Substituted indazole HDAC / Mcl-1 double-target inhibitor as well as preparation method and application thereof

Abstract

The invention discloses a substituted indazole HDAC / Mcl-1 double-target inhibitor and a preparation method and application thereof, the compound has a structure of a general formula (I) and a structure of a general formula (II), # imgabs0 #, in which R is alkyl, aryl or heteroaryl; linker is selected from alkyl groups with 2 to 8 carbon atoms, phenyl groups with different substitutions, naphthenic groups with 3 to 8 carbon atoms or aryl groups with 5 to 10 carbon atoms; and X is carbonyl, sulfonyl or-CH2-. The compound disclosed by the invention has relatively strong inhibitory activity on HDAC and Mcl-1 proteins, and can be used for preventing or treating related mammalian diseases caused by abnormal expression of the HDAC / Mcl-1 proteins. The invention also relates to the pharmaceutical application of the composition of the compounds with the general formulas (I) and (II).

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to a substituted indazole HDAC / Mcl-1 dual-target inhibitor, a preparation method and an application thereof. Background Art

[0002] HDAC inhibitors target histone deacetylases (HDACs), disrupting the dynamic balance between histone acetylases (HATs) and HDACs, increasing histone acetylation levels, loosening chromatin structure, and activating the expression of tumor suppressor genes (such as p21), thereby inhibiting tumor growth. Furthermore, HDAC inhibitors can regulate non-histone substrates (such as α-tubulin and Hsp90), affecting cell migration, protein stability, and inflammatory pathways. With the continuous advancement of HDAC research, HDAC inhibitors have become a hot topic in anti-tumor drug research both domestically and internationally. Mcl-1 (myeloid cell leukemia-1) is a key anti-apoptotic member of the Bcl-2 protein family. It inhibits apoptosis by binding to pro-apoptotic proteins (such as Bak and Bim), thereby maintaining tumor cell survival. Mcl-1 inhibitors competitively bind to its hydrophobic groove, inhibiting its binding to pro-apoptotic proteins. This leads to the release of pro-apoptotic factors (such as cytochrome c) from mitochondria, activating the mitochondrial apoptotic pathway, and restoring normal apoptosis in tumor cells. Mcl-1 inhibitors have become a key research focus in anti-tumor drugs.

[0003] Studies have shown that histone deacetylase (HDAC) inhibitors can significantly upregulate the expression levels of pro-apoptotic genes by blocking the histone deacetylation process, thereby activating the mitochondrial-mediated apoptosis pathway. This mechanism of action can significantly lower the apoptosis resistance threshold of tumor cells, thereby enhancing their programmed cell death effect. However, overexpression of anti-apoptotic proteins such as Mcl-1 in tumor cells antagonizes this apoptotic pathway, resulting in a significant inhibition of the therapeutic effect of HDAC inhibitors. Studies have reported that the combination of the Mcl-1 inhibitor S63845 and the HDAC inhibitors Panobinostat or Ricolinostat can significantly induce apoptosis in multiple myeloma cells, more effectively killing tumor cells and showing a good synergistic anti-tumor effect.

[0004] Compared to traditional single-target drugs, multi-target drugs can simultaneously regulate multiple key signaling nodes, achieving therapeutic effects at lower doses through synergistic interactions between targets, thereby reducing the likelihood of drug-resistant mutations. Compared with multidrug combination regimens, multi-target inhibitors have more optimized pharmacokinetic profiles, avoiding the complex dosage matching issues of combination therapy and eliminating the potential risk of drug interactions between different drugs. This novel drug design strategy not only helps improve anti-tumor efficacy but also effectively overcomes the challenge of tumor resistance, demonstrating significant clinical value. The development of HDAC-based multi-target drugs has become an effective means to improve tumor efficacy and reduce tumor resistance. Currently, there are no reports of HDAC / Mcl-1 dual-target inhibitors. Therefore, organically integrating the structure of Mcl-1 inhibitors into HDAC inhibitors to develop new and effective dual-target inhibitors is a highly challenging and highly valuable research topic in anti-tumor drug research.

[0005] In view of this, the present invention is proposed. Summary of the Invention

[0006] The technical problem to be solved by the present invention is to overcome the deficiencies of the prior art and provide a substituted indazole HDAC / Mcl-1 dual-target inhibitor and a preparation method and application thereof.

[0007] In order to solve the above technical problems, the basic concept of the technical solution adopted by the present invention is:

[0008] A substituted indazole HDAC / Mcl-1 dual-target inhibitor is a compound or a pharmaceutically acceptable salt having the structure of general formula (I) and general formula (II):

[0009]

[0010] In general formulas (I) and (II), R is an alkyl, aryl, or heteroaryl group. Linker is selected from C2-C8 alkyl or phenyl groups with different substitutions, C3-C8 cycloalkyl, or C5-C10 aryl groups. X is a carbonyl, sulfonyl, or -CH2- group.

[0011] R is preferably an optionally substituted C1-C10 alkyl, C3-C10 cycloalkyl, C5-C15 aryl, and a monocyclic heterocyclic aromatic group containing 5 or 6 ring atoms, or a bicyclic heterocyclic aromatic group having 8 to 15 ring atoms, wherein the heterocyclic aromatic group contains 1-4 heteroatoms, and the heteroatoms are independently selected from O, S, N or oxidized S or N; the carbon atom or nitrogen atom is the connection point of the heteroaromatic ring structure, maintaining a stable aromatic ring.

[0012] Preferably, R is a halogen C1-C6 alkyl, C1-C6 alkyl, C3-C8 cycloalkyl, an aromatic group Ar, -NH-R1, substituted by 1-2 hydroxyl, halogen, nitro, cyano substituents or unsubstituted morpholine group, or a piperazine group; Ar is a phenyl group, naphthyl, pyridyl, pyridazinyl, pyrazinyl, indolizine, quinazolinyl, purinyl, indolyl, quinolyl, pyrimidinyl, pyrrolyl, pyrazolyl, thiazolyl, benzothiazolyl, thienyl, benzo[b]thienyl, isoxazolyl, oxathiadiazolyl, isothiazolyl, tetrazolyl, imidazolyl, triazinyl, furyl, benzofuranyl and indolyl containing 1 substituent or no substituent.

[0013] More preferably, the substituted indazole HDAC / Mcl-1 dual-target inhibitor is one of the following:

[0014] 1-(3,4-Dichlorobenzyl)-N-[4-(4-(2-hydroxyamino-2-oxoethyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A1);

[0015] 1-(3,4-Dichlorobenzyl)-N-[4-(4-(4-hydroxyamino-4-oxobutyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A2);

[0016] 1-(3,4-Dichlorobenzyl)-N-[4-(4-(5-hydroxyamino-5-oxopentyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A3);

[0017] 1-(3,4-Dichlorobenzyl)-N-[4-(4-(6-hydroxyamino-6-oxohexyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A4);

[0018] 1-(3,4-Dichlorobenzyl)-N-[4-(4-(7-hydroxyamino-7-oxoheptyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A5);

[0019] 1-(3,4-Dichlorobenzyl)-N-[4-(4-(8-hydroxyamino-8-oxooctyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A6);

[0020] (E)-1-(3,4-dichlorobenzyl)-N-[4-(4-(4-(3-hydroxyamino-3-oxopropen-1-yl)benzyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A7);

[0021] 1-(3,4-Dichlorobenzyl)-N-[4-(4-(4-hydroxycarbamoylbenzyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A8);

[0022] 1-(Naphthalen-2-ylmethyl)-N-[4-(4-(2-hydroxyamino-2-oxoethyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A9);

[0023] 1-(Naphthalen-2-ylmethyl)-N-[4-(4-(4-hydroxyamino-4-oxobutyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A10);

[0024] 1-(Naphthalen-2-ylmethyl)-N-[4-(4-(5-hydroxyamino-5-oxopentyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A11);

[0025] 1-(Naphthalen-2-ylmethyl)-N-[4-(4-(6-hydroxyamino-6-oxohexyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A12);

[0026] 1-(Naphthalen-2-ylmethyl)-N-[4-(4-(7-hydroxyamino-7-oxoheptyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A13);

[0027] 1-(Naphthalen-2-ylmethyl)-N-[4-(4-(8-hydroxyamino-8-oxooctyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A14);

[0028] (E)-1-(naphthalen-2-ylmethyl)-N-[4-(4-(4-(3-hydroxyamino-3-oxoprop-1-en-1-yl)benzyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A15);

[0029] 1-(4'-Methyl-[1,1'-biphenyl]-4-ylmethyl)-N-[4-(4-(2-hydroxyamino-2-oxoethyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A16);

[0030] 1-(4'-Methyl-[1,1'-biphenyl]-4-ylmethyl)-N-[4-(4-(4-hydroxyamino-4-oxobutyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A17);

[0031] 1-(4'-Methyl-[1,1'-biphenyl]-4-ylmethyl)-N-[4-(4-(5-hydroxyamino-5-oxopentyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A18);

[0032] 1-(4'-Methyl-[1,1'-biphenyl]-4-ylmethyl)-N-[4-(4-(2-hydroxyamino-2-oxohexyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A19);

[0033] 1-(4'-Methyl-[1,1'-biphenyl]-4-ylmethyl)-N-[4-(4-(7-hydroxyamino-7-oxoheptyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A20);

[0034] 1-(4'-Methyl-[1,1'-biphenyl]-4-ylmethyl)-N-[4-(4-(8-hydroxyamino-8-oxooctyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A21);

[0035] (E)-1-(4'-Methyl-[1,1'-biphenyl]-4-ylmethyl)-N-[4-(4-(4-(3-hydroxyamino-3-oxopropen-1-en-1-yl)benzyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A22);

[0036] 1-(4'-Methyl-[1,1'-biphenyl]-4-ylmethyl)-N-[4-(4-(4-hydroxycarbamoylbenzyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A23);

[0037] 1-(3,4-Dichlorobenzyl)-N-[4-(4-hydroxyamino-4-oxobutoxy)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (B1);

[0038] 1-(3,4-Dichlorobenzyl)-N-[4-(5-hydroxyamino-5-oxopentyloxy)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (B2);

[0039] 1-(3,4-Dichlorobenzyl)-N-[4-(6-hydroxyamino-6-oxohexyloxy)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (B3);

[0040] 1-(3,4-Dichlorobenzyl)-N-[4-(7-hydroxyamino-7-oxoheptyloxy)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (B4);

[0041] 1-(3,4-Dichlorobenzyl)-N-[4-(8-hydroxyamino-8-oxooctyloxy)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (B5);

[0042] 1-(3,4-dichlorobenzyl)-N-[4-(4-hydroxycarbamoylbenzyloxy)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (B6);

[0043] 1-(Naphthalen-2-ylmethyl)-N-[4-(4-hydroxyamino-4-oxobutoxy)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (B7);

[0044] N-[4-(5-Hydroxyamino-5-oxopentyloxy)-3-nitrobenzenesulfonyl]-1-(naphthalen-2-ylmethyl)-1H-indazole-3-carboxamide (B8);

[0045] 1-(Naphthalen-2-ylmethyl)-N-[4-(6-hydroxyamino-6-oxohexyloxy)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (B9);

[0046] N-[4-(7-hydroxyamino-7-oxoheptyloxy)-3-nitrobenzenesulfonyl]-1-(naphthalen-2-ylmethyl)-1H-indazole-3-carboxamide (B10);

[0047] N-[4-(8-Hydroxyamino-8-oxooctyloxy)-3-nitrobenzenesulfonyl]-1-(naphthalen-2-ylmethyl)-1H-indazole-3-carboxamide (B11);

[0048] (E)-1-(naphthalen-2-ylmethyl)-N-[4-([4-(3-hydroxyamino-3-oxoprop-1-en-1-yl)benzyl]oxy)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (B12);

[0049] N-[4-(4-Hydroxycarbamoylbenzyloxy)-3-nitrobenzenesulfonyl]-1-(naphthalen-2-ylmethyl)-1H-indazole-3-carboxamide (B13);

[0050] N-[4-(4-Hydroxyamino-4-oxobutoxy)-3-nitrobenzenesulfonyl]-1-([1,1'-biphenyl]-4'-yl-4-ylmethyl)-1H-indazole-3-carboxamide (B14);

[0051] N-[4-(5-Hydroxyamino-5-oxopentyloxy)-3-nitrobenzenesulfonyl]-1-[(4'-methyl-[1,1'-biphenyl]-4-yl)methyl]-1H-indazole-3-carboxamide (B15);

[0052] N-[4-(6-hydroxyamino-6-oxohexyloxy)-3-nitrobenzenesulfonyl]-1-[(4'-methyl-[1,1'-biphenyl]-4-yl)methyl]-1H-indazole-3-carboxamide (B16);

[0053] N-[4-(7-hydroxyamino-7-oxoheptyloxy)-3-nitrobenzenesulfonyl]-1-[(4'-methyl-[1,1'-biphenyl]-4-yl)methyl]-1H-indazole-3-carboxamide (B17);

[0054] N-[4-(8-Hydroxyamino-8-oxooctyloxy)-3-nitrobenzenesulfonyl]-1-[(4'-methyl-[1,1'-biphenyl]-4-yl)methyl]-1H-indazole-3-carboxamide (B18).

[0055] The numbers in the brackets following the above preferred compounds are the numbers corresponding to the compound structures in the following reaction scheme and Table 1. Detailed Description of the Invention

[0057] The terms and definitions used in this document have the following meanings:

[0058] "Aryl" refers to an aromatic hydrocarbon containing a ring system, such as phenyl or naphthyl, which is optionally fused to a cycloalkyl group, preferably having 5 to 7 ring atoms, more preferably 5 to 6 ring atoms. Preferred aryl groups contain 5 to 15 carbon atoms.

[0059] "Heteroaryl" is an aromatic heterocycle that can be a monocyclic or bicyclic group. They contain one or more, preferably 1-4, more preferably 1-3, and even more preferably 1-2 heteroatoms, independently selected from O, S, and N. Heteroaryl includes oxidized S or N, such as sulfinyl, sulfonyl, and tricyclic nitrogen N-oxides. The carbon atom or nitrogen atom is the point of attachment of the heteroaromatic ring structure, thereby maintaining a stable aromatic ring. Examples of heteroaryl include, but are not limited to, pyridyl, pyridazinyl, pyrazinyl, indolizinyl, benzothiophenyl, quinazolinyl, purinyl, indolyl, quinolyl, pyrimidinyl, pyrrolyl, oxazolyl, thiazolyl, thienyl, isoxazolyl, oxathiadiazolyl, isothiazolyl, tetrazolyl, imidazolyl, triazinyl, furyl, benzofuranyl, and indolyl.

[0060] "Arylalkyl" refers to an aryl group linked to a C1-C6 alkylene group.

[0061] "Heteroarylalkyl" refers to a heteroaryl group linked to a C1-C6 alkylene group.

[0062] "Arylalkenyl" refers to an aryl group to which an alkenyl group is attached.

[0063] "Heteroarylalkenyl" refers to a heteroaryl group linked to a C2-C6 alkenyl group.

[0064] "Alkyl", alone or in combination, refers to a group derived from an alkane, containing 1 to 20 carbon atoms, preferably 1 to 12 carbon atoms (if not otherwise specified). It is a straight chain alkyl or a branched chain alkyl, and includes straight chain alkyl or branched chain alkyl containing a cycloalkyl portion or interrupted by a cycloalkyl portion. A straight chain alkyl or a branched chain alkyl is connected at any available point to produce a stable compound. Examples include, but are not limited to, 4-(isopropyl)-cyclohexylethyl or 2-methyl-cyclopropylpentyl. In many embodiments, the alkyl group is a straight chain alkyl or a branched chain alkyl group containing 1 to 15 carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbon atoms, 1 to 4 carbon atoms or 1 to 2 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, tert-butyl and similar alkyl groups.

[0065] "Alkylene" is a divalent alkane-derived carbon atom group, which is straight or branched, in which two hydrogen atoms are removed from the same carbon atom or different carbon atoms. Examples of alkylene include, but are not limited to, -CH2-, -CH2CH2-, and -CH2CH(CH3)-.

[0066] "Alkenyl," as used herein, alone or in combination, refers to a straight or branched chain hydrocarbon containing 2-6, preferably 2-4, carbon atoms and containing 1-2, preferably one, carbon-carbon double bonds. Examples of alkenyl groups include, but are not limited to, ethenyl, propenyl, isopropenyl, and butenyl.

[0067] "Cycloalkyl" is a substituted or unsubstituted, saturated or unsaturated cyclic group containing carbon atoms and / or one or more heteroatoms. The ring can be a monocyclic or fused, bridged, or spirocyclic ring system. The number of ring atoms in each ring is 3-8, more preferably 3-6, such as cyclopropyl, cyclopentyl, cyclohexyl, adamantyl, and similar groups.

[0068] "Alkoxy" means the group -O-alkyl.

[0069] "Halogen" alone or in combination refers to all halogens, ie, chlorine (Cl), fluorine (F), bromine (Br) or iodine (I).

[0070] "Pharmaceutically acceptable salts" refer to salt forms of compounds of Formula I that are therapeutically effective and non-toxic. They can form anionic salts with any acidic group (such as a carboxyl group), or cationic salts with any basic group (such as an amino group). Many such salts are known in the art. Cationic salts formed on any acidic group (such as a carboxyl group), or anionic salts formed on any basic group (such as an amino group). Many of these salts are known in the art, such as cationic salts including salts of alkali metals (such as sodium and potassium) and alkaline earth metals (such as magnesium and calcium), as well as organic salts (such as ammonium salts). Anionic salts can also be conveniently obtained by treating the basic form of I with the corresponding acid, including inorganic acids such as sulfuric acid, nitric acid, phosphoric acid, and the like; or organic acids such as acetic acid, propionic acid, glycolic acid, 2-hydroxypropionic acid, 2-oxopropionic acid, oxalic acid, malonic acid, succinic acid, maleic acid, fumaric acid, malic acid, tartaric acid, 2-hydroxy-1,2,3-propanetriol, methanesulfonic acid, ethanesulfonic acid, benzenemethylsulfonic acid, 4-methylbenzenesulfonic acid, cyclohexylsulfinic acid, 2-hydroxybenzoic acid, 4-amino-2-hydroxybenzoic acid, and the like. These salts are well known to those skilled in the art, and those skilled in the art can prepare any salt provided by the knowledge in the art. In addition, those skilled in the art may choose one salt over another based on factors such as solubility, stability, and ease of formulation. The determination and optimization of these salts are within the experience of those skilled in the art.

[0071] As used herein, "stereoisomers" define all possible stereoisomeric forms of the compounds of the present invention or their physiological derivatives. Unless otherwise indicated, the chemical designations of the compounds of the present invention encompass mixtures of all possible stereochemical forms, mixtures containing all diastereomers and enantiomers of the basic structural molecule, as well as substantially pure individual isomeric forms of the compounds of the present invention, i.e., containing less than 10%, preferably less than 5%, particularly less than 2%, and most preferably less than 1% of other isomers. Various stereoisomeric forms of the peptoid compounds of the present invention are clearly encompassed within the scope of the present invention.

[0072] The compound of formula I or formula II may also exist in other protected forms or derivative forms, which are obvious to those skilled in the art and should all be included in the scope of the present invention.

[0073] Substituents as described above themselves may also be substituted with one or more substituents. Such substituents include those listed in C. Hansch and A. Leo, Substituent Constants for Correlation Analysis in Chemistry and Biology (1979). Preferred substituents include alkyl, alkenyl, alkoxy, hydroxy, oxy, nitro, amino, aminoalkyl (such as aminomethyl etc.), cyano, halogen, carboxyl, carbonylalkoxy (such as carbonylethoxy etc.), sulfenyl, aryl, cycloalkyl, heteroaryl, heterocycloalkyl (such as piperidinyl, morpholinyl, pyrrolyl etc.), imino, hydroxyalkyl, aryloxy, arylalkyl and combinations thereof.

[0074] A "pharmaceutical composition" is a preparation containing a therapeutically significant amount of an active pharmaceutical agent, which is prepared in a form suitable for administration to a patient. Thus, the preparation does not contain any component or components in such an amount that a reasonably careful medical practitioner would find the preparation unsuitable for administration to an ordinary subject. In many cases, such pharmaceutical compositions are sterile preparations.

[0075] Room temperature refers to the ambient temperature in which the experiment is performed, which is controlled within the range of 10 to 30°C.

[0076] 2. Preparation of substituted indazole HDAC / Mcl-1 dual-target inhibitors

[0077] A method for preparing a substituted indazole HDAC / Mcl-1 dual-target inhibitor comprises the following steps:

[0078] Starting from indazole-3-carboxylic acid, the carboxyl group is first protected by esterification under acidic conditions to obtain intermediate 1. This is followed by nucleophilic substitution with benzyl bromide of varying substituents at the 1-nitrogen position of the indazole-3-carboxylic acid and removal of the methyl ester to produce key intermediates 2a-2c. On the other hand, 4-fluoro-3-nitrobenzenesulfonamide reacts with N-Boc-piperazine and removes the protecting group, followed by reaction with methyl or ethyl bromocarboxylates of varying chain lengths to produce key intermediates 3a-3h. On the other hand, 4-chloro-3-nitrobenzenesulfonamide is reacted with sodium hydroxide, the amino group is protected, and then reacted with methyl or ethyl bromocarboxylates of varying chain lengths and removed from the protecting group to produce key intermediates 4a-4g. Intermediates 2a-2c are then reacted with 3a-3h and 4a-4g, respectively, to produce key intermediates 5a-5w and 6a-6r. Finally, the prepared potassium hydroxylammonium solution was added and the target compounds A1-A23 and B1-B18 were obtained by acidification.

[0079] The synthetic route is as follows:

[0080]

[0081] Wherein, R and Linker are defined as described in Formulas I and II above;

[0082] Reagents and conditions: a) acetyl chloride, methanol, ice bath-reflux; b) i. substituted benzyl bromide, potassium carbonate, N,N-dimethylformamide (DMF), room temperature; ii. 1 mol / L sodium hydroxide solution, tetrahydrofuran, room temperature; iii. 1 mol / L hydrochloric acid solution, room temperature; c) i. DMF, N-Boc-piperazine, 80°C; ii. ethyl acetate saturated with hydrogen chloride, room temperature; iii. acetone, triethylamine, methyl or ethyl bromocarboxylate, 60°C; d) i. 16.5 mol / L sodium hydroxide solution, reflux; ii. 3 mol / L hydrochloric acid solution, room temperature; iii. DMF, N,N-dimethylformamide dimethyl acetal, 45°C; iv. potassium carbonate, DMF, methyl or ethyl bromocarboxylate, 80°C; v. methanol, hydrochloric acid, 60°C; e) i. thionyl chloride, reflux; ii. dichloromethane, triethylamine, ice bath - room temperature; f) i. hydroxylamine hydrochloride, potassium hydroxide, room temperature; ii. 1 mol / L hydrochloric acid solution, room temperature.

[0083] The structural formula of the target compound in the synthesis route is shown in Table 1 below:

[0084] Table 1 Structural formula of target compound

[0085]

[0086]

[0087] The specific operation steps of the compound will be described in detail in the examples.

[0088] Those skilled in the art may modify the above steps to improve yields. They may determine the synthetic route based on basic knowledge in the art, such as selecting reactants, solvents, and temperatures. They may also utilize various conventional protecting groups to prevent side reactions and thereby improve yields. These conventional protection methods can be found, for example, in T. Greene, Protecting Groups in Organic Synthesis.

[0089] III. Application of novel indazole HDAC / Mcl-1 dual-target inhibitors

[0090] The present invention also provides the use of this series of compounds in the preparation of drugs for preventing or treating mammalian diseases caused by abnormal HDAC / Mcl-1 protein expression. These mammalian diseases associated with abnormal HDAC / Mcl-1 protein expression include cancer, neurodegenerative diseases, viral infections, inflammation, leukemia, malaria, and diabetes.

[0091] In addition, the present invention also includes a pharmaceutical composition suitable for oral administration to mammals, comprising any compound of the above-mentioned general formula I or general formula II, and a pharmaceutically acceptable carrier, optionally comprising one or more pharmaceutically acceptable excipients.

[0092] In addition, the present invention also includes a pharmaceutical composition suitable for parenteral administration to mammals, comprising any compound of the above-mentioned general formula I or general formula II, and a pharmaceutically acceptable carrier, optionally comprising one or more pharmaceutically acceptable excipients.

[0093] Fluorescence analysis is used to evaluate the inhibitory activity of compounds against HDAC. HDAC hydrolyzes the acetyl groups on lysine residues in the fluorescent substrate (Ac-Leu-Gly-Lys(Ac)-AMC), activating the substrate. The activated fluorescent substrate is hydrolyzed by trypsin, releasing the fluorophore, which emits fluorescence at 390 / 460 nm. The HDAC inhibitory activity of the test compound can be calculated by measuring the fluorescence values ​​of the control and experimental groups.

[0094] The inhibitory activity of Mcl-1 protein was tested by fluorescence polarization experiment. In the specific measurement system, 5-FAM-labeled Bid-BH3 peptide was used as the fluorescent labeling molecule. This molecule can specifically bind to Mcl-1 protein, and its dissociation constant (K d ) is around 30-60nM, and the combination of the two produces a higher polarization value. If the target compound being tested can bind to the target protein, it will competitively inhibit the binding of Bid to the protein, resulting in a decrease in the polarization value, and then obtain the dose-effect curve of the target compound's competitive binding, and finally calculate the inhibition constant K i .

[0095] In vitro enzyme inhibition experiments showed that some compounds, such as compounds A5, A6, A7, A14, A15, A17, A19, A23, B5, B11, B16, B17, and B18, exhibited significant HDAC1 inhibitory activity. Compounds A7 and B17 exhibited similar HDAC1 inhibitory activity to the positive control drug, SAHA. Further testing of the target compounds against Mcl-1 protein revealed that compounds A6, A7, A14, A15, A17, A19, A23, B5, B11, B16, B17, and B18 exhibited significant Mcl-1 inhibitory activity. Compounds A7, A15, A23, B17, and B18 exhibited comparable or superior Mcl-1 inhibitory activity to the positive control drug, UMI-77. Most compounds have dual HDAC / Mcl-1 inhibitory activity, especially B17 and B18, whose inhibitory activity against both targets reaches the nanomolar level. This is expected to lead to the further development of HDAC / Mcl-1 protein inhibitors with higher activity, which is of great significance for the preparation of drugs to prevent or treat related mammalian diseases caused by abnormal HDAC / Mcl-1 proteins.

[0096] After adopting the above technical solution, the present invention has the following beneficial effects compared with the prior art.

[0097] The compounds of the present invention have strong inhibitory activity on HDAC and Mcl-1 proteins, and can prevent or treat related mammalian diseases caused by abnormal expression of HDAC / Mcl-1 proteins.

[0098] The specific embodiments of the present invention are described in further detail below. DETAILED DESCRIPTION

[0099] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments will be clearly and completely described below. The following embodiments are used to illustrate the present invention but are not used to limit the scope of the present invention.

[0100] Example 1. Synthesis of indazole-3-carboxylic acid methyl ester (1)

[0101] Acetyl chloride (5.69 mL, 80 mmol) was slowly added dropwise to methanol (100 mL) in an ice bath. Stirring was continued for 30 minutes after the addition was complete. Indazole-3-carboxylic acid (3.24 g, 20 mmol) was added to the reaction flask, and the mixture was refluxed in a 90°C oil bath for 4 hours. After the reaction was complete, heating was stopped and the mixture was cooled. Saturated NaHCO₃ solution was added to the reaction solution to adjust the pH to approximately 7. An off-white solid precipitated and was filtered and dried to obtain 3.47 g of an off-white solid in a 99% yield, mp: 167-169°C. This intermediate was used directly in the next step without purification.

[0102] Synthesis of 1-(3,4-dichlorobenzyl)-1H-indazole-3-carboxylic acid (2a)

[0103] Intermediate 1 (1.76 g, 10 mmol) was dissolved in DMF, and K2CO3 (4.14 g, 30 mmol) was added. After stirring for 30 minutes, 3,4-dichlorobenzyl bromide (1.60 mL, 11 mmol) was added. After completion of the reaction, water was added to the reaction solution, extracted with ethyl acetate, and dried. Purification by silica gel column chromatography (petroleum ether / ethyl acetate) afforded 1.83 g of a white solid in a 51% yield. The white solid (1.78 g, 5 mmol) was dissolved in THF (20 mL) and 1 mol / L NaOH solution (20 mL) was added, followed by stirring overnight. After completion of the reaction, the THF was dried, and 1 mol / L HCl solution was added to adjust the pH to approximately 6. A white solid precipitated, which was filtered and dried to afford 1.02 g of a white solid in a 63% yield, mp: 156-158°C.

[0104] Synthesis of methyl 2-[4-(2-nitro-4-sulfanilylphenyl)piperazin-1-yl]acetate (3a)

[0105] Dissolve N-Boc-piperazine (3.73 g, 20 mmol) in DMF and add K2CO3 (5.52 g, 40 mmol). Stir for 30 minutes, then add 4-fluoro-3-nitrobenzenesulfonamide (4.62 g, 20 mmol) and dissolve. Reflux in an 80°C oil bath for 4 hours. After completion, remove from heat. Pour the reaction mixture into ice water and stir. Extract with ethyl acetate, and wash the extract with saturated brine. Add anhydrous sodium sulfate and dry the extract to obtain a yellow solid. Add 100 mL of saturated hydrochloric acid in ethyl acetate, seal the container, and allow to react for 12 hours. After the reaction is complete, filter and collect the filter cake. Add 1.50 g of the filter cake to acetone and stir evenly. Add triethylamine (3.75 mL, 27 mmol) and stir for 30 minutes. Then, add methyl bromoacetate (1.08 mL, 11.25 mmol) and allow to react for 4 hours. Filter the reaction mixture and spin dry. The product was purified by silica gel column chromatography (petroleum ether / ethyl acetate) to give 0.73 g of a yellow solid with a yield of 56% and mp: 184-186°C. 1 H NMR (400MHz, DMSO-d6), δ8.20(s,1H),7.90(d,J=8.8Hz,1H),7.45-7.43(m,3H),3.62(s,3H),3.33(s,2H),3.16-3.13(m,4H),2.66-2.64(m,4H).

[0106] Synthesis of methyl 2-[4-(4-(N-(1-(3,4-dichlorobenzyl)-1H-indazole-3-carbonyl)sulfonyl)-2-nitrophenyl)piperazin-1-yl]acetate (5a)

[0107] Intermediate 2a (0.32 g, 1 mmol) was added to a reaction flask, followed by the addition of thionyl chloride (6 mL). The reaction was stopped by reflux in an 82°C oil bath for 4 h. After cooling to room temperature, the thionyl chloride was removed by spin drying, resulting in the appearance of a white solid in the reaction flask. Dichloromethane (15 mL) was added to the reaction flask under ice-cooling, and stirring was continued for 15 min. Intermediate 3a (0.29 g, 0.8 mmol) and triethylamine (0.42 mL, 3 mmol) were added sequentially. After reacting for 30 min, the ice-cooling bath was removed and the reaction was continued for 12 h. The reaction solution was spin-dried, reconstituted in dichloromethane, and washed twice with water. The product was then spin-dried and purified by silica gel column chromatography (petroleum ether / ethyl acetate) to obtain 0.19 g of a yellow solid in a 33% yield, mp: 153-155°C. 1 HNMR (400MHz, CDCl3), δ8.56 (d, J=2.0Hz, 1H), 8.26-8.21 (m, 2H), 7.46-7.28 (m, 6H), 7.17 (d, J= 9.2Hz,1H),7.03-7.01(m,1H),5.56(s,2H),3.74(s,3H),3.32-3.28(m,6H),2.78-2.76(m,4H).

[0108] Synthesis of 1-(3,4-dichlorobenzyl)-N-[4-(4-(2-hydroxyamino-2-oxoethyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A1)

[0109] Hydroxylamine hydrochloride (4.67 g, 67 mmol) was dissolved in methanol (24 mL) under ice-cooling conditions to obtain Solution A. KOH (5.6 g, 100 mmol) was dissolved in methanol (14 mL) and stirred in a 40°C oil bath to dissolve, obtaining Solution B. Solution B was slowly added dropwise to Solution A under ice-cooling conditions. After the addition was complete, the mixture was ice-cooled for 30 minutes and then filtered to obtain a clear, transparent potassium hydroxylamine solution. Intermediate 5a (0.092 g, 0.14 mmol) was placed in a reaction flask and 2 mL of potassium hydroxylamine solution was added for reaction. After the reaction was complete, the pH was adjusted to 7 with 1 mol / L NaOH and filtered to obtain 0.024 g of a yellow solid with a yield of 27% and mp: 206-208°C. 1HNMR(DMSO-d6,400MHz), δ10.52(s,1H),8.85(s,1H),8.34-8.26(m,2H),8.01-7.98(m,1H),7.69(d,J=8.4Hz,1H),7.57-7.53(m,2H ),7.38(t,J=7.6Hz,1H),7.30(d,J=8.8Hz,1H),7.21-7.13(m,2H),5.69(s,2H),3.09-3.07(m,4H),2.95(s,2H),2.57-2.55(m,4H). 13 C NMR(DMSO-d6,100MHz),δ166.35,165.59,146.29,143.07,140.28,139.40,138.51,137.95,132.49,131.09,130.8 6,130.24,129.40,127.73,126.22,125.23,123.58,123.02,121.45,119.96,109.72,58.85,52.37,50.68,50.51.

[0110] Example 2. 1-(3,4-dichlorobenzyl)-N-[4-(4-(4-hydroxyamino-4-oxobutyl)piperazin-1-yl)-3-nitro Synthesis of [1-(2-aminophenylsulfonyl)-1H-indazole-3-carboxamide (A2)]

[0111] The intermediate and target compound were prepared as in Example 1. Yellow solid, yield 44%, mp: 150-151°C. 1 HNMR(400MHz, DMSO-d6), δ10.57(s,1H),8.82(s,1H),8.51(d,J=2.4Hz,1H) ,8.17-8.14(m,1H),8.07(d,J=8.0Hz,1H),7.88(d,J=8.4Hz,1H),7.68(s,1 H),7.61-7.56(m,2H),7.51-7.48(m,1H),7.34-7.27(m,2H),5.81(s,2H),3 .54-3.46(m,6H),3.14-3.11(m,4H),2.10-2.07(m,2H),1.97-1.92(m,2H). 13C NMR (100MHz, DMSO-d6), δ167.93,147.23,140.57,138.41,137.47,132.90,131.23,130.95,130.65,129. 67,127.94,127.44,126.98,123.65,121.50,121.40,110.83,55.15,51.34,50.47,47.25,29.25,19.39.

[0112] Example 3. 1-(3,4-dichlorobenzyl)-N-[4-(4-(5-hydroxyamino-5-oxopentyl)piperazin-1-yl)-3-nitro Synthesis of [1-(2-aminophenylsulfonyl)-1H-indazole-3-carboxamide (A3)]

[0113] The intermediate and target compound were prepared as in Example 1. Yellow solid, yield 49%, mp: 184-186°C. 1 HNMR(400MHz,DMSO-d6),δ12.43(s,1H),10.45(s,1H),8.75(s,1H),8.49(s,1H),8.15(s,1H),8.08(s,1H),7.86(s,1H),7.66-7.47(m, 4H),7.30(d,J=12.8Hz,2H),5.79(s,2H),3.5-3.43(m,6H),3.22-2.93(m,4H),2.01-1.91(m,2H),1.69-1.67(m,2H),1.53-1.52(m,2H). 13 C NMR (100MHz, DMSO-d6), δ168.65,145.58,140.55,139.80,134.76,132.72,132.30,128.34,127.67,127.57,126.45,126.28, 126.14,125.92,125.58,125.34,123.18,122.94,121.71,120.80,110.05,55.65,52.36,51.18,48.33,31.59,23.25,22.28.

[0114] Example 4. 1-(3,4-dichlorobenzyl)-N-[4-(4-(6-hydroxyamino-6-oxohexyl)piperazin-1-yl)-3-nitro Synthesis of [1-(2-aminophenylsulfonyl)-1H-indazole-3-carboxamide (A4)]

[0115] The intermediate and target compound were prepared as in Example 1. Yellow solid, yield 62%, mp: 162-164°C. 1HNMR(400MHz, DMSO-d6), δ10.37(s,1H),8.71(s,1H),8.35(d,J=1.6Hz,1H),8.30(d,J= 8.0Hz,1H),8.06(d,J=8.4Hz,1H),7.71(d,J=8.4Hz,1H),7.57(d,J=8.0Hz,1H),7.51(s ,1H),7.39-7.36(m,2H),7.22-7.18(m,1H),7.13(d,J=7.6Hz,1H),5.69(s,2H),3.26-3 .15(m,8H),2.94-2.89(m,2H),1.97-1.92(m,2H),1.63-1.47(m,4H),1.28-1.21(m,2H). 13 C NMR (100MHz, DMSO-d6), δ169.13,166.36,152.56,143.05,140.27,138.47,138.15,137.91,133.16,131.07,130.83,130.22,129 .38,127.70,126.18,124.12,123.54,123.00,121.43,114.26,109.69,69.50,50.67,32.25,28.53,28.31,28.24,25.17,25.06.

[0116] Example 5. 1-(3,4-dichlorobenzyl)-N-[4-(4-(7-hydroxyamino-7-oxoheptyl)piperazin-1-yl)-3-nitro Synthesis of [1-(2-aminophenylsulfonyl)-1H-indazole-3-carboxamide (A5)]

[0117] The intermediate and target compound were prepared as in Example 1. Yellow solid, yield 58%, mp: 203-205°C. 1 HNMR(400MHz,DMSO-d6),δ10.37(s,1H),8.70(s,1H),8.31-8.29(m,2H),8 .03-8.01(m,1H),7.70(d,J=8.4Hz,1H),7.56-7.52(m,2H),7.38-7.31(m,2 H),7.21-7.17(m,2H),7.14-7.11(m,2H),5.69(s,2H),3.17-3.15(m,4H),2 .90-2.59(m,6H),1.96-1.92(m,2H),1.52-1.46(m,4H),1.25-1.24(m,4H). 13C NMR (100MHz, DMSO-d6), δ169.11,166.29,145.90,142.97,140.34,139.67,138.50,132.62,131.10,130.86,130.24,129 .37,127.69,126.25,125.27,123.54,122.98,121.49,120.27,109.73,56.94,51.93,50.66,32.22,28.37,26.33,25.04.

[0118] Example 6. 1-(3,4-dichlorobenzyl)-N-[4-(4-(8-hydroxyamino-8-oxooctyl)piperazin-1-yl)-3-nitro Synthesis of [1-(2-aminophenylsulfonyl)-1H-indazole-3-carboxamide (A6)]

[0119] The intermediate and target compound were prepared as in Example 1. Yellow solid, yield 48%, mp: 175-177°C. 1 HNMR (400MHz, DMSO-d6), δ10.35(s,1H),8.67(s,1H),8.35(s,1H),8.31(d,J=8. 0Hz,1H),8.07(d,J=6.8Hz,1H),7.72(d,J=8.0Hz,1H),7.57(d,J=7.6Hz,1H),7.5 1(s,1H),7.40-7.37(m,2H),7.23-7.19(m,1H),7.12(d,J=6.4Hz,1H),5.70(s,2 H),3.27-3.00(m,10H),1.95-1.92(m,2H),1.59-1.43(m,4H),1.37-1.10(m,6H). 13 C NMR (100MHz, DMSO-d6), δ169.02,165.60,145.60,140.50,139.83,139.41,136.79,136.74,136.04,132.81,129.48,127.85,126.61,12 6.41,126.27,125.58,123.10,122.90,121.72,120.89,110.07,55.80,51.80,51.04,48.16,32.10,28.05,25.81,24.83,23.32,20.66.

[0120] Example 7. (E)-1-(3,4-dichlorobenzyl)-N-[4-(4-(4-(3-hydroxyamino-3-oxopropen-1-yl)benzyl]-1-(3,4-dichlorobenzyl)-N-[ ... Synthesis of [3-[(1-[(1-[(1-[(1-[(1-piperazin-1-yl)-3-nitrobenzenesulfonyl)-1H-indazole-3-carboxamide (A7)] ...

[0121] The intermediate and target compound were prepared as in Example 1. Yellow solid, yield 53%, mp: 202-205°C. 1HNMR(400MHz,DMSO-d6),δ10.80(s,1H),9.08(s,1H),8.36(s,1H),8.21(d, J=7.2Hz,1H),8.07(d,J=8.4Hz,1H),7.76(d,J=8.4Hz,1H),7.62-7.55(m,4 H),7.49-7.38(m,5H),7.26-7.22(m,1H),7.18(d,J=8.0Hz,1H),6.51(d,J= 16Hz,1H),5.72(s,2H),3.94(s,2H),3.24-3.22(m,4H),2.87-2.83(m,4H). 13 C NMR (100MHz, DMSO-d6), δ162.66,140.43,138.06,137.86,132.68,131.14,130.99,130.88,130.55,130.39,129.46,127.7 5,127.61,126.70,126.69,125.98,125.97,122.89,122.68,122.31,120.63,119.43,110.12,60.18,51.57,50.91,49.27.

[0122] Example 8. 1-(3,4-dichlorobenzyl)-N-[4-(4-(4-hydroxycarbamoylbenzyl)piperazin-1-yl)-3-nitro Synthesis of [1-(2-aminophenylsulfonyl)-1H-indazole-3-carboxamide (A8)]

[0123] The intermediate and target compound were prepared as in Example 1. Yellow solid, yield 30%, mp: 155-156°C. 1 HNMR (400MHz, DMSO-d6), δ11.25 (s, 1H), 9.08 (s, 1H), 8.37 (d, J = 2.0Hz, 1H) ,8.20(d,J=8.0Hz,1H),8.08-8.05(m,1H),7.78-7.76(m,3H),7.59-7.55(m ,2H),7.49(d,J=8.0Hz,2H),7.44-7.38(m,2H),7.27-7.23(m,1H),7.20-7. 18(m,1H),5.73(s,2H),3.91(s,2H),3.23-3.21(m,4H),2.81-2.79(m,4H). 13C NMR (100MHz, DMSO-d6), δ163.92,146.69,140.44,139.00,137.96,132.63,132.39,131.14,130.87,130.41,1 29.70,129.47,127.75,127.03,126.77,126.08,122.86,122.47,120.61,110.19,60.30,51.67,50.96,49.47.

[0124] Example 9. 1-(Naphthalen-2-ylmethyl)-N-[4-(4-(2-hydroxyamino-2-oxoethyl)piperazin-1-yl)-3-nitro Synthesis of [1-(2-aminophenylsulfonyl)-1H-indazole-3-carboxamide (A9)]

[0125] The intermediate and target compound were prepared as in Example 1. Yellow solid, yield 48%, mp: 229-230°C. 1 HNMR(400MHz, DMSO-d6), δ10.58(s,1H),8.89(s,1H),8.34-8.33(m,1H),8.24(d,J=8.4Hz,1H),8.05-8.02(m,1H),7.87-7.84(m,3H),7.81(s,1H) ,7.74(d,J=8.4Hz,1H),7.51-7.46(m,2H),7.39-7.35(m,3H),7.23-7.19 (m,1H),5.87(s,2H),3.17-3.15(m,4H),3.08(s,2H),2.68-2.66(m,4H). 13 C NMR (100MHz, DMSO-d6), δ164.90,146.68,140.38,138.90,134.56,132.74,132.56,132.35,128.33,127.72,127.5 9,126.44,126.16,126.09,125.89,125.48,123.04,122.78,122.01,120.28,110.24,58.22,52.58,52.18,49.98.

[0126] Example 10. 1-(Naphthalen-2-ylmethyl)-N-[4-(4-(4-hydroxyamino-4-oxobutyl)piperazin-1-yl)-3-nitro Synthesis of [1-(2-aminophenylsulfonyl)-1H-indazole-3-carboxamide (A10)]

[0127] The intermediate and target compound were prepared as in Example 1. Yellow solid, yield 72%, mp: 199-200°C. 1HNMR(400MHz, DMSO-d6), δ10.49(s,1H),8.81(s,1H),8.36(s,1H),8.29(d,J=8.4Hz,1H),8.05(d,J=8.8Hz,1H),7.85-7.71(m,5H),7.48(d,J =0.8Hz,2H),7.38-7.30(m,3H),7.23-7.19(m,1H),5.85(s,2H),3.20- 3.13(m,8H),2.94-2.89(m,2H),2.05-2.02(m,2H),1.85-1.81(m,2H). 13 C NMR (100MHz, DMSO-d6), δ168.20,145.71,140.52,140.50,139.69,134.73,132.73,132.31,128.34,127.69,127.58,126.45, 126.27,126.14,125.96,125.62,125.38,123.14,122.97,121.74,120.77,110.07,55.78,52.42,51.33,48.64,29.38,20.17.

[0128] Example 11. 1-(Naphthalen-2-ylmethyl)-N-[4-(4-(5-hydroxyamino-5-oxopentyl)piperazin-1-yl)-3-nitro Synthesis of [1-(2-aminophenylsulfonyl)-1H-indazole-3-carboxamide (A11)]

[0129] The intermediate and target compound were prepared as in Example 1. Yellow solid, yield 82%, mp: 185-186°C. 1 HNMR(400MHz, DMSO-d6), δ10.40(s,1H),8.73(s,1H),8.36(s,1H),8.31(d,J=8.4Hz,1H),8.04(d,J=8.8Hz,1H),7.84 -7.72(m,5H),7.47(s,2H),7.39-7.19(m,4H),5.85(s,2H),3.19-3.01(m,10H),1.97-1.92(m,2H),1.58-1.47(m,4H). 13C NMR (100MHz, DMSO-d6), δ168.65,145.57,140.55,139.80,134.76,132.74,132.30,128.34,127.67,127.57,126.45, 126.14,125.92,125.34,123.18,122.94,121.71,120.80,110.05,55.66,52.35,51.18,48.33,31.59,23.27,22.28.

[0130] Example 12. 1-(Naphthalen-2-ylmethyl)-N-[4-(4-(6-hydroxyamino-6-oxohexyl)piperazin-1-yl)-3-nitro Synthesis of [1-(2-aminophenylsulfonyl)-1H-indazole-3-carboxamide (A12)]

[0131] The intermediate and target compound were prepared as in Example 1. Yellow solid, yield 86%, mp: 178-180°C. 1 HNMR(400MHz, DMSO-d6), δ10.36(s,1H),8.71(s,1H),8.35-8.31(m,2H),8.03(d,J=8.4Hz,1H),7.85-7.72(m,5H ),7.47(s,2H),7.37-7.19(m,4H),5.85(s,2H),3.17-2.95(m,10H),1.92(s,2H),1.56-1.46(m,4H),1.18(s,2H). 13 C NMR (100MHz, DMSO-d6), δ168.91,145.49,141.79,140.61,139.88,134.81,132.72,132.30,128.34,127.66,127.57,126.45,126 .26,125.88,125.50,125.31,123.29,122.89,121.64,120.72,109.99,55.82,52.29,51.20,48.39,31.99,25.68,24.61,23.40.

[0132] Example 13. 1-(Naphthalen-2-ylmethyl)-N-[4-(4-(7-hydroxyamino-7-oxoheptyl)piperazin-1-yl)-3-nitro Synthesis of [1-(2-aminophenylsulfonyl)-1H-indazole-3-carboxamide (A13)]

[0133] The intermediate and target compound were prepared as in Example 1. Yellow solid, yield 22%, mp: 185-186°C. 1HNMR(400MHz,DMSO-d6),δ10.38(s,1H),8.69(s,1H),8.45(s,1H),8.20(s,1H),8.11(s,1H),7.86-7.85(m,5H ),7.50-7.26(m,6H),5.91(s,2H),3.39-3.36(m,8H),3.08-2.90(m,2H),1.95-1.49(m,6H),1.25-1.24(m,4H). 13 C NMR (100MHz, DMSO-d6), δ169.04,146.32,140.58,139.22,134.40,132.89,132.73,132.36,128.40,127.73,127.61,126.73 ,126.50,126.23,126.13,125.41,122.92,121.18,110.50,55.59,52.66,50.72,47.67,32.12,28.04,25.77,24.84,23.05.

[0134] Example 14. 1-(Naphthalen-2-ylmethyl)-N-[4-(4-(8-hydroxyamino-8-oxooctyl)piperazin-1-yl)-3-nitro Synthesis of [1-(2-aminophenylsulfonyl)-1H-indazole-3-carboxamide (A14)]

[0135] The intermediate and target compound were prepared as in Example 1. Yellow solid, yield 76%, mp: 178-179°C. 1 HNMR(400MHz,DMSO-d6),δ10.34(s,1H),8.68(s,1H),8.34-8.31(m,2H),8.03(d ,J=8.0Hz,1H),7.85-7.83(m,3H),7.75-7.73(m,2H),7.47(d,J=0.4Hz,2H),7.39 -7.35(m,1H),7.31-7.20(m,3H),5.85(s,2H),3.26-3.07(m,8H),2.96-2.89(m, 2H),1.93-1.90(m,2H),1.54-1.53(m,2H),1.43-1.41(m,2H),1.24-1.09(m,6H). 13C NMR (100MHz, DMSO-d6), δ169.08,145.47,140.59,139.91,134.80,132.72,132.29,128.32,127.65,127.56,126.25,12 6.11,125.86,125.29,122.86,121.63,120.73,109.97,55.94,52.28,48.33,32.20,28.32,28.16,25.92,24.95,23.51.

[0136] Example 15. (E)-1-(Naphthalen-2-ylmethyl)-N-[4-(4-(3-hydroxyamino-3-oxoprop-1-en-1-yl) Synthesis of [(1-[(2-[( ...

[0137] The intermediate and target compound were prepared as in Example 1. Yellow solid, yield 72%, mp: 159-161°C. 1 HNMR(400MHz,DMSO-d6),δ10.79(s,1H),9.08(s,1H),8.36-8.35(m,1H),8.24-8 .20(m,1H),8.06-8.04(m,1H),7.86-7.80(m,4H),7.76(d,J=8.4Hz,1H),7.58(d ,J=7.2Hz,2H),7.48-7.40(m,6H),7.38-7.35(m,3H),7.24-7.21(m,1H),6.50(d ,J=15.6Hz,1H),5.86(s,2H),3.88(s,2H),3.20-3.17(m,4H),2.81-2.78(m,4H). 13 C NMR (100MHz, DMSO-d6), δ168.86,152.58,140.24,139.37,137.90,136.83,136.74,136.14,133.21,129.47,127.92, 126.58,126.44,125.97,124.19,123.36,123.07,121.31,114.31,109.88,69.11,51.78,31.74,27.81,21.52,20.65.

[0138] Example 16. 1-(4'-Methyl-[1,1'-biphenyl]-4-ylmethyl)-N-[4-(4-(2-hydroxyamino-2-oxoethyl)- Synthesis of [3-[(1-[(1-[(1-[(1-nitrophenylsulfonyl)piperazin-1-yl)-3-nitrobenzenesulfonyl)-1H-indazole-3-carboxamide (A16)]]]

[0139] The intermediate and target compound were prepared as in Example 1. Yellow solid, yield 42%, mp: 196-198°C. 1HNMR(400MHz,DMSO-d6),δ11.15(s,1H),9.26(s,1H),8.51-8.49(m,1H),8.16-8 .13(m,1H),8.08(d,J=8.0Hz,1H),7.88(d,J=8.8Hz,1H),7.60(d,J=8.4Hz,2H), 7.55-7.46(m,4H),7.41(d,J=8.0Hz,2H),7.33-7.29(m,1H),7.25(d,J=7.6Hz,2 H),5.83(s,2H),3.70(s,2H),3.47-3.44(m,4H),3.22-3.20(m,4H),2.32(s,3H). 13 C NMR (100MHz, DMSO-d6), δ166.28,164.03,152.13,140.26,138.84,138.10,134.80,133.25,132.74,132.63,132.31,128.26,127.70,127. 57,127.22,127.18,126.39,126.09,126.02,125.50,124.36,123.35 ,123.14,121.38,114.80,109.92,70.23,59.77,52.42,20.78,14.10.

[0140] Example 17. 1-(4'-Methyl-[1,1'-biphenyl]-4-ylmethyl)-N-[4-(4-(4-hydroxyamino-4-oxobutane)- Synthesis of [3-[(1-[(1-[(1-[(1-[(1-piperazin-1-yl)-3-nitrobenzenesulfonyl)-1H-indazole-3-carboxamide (A17)] ...

[0141] The intermediate and target compound were prepared as in Example 1. Yellow solid, yield 97%, mp: 190-192°C. 1 HNMR(400MHz,DMSO-d6),δ10.49(s,1H),8.80(s,1H),8.37(s,1H),8.29(d,J=8.0Hz,1 H),8.06(d,J=8.4Hz,1H),7.74(d,J=8.8Hz,1H),7.57(d,J=7.6Hz,2H),7.49(d,J=7.6H z,2H),7.40-7.33(m,2H),7.28(d,J=7.6Hz,2H),7.23(d,J=7.6Hz,3H),5.72(s,2H),3. 21-3.14(m,8H),2.95-2.93(m,2H),2.31(s,3H),2.06-2.02(m,2H),1.86-1.83(m,2H). 13C NMR (100MHz, DMSO-d6), δ168.23,162.24,145.73,140.49,139.71,139.42,136.81,136.78,136.08,132.79,129.51,127.88, 126.65,126.45,126.27,125.62,123.15,122.94,121.71,120.80,110.09,55.81,51.81,51.36,48.66,29.42,20.69,20.21.

[0142] Example 18. 1-(4'-Methyl-[1,1'-biphenyl]-4-ylmethyl)-N-[4-(4-(5-hydroxyamino-5-oxopentanyl)- Synthesis of [3-[(1-[(1-[(1-[(1-[(1-piperazin-1-yl)-3-nitrobenzenesulfonyl)-1H-indazole-3-carboxamide (A18)] ...

[0143] The intermediate and target compound were prepared as in Example 1. Yellow solid, yield 57%, mp: 201-203°C. 1 HNMR(400MHz, DMSO-d6), δ10.40(s,1H),8.73(s,1H),8.36(s,1H),8.30(d,J=0.8Hz,1H),8.06(d,J=8.8Hz,1H),7.74(d,J=8.4Hz,1H),7.5 6-7.48(m,4H),7.40-7.32(m,2H),7.27-7.21(m,5H),5.72(s,2H),3. 21-2.99(m,10H),2.31(s,3H),1.98-1.93(m,2H),1.59-1.51(m,4H). 13 C NMR (100MHz, DMSO-d6), δ168.68,145.60,140.50,139.85,139.41,136.79,136.75,136.08,132.77,129.49,127.84,126.62 ,126.43,126.23,125.53,123.19,122.92,121.64,120.81,110.03,55.74,51.78,51.25,48.45,31.63,23.37,22.33,20.66.

[0144] Example 18. 1-(4'-Methyl-[1,1'-biphenyl]-4-ylmethyl)-N-[4-(4-(2-hydroxyamino-2-oxohexyl)- Synthesis of [3-nitrophenylsulfonyl]-1H-indazole-3-carboxamide (A19)

[0145] The intermediate and target compound were prepared as in Example 1. Yellow solid, yield 52%, mp: 151-152°C. 1HNMR (400MHz, DMSO-d6), δ10.37(s,1H),8.70(s,1H),8.37(s,1H),8.31(d,J=8. 0Hz,1H),8.06(d,J=8.4Hz,1H),7.74(d,J=8.8Hz,1H),7.57(d,J=8.0Hz,2H),7.4 9(d,J=7.6Hz,2H),7.40-7.33(m,2H),7.27-7.14(m,5H),5.71(s,2H),3.25-2.9 9(m,10H),2.31(s,3H),1.96-1.93(m,2H),1.61-1.48(m,4H),1.25-1.22(m,2H). 13 C NMR (100MHz, DMSO-d6), δ168.91,145.58,140.46,139.88,139.40,136.78,136.08,132.80,129.48,127.85,126.61,126.43 ,126.20,125.52,123.19,122.93,121.62,120.84,110.01,55.76,51.77,51.11,48.32,32.00,25.70,24.63,23.35,20.66.

[0146] Example 20. 1-(4'-Methyl-[1,1'-biphenyl]-4-ylmethyl)-N-[4-(4-(7-hydroxyamino-7-oxoheptyl)- Synthesis of [3-[(1-[(1-[(1-[(1-nitrophenylsulfonyl)piperazin-1-yl)-3-nitrobenzenesulfonyl)-1H-indazole-3-carboxamide (A20)]]]

[0147] The intermediate and target compound were prepared as in Example 1. Yellow solid, yield 52%, mp: 187-188°C. 1 HNMR(400MHz, DMSO-d6), δ10.36(s,1H),8.69(s,1H),8.37(s,1H),8.30(d,J=8.4Hz, 1H),8.07(d,J=8.4Hz,1H),7.75(d,J=8.4Hz,1H),7.57(d,J=7.6Hz,2H),7.49(d,J=8. 0Hz,2H),7.40-7.34(m,2H),7.28-7.20(m,5H),5.72(s,2H),3.26-3.00(m,10H),2.3 1(s,3H),1.94-1.91(m,2H),1.62-1.59(m,2H),1.48-1.45(m,2H),1.24-1.22(m,4H). 13C NMR (100MHz, DMSO-d6), δ169.02,165.60,145.61,140.50,139.83,139.41,136.79,136.74,136.04,132.81,129.48,127.85,126.61,12 6.41,126.27,125.59,123.10,122.90,121.72,120.89,110.07,55.80,51.79,51.03,48.16,32.10,28.05,25.81,24.83,23.32,20.66.

[0148] Example 21. 1-(4'-Methyl-[1,1'-biphenyl]-4-ylmethyl)-N-[4-(4-(8-hydroxyamino-8-oxooctane)] Synthesis of [3-[(1-[(1-[(1-[(1-nitrophenylsulfonyl)piperazin-1-yl)-3-nitrobenzenesulfonyl)-1H-indazole-3-carboxamide (A21)]]]

[0149] The intermediate and target compound were prepared as in Example 1. Yellow solid, yield 47%, mp: 228-230°C. 1 HNMR(400MHz,DMSO-d6),δ10.38(s,1H),8.69(s,1H),8.38-8.36(m,1H),8.30-8.28(m,1H),8. 07-8.04(m,1H),7.74(d,J=8.4Hz,1H),7.57(d,J=8.0Hz,2H),7.49(d,J=8.0Hz,2H),7.40-7.34 (m,2H),7.28(d,J=8.0Hz,2H),7.24-7.19(m,3H),5.72(s,2H),3.30-3.24(m,8H),3.03-2.99(m ,2H),2.31(s,3H),1.94-1.91(m,2H),1.63-1.60(m,2H),1.47-1.45(m,2H),1.23-1.21(m,6H). 13 C NMR (101MHz, DMSO-d6), δ169.08,145.58,140.46,139.85,139.39,136.79,136.76,136.09,132.82,129.49,127.88,126.61,126.43,1 26.22,125.52,123.20,122.93,121.63,120.85,110.03,55.78,51.77,51.00,48.15,32.21,28.33,28.21,25.99,24.98,23.36,20.67.

[0150] Example 22. (E)-1-(4'-Methyl-[1,1'-biphenyl]-4-ylmethyl)-N-[4-(4-(3-hydroxyamino-3- Synthesis of [1-(2-nitropropene-1-en-1-yl)benzyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A22) become

[0151] The intermediate and target compound were prepared as in Example 1. Yellow solid, yield 61%, mp: 189-191°C. 1 HNMR(400MHz,DMSO-d6),δ10.79(s,1H),9.07(s,1H),8.34-8.32(m,1H)),8.26-8.22 (m,1H),8.04-8.02(m,1H),7.74(d,J=8.4Hz,1H),7.56(d,J=8.0Hz,4H),7.50-7.44( m,3H),7.42(d,J=7.2Hz,2H),7.38-7.29(m,4H),7.24-7.19(m,3H),6.47(d,J=15.6H z,1H),5.72(s,2H),3.78(s,2H),3.16-3.15(m,4H),2.73-2.69(m,4H),2.31(s,3H). 13 C NMR (100MHz, DMSO-d6), δ162.71,162.32,146.43,140.37,139.45,139.21,137.96,136.78,135.90,132.59,130.13,129.49,127. 96,127.53,126.62,126.44,126.33,125.76,122.97,122.86,121.90,120.40,119.14,110.18,60.65,51.92,51.82,49.80,20.67.

[0152] Example 23. 2-(4'-Methyl-[1,1'-biphenyl]-4-ylmethyl)-N-[4-(4-(4-hydroxycarbamoylbenzyl)piperazin-1-yl)-3-nitro Synthesis of [1-(2-aminophenylsulfonyl)-1H-indazole-3-carboxamide (A23)]

[0153] The intermediate and target compound were prepared as in Example 1. Yellow solid, yield 49%, mp: 206-208°C. 1HNMR(400MHz,DMSO-d6),δ11.26(s,1H),9.08(s,1H),8.40(s,1H),8.16(d,J=8.0Hz,1H),8.10(d,J=8.4Hz,1H),7.81-7.77(m,3H),7.58(d,J=8.0Hz,2H),7.51-7.48(m,4H),7.45-7.41(m,2H),7.35(d,J=8.0Hz,2H),7.28-7.22(m,3H),5.76(s,2H),3.97(s,2H),3.27-3.25(m,4H),2.89-2.83(m,4H),2.31(s,3H). 13 C NMR(100MHz,DMSO-d6),δ163.85,146.87,140.49,139.57,138.81,136.85,136.75,135.62,132.73,132.62,129.52,128.04,127.09,126.76,126.68,126.37,122.91,122.68,120.78,110.60,59.96,52.17,51.46,49.08,20.68.

[0154] Example 24. Synthesis of methyl 4-(2-nitro-4-sulfonamidophenoxy)butyrate (4a)

[0155] To 3-nitro-4-chlorobenzenesulfonamide (4.73 g, 20 mmol), add water until stirring is sufficient. Saturated NaOH solution (20 mL) is slowly added to the mixture at 95°C. After the addition is complete, heat the mixture to 105°C and continue reacting for approximately 30 minutes. Once the reaction is complete, cool the mixture. Add 3 mol / L HCl solution to the reaction mixture, adjusting the pH to approximately 4. A light green solid precipitates, which is filtered and dried to yield 4.00 g of a light green solid in a 92% yield. Dissolve the light green solid (2.18 g, 10 mmol) in DMF (10 mL), add N,N-dimethylformamide dimethyl acetal (15 mL), and place in a 45°C oil bath. After the reaction is complete, add water to the reaction mixture, wash three times with ethyl acetate, and then adjust the pH of the aqueous phase to approximately 6 with 1 mol / L HCl solution. Wash and extract with ethyl acetate, then spin dry to yield 2.44 g of a light green solid in an 89% yield. A pale green solid (0.82 g, 3 mmol) was dissolved in DMF (15 mL). K2CO3 (1.24 g, 9 mmol) and methyl 4-bromobutyrate (0.63 mL, 4.5 mmol) were added, and the mixture was transferred to an 80°C oil bath and stirred overnight. After the reaction was complete, the mixture was cooled and ice water was added. Extraction with ethyl acetate and spin-drying gave a pale yellow oil. Methanol (15 mL) was stirred in an ice bath for 30 minutes. Acetyl chloride (3 mL) was slowly added dropwise. Stirring was continued for 30 minutes after the addition was complete. The pale yellow oil was transferred to a sealed tube and the prepared methanol solution was added. The mixture was sealed and stirred in a 60°C oil bath overnight. After the reaction was complete, the mixture was spin-dried and purified by silica gel column chromatography (petroleum ether / ethyl acetate) to give 0.73 g of a white solid in a 77% yield, mp: 203-205°C. 1 HNMR(400MHz, DMSO-d6), δ8.28(d,J=2.4Hz,1H),8.05-8.02(m,1H),7.57(d,J=9.2Hz,1H),7.52(s,2H),4. 26-4.23(m,2H),3.58(s,3H),2.34-2.31(m,2H),1.78-1.72(m,2H),1.62-1.55(m,2H),1.46-1.38(m,2H).

[0156] Synthesis of 4-[4-(N-[1-(3,4-dichlorobenzyl)-1H-indazole-3-carbonyl]sulfonyl)-2-nitrophenoxy]butyrate (6a)

[0157] 2a (0.32 g, 1 mmol) was added to a reaction flask, followed by the addition of thionyl chloride (6 mL). The reaction was stopped by reflux in an 82°C oil bath for 4 h. After cooling to room temperature, the mixture was spin-dried to dryness, and a white solid appeared in the reaction flask. Dichloromethane (15 mL) was added to the reaction flask under ice-cooling, and stirring was continued for 15 min. 4a (0.24 g, 0.75 mmol) was added, followed by triethylamine (0.42 mL, 3 mmol). After reacting for 30 min, the ice-cooling bath was removed and the reaction was continued at room temperature for 12 h. The reaction solution was spin-dried to dryness, reconstituted in dichloromethane, and washed twice with water. The residue was spin-dried and purified by silica gel column chromatography (petroleum ether:ethyl acetate) to afford 0.11 g of a white solid in a 23% yield, mp: 102-104°C. 1 H NMR(400MHz,DMSO-d6),δ12.62(s,1H),8.55-8.54(m,1H),8.32-8.29(m,1H),8.0 5-8.02(m,1H),7.89(d,J=8.0Hz,1H),7.71(d,J=2.0Hz,1H),7.65(d,J=8.8Hz,1H ),7.61-7.59(m,1H),7.53-7.49(m,1H),7.36-7.32(m,1H),7.31-7.28(m,1H),5. 83(s,2H),4.34-4.31(m,2H),3.60(s,3H),2.51-2.49(m,2H),2.06-2.00(m,2H).

[0158] Synthesis of 1-(3,4-dichlorobenzyl)-N-[4-(4-hydroxyamino-4-oxobutoxy)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (B1)

[0159] Hydroxylamine hydrochloride (4.67 g, 67 mmol) was dissolved in methanol (24 mL) under ice-cooling conditions to obtain Solution A. KOH (5.6 g, 100 mmol) was dissolved in methanol (14 mL) and stirred in a 40°C oil bath to dissolve, obtaining Solution B. Solution B was slowly added dropwise to Solution A under ice-cooling conditions. After the addition was complete, the mixture was ice-cooled for 30 minutes and then filtered to obtain a clear, transparent potassium hydroxylamine solution. Intermediate 6a (87 mg, 0.14 mmol) was placed in a reaction flask, and 2 mL of potassium hydroxylamine solution was added to react for 30 minutes. After adjusting the pH to 7 with 1 mol / L NaOH solution, the mixture was filtered to obtain a pale yellow solid with a yield of 60% and mp of 178-179°C. 1H NMR (400MHz, DMSO-d6), δ10.45(s,1H),8.70(s,1H),8.43(s,1H),8.20-8.16(m,2H),7.78(d,J=8.4Hz,1H ),7.61-7.41(m,4H),7.28-7.19(m,2H),5.75(s,2H),4.24(s,2H),2.16-2.13(m,2H),1.98-1.95(m,1H). 13 C NMR (100MHz, DMSO-d6), δ168.42,140.42,138.02,137.93,133.61,131.15,130.88,130.4 3,129.50,127.78,126.83,124.80,122.89,115.01,110.25,69.16,51.01,28.28,24.42.

[0160] Example 25. 1-(3,4-dichlorobenzyl)-N-[4-(5-hydroxyamino-5-oxopentyloxy)-3-nitrobenzenesulfonyl] Synthesis of [3-amino]-1H-indazole-3-carboxamide (B2)

[0161] The intermediate and target compound were prepared as in Example 24. Pale yellow solid, yield 42%, mp: 212-213°C. 1 H NMR (400MHz, DMSO-d6), δ12.61(s,1H),10.38(s,1H),8.53(s,1H),8.31-8. 28(m,1H),8.05(d,J=8.0Hz,1H),7.89(d,J=8.4Hz,1H),7.71(d,J=2.0Hz,1 H),7.65-7.59(m,2H),7.53-7.49(m,1H),7.36-7.28(m,2H),5.83(s,2H),4 .31-4.28(m,2H),2.04-2.01(m,2H),1.78-1.72(m,2H),1.69-1.62(m,2H). 13 C NMR (100MHz, DMSO-d6), δ168.82,160.63,155.11,140.60,138.34,137.35,135.42,133.90,131.26,130.98,130. 70,129.70,127.95,127.59,125.31,123.90,122.76,121.18,115.82,110.94,69.78,51.42,31.73,27.73,21.51.

[0162] Example 26. 1-(3,4-dichlorobenzyl)-N-[4-(6-hydroxyamino-6-oxohexyloxy)-3-nitrobenzenesulfonyl] Synthesis of [3-amino]-1H-indazole-3-carboxamide (B3)

[0163] The intermediate and target compound were prepared as in Example 24. Pale yellow solid, yield 69%, mp: 125-127°C. 1 H NMR(400MHz, DMSO-d6), δ10.36(s,1H),8.68(s,1H),8.31-8.27(m,2H),8.12-8.09(m,1H),7.69(d,J=8.0Hz,1H),7.57-7.53(m,2H),7.42-7 .35(m,2H),7.21-7.14(m,2H),5.69(s,2H),4.19-4.04(m,2H),1.98- 1.94(m,2H),1.73-1.71(m,2H),1.56-1.52(m,2H),1.40-1.36(m,2H). 13 C NMR (100MHz, DMSO-d6), δ168.97,140.30,138.41,137.88,133.23,131.07,130.83,130.23,129.36,127. 67,126.22,124.23,123.38,122.99,121.53,114.36,109.72,69.46,50.69,32.19,28.01,24.87,24.74.

[0164] Example 27. 1-(3,4-dichlorobenzyl)-N-[4-(7-hydroxyamino-7-oxoheptyloxy)-3-nitrobenzenesulfonyl]-1H-indazole-3- Synthesis of formamide (B4)

[0165] The intermediate and target compound were prepared as in Example 24. Pale yellow solid, yield 45%, mp: 118-120°C. 1 H NMR (400MHz, DMSO-d6), δ10.34(s,1H),8.66(s,1H),8.33-8.29(m,2H),8. 13-8.09(m,1H),7.68(d,J=8.4Hz,1H),7.56-7.52(m,2H),7.40-7.34(m,2H ),7.20-7.12(m,2H),5.69(s,2H),4.19-4.17(m,2H),1.96-1.93(m,2H),1 .75-1.68(m,2H),1.54-1.48(m,2H),1.44-1.36(m,2H),1.32-1.24(m,2H). 13C NMR (100MHz, DMSO-d6), δ169.13,166.36,152.58,143.03,140.28,138.48,138.14,137.90,133.20,131.08,130.83,130. 23,129.37,127.69,126.19,124.15,123.52,123.01,121.44,114.28,109.69,69.46,50.68,32.24,28.16,25.10,24.97.

[0166] Example 28. 1-(3,4-dichlorobenzyl)-N-[4-(8-hydroxyamino-8-oxooctyloxy)-3-nitrobenzenesulfonyl] Synthesis of [3-amino]-1H-indazole-3-carboxamide (B5)

[0167] The intermediate and target compound were prepared as in Example 24. Pale yellow solid, yield 75%, mp: 155-156°C. 1 H NMR(400MHz, DMSO-d6), δ10.34(s,1H),8.67(s,1H),8.31-8.30(m,2H),8.11-8.09(m,1H),7.68(d,J=8.4Hz,1H),7.56-7.53(m,2H) ),7.40-7.34(m,2H),7.21-7.13(m,2H),5.69(s,2H),4.20-4.17(m,2H),1.96-1.92(m,2H),1.75-1.68(m,2H),1.52-1.22(m,2H). 13 C NMR (100MHz, DMSO-d6), δ169.13,166.36,152.56,143.05,140.27,138.47,138.16,137.91,133.16,131.07,130.83,130.22,129 .38,127.70,126.19,124.12,123.54,123.00,121.43,114.26,109.69,69.51,50.66,32.25,28.53,28.32,28.24,25.17,25.07.

[0168] Example 29. 1-(3,4-dichlorobenzyl)-N-[4-(4-hydroxycarbamoylbenzyloxy)-3-nitrobenzenesulfonyl] Synthesis of [3-amino]-1H-indazole-3-carboxamide (B6)

[0169] The intermediate and target compound were prepared as in Example 24. Pale yellow solid, yield 47%, mp: 181-183°C. 1H NMR(400MHz, DMSO-d6), δ11.23(s,1H),9.06(s,1H),8.40-8.39(m,1H),8.28(d,J=8.0Hz,1H),8.17-8.13(m,2H),7.79- 7.77(m,2H),7.71(d,J=8.4Hz,1H),7.57-7.51(m,5H),7.40-7.36(m,1H),7.23-7.14(m,2H),5.70(s,2H),5.43(s,2H). 13 C NMR (100MHz, DMSO-d6), δ164.01,152.37,140.33,138.80,138.34,138.10,133.34,132.64,131.11,130.86 ,130.29,129.41,127.72,127.25,127.19,126.36,124.49,123.00,121.75,114.93,109.85,70.29,50.77.

[0170] Example 30. 1-(Naphthalen-2-ylmethyl)-N-[4-(4-hydroxyamino-4-oxobutoxy)-3-nitrobenzenesulfonyl] Synthesis of [3-amino]-1H-indazole-3-carboxamide (B7)

[0171] The intermediate and target compound were prepared as in Example 24. Pale yellow solid, 82% yield, mp: 203-205°C. 1 H NMR (400MHz, DMSO-d6), δ10.45(s,1H),8.70(s,1H),8.35(d,J=1.2Hz,1H) ,8.30(d,J=8.0Hz,1H),8.14-8.11(m,1H),7.86-7.83(m,3H),7.79(s,1H), 7.68(d,J=8.4Hz,1H),7.49-7.47(m,2H),7.41-7.31(m,3H),7.19-7.16(m, 1H),5.84(s,2H),4.21-4.18(m,2H),2.16-2.12(m,2H),1.99-1.92(m,2H). 13 C NMR (100MHz, DMSO-d6), δ168.46,152.65,140.26,137.84,134.78,133.29,132.73,132.30,128.25,127.68,127.5 5,126.38,126.07,125.98,125.46,124.29,123.34,123.10,121.36,114.35,109.89,68.90,52.40,28.32,24.47.

[0172] Example 31. N-[4-(5-hydroxyamino-5-oxopentyloxy)-3-nitrobenzenesulfonyl]-1-(naphthalen-2-ylmethyl Synthesis of 1H-indazole-3-carboxamide (B8)

[0173] The intermediate and target compound were prepared as in Example 24. Pale yellow solid, yield 89%, mp: 195-196°C. 1 H NMR(400MHz, DMSO-d6), δ10.39(s,1H),8.72(s,1H),8.40-8.39(m,1H),8.24(d,J=8.0Hz,1H),8.18-8.16(m,1H),7.87-7.82(m,4H),7.74(d, J=8.4Hz,1H),7.50-7.45(m,3H),7.39-7.35(m,2H),7.23-7.20(m,1H) ,5.88(s,2H),4.23-4.20(m,2H),2.03-2.00(m,2H),1.74-1.61(m,4H). 13 C NMR (100MHz, DMSO-d6), δ168.86,153.28,140.36,138.02,134.53,133.39,132.73,132.34,128.31,127.71,127.57,1 26.42,126.36,126.14,126.09,125.47,124.50,123.04,121.99,114.72,110.22,69.28,52.59,31.73,27.78,21.52.

[0174] Example 32. 1-(Naphthalen-2-ylmethyl)-N-[4-(6-hydroxyamino-6-oxohexyloxy)-3-nitrobenzenesulfonyl] Synthesis of [3-amino]-1H-indazole-3-carboxamide (B9)

[0175] The intermediates and target compounds were prepared as in Example 24. Pale yellow solid, yield 90%, mp: 165-167°C. 1 H NMR(400MHz,DMSO-d6),δ10.36(s,1H),8.69(s,1H),8.33-8.29(m,2H),8.1 3(d,J=8.0Hz,1H),7.85-7.79(m,4H),7.68(d,J=8.0Hz,1H),7.49-7.47(m, 2H),7.41-7.31(m,3H),7.19-7.15(m,4H),5.84(s,2H),4.19-4.16(m,2H), 1.98-1.95(m,2H),1.76-1.69(m,2H),1.57-1.51(m,2H),1.42-1.36(m,2H). 13C NMR (100MHz, DMSO-d6), δ169.00,152.62,140.25,137.91,134.80,133.18,132.73,132.30,128.25,127.69,127.56,126. 38,126.07,126.00,125.49,124.16,123.41,123.10,121.33,114.33,109.88,69.44,52.39,32.20,28.02,24.88,24.74.

[0176] Example 33. N-[4-(7-hydroxyamino-7-oxoheptyloxy)-3-nitrobenzenesulfonyl]-1-(naphthalen-2-ylmethyl Synthesis of 1H-indazole-3-carboxamide (B10)

[0177] The intermediate and target compound were prepared as in Example 24. Pale yellow solid, yield 91%, mp: 168-170°C. 1 H NMR(400MHz, DMSO-d6), δ10.35(s,1H),8.68(s,1H),8.32-8.30(m,2H),8.12(d,J=8.0Hz,1H),7.87-7.79(m,4H),7.67(d,J=7.6Hz,1H),7.48(d ,J=0.8Hz,2H),7.40-7.31(m,3H),7.18-7.15(m,1H),5.83(s,2H),4.19 -4.16(m,2H),1.96-1.93(m,2H),1.73-1.69(m,2H),1.50-1.28(m,6H). 13 C NMR (100MHz, DMSO-d6), δ169.12,166.45,152.55,142.56,140.24,138.18,137.92,134.83,133.16,132.73,132.30,128.24,127.68,127. 55,126.37,126.06,125.99,125.94,125.49,124.12,123.46,123.11 ,121.25,114.27,109.84,69.45,52.37,32.22,28.13,25.06,24.93.

[0178] Example 34. N-[4-(8-hydroxyamino-8-oxooctyloxy)-3-nitrobenzenesulfonyl]-1-(naphthalen-2-ylmethyl Synthesis of 1H-indazole-3-carboxamide (B11)

[0179] The intermediate and target compound were prepared as in Example 24. Pale yellow solid, yield 80%, mp: 172-173°C. 1H NMR (400MHz, DMSO-d6), δ10.34(s,1H),8.68(s,1H),8.33-8.30(m,2H),8.12(d,J=8 .4Hz,1H),7.86-7.79(m,4H),7.67(d,J=8.4Hz,1H),7.49-7.47(m,2H),7.40-7.31( m,3H),7.19-7.15(m,2H),5.84(s,2H),4.19-4.17(m,2H),3.36(s,2H),1.96-1.92( m,2H),1.74-1.67(m,2H),1.50-1.47(m,2H),1.41-1.37(m,2H),1.30-1.24(m,4H). 13 C NMR (100MHz, DMSO-d6), δ169.14,166.43,152.59,140.25,138.09,137.93,134.83,133.16,132.74,132.30,128.25,127.69,127.56,126.39 ,126.08,126.00,125.97,125.49,124.13,123.45,123.10,121.29,11 4.29,109.87,69.51,52.38,32.25,28.52,28.30,28.23,25.16,25.06.

[0180] Example 35. (E)-1-(Naphthalen-2-ylmethyl)-N-[4-([4-(3-hydroxyamino-3-oxoprop-1-en-1-yl)benzyl] ...3-hydroxyamino- Synthesis of [3-nitrophenylsulfonyl]-1H-indazole-3-carboxamide (B12)

[0181] The intermediate and target compound were prepared as in Example 24. Pale yellow solid, yield 48%, mp: 173-175°C. 1 H NMR (400MHz, DMSO-d6), δ10.81(s,1H),9.08(s,1H),8.43(s,1H),8.25-8.18(m,2H),7.87-7.81(m,4H),7.73(d,J=8.4Hz,1H),7.62(d,J=8. 0Hz,2H),7.56(d,J=9.2Hz,1H),7.51-7.45(m,5H),7.39-7.34(m,2H),7.22-7.19(m,1H)),6.51(d,J=15.6Hz,1H),5.87(s,2H),5.39(s,2H). 13C NMR (100MHz, DMSO-d6), δ163.13,140.81,138.64,138.32,137.34,135.16,135.05,133.86,133.19,132.79,128.77 ,128.40,128.17,128.04,126.88,126.60,126.54,125.94,125.05,123.53,119.88,115.60,110.64,71.01,53.02.

[0182] Example 36. N-[4-(4-Hydroxycarbamoylbenzyloxy)-3-nitrobenzenesulfonyl]-1-(naphthalen-2-yl)methyl Synthesis of 1H-indazole-3-carboxamide (B13)

[0183] The intermediate and target compound were prepared as in Example 24. Pale yellow solid, yield 98%, mp: 182-184°C. 1 H NMR(400MHz, DMSO-d6), δ11.23(s,1H),9.07(s,1H),8.39(d,J=2.0Hz,1H),8.30(d,J=8.4Hz,1H),8.16-8.13(m,1H),7.87-7.83(m ,3H),7.79-7.77(m,3H),7.68(d,J=8.4Hz,1H),7.55-7.47(m,5H),7.37-7.31(m,2H),7.19-7.15(m,1H),5.84(s,2H),5.41(s,2H). 13 CNMR(100MHz,DMSO-d6),δ162.68,140.35,138.18,137.87,136.89,134.71,134.60,133.41,132.74,132.34,128.3 2,127.95,127.72,127.59,126.43,126.15,126.09,125.49,124.60,123.08,119.43,115.14,110.19,70.55,52.57.

[0184] Example 37. N-[4-(4-Hydroxyamino-4-oxobutoxy)-3-nitrobenzenesulfonyl]-1-([1,1'-biphenyl]- Synthesis of 4'-amino-4-ylmethyl)-1H-indazole-3-carboxamide (B14)

[0185] The intermediate and target compound were prepared as in Example 24. Pale yellow solid, yield 84%, mp: 183-184°C. 1H NMR (400MHz, DMSO-d6), δ10.45(s,1H),8.71(s,1H),8.34-8.29(m,2H),8.13(d,J=8.4Hz,1H),7.69(d,J=8.4Hz,1H),7.56(d,J=8.0H z,2H),7.50(d,J=7.6Hz,2H),7.40-7.16(m,7H),5.70(s,2H),4.21-4.18(m,2H),2.31(s,3H),2.16-2.13(m,2H),1.98-1.92(m,2H). 13 C NMR (100MHz, DMSO-d6), δ168.48,166.40,152.56,140.24,139.37,137.82,136.84,136.74,136.16,133.28,129.48,1 27.93,126.58,126.44,125.95,124.26,123.43,123.07,121.26,114.30,109.87,68.88,51.76,28.33,24.48,20.66.

[0186] Example 38. N-[4-(5-hydroxyamino-5-oxopentyloxy)-3-nitrobenzenesulfonyl]-1-[(4'-methyl-[1, Synthesis of [1'-biphenyl]-4-yl]methyl]-1H-indazole-3-carboxamide (B15)

[0187] The intermediate and target compound were prepared as in Example 24. Pale yellow solid, yield 94%, mp: 186-188°C. 1 H NMR (400MHz, DMSO-d6), δ10.38(s,1H),8.71(s,1H),8.33-8.28(m,2H),8.13-8.11(m,1H),7.69(d,J=7.6Hz,1H),7.5 6-7.48(m,4H),7.41-7.18(m,7H),5.70(s,2H),4.21-4.19(m,2H),2.31(s,3H),2.03-1.99(m,2H),1.71-1.65(m,4H). 13 C NMR (100MHz, DMSO-d6), δ168.86,152.58,140.24,139.37,137.90,136.83,136.74,136.14,133.20,129.47,127.92, 126.58,126.44,125.96,124.19,123.37,123.07,121.30,114.31,109.88,69.11,51.78,31.74,27.80,21.52,20.65.

[0188] Example 39. N-[4-(6-hydroxyamino-6-oxohexyloxy)-3-nitrobenzenesulfonyl]-1-[(4'-methyl-[1, Synthesis of [1'-biphenyl]-4-yl]methyl]-1H-indazole-3-carboxamide (B16)

[0189] The intermediate and target compound were prepared as in Example 24. Pale yellow solid, yield 55%, mp: 162-164°C. 1 H NMR (400MHz, DMSO-d6) δ10.36(s,1H),8.68(s,1H),8.33-8.28(m,2H),8.13(d,J=8.0Hz,1H),7.69(d,J=8.0Hz,1H),7.56-7.48(m,4H),7. 42-7.16(m,7H),5.70(s,2H),4.20-4.17(m,2H),2.31(s,3H),1.98-1 .94(m,2H),1.76-1.71(m,2H),1.56-1.53(m,2H),1.42-1.35(m,2H). 13 C NMR (100MHz, DMSO-d6), δ168.99,152.66,140.25,139.37,137.89,136.83,136.74,136.13,133.21,129.48,127.93,126 .58,126.44,125.98,124.20,123.34,123.07,121.33,114.35,109.90,69.46,51.79,32.20,28.02,24.88,24.75,20.66.

[0190] Example 40. N-[4-(7-hydroxyamino-7-oxoheptyloxy)-3-nitrobenzenesulfonyl]-1-[(4'-methyl-[1, Synthesis of [1'-biphenyl]-4-yl]methyl]-1H-indazole-3-carboxamide (B17)

[0191] The intermediate and target compound were prepared as in Example 24. Pale yellow solid, yield 52%, mp: 213-215°C. 1 H NMR (400MHz, DMSO-d6), δ10.34(s,1H),8.67(s,1H),8.32-8.29(m,2H),8.12(d,J=8.4Hz,1H),7.68(d,J=8.0Hz,1H),7.56-7.48( m,4H),7.40-7.15(m,7H),5.69(s,2H),4.19-4.17(m,2H),2.31(s,3H),1.96-1.93(m,2H),1.74-1.68(m,2H),1.51-1.24(m,6H). 13CNMR(100MHz,DMSO-d6),δ169.13,152.56,140.23,139.35,137.89,136.83,136.73,136.17,133.18,129.47,127.90,126 .57,126.43,125.90,124.14,123.44,123.08,121.21,114.26,109.83,69.45,51.75,32.22,28.14,25.06,24.93,20.65.

[0192] Example 41. N-[4-(8-hydroxyamino-8-oxooctyloxy)-3-nitrobenzenesulfonyl]-1-[(4'-methyl-[1, Synthesis of [1'-biphenyl]-4-yl]methyl]-1H-indazole-3-carboxamide (B18)

[0193] The intermediate and target compound were prepared as in Example 24. Pale yellow solid, yield 82%, mp: 195-197°C. 1 H NMR(400MHz,DMSO-d6),δ10.34(s,1H),8.68(s,1H),8.32-8.29(m,2H),8.12- 8.09(m,1H),7.68(d,J=8.4Hz,1H),7.56-7.48(m,4H),7.40-7.28(m,4H),7.2 4-7.15(m,3H),5.70(s,2H),4.19-4.16(m,2H),2.31(s,3H),1.95-1.92(m,2H ),1.73-1.68(m,2H),1.50-1.47(m,2H),1.41-1.38(m,2H),1.29-1.24(m,4H). 13 C NMR (100MHz, DMSO-d6), δ169.14,166.52,152.54,142.63,140.23,139.36,138.24,137.92,136.84,136.74,136.17,133.15,129.47,127.93 ,126.57,126.44,125.92,124.10,123.47,123.07,121.22,114.26,10 9.84,69.51,51.75,32.25,28.51,28.30,28.23,25.16,25.06,20.66.

[0194] Activity evaluation of target compounds

[0195] Experimental Example 1: HDAC1 enzyme inhibition test of target compound

[0196] Experimental reagents:

[0197] HDAC1 enzyme solution, HDAC buffer, fluorescent test substrate (Ac-Leu-Gly-Lys(Ac)-AMC), trypsin stop solution, etc.

[0198] Experimental instruments:

[0199] Thermo Varioskan Flash full wavelength multifunctional microplate reader.

[0200] Experimental steps:

[0201] (1) Set up experimental group, 100% group and blank group

[0202] Experimental group: 10 μL compound solution + 50 μL enzyme solution + 40 μL fluorescent substrate;

[0203] 100% group: 10 μL HDAC buffer + 50 μL enzyme solution + 40 μL fluorescent substrate;

[0204] Blank group: 10 μL HDAC buffer + 50 μL HDAC buffer + 40 μL fluorescent substrate;

[0205] (2) 10 μL of compound solution or HDAC buffer at different concentrations was incubated with 50 μL of enzyme solution or HDAC buffer for 10 min (experimental group: 10 μL compound solution + 50 μL enzyme solution; 100% group: 10 μL HDAC buffer + 50 μL enzyme solution; blank group: 10 μL HDAC buffer + 50 μL HDAC buffer). 40 μL of fluorescent substrate was added to all wells and incubated at 37°C for 60-120 min. Subsequently, 100 μL of trypsin stop solution was added to each well and incubated for another 30 min before measuring the fluorescence value of the sample.

[0206] (3) Calculate the inhibition rate of the compound at each concentration according to the following formula, and then fit the standard curve using Graphpad Prism software to obtain the IC of the compound. 50 value.

[0207]

[0208] The experimental results are shown in Table 2

[0209] Table 2. Results of the target compounds' inhibition test on HDAC1 enzyme

[0210]

[0211]

[0212] The values ​​in the table are the average values ​​of three test results.

[0213] As can be seen from the table, some compounds exhibited significant HDAC1 inhibitory activity. These included compounds A5, A6, A7, A14, A15, A17, A19, A23, B5, B11, B16, B17, and B18. Compounds A7 and B17 exhibited similar HDAC1 inhibitory activity to that of the positive control drug, SAHA. Therefore, the highly active compounds A5, A6, A7, A14, A15, A17, A19, A23, B5, B11, B16, B17, and B18 were selected for testing their Mcl-1 inhibitory activity.

[0214] Experimental Example 2: Inhibition test of active target compounds on Mcl-1 protein (Invitro)

[0215] Experimental reagents:

[0216] Bid-BH3 peptide fluorescently labeled with 5-FAM at the N-terminus (5-FAM-QEDIIRNIARHLAQVGDSMDRSIPPG) was dissolved in 1× PBS;

[0217] Assay buffer: 1× PBS;

[0218] Calibration solution: 1 nM fluorescein, 10 mM NaOH.

[0219] Experimental instruments:

[0220] TECAN Genios Pro multi-function microplate reader.

[0221] Experimental steps:

[0222] (1) Add the target protein and the small molecule compound to be tested into the test buffer, mix well and incubate at room temperature in the dark for 30 minutes.

[0223] The total volume of each solution was 200 μL, mixed and incubated at room temperature in the dark for 20 min. (2) 60 μL of each of the above solution and calibration solution was transferred to a black 384-well plate (three parallel groups) and immediately detected on a microplate reader.

[0224] Fluorescence polarization detection was performed with an excitation wavelength of 485 nm and an emission wavelength of 535 nm, and the fluorescence polarization value of the calibration solution was set to 20 mP.

[0225] (3) All compounds were first screened at three typical concentrations (1 μM, 10 μM, and 50 μM). Each compound was tested in parallel in three replicate wells on the same plate, and the polarization value was averaged. The inhibition rate was calculated based on the results of the polarization values ​​of the negative control, positive control, and the tested compound. The target protein concentration usually used in the assay is 300-500 nM, the fluorescently labeled peptide is 5-FAM-Bid-BH3 peptide, and the positive compound is AT-101. If the test results show that the inhibition rate of the compound at a concentration of 50 μM is greater than 50%, and its inhibition rate shows a clear dose-dependent relationship at the three tested concentrations, it is considered that the compound has specific binding to the target protein, and further determination of a more accurate IC is required. 50 Numeric value.

[0226] (4) For compounds that showed significant activity in the initial screening, complete binding curves were determined at 7 different concentrations (1nM, 10nM, 100nM, 1μM, 10μM, 50μM, 100μM). Each compound was tested in three replicate wells on the same plate, and the polarization value was averaged. GraphPad Prism software was used to process the data and plot the graphs to obtain the IC value of the compound. 50 value.

[0227] (5) Based on the total protein concentration, total fluorescent peptide concentration, protein-peptide complex dissociation constant, and IC of the detection compound used in the measurement 50 The competitive inhibition constant K of the test compound was calculated using the calculation method in the following literature. i

[0228] (Nikolovska-Coleska, Z.; et al. Development and optimization of a binding assay for the XIAP BIR3 domain using fluorescence polarization. Anal Biochem. 2004, 332, 261-273).

[0229] The experimental results are shown in Table 3.

[0230] Table 3. In vitro inhibition test results of target compounds on Mcl-1 protein

[0231]

[0232] The values ​​in the table are the average values ​​of three test results.

[0233] As can be seen from the table, most compounds exhibit submicromolar inhibitory activity against Mcl-1. Compounds A6, A7, A14, A15, A17, A19, A23, B5, B11, B16, B17, and B18 exhibited significant inhibitory activity against Mcl-1. Compounds A7, A15, A23, B17, and B18 exhibited comparable or superior Mcl-1 inhibitory activity to the positive-acting drug UMI-77.

[0234] In summary, most compounds have dual HDAC / Mcl-1 inhibitory activity, especially B17 and B18, whose inhibitory activities against both targets have reached the nanomolar level. This is of great significance for the further development of more active HDAC / Mcl-1 protein inhibitors and the preparation of drugs for the prevention or treatment of related mammalian diseases caused by HDAC / Mcl-1 protein abnormalities.

[0235] The above description is merely a preferred embodiment of the present invention and does not constitute any form of limitation to the present invention. Although the present invention has been disclosed as above in terms of a preferred embodiment, it is not intended to limit the present invention. Any technician familiar with the present invention can make slight changes or modifications to equivalent embodiments using the above-mentioned technical contents without departing from the scope of the technical solution of the present invention. However, any simple modifications, equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention without departing from the content of the technical solution of the present invention are still within the scope of the solution of the present invention.

Claims

1. A substituted indazole HDAC / Mcl-1 dual-target inhibitor, characterized by: A compound or a pharmaceutically acceptable salt thereof having the structure of general formula (I) or general formula (II); In the general formulas (I) and (II), R is an alkyl group, an aryl group or a heteroaryl group; Linker is selected from a C2-C8 alkyl group, a differently substituted phenyl group, a C3-C8 cycloalkyl group or a C5-C10 aryl group; and X is a carbonyl group, a sulfonyl group or -CH2-.

2. The substituted indazole HDAC / Mcl-1 dual-target inhibitor according to claim 1, characterized in that: R is C1-C10 alkyl, C3-C10 cycloalkyl, C5-C15 aryl, a monoheterocyclic aryl containing 5 or 6 ring atoms, or a biheterocyclic aryl having 8 to 15 ring atoms; The heterocyclic aromatic group contains 1-4 heteroatoms, which are independently selected from O, S, N, oxidized S or oxidized N. The carbon atom or nitrogen atom is the connection point of the heteroaromatic ring structure to maintain a stable aromatic ring.

3. The substituted indazole HDAC / Mcl-1 dual-target inhibitor according to claim 2, characterized in that: R is a halogen C1-C6 alkyl, a C1-C6 alkyl, a C3-C8 cycloalkyl, an aromatic group Ar or -NH-R1 connected to a morpholine group or a piperazine group substituted or unsubstituted by 1-2 hydroxyl, halogen, nitro, or cyano substituents; wherein Ar is phenyl, naphthyl, pyridyl, pyridazinyl, pyrazinyl, indolizinyl, quinazolinyl, purinyl, indolyl, quinolyl, pyrimidinyl, pyrrolyl, pyrazolyl, thiazolyl, benzothiazolyl, thienyl, benzo[b]thienyl, isoxazolyl, oxathiadiazolyl, isothiazolyl, tetrazolyl, imidazolyl, triazinyl, furyl, benzofuranyl, or indolyl, which may contain one substituent or no substituent.

4. The substituted indazole HDAC / Mcl-1 dual-target inhibitor according to claim 1, characterized in that It is selected from: 1-(3,4-Dichlorobenzyl)-N-[4-(4-(2-hydroxyamino-2-oxoethyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A1); 1-(3,4-Dichlorobenzyl)-N-[4-(4-(4-hydroxyamino-4-oxobutyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A2); 1-(3,4-Dichlorobenzyl)-N-[4-(4-(5-hydroxyamino-5-oxopentyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A3); 1-(3,4-Dichlorobenzyl)-N-[4-(4-(6-hydroxyamino-6-oxohexyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A4); 1-(3,4-Dichlorobenzyl)-N-[4-(4-(7-hydroxyamino-7-oxoheptyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A5); 1-(3,4-Dichlorobenzyl)-N-[4-(4-(8-hydroxyamino-8-oxooctyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A6); (E)-1-(3,4-dichlorobenzyl)-N-[4-(4-(4-(3-hydroxyamino-3-oxopropen-1-yl)benzyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A7); 1-(3,4-Dichlorobenzyl)-N-[4-(4-(4-hydroxycarbamoylbenzyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A8); 1-(Naphthalen-2-ylmethyl)-N-[4-(4-(2-hydroxyamino-2-oxoethyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A9); 1-(Naphthalen-2-ylmethyl)-N-[4-(4-(4-hydroxyamino-4-oxobutyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A10); 1-(Naphthalen-2-ylmethyl)-N-[4-(4-(5-hydroxyamino-5-oxopentyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A11); 1-(Naphthalen-2-ylmethyl)-N-[4-(4-(6-hydroxyamino-6-oxohexyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A12); 1-(Naphthalen-2-ylmethyl)-N-[4-(4-(7-hydroxyamino-7-oxoheptyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A13); 1-(Naphthalen-2-ylmethyl)-N-[4-(4-(8-hydroxyamino-8-oxooctyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A14); (E)-1-(naphthalen-2-ylmethyl)-N-[4-(4-(4-(3-hydroxyamino-3-oxoprop-1-en-1-yl)benzyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A15); 1-(4'-Methyl-[1,1'-biphenyl]-4-ylmethyl)-N-[4-(4-(2-hydroxyamino-2-oxoethyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A16); 1-(4'-Methyl-[1,1'-biphenyl]-4-ylmethyl)-N-[4-(4-(4-hydroxyamino-4-oxobutyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A17); 1-(4'-Methyl-[1,1'-biphenyl]-4-ylmethyl)-N-[4-(4-(5-hydroxyamino-5-oxopentyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A18); 1-(4'-Methyl-[1,1'-biphenyl]-4-ylmethyl)-N-[4-(4-(2-hydroxyamino-2-oxohexyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A19); 1-(4'-Methyl-[1,1'-biphenyl]-4-ylmethyl)-N-[4-(4-(7-hydroxyamino-7-oxoheptyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A20); 1-(4'-Methyl-[1,1'-biphenyl]-4-ylmethyl)-N-[4-(4-(8-hydroxyamino-8-oxooctyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A21); (E)-1-(4'-Methyl-[1,1'-biphenyl]-4-ylmethyl)-N-[4-(4-(4-(3-hydroxyamino-3-oxopropen-1-en-1-yl)benzyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A22); 1-(4'-Methyl-[1,1'-biphenyl]-4-ylmethyl)-N-[4-(4-(4-hydroxycarbamoylbenzyl)piperazin-1-yl)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (A23); 1-(3,4-Dichlorobenzyl)-N-[4-(4-hydroxyamino-4-oxobutoxy)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (B1); 1-(3,4-Dichlorobenzyl)-N-[4-(5-hydroxyamino-5-oxopentyloxy)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (B2); 1-(3,4-Dichlorobenzyl)-N-[4-(6-hydroxyamino-6-oxohexyloxy)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (B3); 1-(3,4-Dichlorobenzyl)-N-[4-(7-hydroxyamino-7-oxoheptyloxy)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (B4); 1-(3,4-Dichlorobenzyl)-N-[4-(8-hydroxyamino-8-oxooctyloxy)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (B5); 1-(3,4-dichlorobenzyl)-N-[4-(4-hydroxycarbamoylbenzyloxy)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (B6); 1-(Naphthalen-2-ylmethyl)-N-[4-(4-hydroxyamino-4-oxobutoxy)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (B7); N-[4-(5-Hydroxyamino-5-oxopentyloxy)-3-nitrobenzenesulfonyl]-1-(naphthalen-2-ylmethyl)-1H-indazole-3-carboxamide (B8); 1-(Naphthalen-2-ylmethyl)-N-[4-(6-hydroxyamino-6-oxohexyloxy)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (B9); N-[4-(7-hydroxyamino-7-oxoheptyloxy)-3-nitrobenzenesulfonyl]-1-(naphthalen-2-ylmethyl)-1H-indazole-3-carboxamide (B10); N-[4-(8-Hydroxyamino-8-oxooctyloxy)-3-nitrobenzenesulfonyl]-1-(naphthalen-2-ylmethyl)-1H-indazole-3-carboxamide (B11); (E)-1-(naphthalen-2-ylmethyl)-N-[4-([4-(3-hydroxyamino-3-oxoprop-1-en-1-yl)benzyl]oxy)-3-nitrobenzenesulfonyl]-1H-indazole-3-carboxamide (B12); N-[4-(4-Hydroxycarbamoylbenzyloxy)-3-nitrobenzenesulfonyl]-1-(naphthalen-2-ylmethyl)-1H-indazole-3-carboxamide (B13); N-[4-(4-Hydroxyamino-4-oxobutoxy)-3-nitrobenzenesulfonyl]-1-([1,1'-biphenyl]-4'-yl-4-ylmethyl)-1H-indazole-3-carboxamide (B14); N-[4-(5-Hydroxyamino-5-oxopentyloxy)-3-nitrobenzenesulfonyl]-1-[(4'-methyl-[1,1'-biphenyl]-4-yl)methyl]-1H-indazole-3-carboxamide (B15); N-[4-(6-hydroxyamino-6-oxohexyloxy)-3-nitrobenzenesulfonyl]-1-[(4'-methyl-[1,1'-biphenyl]-4-yl)methyl]-1H-indazole-3-carboxamide (B16); N-[4-(7-hydroxyamino-7-oxoheptyloxy)-3-nitrobenzenesulfonyl]-1-[(4'-methyl-[1,1'-biphenyl]-4-yl)methyl]-1H-indazole-3-carboxamide (B17); N-[4-(8-Hydroxyamino-8-oxooctyloxy)-3-nitrobenzenesulfonyl]-1-[(4'-methyl-[1,1'-biphenyl]-4-yl)methyl]-1H-indazole-3-carboxamide (B18).

5. The method for preparing the substituted indazole HDAC / Mcl-1 dual-target inhibitor according to any one of claims 1 to 4, characterized in that: The reaction route is as follows: Reagents and conditions: a) acetyl chloride, methanol, ice bath-reflux; b) i. substituted benzyl bromide, potassium carbonate, N,N-dimethylformamide (DMF), room temperature; ii. 1 mol / L sodium hydroxide solution, tetrahydrofuran, room temperature; iii. 1 mol / L hydrochloric acid solution, room temperature; c) i. DMF, N-Boc-piperazine, 80°C; ii. ethyl acetate saturated with hydrogen chloride, room temperature; iii. acetone, triethylamine, methyl or ethyl bromocarboxylate, 60°C; d) i.16.5 mol / L sodium hydroxide solution, reflux; ii. 3 mol / L hydrochloric acid solution, room temperature; iii. DMF, N,N-dimethylformamide dimethyl acetal, 45°C; iv. potassium carbonate, DMF, methyl or ethyl bromocarboxylate, 80°C; v. methanol, hydrochloric acid, 60°C; e) i. thionyl chloride, reflux; ii. dichloromethane, triethylamine, ice bath-room temperature; f) i. hydroxylamine hydrochloride, potassium hydroxide, room temperature; ii. 1 mol / L hydrochloric acid solution, room temperature.

6. A pharmaceutical composition suitable for oral administration to a mammal, characterized in that: The invention comprises an inhibitor and one or more pharmaceutically acceptable carriers or excipients, wherein the inhibitor is the inhibitor according to any one of claims 1 to 4 or the inhibitor prepared by the method according to claim 5.

7. A pharmaceutical composition suitable for parenteral administration to mammals, characterized in that The invention comprises an inhibitor and one or more pharmaceutically acceptable carriers or excipients, wherein the inhibitor is the inhibitor according to any one of claims 1 to 4 or the inhibitor prepared by the method according to claim 5.

8. Use of the inhibitor according to any one of claims 1 to 4, the inhibitor prepared by the method according to claim 5, the pharmaceutical composition according to claim 6, or the pharmaceutical composition according to claim 7 in the preparation of a medicament for preventing or treating mammalian diseases caused by abnormal expression of HDAC / Mcl-1 protein.

9. The use according to claim 8, characterized in that The mammalian diseases include cancer, neurodegenerative diseases, viral infections, inflammation, leukemia, malaria, and diabetes.

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