mL of water and extracted with ethyl acetate (50 mL×2). The ethyl acetate phase was combined, and the ethyl acetate phase was dried, filtered and concentrated to give a crude product. White solid N-methoxycarbonyl-2,5-dihydro-1H-pyrrole-3-(tetrahydropyran-2-oxo)-carboxamide (12 g, 48% yield).

MS (ESI) m / z435 ( M + 1) +.

Preparation of 4,5,5-dihydro-1H-pyrrole-3-(tetrahydropyran-2-oxo)-carboxamide

N-Methoxycarbonyl-2,5-dihydro-1H-pyrrole-3-(tetrahydropyran-2-oxo)-carboxamide (10 g, 23 mmol) was dissolved in 100 mL of DMF then 20 mL of piperidine The reaction was stirred at 25 ° C for 4 hours, then diluted with 800 mL of water, extracted with ethyl acetate, and the organic phase was combined. The organic phase was dried, filtered and concentrated to give a white solid 2,5-dihydro-1H-pyrrole-3- (tetrahydropyran-2-oxo)-carboxamide (4.5 g, 92% yield).

MS (ESI) m / z213 ( M + 1) +.

Preparation of 1-((4-benzonitrile)sulfonyl)-N-hydroxy-2,5-dihydro-1H-pyrrole-3-carboxamide

2,5-Dihydro-1H-pyrrole-3-(tetrahydropyran-2-oxo)-carboxamide (100 mg, 0.5 mmol) and triethylamine (66 mg, 0.7 mmol) were dissolved in dichloromethane (8 mL) In the reaction solution, 4'-methoxy-[1,1'-biphenyl]-4-sulfonyl chloride (113 mg, 0.6 mmol; manufacturer: BEHRINGER TECHNOLOGY CO., LTD.) was added to the reaction solution at 25 ° C, 25 After stirring at °C for 2 hours, thick The solvent is removed to give a crude material; the crude material is purified by column chromatography to give N-((tetrahydro-2H-pyran-2-yl)oxy)-1-(4'-methoxy-[1,1'- Dibiphenyl]-4-sulfonyl) 2,5-dihydro-1H-pyrrole-3-carboxamide, white solid.

N-((tetrahydro-2H-pyran-2-yl)oxy)-1-(4'-methoxy-[1,1'-biphenyl]-4-sulfonyl group under ice bath 2,5-Dihydro-1H-pyrrole-3-carboxamide (80 mg, 0.3 mmol) was dissolved in 5 mL of dichloromethane solution, then 2 mL of trifluoroacetic acid was added dropwise, stirred, and slowly raised to 25 ° C to continue stirring. After 2 h, the solvent was concentrated to give a crude material. EtOAc m. m. 2,5-Dihydro-1H-pyrrole-3-carboxamide (29 mg, 7% yield).

MS (ESI) m / z 375 (M + 1) +.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.81 (br s, 1H), 9.05 (br s, 1H), 7.90-7.85 (m, 4H), 7.73-7.71 (m, 2H), 7.08-7.06 ( m, 2H), 6.34 (s, 1H), 4.20 (s, 4H), 3.82 (s, 3H).

Example 2 Preparation of N-Hydroxy-1-((4-(4-fluorophenyl)phenyl)sulfonyl) 2,5-dihydro-1H-pyrrole-3-carboxamide

2,5-Dihydro-1H-pyrrole-3-(tetrahydropyran-2-oxo)-carboxamide (100 mg, 0.5 mmol) and 4-(4-fluorophenyl)benzenesulfonyl chloride (162 mg, 0.6) Methyl acetate N-hydroxy-1-((4-(4-fluorophenyl)phenyl)sulfonyl) 2 was obtained as a starting material according to a similar procedure as in Example 1. 5-Dihydro-1H-pyrrole-3-carboxamide (32 mg, 18% yield).

MS (ESI) m / z 363 (M + 1) +.

¹ H NMR (400 Hz, DMSO-D ₆ ) δ = 10.83 (s, 1H), 9.02 (s, 1H), 7.94-7.89 (m, 4H), 7.84-7.80 (m, 2H), 7.38-7.33 (m, 2H), 6.35 (s, 1H), 4.22 (s, 4H).

Example 3 Preparation of N-Hydroxy-1-((4-(4-tert-butylphenyl)phenyl)sulfonyl) 2,5-dihydro-1H-pyrrole-3-carboxamide

2,5-Dihydro-1H-pyrrole-3-(tetrahydropyran-2-oxo)-carboxamide (100 mg, 0.5 mmol) and 4-(4-tert-butylphenyl)benzenesulfonyl chloride (184 mg) , 0.6 mmol; manufacturer: Belling Technology Co., Ltd.) as a raw material, according to the similar procedure in Example 1 to obtain a white solid N-hydroxy-1-((4-(4-tert-butylphenyl)phenyl)sulfonate Acyl) 2,5-dihydro-1H-pyrrole-3-carboxamide (23 mg, 12% yield).

MS (ESI) m / z 401 (M + 1) +.

¹ H NMR (400 Hz, DMSO-D ₆ ) δ = 10.83 (s, 1H), 9.03 (s, 1H), 7.92-7.87 (m, 4H), 7.77 (d, J = 8.4 Hz, 2H), 7. , J = 8.4 Hz, 2H), 6.35 (s, 1H), 4.22 (s, 4H).

Example 4 Preparation of N-Hydroxy-1-((4-(4-morpholine-N-phenyl)phenyl)sulfonyl) 2,5-dihydro-1H-pyrrole-3-carboxamide

2,5-Dihydro-1H-pyrrole-3-(tetrahydropyran-2-oxo)-carboxamide (100 mg, 0.5 mmol) and 4-(4-morpholine-N-phenyl)benzenesulfonyl chloride (202mg, 0.6mmol; manufacturer: Belling Technology Co., Ltd.) as a raw material, according to the similar procedure in Example 1 to obtain a white solid N-hydroxy-1-((4-(4-morpholine-N-phenyl)) Phenyl)sulfonyl) 2,5-dihydro-1H-pyrrole-3-carboxamide (45 mg, 21% yield).

MS (ESI) m / z 430 (M + 1) +.

¹ H NMR (400 Hz, DMSO-D ₆ ) δ = 10.83 (s, 1H), 9.03 (s, 1H), 7.87-7.82 (m, 4H), 7.66 (d, J = 8.4 Hz, 2H), 7.05 (d) , J = 8.4 Hz, 2H), 6.35 (s, 1H), 4.18 (s, 4H), 3.76-3.74 (m, 4H), 3.51-3.50 (m, 4H).

Example 5 Preparation of N-Hydroxy-1-(((3-methyl-4-phenyl)phenyl)sulfonyl) 2,5-dihydro-1H-pyrrole-3-carboxamide

2,5-Dihydro-1H-pyrrole-3-(tetrahydropyran-2-oxo)-carboxamide (100 mg, 0.5 mmol) and (3-methyl-4-phenyl)benzenesulfonyl chloride (177 mg) , 0.6 mmol; manufacturer: Belling Technology Co., Ltd.) as a raw material, according to the similar procedure in Example 1, to obtain a white solid N-hydroxy-1-(((3-methyl-4-phenyl)phenyl)sulfonate Preparation of acyl) 2,5-dihydro-1H-pyrrole-3-carboxamide (33 mg, 18% yield).

MS (ESI) m / z 359 (M + 1) +.

¹ H NMR (400 Hz, DMSO-D ₆ ) δ = 10.85 (s, 1H), 9.04 (s, 1H), 7.78 (s, 1H), 7.72-7.70 (m, 1H), 7.50-7.39 (m, 6H) , 6.37 (s, 1H), 4.23 (s, 4H), 2.33 (s, 3H).

Example 6. Preparation of N-hydroxy-1-(((3-methyl-4-p-fluorophenyl)phenyl)sulfonyl) 2,5-dihydro-1H-pyrrole-3-carboxamide

2,5-Dihydro-1H-pyrrole-3-(tetrahydropyran-2-oxo)-carboxamide (100 mg, 0.5 mmol) and (3-methyl-4-p-fluorophenyl)benzenesulfonyl chloride (177 mg, 0.6 mmol; manufacturer: Belling Technology Co., Ltd.) as a raw material, according to the similar procedure in Example 1, a white solid N-hydroxy-1-(((3-methyl-4-phenyl)phenyl) Preparation of sulfonyl) 2,5-dihydro-1H-pyrrole-3-carboxamide (29 mg, 15% yield).

MS (ESI) m / z 377 (M + 1) +.

¹ H NMR (400 Hz, DMSO-D ₆ ) δ = 10.85 (s, 1H), 9.04 (s, 1H), 7.78 (s, 1H), 7.71-7.69 (m, 1H), 7.48-7.45 (m, 3H) , 7.33 - 7.29 (d, J = 8.0 Hz, 2H), 6.37 (s, 1H), 4.23 (s, 4H), 2.32 (s, 3H).

Example 7. N-Hydroxy-1-((3'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl)sulfonyl)-2,5 -Preparation of dihydro-1H-pyrrole-3-carboxamide

1. Preparation of 1-((4-bromophenyl)sulfonyl)-2,5-dihydro-1H-pyrrole-3-(tetrahydropyran-2-oxo)-carboxamide

2,5-Dihydro-1H-pyrrole-3-(tetrahydropyran-2-oxo)-carboxamide (2.1 g, 10 mmol) was dissolved in a mixed solution of 50 mL of tetrahydrofuran and 50 mL of water, then p-bromobenzene was added. Sulfonyl chloride (2.6 g, 10 mmol). Additional sodium bicarbonate (2.6 g, 30 mmol) was added. The solution was stirred at room temperature for 1 hour. After the end of the reaction, the organic solvent was evaporated in vacuo. EtOAc m. 5-Dihydro-1H-pyrrole-3-(tetrahydropyran-2-oxo)-carboxamide (3.8 g, yield 88.2%).

MS (ESI) m / z 431 (M + 1) +.

¹ H NMR (400 MHz, CD 3 OD) δ = 11.38 (br, 1 H), 7.86 (d, J = 4.0 Hz, 2H), 7.84 (d, J = 4.0 Hz, 2H), 6.42 (s, 1H), 4.82 (s) , 1H), 4.21-4.19 (m, 4H), 4.03-4.01 (m, 1H), 3.49 (s, 1H), 1.62 (s, 3H), 1.50 (s, 3H).

2. N-Hydroxy-1-((3'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl)sulfonyl)-2,5-di Preparation of hydrogen-1H-pyrrol-3-(tetrahydropyran-2-oxo)-carboxamide

Dissolving 1-((4-bromophenyl)sulfonyl)-2,5-dihydro-1H-pyrrole-3-(tetrahydropyran-2-oxo)-carboxamide (431 g, 1.0 mmol) 10 mL of dioxane and 2 mL of water, then sodium carbonate (212 mg, 2 mmol,) and 1-((3'-(4-methylpiperazin-1-yl)-phenylboronic acid pinacol ester (302 mg) , 1.0 mmol, manufacturer: BEHRINGER TECHNOLOGY CO., LTD.), followed by [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (50 mg). The solution was replaced with nitrogen three times and heated to The reaction was carried out at 80 ° C for 2 hours. After the reaction was completed, the reaction mixture was evaporated to dryness with EtOAc. N-hydroxy-1-((3'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl)sulfonyl)-2,5-dihydro- 1H-pyrrole-3-(tetrahydropyran-2-oxo)-carboxamide (250.00 mg, 47.5% yield).

MS (ESI) m / z 527 (M + 1) +.

3. N-Hydroxy-1-((3'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl)sulfonyl)-2,5-di Preparation of hydrogen-1H-pyrrole-3-carboxamide

N-Hydroxy-1-((3'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl)sulfonyl)-2,5-dihydro -1H-pyrrole-3-(tetrahydropyran-2-oxo)-carboxamide (250 mg, 0.48 mmol) was dissolved in 5 mL of methanol, and 1N HCl solution (1.0 ml) was added dropwise to the solution. After stirring for 2 hours, the reaction was completed and filtered to give a white solid N-hydroxy-1-((3'-(4-methylpiperazin-1-yl)-[1,1'-biphenyl]-4-yl Sulfonyl)-2,5-dihydro-1H-pyrrole-3-carboxamide (46 mg, yield 13.3%).

MS (ESI) m / z 443 (M + 1) +.

¹ H NMR (400 Hz, DMSO-D ₆ ) δ = 10.98 (br, 1H), 7.95-7.88 (m, 4H), 7.42 - 7.38 (t, J = 8.0 Hz 1H), 7.32 (s, 1H), 7.23 - 7.21. (d, J=8.0 Hz, 1H), 7.09-7.07 (m, 1H), 6.36 (s, 1H), 4.21 (s, 4H), 3.98-3.93 (m,, 2H), 3.50-3.43 (m, 2H), 3, 22-3.148 (m, 4H), 2.81 (s, 3H).

Example 8. N-Hydroxy-1-((4'-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-[1,1'-biphenyl]-4 Of -yl)sulfonyl)-2,5-dihydro-1H-pyrrole-3-carboxamide

2,5-Dihydro-1H-pyrrole-3-(tetrahydropyran-2-oxo)-carboxamide (300 mg, 1.41 mmol) and 1-((4'-(1-methyl-1,2) , 3,6-tetrahydropyridin-4-yl)phenylboronic acid pinacol ester (321 mg, 1.0 mmol, manufacturer: Belling Technology Co., Ltd.) as a raw material, according to the similar procedure in Example 7 to obtain a white solid N- Hydroxy-1-((4'-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-[1,1'-biphenyl]-4-yl)sulfonyl) -2,5-Dihydro-1H-pyrrole-3-carboxamide hydrochloride (22 mg, yield 6.6%).

MS (ESI) m / z 440 (M + 1) +.

¹ H NMR (400 Hz, DMSO-D ₆ ) δ = 10.86 (s, 1H), 10.84-10.72 (br, 1H), 9.01 (s, 1H), 7.98-7.96 (d, J = 8.8 Hz, 2H), 7.92 - 7.90 (d, J = 8.4 Hz, 2H), 7.81 - 7.79 (d, J = 8.4 Hz, 2H), 7.65 - 7.63 (d, J = 8.8 Hz, 2H) 6.37 (s, 1H), 6.32 (s , 1H), 4.23 (s, 4H), 4.02-3.82 (br, 2H), 3.75-3.52 (br, 2H), 2.86 (s, 5H).

Example 9. N-Hydroxy-1-((4'-(1-methylpiperidin-4-yl)-[1,1'-biphenyl]-4-yl)sulfonyl)-2,5 -Preparation of dihydro-1H-pyrrole-3-carboxamide

2,5-Dihydro-1H-pyrrole-3-(tetrahydropyran-2-oxo)-carboxamide (300 mg, 1.4 mmol) and 1-((4'-(1-methylpiperidin-4) -Based on methyl phenylboronic acid pinacol ester (321 mg, 1.0 mmol, manufacturer: Belling Technology Co., Ltd.) as a starting material, according to a similar procedure in Example 7 to obtain a white solid N-hydroxy-1-(4 '-(1-Methylpiperidin-4-yl)-[1,1'-biphenyl]-4-yl)sulfonyl)-2,5-dihydro-1H-pyrrole-3-carboxamide ( 24 mg, yield 10.3%).

MS (ESI) m / z 442 (M + 1) +.

¹ H NMR (400 Hz, DMSO-D ₆ ) δ = 10.82 (br, 1 H), 7.91 - 7.90 (d, J = 2.8 Hz, 4H), 7.75.7.73 (d, J = 8.4 Hz 2H), 7.40 - 7.38 ( d, J = 8.4 Hz, 2H), 6.36 (s, 1H), 4.22 (s, 4H), 3.20-3.02 (m, 2H), 2.90-2.84 (m, 1H), 2.75 (s, 3H), 2.20 -1.95 (m, 4H).

Example 10 N-Hydroxy-1-((4'-(4-methyl-2-oxopiperin-1-yl)-[1,1'-biphenyl]-4-yl)sulfonyl Preparation of-2,5-dihydro-1H-pyrrole-3-carboxamide

2,5-Dihydro-1H-pyrrole-3-(tetrahydropyran-2-oxo)-carboxamide (300 mg, 1.4 mmol) and 1-((4'-(4-methyl-2-oxo) The p-piperazin-1-yl)phenylboronic acid pinacol ester (321 mg, 1.0 mmol, manufacturer: Belling Technology Co., Ltd.) was used as a raw material, and a white solid N-hydroxy-1-(#) was obtained according to the similar procedure of Example 7. (4'-(4-Methyl-2-oxopiperin-1-yl)-[1,1'-biphenyl]-4-yl)sulfonyl)-2,5-dihydro-1H- Pyrrole-3-carboxamide hydrochloride (32 mg, yield 10.6%).

MS (ESI) m / z 457 (M + 1) +.

¹ H NMR (400 Hz, DMSO-D ₆ ) δ=11.82-11.70 (br, 1H), 10.86 (s, 1H), 9.10-8.92 (br, 1H), 9.01 (s, 1H), 7.97-7.91 (m, 4H), 7.85-7.83 (m, 2H), 7.52-7.49 (m, 2H), 6.36 (s, 1H), 4.23 (s, 4H), 4.02 (s, 2H), 3.70-3.45 (m, 2H) , 3.43 - 3.41 (m, 2H), 2.91 (s, 3H).

Example 11. N-Hydroxy-1-((4'-(4-methyl-3-oxopiperin-1-yl)-[1,1'-biphenyl]-4-yl)sulfonyl Preparation of-2,5-dihydro-1H-pyrrole-3-carboxamide

2,5-Dihydro-1H-pyrrole-3-(tetrahydropyran-2-oxo)-carboxamide (300 mg, 1.4 mmol) and 1-((4'-(4-methyl-3-oxo) The p-piperazin-1-yl)phenylboronic acid pinacol ester (321 mg, 1.0 mmol, manufacturer: Belling Technology Co., Ltd.) was used as a raw material, and a white solid N-hydroxy-1-(#) was obtained according to the similar procedure of Example 7. (4'-(4-Methyl-3-oxopiperin-1-yl)-[1,1'-biphenyl]-4-yl)sulfonyl)-2,5-dihydro-1H- Pyrrole-3-carboxamide hydrochloride (38 mg, yield 11.5%).

MS (ESI) m / z 457 (M + 1) +.

¹ H NMR (400 Hz, DMSO-D ₆ ) δ = 10.84 (s, 1H), 10.61-10.53 (br, 1H), 9.0 (s, 1H), 7.90-7.84 (m, 4H), 7.71-7.69 (d, J=9.2 Hz, 2H), 7.15-7.13 (d, J=8.8 Hz, 2H), 6.36 (s, 1H), 4.21 (s, 4H), 3.98-3.95 (d, J = 12.4 Hz, 2H), 3.58-3.55 (d, J = 11.6 Hz, 2H), 3.21-3.08 (m, 6H), 1.31-1.27 (t, J = 7.2 Hz, 3H).

In order to illustrate the beneficial effects of the present invention, the present invention provides the following test examples:

Test Example 1 Biological Activity Test

The HDA inhibitory activity of the compounds of the invention is tested in a substrate deacetylation assay.

A: Detection of enzymatic activity of deacetylase 6 (#50076, BPS Bioscience):

HDAC 6 removes the acetyl group on the substrate, allowing the substrate to activate, being able to act on the subsequently added chromogenic solution and releasing the fluorophore, the magnitude of which reflects the activity of HDAC 6. The IC50 detection method for this enzyme is in Chuping Xu, Elisabetta Soragni Improved Histone Deacetylase Inhibitors as Therapeutics for the Neurdegenerative DiseaseFriedreich’s Ataxia: A New Synthetic Route is disclosed. The total reaction system (100 μL/well) contained 0.35 ng/μL of HDAC 6, 20 μM substrate and various concentrations of compound. After incubating at 37 ° C for 30 minutes, the fluorescence signal was measured, and the inhibition of the compound was determined from the obtained data and plotted against the compound concentration to obtain a concentration response curve, and the IC50 value was fitted according to a four-parameter model.

B: Deacetylase 3 enzyme activity assay (#50003, BPS Bioscience):

HDAC 3 removes the acetyl group on the substrate, activates the substrate, acts on the chromogenic solution and releases the fluorophore, and the size of the fluorescent signal reflects the activity of HDAC 3. The IC50 detection method for this enzyme is disclosed in Chuping Xu, Elisabetta Soragni Improved Histone Deacetylase Inhibitors as Therapeutics for the Neurdegenerative Disease Friedreich's Ataxia: A New Synthetic Route. The total reaction system (100 μL/well) contained 0.16 ng/μL of HDAC 3, 10 μM substrate and various concentrations of compounds. Fluorescence signals were detected online at Ex/Em=360/460. The inhibition of the compound was determined from the obtained data and plotted against the compound concentration to obtain a concentration response curve, and the IC50 value was fitted according to a four-parameter model.

The enzymatic activities of deacetylase 6 (i.e., HDAC6) were tested on the compounds 1 to 11 prepared in the examples according to the above methods. The test results are shown in Table 2, wherein the IC50 of each compound was determined according to the description, in Table 2:

"+" indicates that the IC50 of HDAC6 is determined to be greater than 500 nM;

"++" means that the IC50 of HDAC6 is less than 300nM greater than 100nM;

“+++” means that the IC50 of HDAC6 is less than 100nM

The enzymatic activities of deacetylase 3 (i.e., HDAC3) were tested on the compounds 1 to 11 prepared in the examples according to the above methods. The test results are shown in Table 2, wherein the IC50 of each compound was determined according to the description, in Table 2:

"+" indicates that the IC50 of HDAC3 is determined to be greater than 1000 nM;

"++" means that the IC50 of HDAC3 is greater than 100nM and less than 1000nM;

"+++" means that the IC50 of HDAC3 is less than 100nM.

Table 2. Inhibitory activities of compounds 1 to 11 of the present invention against HDAC6 & HDAC3

Compound	Activity (HDAC6)	Activity (HDAC3)
1	+++	+++
2	+++	++
3	++	+
4	+	+
5	+++	++
6	+++	+

7	+++	+++
8	+++	+++
9	+++	+++
10	++	++
11	+++	++

The test results indicate that the compounds 1 to 11 of the present invention have good deacetylase inhibitory activity and have the potential of preventing and/or treating diseases caused by abnormal histone deacetylase activity.

Test Example 2 Cell assay - Cell growth inhibition assay

Materials and reagents

HepG2 cell line, Hep3B cell line, Huh7 cell line and Li7 cell line were purchased from Shanghai Institute of Biological Sciences, Chinese Academy of Sciences; DMEM high glucose medium and MEM medium were purchased from Hyclone; fetal bovine serum was purchased from Gibco; trypsin was purchased. From Invitrogen Shanghai; CCK-8 kit was purchased from Biyuntian Biotechnology Research Institute (beyotime); other cell culture dishes and other consumables were purchased from Corning China.

Cell preparation before compound action

HepG2 cells, Hep3B cells, Huh7 cells and Li7 cells in the logarithmic growth phase were digested with trypsin, and the cell suspension was counted as a uniform cell suspension. The cell density was adjusted to 1500 cells/well in a medium containing 10% serum, and re-inoculated into 96 cells. In a well cell culture plate, a culture volume of 200 μL was cultured at 37 ° C in a 5% CO ₂ incubator; the culture was carried out for 24 hours and used for the experiment.

Compound action

The cells cultured for 24 hours were taken out from the incubator, and the culture medium in the well plate was aspirated, and 200 μL of a compound solution prepared in a medium containing 10% fetal bovine serum was added to each well, and each concentration was 5 parallel, and DMSO was set as a negative. Control, CCK-8 detection was carried out by culturing at 37 ° C, 5% CO 2 culture for 72 hours.

CCK-8 detection

Take serum-free medium and CCK-8 solution and prepare CCK-8 working solution in a ratio of 10:1 (this process needs to be protected from light).

The cells cultured for 72 hours were taken out from the incubator, the culture medium in the well plate was aspirated, 120 μL of LCCK-8 working solution was added to each well, and 120 μL of LCCK-8 working solution was added to the cell-free well plate as a blank control at 37 ° C. Incubate for 1 hour in a 5% CO2 incubator (this process needs to be protected from light).

The well plates were removed from the incubator, and 100 μL of each solution was pipetted into a new 96-well plate, and the absorbance was read at 450 nm (this process needs to be protected from light).

data processing:

Tx: absorbance measured by CCK-8 after 72 hours of compound action

C: Negative control wells were cultured for 72 hours, and the absorbance measured by CCK-8

B: blank control well, absorbance measured by CCK-8

The compounds 1-11 prepared in the examples were run in the above assay. The test results are shown in Table 3, wherein the highest IC50 of one or more runs of each compound determined was classified according to the description, in Table 3:

"+" means that the IC50 of the compound in cancer cells is greater than 10 [mu]M;

"++" means that the IC50 of the compound in cancer cells is less than 10 [mu]M;

Table 3. Inhibitory activities of the compounds of the present invention 1 to 11 on different liver cancer cells

Compound	HepG2	Huh-7	Li-7	Hep3B
1	+	+	+	+
2	++	++	+	++
3	+	+	+	+
4	++	++	++	++
5	+	+	++	++
6	+	+	+	+
7	++	++	++	++
8	++	++	++	++
9	++	++	++	++
10	++	+	+	+
11	++	+	+	+

The test results show that the compounds 1-11 of the present invention have good inhibitory activity against different liver cancer cells (HepG2, Huh-7, Li-7, Hep3B), and have a prospect of clinical application.

In summary, the novel compound of Formula I provided by the present invention exhibits good deacetylase inhibitory activity and has the potential to prevent and/or treat diseases caused by abnormal histone deacetylase activity; At the same time, the novel compound of the invention has good inhibitory activity against different liver cancer cells, and has a prospect of clinical application.

Claims (24)

1. A pyrrolamide compound of the formula I or a pharmaceutically acceptable salt, crystal form, hydrate or solvate thereof:

   

   among them,

   R _{1 is} selected from the group consisting of hydrogen, hydroxy, cyano, halogen, carboxyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₂ -C ₆ amide, C ₁ -C ₆ amino An alkyl group, a C ₂ -C ₆ aminoacyl group, a C ₃ -C ₆ heterocyclic group, a C ₃ -C ₆ heterocycloalkenyl group, a phenoxy group, a phenyl group or a substituted phenyl group;

   R ₂ and R ₃ are respectively or simultaneously selected from the group consisting of hydrogen, hydroxy, cyano, halogen, carboxyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ aminoalkyl, C ₂ -C ₆ amide group, C ₂ -C ₆ aminoacyl group, C ₃ -C ₆ heterocyclic group, C ₃ -C ₆ heterocycloalkenyl group, phenoxy group, phenyl group, substituted phenyl group , piperazinyl or substituted piperazinyl;

   R _{4 is} selected from a hydroxy, thiol, amino substituted phenyl or epoxy ketone group.
2. The pyrrole amide compound according to claim 1, or a pharmaceutically acceptable salt, crystal form, hydrate or solvate thereof, which is characterized in that:

   R _{1 is} selected from the group consisting of hydrogen, hydroxy, cyano, fluoro, chloro, bromo, carboxy, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentylene, hexyl Alkyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentyloxy, hexyloxy, aminomethyl, Aminoethyl, aminopropyl, aminobutylalkyl, aminopentyl, aminohexane, carboxamido, acetylamino, n-propionylamino, isopropylamide, n-butyryl, isobutyryl, Tert-butylamide, pentanoamide, hexane amide, methyl, acetyl, n-alanyl, isopropylyl, n-butyryl, isobutyryl, tert-butyryl, pentylamine Acyl group, hexyl group, nitrogen heterocyclic group of C ₃ , nitrogen heterocyclic group of C ₄ , nitrogen heterocyclic group of C ₅ , nitrogen heterocyclic group of C ₆ , azacycloheteroyl group of C ₃ , C ₄ Azacycloheteroenyl, C ₅ azacycloalkenyl, C ₆ azacycloalkenyl, phenoxy, phenyl or substituted phenyl;

   R ₂ and R ₃ are respectively or simultaneously selected from the group consisting of hydrogen, hydroxyl, cyano, fluorine, chlorine, bromine, carboxyl, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl , pentyl, hexane, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentaneoxy, hexaneoxy Base, aminomethyl, aminoethyl, aminopropyl, aminobutylalkyl, aminopentyl, aminohexane, carboxamide, acetylamino, n-propionyl, isopropylamide, n-butyryl , isobutyryl amide, tert-butyryl amide, pentane amide, hexane amide, carbamoyl, ethoxyyl, n-alanyl, isopropyl, n-butyryl, isobutyryl, uncle Butyryl group, valyl group, hexyl group, nitrogen heterocyclic group of C ₃ , nitrogen heterocyclic group of C ₄ , nitrogen heterocyclic group of C ₅ , nitrogen heterocyclic group of C ₆ , nitrogen heterocyclic ring of C ₃ Alkenyl, C ₄ azacycloalkenyl, C ₅ azacycloalkenyl, C ₆ azacycloalkenyl, phenoxy, phenyl, substituted phenyl, piperazinyl, methylpiperazine Base, ethyl piperazinyl, propyl piperazine , D piperazinyl, piperazinyl amyl or hexyl alkyl piperazinyl group;

   R _{4 is} selected from hydroxy, decyl or amino substituted phenyl.
3. The pyrrole amide compound according to claim 1 or 2, or a pharmaceutically acceptable salt, crystal form, hydrate or solvate thereof, wherein the compound of the formula I is:

   
4. A method for preparing a pyrrole amide compound of the formula I, characterized in that:

   When R ₄ is a hydroxyl group, its synthetic route is:

   

   Wherein, Boc represents tert-butoxycarbonyl; TFA represents trifluoroacetic acid; Fmoc-Cl represents fluorenylmethoxycarbonyl chloride; and HATU represents 2-(7-azobenzotriazole)-N,N,N',N' -tetramethylurea hexafluorophosphate; DIEA stands for N,N-diisopropylethylamine; DCM stands for dichloromethane;

   R _{1 is} selected from the group consisting of hydrogen, hydroxy, cyano, halogen, carboxyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₂ -C ₆ amide, C ₁ -C ₆ amino An alkyl group, a C ₂ -C ₆ aminoacyl group, a C ₃ -C ₆ heterocyclic group, a C ₃ -C ₆ heterocycloalkenyl group, a phenoxy group, a phenyl group or a substituted phenyl group;

   R ₂ and R ₃ are respectively or simultaneously selected from the group consisting of hydrogen, hydroxy, cyano, halogen, carboxyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ aminoalkyl, C ₂ -C ₆ amide group, C ₂ -C ₆ aminoacyl group, C ₃ -C ₆ heterocyclic group, C ₃ -C ₆ heterocycloalkenyl group, phenoxy group, phenyl group, substituted phenyl group , piperazinyl or substituted piperazinyl;

   R ₅ is a halogen;

   Includes the following steps:

   a, compound IM-1a, lithium hydroxide and ether solvent / water mixed solvent, stirred at 20 ° C ~ 30 ° C for 1 h ~ 6h, remove the organic solvent, diluted with water, adjust the pH = 3 ~ 6, precipitated solid, Filtration to obtain a solid; washing the solid with water and drying to give N-tert-butoxycarbonyl-2,5-dihydro-1H-pyrrole-3-carboxylic acid;

   The molar ratio of the compound IM-1a to lithium hydroxide is 1:1 to 10; the compound IM-1a and the mixed solvent a mass to volume ratio of 1: 7 ~ 20g / ml; in the mixed solvent, the ether solvent and water volume ratio of 1 ~ 2: 1;

   b, 0 ° C ~ 5 ° C, the N-tert-butoxycarbonyl-2,5-dihydro-1H-pyrrole-3-carboxylic acid obtained in step a is dissolved in a halogen hydrocarbon solvent, added trifluoroacetic acid, at 20 After stirring at °C ~ 30 ° C for 2h ~ 12h, the reaction liquid is obtained; the reaction liquid is concentrated to obtain a yellow oil, which is the compound IM-2a;

   a mass to volume ratio of the N-tert-butoxycarbonyl-2,5-dihydro-1H-pyrrole-3-carboxylic acid to a halocarbon solvent: 1:5 to 20 g/ml; the N-tert-butoxycarbonyl-2 , the mass to volume ratio of 5-dihydro-1H-pyrrole-3-carboxylic acid to trifluoroacetic acid is 1:2 to 10 g/ml;

   c. The compound IM-2a obtained in step b, a mixed solvent of sodium carbonate, decanoyl chloride and ether solvent/water is stirred at 20 ° C to 30 ° C for 12 h to 16 h, diluted with water to adjust pH = 1 to 3 , ester solvent extraction, the organic phase is combined, the organic phase is dried, filtered, concentrated to give the compound IM-3a;

   The molar ratio of the compound IM-2a, sodium carbonate and decanoyl chloride is 1:1 to 5: 0.9 to 1.5; the mass ratio of the compound IM-2a to the mixed solvent is 1:10 to 25 g/ml; The volume ratio of the ether solvent to water in the mixed solvent is 1 to 2:1;

   d, compound IM-3a, O-(tetrahydro-2H-pyran-2-yl)hydroxylamine, 2-(7-azobenzotriazole)-N,N,N' obtained in step c, N'-tetramethylurea hexafluorophosphate, N,N-diisopropylethylamine and halogen hydrocarbon solvent, stirred at 25 ° C ~ 30 ° C for 12h ~ 16h, diluted with water, ester solvent extraction, combined The organic phase is dried, filtered and concentrated to obtain a crude product; the crude product is purified by column chromatography to give compound IM-4a;

   The compound IM-3a, O-(tetrahydro-2H-pyran-2-yl)hydroxylamine, 2-(7-azobenzotriazole)-N,N,N',N'-four The molar ratio of methyl urea hexafluorophosphate to N,N-diisopropylethylamine is 1:1 to 2:1 to 2:2 to 4; the mass volume of the compound IM-3a and the halogen hydrocarbon solvent The ratio is 1:9-20g/ml;

   e, the compound IM-4a obtained in the step d, the piperidine and the nitrogen-containing solvent are stirred at 25 ° C to 30 ° C for 4 h to 6 h, diluted with water, extracted with an ester solvent, and the organic phase is combined to dry the organic phase. Filtration and concentration to obtain compound IM-5a;

   The mass ratio of the compound IM-4a to piperidine is 1:1 to 4 g / ml; the mass ratio of the compound IM-4a to the nitrogen-containing solvent is 1: 5 to 20 g / ml;

   f, the compound IM-5a, the triethylamine, the compound IM-6a, and the halocarbon solvent in the step e are stirred at 25 ° C to 30 ° C for 1 h to 10 h, and then the solvent is removed to obtain a crude product; the crude product is purified by column chromatography. , obtaining compound TM-1a;

   The molar ratio of the compound IM-5a, triethylamine, and compound IM-6a is 1:1 to 5:1 to 2; the mass to volume ratio of the compound IM-5a to the halocarbon solvent is 1:50 to 100 g. /ml;

   g, 0 ° C ~ 5 ° C, the compound TM-1a obtained in step f is dissolved in a halocarbon solvent, trifluoroacetic acid, stirred at 25 ° C ~ 30 ° C for 1 h ~ 12h, the solvent is removed, to obtain a crude product; The crude product is purified by preparative high performance liquid chromatography to give the compound of formula I;

   The mass to volume ratio of the compound TM-1a to the halocarbon solvent is 1:50 to 100 g/ml; and the mass ratio of the compound TM-1a to trifluoroacetic acid is 1:10 to 50 g/ml.
5. The preparation method according to claim 4, wherein:

   a, compound IM-1a, lithium hydroxide and ether solvent / water mixed solvent, stirred at 25 ° C for 2h, remove the organic solvent, diluted with water, adjust the pH = 5, precipitate a solid, filtered to give a solid; Washing with water and drying to obtain N-tert-butoxycarbonyl-2,5-dihydro-1H-pyrrole-3-carboxylic acid;

   The molar ratio of the compound IM-1a to lithium hydroxide is 1:4.5 to 5; the mass to volume ratio of the compound IM-1a to the mixed solvent is 1:10 to 12 g/ml; in the mixed solvent, the ether The volume ratio of solvent to water is 2:1;

   b, at 0 ° C, the N-tert-butoxycarbonyl-2,5-dihydro-1H-pyrrole-3-carboxylic acid obtained in step a is dissolved in a halogen hydrocarbon solvent, trifluoroacetic acid is added, and the reaction is stirred at 25 ° C After 2h, the reaction liquid is obtained; the reaction liquid is concentrated to obtain a yellow oil, which is the compound IM-2a;

   a mass to volume ratio of the N-tert-butoxycarbonyl-2,5-dihydro-1H-pyrrole-3-carboxylic acid to a halocarbon solvent: 1:10 g/ml; the N-tert-butoxycarbonyl-2,5 - a mass to volume ratio of dihydro-1H-pyrrole-3-carboxylic acid to trifluoroacetic acid of 1:4 to 5 g/ml;

   c. The compound IM-2a obtained in step b, a mixed solvent of sodium carbonate, decanoyl chloride and ether solvent/water is stirred at 25 ° C for 12 h to 16 h, diluted with water, adjusted to pH = 1, ester solvent extraction , the organic phase is combined, the organic phase is dried, filtered, and concentrated to obtain the compound IM-3a;

   The molar ratio of the compound IM-2a, sodium carbonate and decyloxycarbonyl chloride is 1:3:1; the mass to volume ratio of the compound IM-2a to the mixed solvent is 1:20 g/ml; in the mixed solvent, The volume ratio of the ether solvent to water is 5:3;

   d, compound IM-3a, O-(tetrahydro-2H-pyran-2-yl)hydroxylamine, 2-(7-azobenzotriazole)-N,N,N' obtained in step c, N'-tetramethyluronium hexafluorophosphate, N,N-diisopropylethylamine and a halogenated hydrocarbon solvent are stirred at 25 ° C for 12 h to 16 h, diluted with water, extracted with an ester solvent, and combined with an organic phase. Drying the organic phase, filtering and concentrating to obtain a crude product; the crude product is purified by column chromatography to give compound IM-4a;

   The compound IM-3a, O-(tetrahydro-2H-pyran-2-yl)hydroxylamine, 2-(7-azobenzotriazole)-N,N,N',N'-four The molar ratio of methyl urea hexafluorophosphate to N,N-diisopropylethylamine is 1:1.1:1.2:3; the mass to volume ratio of the compound IM-3a to the halocarbon solvent is 1:9~ 10g/ml;

   e, the compound IM-4a obtained in step d, piperidine and a nitrogen-containing solvent are stirred at 25 ° C for 4 h to 6 h, diluted with water, extracted with an ester solvent, and the organic phase is combined, and the organic phase is dried, filtered and concentrated. , obtaining compound IM-5a;

   The mass ratio of the compound IM-4a to piperidine is 1:2g / ml; the mass ratio of the compound IM-4a to the nitrogen-containing solvent is 1:10g / ml;

   f, the compound IM-5a, the triethylamine, the compound IM-6a, and the halocarbon solvent in the step e are stirred at 25 ° C for 2 h, and then the solvent is removed to obtain a crude product; the crude product is purified by column chromatography to give the compound TM- 1a;

   The molar ratio of the compound IM-5a, triethylamine, and compound IM-6a is 1:1.4:1 to 1.2; the mass to volume ratio of the compound IM-5a to the halocarbon solvent is 1:80 g/ml;

   g, at 0 ° C, the compound TM-1a obtained in the step f is dissolved in a halogen hydrocarbon solvent, trifluoroacetic acid is added, and the reaction is stirred at 25 ° C for 2 h, then the solvent is removed to obtain a crude product; the crude product is purified by preparative high performance liquid chromatography. Obtaining a compound of formula I;

   The mass-to-volume ratio of the compound TM-1a to the halocarbon solvent is 1:60 to 65 g/ml; and the mass ratio of the compound TM-1a to trifluoroacetic acid is 1:25 g/ml.
6. The preparation method according to claim 4 or 5, characterized in that:

   R _{1 is} selected from the group consisting of hydrogen, hydroxy, cyano, fluoro, chloro, bromo, carboxy, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentylene, hexyl Alkyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentyloxy, hexyloxy, aminomethyl, Aminoethyl, aminopropyl, aminobutylalkyl, aminopentyl, aminohexane, carboxamido, acetylamino, n-propionylamino, isopropylamide, n-butyryl, isobutyryl, Tert-butylamide, pentanoamide, hexane amide, methyl, acetyl, n-alanyl, isopropylyl, n-butyryl, isobutyryl, tert-butyryl, pentylamine Acyl group, hexyl group, nitrogen heterocyclic group of C ₃ , nitrogen heterocyclic group of C ₄ , nitrogen heterocyclic group of C ₅ , nitrogen heterocyclic group of C ₆ , azacycloheteroyl group of C ₃ , C ₄ Azacycloheteroenyl, C ₅ azacycloalkenyl, C ₆ azacycloalkenyl, phenoxy, phenyl or substituted phenyl;

   R ₂ and R ₃ are respectively or simultaneously selected from the group consisting of hydrogen, hydroxyl, cyano, fluorine, chlorine, bromine, carboxyl, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl , pentyl, hexane, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentaneoxy, hexaneoxy Base, aminomethyl, aminoethyl, aminopropyl, aminobutylalkyl, aminopentyl, aminohexane, carboxamide, acetylamino, n-propionyl, isopropylamide, n-butyryl , isobutyryl amide, tert-butyryl amide, pentane amide, hexane amide, carbamoyl, acetyl, n-alanyl, isopropyl, n-butyryl, isobutyryl, uncle Butyryl group, valyl group, hexyl group, nitrogen heterocyclic group of C ₃ , nitrogen heterocyclic group of C ₄ , nitrogen heterocyclic group of C ₅ , nitrogen heterocyclic group of C ₆ , nitrogen heterocyclic ring of C ₃ Alkenyl, C ₄ azacycloalkenyl, C ₅ azacycloalkenyl, C ₆ azacycloalkenyl, phenoxy, phenyl, substituted phenyl, piperazinyl, methylpiperazine Base, ethyl piperazinyl, propyl piperazinyl , butyryl piperazinyl, pentyl piperazinyl or hexyl piperazinyl;

   R _{5 is} selected from the group consisting of fluorine, chlorine, bromine or iodine.
7. The preparation method according to claim 6, wherein in the step f, the compound IM-6a is:

   

   
8. The preparation method according to claim 4 or 5, wherein in the steps a to g, the ether solvent is selected from any one of tetrahydrofuran, diethyl ether, tert-butyl methyl ether, diisopropyl ether, and dibutyl ether. Two or more; the halogen hydrocarbon solvent is selected from any one or two or more selected from the group consisting of dichloromethane, ethyl chloride, dichloroethane, chloroform, and carbon tetrachloride; and the ester solvent is selected from the group consisting of Any one or two or more of ethyl acetate and ethyl formate; the nitrogen-containing solvent is selected from the group consisting of N,N-dimethylformamide, N,N-dimethylacetamide, acetonitrile, and pyridine Any one or two or more.
9. A method for preparing a pyrrole amide compound of the formula I, characterized in that:

   When R ₄ is a hydroxyl group, its synthetic route is:

   

   among them,

   R _{1 is} selected from the group consisting of hydrogen, hydroxy, cyano, halogen, carboxyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₂ -C ₆ amide, C ₁ -C ₆ amino An alkyl group, a C ₂ -C ₆ aminoacyl group, a C ₃ -C ₆ heterocyclic group, a C ₃ -C ₆ heterocycloalkenyl group, a phenoxy group, a phenyl group or a substituted phenyl group;

   R ₂ and R ₃ are respectively or simultaneously selected from the group consisting of hydrogen, hydroxy, cyano, halogen, carboxyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ aminoalkyl, C ₂ -C ₆ amide group, C ₂ -C ₆ aminoacyl group, C ₃ -C ₆ heterocyclic group, C ₃ -C ₆ heterocycloalkenyl group, phenoxy group, phenyl group, substituted phenyl group , piperazinyl or substituted piperazinyl;

   R ₅ and R ₆ are each a halogen;

   Includes the following steps:

   i, a mixed solvent of the compound IM-5b, the compound IM-6b, sodium hydrogencarbonate and an ether solvent/water at 25 ° C to 30 ° C After stirring for 1 h to 10 h, the solvent is removed, water is added, and the solvent is extracted with ethyl acetate. The organic phase is combined, washed with saturated brine, and the organic phase is dried, filtered and concentrated to give a crude product. -7b;

   The molar ratio of the compound IM-5b, the compound IM-6b, and the sodium hydrogencarbonate is 1:1 to 2:1 to 5; the mass to volume ratio of the mixed solvent of the compound IM-5b and the ether solvent/water is 1 : 20～100g/ml; in the mixed solvent, the volume ratio of the ether solvent to water is 1 to 5:1;

   Ii, a mixed solvent of the compound IM-7b, the compound IM-8b, sodium carbonate, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride and an ether solvent/water obtained in the step i After the reaction is stirred at 50 ° C to 100 ° C for 1 h to 10 h under an inert gas atmosphere, the solvent is removed, water is added, and the organic phase is extracted with a halogenated solvent, and the organic phase is dried, filtered, and concentrated. Crude; crude product is purified by column chromatography to give compound TM-1b;

   The molar ratio of the compound IM-7b, the compound IM-8b, and the sodium carbonate is 1:1 to 2:1 to 5; the compound IM-7b and [1,1'-bis(diphenylphosphino)2 The mass ratio of the ferromagnetic] palladium dichloride is 1:0.05 to 0.2; the mass ratio of the mixed solvent of the compound IM-7b to the ether solvent/water is 1:20 to 100 g/ml; in the mixed solvent , the volume ratio of the ether solvent to water is from 1 to 10:1;

   Iii, the compound TM-1b of the step II is dissolved in an alcohol solvent, hydrochloric acid is added, and the reaction is stirred at 25 ° C to 30 ° C for 1 h to 10 h, then separated and purified to obtain a compound of the formula I;

   The mass to volume ratio of the compound TM-1b to the alcohol solvent is 1:18 to 100 g/ml; the mass ratio of the compound TM-1b to hydrochloric acid is 1:3 to 20 g/ml; The concentration ranges from 0.5N to 2N.
10. The preparation method according to claim 9, wherein:

    i, compound IM-5b, compound IM-6b, sodium hydrogencarbonate and ether solvent / water mixed solvent, stirred at 25 ° C for 2h, remove the solvent, add water, extract the ester solvent, combine the organic phase, with saturated salt Washing with water, drying the organic phase, filtering and concentrating to obtain a crude product; the crude product is purified by column chromatography to give compound IM-7b;

    The molar ratio of the compound IM-5b, the compound IM-6b, and the sodium hydrogencarbonate is 1:1:2 to 3; the mass ratio of the compound IM-5b to the ether solvent/water mixed solvent is 1:20. ～40g / ml; in the mixed solvent, the volume ratio of the ether solvent to water is 1 to 2:1;

    Ii, a mixed solvent of the compound IM-7b, the compound IM-8b, sodium carbonate, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride and an ether solvent/water obtained in the step i After stirring at 80 ° C for 2 h under inert gas atmosphere, the solvent is removed, water is added, and the organic solvent is extracted with a halogenated solvent. The organic phase is washed with saturated brine, and the organic phase is dried, filtered and concentrated to give a crude product. Purification by chromatography to give the compound TM-1b;

    The molar ratio of the compound IM-7b, the compound IM-8b, and the sodium carbonate is 1:1:2 to 3; the compound IM-7b and [1,1'-bis(diphenylphosphino)ferrocene The mass ratio of palladium dichloride to 1:0.08 to 0.12; the mass ratio of the mixed solvent of the compound IM-7b to the ether solvent/water is 1:20 to 40 g/ml; in the mixed solvent, the ether The volume ratio of solvent to water is 4 to 6:1;

    Iii, the compound TM-1b of the step II is dissolved in an alcohol solvent, hydrochloric acid is added, and the reaction is stirred at 25 ° C for 2 h, then separated and purified to obtain a compound of the formula I;

    The mass to volume ratio of the compound TM-1b to the alcohol solvent is 1:18-40 g/ml; the compound The mass to volume ratio of TM-1b to hydrochloric acid is 1:3 to 10 g/ml; and the concentration of the hydrochloric acid is in the range of 0.5 N to 2 N.
11. The preparation method according to claim 9 or 10, characterized in that:

    R _{1 is} selected from the group consisting of hydrogen, hydroxy, cyano, fluoro, chloro, bromo, carboxy, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentylene, hexyl Alkyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentyloxy, hexyloxy, aminomethyl, Aminoethyl, aminopropyl, aminobutylalkyl, aminopentyl, aminohexane, carboxamido, acetylamino, n-propionylamino, isopropylamide, n-butyryl, isobutyryl, Tert-butylamide, pentanoamide, hexane amide, methyl, acetyl, n-alanyl, isopropylyl, n-butyryl, isobutyryl, tert-butyryl, pentylamine Acyl group, hexyl group, nitrogen heterocyclic group of C ₃ , nitrogen heterocyclic group of C ₄ , nitrogen heterocyclic group of C ₅ , nitrogen heterocyclic group of C ₆ , azacycloheteroyl group of C ₃ , C ₄ Azacycloheteroenyl, C ₅ azacycloalkenyl, C ₆ azacycloalkenyl, phenoxy, phenyl or substituted phenyl;

    R ₂ and R ₃ are respectively or simultaneously selected from the group consisting of hydrogen, hydroxyl, cyano, fluorine, chlorine, bromine, carboxyl, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl , pentyl, hexane, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentaneoxy, hexaneoxy Base, aminomethyl, aminoethyl, aminopropyl, aminobutylalkyl, aminopentyl, aminohexane, carboxamide, acetylamino, n-propionyl, isopropylamide, n-butyryl , isobutyryl amide, tert-butyryl amide, pentane amide, hexane amide, carbamoyl, acetyl, n-alanyl, isopropyl, n-butyryl, isobutyryl, uncle Butyryl group, valyl group, hexyl group, nitrogen heterocyclic group of C ₃ , nitrogen heterocyclic group of C ₄ , nitrogen heterocyclic group of C ₅ , nitrogen heterocyclic group of C ₆ , nitrogen heterocyclic ring of C ₃ Alkenyl, C ₄ azacycloalkenyl, C ₅ azacycloalkenyl, C ₆ azacycloalkenyl, phenoxy, phenyl, substituted phenyl, piperazinyl, methylpiperazine Base, ethyl piperazinyl, propyl piperazinyl , butyryl piperazinyl, pentyl piperazinyl or hexyl piperazinyl;

    R ₅ and R _{6 are} selected from fluorine, chlorine, bromine or iodine.
12. The preparation method according to claim 11, wherein:

    In step i, the compound IM-6b is:

    

    In step ii, the compound IM-8b is:

    
13. The preparation method according to claim 9 or 10, wherein in the steps i to iii, the ether solvent is selected from the group consisting of tetrahydrofuran, dioxane, diethyl ether, tert-butyl methyl ether, diisopropyl ether and dibutyl ether. Any one or two or more kinds; the halogen hydrocarbon solvent is selected from any one or more selected from the group consisting of dichloromethane, ethyl chloride, dichloroethane, chloroform, and carbon tetrachloride; The ester solvent is selected from any one or two or more selected from the group consisting of ethyl acetate and ethyl formate; and the alcohol solvent is selected from any one or two or more selected from the group consisting of methanol, ethanol, n-propanol and isopropanol.
14. A method for preparing a pyrrole amide compound of the formula I, characterized in that:

    When R ₄ is an amino-substituted phenyl group, its synthetic route is:

    

    Wherein, Boc represents a tert-butoxycarbonyl group; TFA represents trifluoroacetic acid; and HATU represents 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate; DIEA stands for N,N-diisopropylethylamine; LiOH stands for lithium hydroxide;

    R _{1 is} selected from the group consisting of hydrogen, hydroxy, cyano, halogen, carboxyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₂ -C ₆ amide, C ₁ -C ₆ amino An alkyl group, a C ₂ -C ₆ aminoacyl group, a C ₃ -C ₆ heterocyclic group, a C ₃ -C ₆ heterocycloalkenyl group, a phenoxy group, a phenyl group or a substituted phenyl group;

    R ₂ and R ₃ are respectively or simultaneously selected from the group consisting of hydrogen, hydroxy, cyano, halogen, carboxyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ aminoalkyl, C ₂ -C ₆ amide group, C ₂ -C ₆ aminoacyl group, C ₃ -C ₆ heterocyclic group, C ₃ -C ₆ heterocycloalkenyl group, phenoxy group, phenyl group, substituted phenyl group , piperazinyl or substituted piperazinyl;

    R _{5 is} selected from halogen;

    Includes the following steps:

    1. At 0 ° C to 5 ° C, the compound IM-1c is dissolved in a halogen hydrocarbon solvent, trifluoroacetic acid is added, and the reaction is stirred at 20 ° C to 30 ° C for 2 h to 12 h to obtain a reaction liquid; the reaction liquid is concentrated to obtain a yellow oil, which is the compound IM-2c;

    The mass ratio of the compound IM-1c to the halocarbon solvent is 1:5-20 g/ml; the mass ratio of the compound IM-1c to trifluoroacetic acid is 1:2-10 g/ml;

    2. The compound IM-2c, the compound IM-3c, the triethylamine and the halocarbon solvent obtained in the step 1 are stirred at 25 ° C to 30 ° C for 1 h to 10 h, and then concentrated to obtain a crude product; the crude product is purified by column chromatography. Obtaining compound IM-4c;

    The molar ratio of the compound IM-2c, the compound IM-3c, and the triethylamine is 1:1 to 2:1 to 5; the mass to volume ratio of the compound IM-2c to the halocarbon solvent is 1:50 to 100 g. /ml;

    3. The compound IM-4c obtained in the second step, a mixed solvent of lithium hydroxide and an ether solvent/water, and stirred at 20 ° C to 30 ° C for 2 h to 16 h to obtain a reaction liquid; separated and purified to obtain a compound IM-5c. ;

    The molar ratio of the compound IM-4c to lithium hydroxide is 1:1 to 10; the mass ratio of the compound IM-4c to the mixed solvent is 1:55 to 60 g/ml; in the mixed solvent, the ether The volume ratio of solvent to water is 1 to 5:1;

    4. Compound IM-5c, 1,2-diaminobenzene, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate obtained in step 3. The ester, N,N-diisopropylethylamine and a halogenated hydrocarbon solvent are stirred at 25 ° C to 30 ° C for 12 h to 16 h to obtain a reaction liquid; isolated and purified to obtain a compound of the formula I;

    The compound IM-5c, 1,2-diaminobenzene, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluron hexafluorophosphate, N, The molar ratio of N-diisopropylethylamine is 1:0.8 to 2:0.8 to 2:1.5 to 4; and the mass ratio of the compound IM-5c to the halogen hydrocarbon solvent is 1:40 to 100 g/ml.
15. The preparation method according to claim 14, wherein:

    1. At 0 ° C, the compound IM-1c is dissolved in a halogen hydrocarbon solvent, trifluoroacetic acid is added, and the reaction mixture is stirred at 25 ° C for 2 h to obtain a reaction liquid; the reaction liquid is concentrated to obtain a yellow oil, which is a compound IM. -2c;

    The mass to volume ratio of the compound IM-1c to the halocarbon solvent is 1:20 g/ml; the mass to volume ratio of the compound IM-1c to trifluoroacetic acid is 1:8 g/ml;

    2. The compound IM-2c, the compound IM-3c, the triethylamine and the halogen hydrocarbon solvent obtained in the step 1 are stirred and reacted at 25 ° C for 2 hours, and then concentrated to obtain a crude product; the crude product is purified by column chromatography to give the compound IM-4c. ;

    The molar ratio of the compound IM-2c, the compound IM-3c, and the triethylamine is 1:1.1 to 1.2:2.5 to 3; the mass to volume ratio of the compound IM-2c to the halocarbon solvent is 1:65 to 70 g. /ml;

    3, the compound IM-4c obtained in step 2, lithium hydroxide and ether solvent / water mixed solvent, stirring reaction at 25 ° C for 2h ~ 16h, the reaction liquid is obtained; isolated, purified, to obtain the compound IM-5c;

    The molar ratio of the compound IM-4c to lithium hydroxide is 1:4.5; the mass to volume ratio of the compound IM-4c to the mixed solvent is 1:55-60 g/ml; in the mixed solvent, the ether solvent and The volume ratio of water is 3:1;

    4. Compound IM-5c, 1,2-diaminobenzene, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate obtained in step 3. The ester, N,N-diisopropylethylamine and a halogenated hydrocarbon solvent are stirred at 25 ° C for 12 h to 16 h to obtain a reaction liquid; isolated and purified to obtain a compound of the formula I;

    The compound IM-5c, 1,2-diaminobenzene, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluron hexafluorophosphate, N, The molar ratio of N-diisopropylethylamine is 1:0.8 to 1.2:0.8 to 1.2:1.5 to 2; the mass to volume ratio of the compound IM-5c to the halocarbon solvent is 1:40 to 45 g/ml.
16. The preparation method according to claim 14 or 15, wherein:

    R _{1 is} selected from the group consisting of hydrogen, hydroxy, cyano, fluoro, chloro, bromo, carboxy, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentylene, hexyl Alkyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentyloxy, hexyloxy, aminomethyl, Aminoethyl, aminopropyl, aminobutylalkyl, aminopentyl, aminohexane, carboxamido, acetylamino, n-propionylamino, isopropylamide, n-butyryl, isobutyryl, Tert-butylamide, pentanoamide, hexane amide, carbamoyl, ethoxyyl, n-alanyl, isopropylyl, n-butyryl, isobutyryl, tert-butyryl, pentylamine Acyl group, hexyl group, nitrogen heterocyclic group of C ₃ , nitrogen heterocyclic group of C ₄ , nitrogen heterocyclic group of C ₅ , nitrogen heterocyclic group of C ₆ , azacycloheteroyl group of C ₃ , C ₄ Azacycloheteroenyl, C ₅ azacycloalkenyl, C ₆ azacycloalkenyl, phenoxy, phenyl or substituted phenyl;

    R ₂ and R ₃ are respectively or simultaneously selected from the group consisting of hydrogen, hydroxyl, cyano, fluorine, chlorine, bromine, carboxyl, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl , pentyl, hexane, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentaneoxy, hexaneoxy Base, aminomethyl, aminoethyl, aminopropyl, aminobutylalkyl, aminopentyl, aminohexane, carboxamide, acetylamino, n-propionyl, isopropylamide, n-butyryl , isobutyryl amide, tert-butyryl amide, pentane amide, hexane amide, carbamoyl, acetyl, n-alanyl, isopropyl, n-butyryl, isobutyryl, uncle Butyryl group, valyl group, hexyl group, nitrogen heterocyclic group of C ₃ , nitrogen heterocyclic group of C ₄ , nitrogen heterocyclic group of C ₅ , nitrogen heterocyclic group of C ₆ , nitrogen heterocyclic ring of C ₃ Alkenyl, C ₄ azacycloalkenyl, C ₅ azacycloalkenyl, C ₆ azacycloalkenyl, phenoxy, phenyl, substituted phenyl, piperazinyl, methylpiperazine Base, ethyl piperazinyl, propyl piperazinyl , butyryl piperazinyl, pentyl piperazinyl or hexyl piperazinyl;

    R _{5 is} selected from the group consisting of fluorine, chlorine, bromine or iodine.
17. The preparation method according to claim 16, wherein in the step 2, the compound IM-3c is:

    
18. The preparation method according to claim 9 or 10, wherein in the steps 1 to 4, the halocarbon The solvent is selected from any one or more selected from the group consisting of dichloromethane, ethyl chloride, dichloroethane, chloroform, and carbon tetrachloride; and the ether solvent is selected from the group consisting of tetrahydrofuran, dioxane, and diethyl ether. Any one or two or more of tert-butyl methyl ether, diisopropyl ether and dibutyl ether.
19. The use of the pyrrole amide compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt, crystal form, hydrate or solvate thereof, for producing a histone deacetylase inhibitor drug.
20. The use according to claim 19, wherein the histone deacetylase inhibitor drug is a drug for preventing and/or treating a disease caused by abnormal histone deacetylase activity.
21. The use according to claim 20, wherein the disease is any one or more of a cell proliferative disease, an autoimmune disease, an inflammation, a neurodegenerative disease, or a viral disease.
22. The use according to claim 21, wherein the disease is cancer.
23. A pharmaceutical composition for inhibiting histone deacetylase activity, which is characterized by being a pyrrole amide compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, a crystal form, or a hydrated product The preparation or the solvate is an active ingredient, and a preparation prepared by adding a pharmaceutically acceptable adjuvant or auxiliary ingredient.
24. The pharmaceutical composition according to claim 23, wherein the preparation comprises an orally administered preparation, a sublingual preparation, a buccal preparation, a transdermal absorption preparation or an injection preparation.