CN113735830A

CN113735830A - Hydroximic acid derivatives and application thereof

Info

Publication number: CN113735830A
Application number: CN202111183961.7A
Authority: CN
Inventors: 周鑫; 叶兵
Original assignee: Chengdu Zihao Pharmaceutical Co ltd
Current assignee: Chengdu Zihao Pharmaceutical Co ltd
Priority date: 2021-10-11
Filing date: 2021-10-11
Publication date: 2021-12-03
Also published as: WO2023061368A1

Abstract

The invention provides hydroximic acid derivatives and application thereof, and relates to the field of biological medicines. The hydroximic acid derivative is a compound represented by formula I, or a salt thereof, or a stereoisomer thereof, or a solvate thereof, or a hydrate thereof, or a prodrug thereof. The invention provides a compound with good inhibitory activity on HDAC and PARP, which has good inhibitory effect on HDAC and PARP and can be used for preparing PARP/HDAC double-target inhibitor and preventing and/or treating PARP and/or HDAC activity inhibitionDiseases to be improved, such as tumor, ischemic diseases, diabetes, and inflammatory diseases. Meanwhile, the compound has synthetic lethal effect, can be used as a chemo-therapeutic drug sensitizer and a radiotherapy sensitizer and is used together with other anti-tumor drugs, so that the treatment effect is improved, and the adverse reaction is reduced. In addition, the compound improves the compliance of patients, has more medicament economy advantages and has good application prospect.

Description

Hydroximic acid derivatives and application thereof

Technical Field

The invention relates to the field of biological medicine, in particular to hydroximic acid derivatives and application thereof.

Background

Poly (adenosine diphosphate ribose) polymerase (PARP) is an important DNA repair enzyme, and is present in eukaryotic cells as a protease that catalyzes the ribosylation of poly (adenosine diphosphate) (poly (ADP)). It can recognize DNA single-strand break point and start repair. The repair acts on a plurality of proteins, relates to the aspects of chromosome stabilization, DNA damage repair, gene transcription, cell growth, death, apoptosis and the like, and is closely related to diseases such as inflammation, tumor, aging and the like in the aspects of physiology and pathology.

The PARP inhibitor can inhibit the activity of PARP, has the effect of enhancing the effects of radiotherapy and DNA damage chemotherapeutic drugs, and can selectively kill tumor cells with DNA repair defects when used alone. The application of PARP inhibitors in tumor therapy is mainly based on two mechanisms: first, PARP inhibitors inhibit the repair process of DNA single strand damage, but this DNA single strand damage can be converted into double strand Damage (DSB) during DNA replication to form replication forks, and this DSB can still be repaired by Homologous Recombination (HR) pathways. If the tumor cells have a defect in homologous recombination repair (including the BRCA1/2 mutation) such that DSB damage cannot be repaired, the effect of PARP inhibitors and the defect in homologous recombination repair on synthetic lethality of the tumor cells can be caused. The PARP inhibitor has obvious inhibition effect on breast cancer and ovarian cancer cells with BRCA1 and BRCA2 mutation. Secondly, BRCA1/2 is only part of HR repair, and other proteins such as EMSY and PTEN are equally important for the HR pathway, and PARP inhibitors may produce single drug anti-tumor activity through synthetic lethality if these genes are mutated or expression silenced in the HR repair pathway.

Histone Deacetylases (HDACs) are a class of proteases and play an important role in the structural modification of chromosomes and the regulation of gene expression. Generally, acetylation of histones facilitates dissociation of DNA from histone octamers and relaxation of nucleosome structure, thereby allowing specific binding of various transcription factors and co-transcription factors to DNA binding sites, activating gene transcription. In the nucleus, histone acetylation and histone deacetylation processes are in dynamic equilibrium and are regulated by Histone Acetyltransferase (HAT) and Histone Deacetylase (HDAC) together. HAT transfers acetyl group of acetyl-CoA to specific lysine residue at the amino terminal of histone, HDAC deacetylates histone, tightly binds to DNA with negative charge, chromatin is densely coiled, and gene transcription is inhibited.

In cancer cells, the abnormal allosteric conformation of the chromosome is caused by the increased binding of histones to DNA as a result of overexpression of HDACs. Inhibiting HDAC activity is effective in inhibiting cancer cell proliferation, inducing cell cycle arrest and promoting apoptosis. In addition, HDAC inhibitors can increase DNA damage, interfere with double strand damage repair (DSB) recognition, inhibit DNA repair. Studies have shown that HDACs are closely related to homologous recombination repair, and inhibition of HDAC activity affects a range of protein functions, including RAD51 and its analogs, RAD52, RAD54, BRCA1, BRCA2.mrn, and the damage detection molecules ATM, ATR, etc., which initiate DNA repair reactions.

Timothy AY et al indicate (Yap T A et al, CA Cancer J Clin.2011; 61(1):31-49.) that PARP not only exhibits a synthetic lethal effect on BRCA defects, but also may interact with genes associated with homologous recombination defects, including ATM.Rad51.CHK et al. The literature reports that the PARP inhibitor and the HDAC inhibitor have synergistic effect in combination, can promote the accumulation of DNA damage and reduce the response of a homologous recombination pathway in tumor cells, thereby promoting the death of various tumor cells and having good killing effect on the tumor cells without deletion of the homologous recombination repair pathway. However, the combined medication has the defects of complex pharmacokinetics, possibility of drug interaction and more toxic and side effects. While a single small molecule with multi-target inhibitory activity promises to avoid these problems. Therefore, the research on a small molecule compound with PARP/HDAC double-target point inhibitory activity is of great significance.

Disclosure of Invention

In order to solve the problems, the invention provides a hydroximic acid derivative and application thereof, and the technical scheme is as follows:

the invention provides a compound shown as a formula I, or a salt, a stereoisomer, a solvate, a hydrate or a prodrug thereof:

wherein the content of the first and second substances,

X₁selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

X₂Selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

Z₁Selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

Z₂Selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

Y₁、Y₂Each independently selected from N, CR₅；

R₂、R₃、R₄、R₅Are respectively and independently selected from hydrogen and C₁～C₈An alkyl group;

the ring A is selected from 6-10 membered aryl and 5-10 membered heteroaryl;

R₁is a substituent on the A ring and is respectively and independently selected from substituted or unsubstituted C₁～C₈Alkyl radical, C₁～C₈Alkoxy, halogen, substituted or unsubstituted 3-to 10-membered cycloalkyl, substituted or unsubstituted 3-to 10-membered saturated heterocyclic group, substituted or unsubstituted 3-to 10-membered unsaturated heterocyclic group;

the substituent of the alkyl is selected from substituted or unsubstituted 5-10-membered heteroaryl;

the substituents of the cycloalkyl and the heterocyclic radical are selected from C₁～C₈Alkyl radical, C₁～C₈Alkoxy, halogen, hydroxy, amino, carboxyl, nitro, cyano; or two substituents on the same carbon atom form a double bond with O;

the substituent of the heteroaryl is selected from C₁～C₈Alkyl radical, C₁～C₈Alkoxy, halogen, hydroxy, amino, carboxyl, nitro, cyano; or two substituents on the same carbon atom form a double bond with O;

n represents a substituent R on the A ring₁The number of (b) is an integer of 0 to 5, and when n is 0, it means that the ring A has no substituent.

Further, the air conditioner is provided with a fan,

X₁selected from none, O or NR₂；

X₂Selected from none or CR₃R₄；

Z₁Selected from none or CR₃R₄；

Z₂Selected from none, O or NR₂；

Y₁、Y₂Each independently selected from N, CR₅；

R₂、R₃、R₄、R₅Are respectively and independently selected from hydrogen and C₁～C₄An alkyl group;

the ring A is selected from 6-10 membered aryl and 5-10 membered heteroaryl; the heteroatom in the heteroaryl is N, O or S, and the number of the heteroatoms is 1, 2 or 3;

R₁is a substituent on the A ring and is respectively and independently selected from substituted or unsubstituted C₁～C₄Alkyl radical, C₁～C₄Alkoxy, halogen, substituted or unsubstituted 3-to 6-membered cycloalkyl, substituted or unsubstituted 3-to 6-membered saturated heterocyclic group; the heteroatom in the heterocyclic group is N, O or S, and the number of the heteroatoms is 1, 2 or 3;

the substituent of the alkyl is selected from substituted or unsubstituted 5-10-membered heteroaryl; the heteroatom in the heteroaryl is N, O or S, and the number of the heteroatoms is 1, 2 or 3;

the substituents of the cycloalkyl and the heterocyclic radical are selected from C₁～C₄Alkyl radical, C₁～C₄Alkoxy, halogen, hydroxy, amino, carboxyl, nitro, cyano; or two substituents on the same carbon atom form a double bondO；

The substituent of the heteroaryl is selected from C₁～C₄Alkyl radical, C₁～C₄Alkoxy, halogen, hydroxy, amino, carboxyl, nitro, cyano; or two substituents on the same carbon atom form a double bond with O;

n represents a substituent R on the A ring₁The number of (b) is an integer selected from 0, 1, 2 or 3, and when n is 0, it means that the ring A has no substituent.

Further, the air conditioner is provided with a fan,

X₁selected from none, O or NR₂；

X₂Selected from none or CR₃R₄；

Z₁Selected from none or CR₃R₄；

Z₂Selected from none, O or NR₂；

And X₂And Z₁Cannot be simultaneously CR₃R₄Or simultaneously none;

Y₁、Y₂each independently selected from N, CR₅；

And Y is₁And Y₂Can not be simultaneously selected from N, but can be simultaneously selected from CR₅；

preferably, the first and second electrodes are formed of a metal,

X₁and X₂Simultaneously absent or not simultaneously absent;

Z₁and Z₂Simultaneously absent or not simultaneously absent;

Y₁and Y₂Are not identical.

Further, the air conditioner is provided with a fan,

ring A is selected from phenyl, naphthyl, indazolyl, indolyl, thienyl, furyl, pyrrolyl, pyridyl, pyrazinyl, pyridazinyl, triazinyl;

R₁is a substituent on the A ring and is respectively and independently selected from substituted or unsubstituted C₁～C₄Alkyl radical, C₁～C₄Alkoxy, halogen, substituted or unsubstituted 3-to 6-membered cycloalkyl, substituted or unsubstituted 3-to 6-membered saturated heterocyclic group; the hetero atom in the heterocyclic group is N, and the number of the hetero atoms is 1, 2 or 3;

the alkyl group has a substituent selected from the group consisting of substituted or unsubstituted indazolyl, substituted or unsubstituted indolyl, substituted or unsubstituted thienyl, substituted or unsubstituted furyl, substituted or unsubstituted pyrrolyl, substituted or unsubstituted pyridyl, substituted or unsubstituted pyrazinyl, substituted or unsubstituted pyridazinyl, substituted or unsubstituted triazinyl, and substituted or unsubstituted dihydrophthalazinyl;

the substituents of the cycloalkyl and the heterocyclic radical are selected from C₁～C₄Alkyl radical, C₁～C₄Alkoxy, halogen, hydroxy, amino, carboxyl, nitro, cyano; or two substituents on the same carbon atom form a double bond with O;

the substituent of the indazolyl, indolyl, thienyl, furyl, pyrrolyl, pyridyl, pyrazinyl, pyridazinyl, triazinyl and dihydrophthalazinyl is selected from C₁～C₄Alkyl radical, C₁～C₄Alkoxy, halogen, hydroxy, amino, carboxyl, nitro, cyano; or two substituents on the same carbon atom form a double bond with O;

n represents a substituent R on the A ring₁The number of (a) is an integer selected from 0, 1, 2 or 3, and when n is 0, it means that the ring A has no substituent;

preferably, the first and second electrodes are formed of a metal,

a ring is selected from

R₁' is selected from halogen.

Further, the compound is represented by formula II:

wherein the content of the first and second substances,

X₁selected from O or NR₂；

X₂Selected from the group consisting of CR₃R₄；

Y₁、Y₂Each independently selected from N, CR₅(ii) a And Y is₁And Y₂Can not be simultaneously selected from N, but can be simultaneously selected from CR₅；

preferably, the first and second electrodes are formed of a metal,

more preferably still, the first and second liquid crystal compositions are,

a ring is selected from

R₁' is selected from halogen.

Further, the compound is represented by formula III:

wherein the content of the first and second substances,

X₁selected from O or NR₂；

R₂、R₅Are respectively and independently selected from hydrogen and C₁～C₄An alkyl group;

n represents a substituent R on the A ring₁The number of (b) is an integer selected from 0, 1, 2 or 3, and when n is 0, it means that there is no substituent on the A ring；

Preferably, the first and second electrodes are formed of a metal,

more preferably still, the first and second liquid crystal compositions are,

a ring is selected from

R₁' is selected from halogen.

Further, the compound is represented by formula IV:

wherein the content of the first and second substances,

X₁selected from O or NR₂；

R₂Selected from hydrogen, C₁～C₄An alkyl group;

preferably, the first and second electrodes are formed of a metal,

more preferably still, the first and second liquid crystal compositions are,

a ring is selected from

R₁' is selected from halogen.

Further, the compound is represented by formula V:

wherein the content of the first and second substances,

Z₁selected from the group consisting of CR₃R₄；

Z₂Selected from O or NR₂；

the substituent of the heteroaryl is selected from C₁～C₄Alkyl radical, C₁～C₄Alkoxy, halogen, hydroxy, amino, carboxyl, nitro, cyano; or two substituents on the same carbon atom form a double bondO；

preferably, the first and second electrodes are formed of a metal,

more preferably still, the first and second liquid crystal compositions are,

a ring is selected from

R₁' is selected from halogen.

Further, the compound is represented by formula VI:

wherein the content of the first and second substances,

Z₂selected from O or NR₂；

preferably, the first and second electrodes are formed of a metal,

n represents a substituent R on the A ring₁The number of (2) is an integer selected from 0, 1, 2 or 3And when n is 0, it means that there is no substituent on the A ring;

more preferably still, the first and second liquid crystal compositions are,

a ring is selected from

R₁' is selected from halogen.

Further, the compound is represented by formula VII:

wherein the content of the first and second substances,

Z₂selected from O or NR₂；

R₂Selected from hydrogen, C₁～C₄An alkyl group;

the substituent of the heteroaryl is selected from C₁～C₄Alkyl radical, C₁～C₄Alkoxy, halogen, hydroxy, amino, carboxyl, nitro, cyano; or the same carbon atomTwo substituents on the molecule form a double bond with O;

preferably, the first and second electrodes are formed of a metal,

more preferably still, the first and second liquid crystal compositions are,

a ring is selected from

R₁' is selected from halogen.

Further, the compound is represented by formula VIII:

wherein the content of the first and second substances,

X₁selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

X₂Selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

Z₁Selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

Z₂Selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

Y₁、Y₂Each independently selected from N, CR₅；

preferably, the first and second electrodes are formed of a metal,

X₁selected from none, O or NR₂；

X₂Selected from none or CR₃R₄；

Z₁Selected from none or CR₃R₄；

Z₂Selected from none, O or NR₂；

And X₂And Z₁Cannot be simultaneously CR₃R₄Or simultaneously none;

Y₁、Y₂each independently selected from N, CR₅；

more preferably, the compound is of formula VIIIa:

wherein the content of the first and second substances,

X₁selected from O or NR₂；

R₂Selected from hydrogen, C₁～C₄An alkyl group;

alternatively, the compound is of formula VIIIb:

wherein the content of the first and second substances,

Z₂selected from O or NR₂；

R₂Selected from hydrogen, C₁～C₄An alkyl group.

Further, the compound is represented by formula IX:

wherein the content of the first and second substances,

X₁selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

X₂Selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

Z₁Selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

Z₂Selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

Y₁、Y₂Each independently selected from N, CR₅；

preferably, the first and second electrodes are formed of a metal,

X₁selected from none, O or NR₂；

X₂Selected from none or CR₃R₄；

Z₁Selected from none or CR₃R₄；

Z₂Selected from none, O or NR₂；

And X₂And Z₁Cannot be simultaneously CR₃R₄Or simultaneously none;

Y₁、Y₂each independently selected from N, CR₅；

more preferably, the compound is of formula IXa:

wherein the content of the first and second substances,

X₁selected from O or NR₂；

R₂Selected from hydrogen, C₁～C₄An alkyl group;

alternatively, the compound is of formula IXb:

wherein the content of the first and second substances,

Z₂selected from O or NR₂；

R₂Selected from hydrogen, C₁～C₄An alkyl group.

Further, the compound is represented by formula X:

wherein the content of the first and second substances,

X₁selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

X₂Selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

Z₁Selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

Z₂Selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

Y₁、Y₂Each independently selected from N, CR₅；

R₁' is selected from halogen;

preferably, the first and second electrodes are formed of a metal,

X₁selected from none, O or NR₂；

X₂Selected from none or CR₃R₄；

Z₁Selected from none or CR₃R₄；

Z₂Selected from none, O or NR₂；

And X₂And Z₁Cannot be simultaneously CR₃R₄Or simultaneously none;

Y₁、Y₂each independently selected from N, CR₅；

R₁' is selected from halogen;

more preferably, the compound is of formula Xa:

wherein the content of the first and second substances,

X₁selected from O or NR₂；

R₂Selected from hydrogen, C₁～C₄An alkyl group;

R₁' is selected from halogen;

alternatively, the compound is of formula Xb:

wherein the content of the first and second substances,

Z₂selected from O or NR₂；

R₂Selected from hydrogen, C₁～C₄An alkyl group;

R₁' is selected from halogen.

Further, the compound is one of the following compounds:

the invention also provides the use of the aforementioned compound, or a salt thereof, or a stereoisomer thereof, or a solvate thereof, or a hydrate thereof, or a prodrug thereof, in the manufacture of a medicament for inhibiting the activity of PARP and/or HDAC.

Further, the drug is a drug for treating ischemic diseases, diabetes, or inflammatory diseases.

Further, the medicine is a chemo-therapeutic medicine sensitizer, a radiotherapy sensitizer or a medicine for treating tumor;

preferably, the tumor is breast, ovarian, pancreatic or prostate cancer.

The invention also provides a pharmaceutical preparation which is prepared by taking one or more of the compounds, or the salts, or the stereoisomers, or the solvates, or the hydrates, or the prodrugs thereof as active ingredients and adding pharmaceutically acceptable auxiliary materials or auxiliary ingredients.

The invention also provides a pharmaceutical composition which comprises the compound, or the salt, the stereoisomer, the solvate, the hydrate or the prodrug thereof and a chemotherapeutic drug or a radiotherapeutic drug.

The invention also provides application of the compound, or a salt, a stereoisomer, a solvate, a hydrate or a prodrug thereof and a chemotherapeutic drug or a radiotherapeutic drug in preparation of combined drugs.

The preparation is prepared by taking one or more of the compounds, the salts, the stereoisomers, the solvates, the hydrates and the prodrugs thereof, or the pharmaceutical composition as an active ingredient, adding pharmaceutically acceptable auxiliary materials or auxiliary ingredients and preparing the preparation by a conventional method. The dosage forms include but are not limited to: tablet, granule, capsule, oral liquid and other pharmaceutically acceptable dosage forms. The pharmaceutical compositions and formulations thereof are useful in the treatment of diseases where the clinical benefit is directly or indirectly produced by inhibition of PARP and/or HDAC.

The diseases ameliorated by the inhibition of PARP and/or HDAC activity of the present invention include, but are not limited to, prevention and/or treatment of ischemic diseases, diabetes, inflammatory diseases.

The aforementioned compound, or a salt thereof, or a stereoisomer thereof, or a solvate thereof, or a hydrate thereof, or a prodrug thereof of the present invention is used as an auxiliary means for cancer therapy including, but not limited to, breast cancer, ovarian cancer, pancreatic cancer, prostate cancer, or the like, or for enhancing the inhibition and/or killing of tumor cells by radiotherapy or chemotherapeutic agents.

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) naming system.

Definitions of terms used in connection with the present invention: the initial definitions provided herein for a group or term apply to that group or term throughout the specification unless otherwise indicated; for terms not specifically defined herein, the meanings that would be given to them by a person skilled in the art are to be given in light of the disclosure and the context.

In the present invention, the structures of the compounds are all structures capable of stably existing.

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

In the present invention, the minimum and maximum values of the carbon atom content in the hydrocarbon group are indicated by a prefix, for example, the prefix (C)_a～C_b) Alkyl means any alkyl group containing from "a" to "b" carbon atoms. Thus, for example, C₁～C₈The alkyl group is a straight-chain or branched alkyl group having 1 to 8 carbon atoms, and includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, and the like.

Halogen is fluorine, chlorine, bromine or iodine.

"substituted or unsubstituted" means that the group may be substituted with or without any substituent.

"aryl" refers to an all-carbon monocyclic or fused polycyclic (i.e., rings which share adjacent pairs of carbon atoms) groups having a conjugated pi-electron system, such as phenyl and naphthyl. The aryl ring may be fused to other cyclic groups (including saturated and unsaturated rings) but must not contain heteroatoms such as nitrogen, oxygen, or sulfur, and the point of attachment to the parent must be at a carbon atom on the ring which has a conjugated pi-electron system. The 6-to 10-membered aryl group means that the aryl group has 6 to 10 carbon atoms.

"heteroaryl" means that one or more carbon atoms in an aryl group are replaced with a heteroatom (e.g., nitrogen, oxygen, or sulfur, etc.). The 5-to 10-membered heteroaryl group means that the number of atoms in the heteroaryl skeleton is 5 to 10.

"cycloalkyl" means an all-carbon monocycloalkyl or bridged, fused, spirocycloalkyl group. 3 to 10 membered means that the cycloalkyl group is composed of 3 to 10 carbon atoms.

"saturated heterocyclyl" means that one or more carbon atoms in a cycloalkyl group have been replaced with a heteroatom (e.g., nitrogen, oxygen, or sulfur, etc.).

"unsaturated heterocyclyl" partially unsaturated cycloalkyl groups have one or more of the carbon atoms replaced with a heteroatom (e.g., nitrogen, oxygen, or sulfur, etc.).

"two substituents on the same carbon atom form a double bond O" -means

And (5) structure. "NR₂' finger shaped formation

And (5) structure. "CR₃R₄' finger shaped formation

And (5) structure.

The "salt" in the present invention means "pharmaceutically acceptable salt". "pharmaceutically acceptable salts" refers to those salts that retain the biological effectiveness and properties of the parent compound. Such salts include:

(1) salifying with an acid by reacting the free base of the parent compound with an inorganic acid or an organic acid, the inorganic acid including hydrochloric acid, hydrogalactic acid, hydroiodic acid, nitric acid, nitrous acid, phosphoric acid, metaphosphoric acid, sulfuric acid, sulfurous acid, perchloric acid, and the like, and the organic acid including acetic acid, propionic acid, acrylic acid, oxalic acid, (D) or (L) malic acid, fumaric acid, maleic acid, hydroxybenzoic acid, crotonic acid, methoxybenzoic acid, phthalic acid, methanesulfonic acid, ethanesulfonic acid, naphthalene-1-sulfonic acid, naphthalene-2-sulfonic acid, p-toluenesulfonic acid, salicylic acid, tartaric acid, citric acid, lactic acid, mandelic acid, succinic acid, malonic acid, and the like.

(2) The acidic proton present in the parent compound is replaced with a metal ion such as an alkali metal ion, an alkaline earth metal ion or an aluminum ion, or is complexed with an organic base such as ethylamine, diethylamine, dicyclohexylamine, triethylamine, butylamine, ethylenediamine, ethanolamine, diethanolamine, triethanolamine, piperazine, mepiquat, phenylmepiquat chloride, choline, meglumine, tromethamine, N-methylglucamine or the like.

The term "solvate" refers to a compound of the present invention that forms a solvate with a pharmaceutically acceptable solvent, wherein the pharmaceutically acceptable solvent includes, but is not limited to, water, ethanol, methanol, isopropanol, propylene glycol, tetrahydrofuran, dichloromethane.

The term "stereoisomer" refers to a compound of the invention involving chiral carbon atoms that may be in the R configuration, or in the S configuration, or a combination thereof.

PARP plays a key role in cellular DNA damage repair, and is also involved in DNA methylation modification and transcription, cell signaling, cell cycle regulation, and cell mitosis. Studies have shown that PARP inhibitors have anti-cancer effects, and more importantly, that the combined use of PARP inhibitors with other anti-cancer drugs can cause synthetic lethal effects. For example, PARP inhibitors are used in combination with chemotherapeutic agents to both reduce the adverse effects of the chemotherapeutic agents and to improve their therapeutic efficacy. Therefore, the PARP inhibitor can treat the diseases related to the PARP, is used as a medicament for treating tumors, is a chemotherapeutic medicament sensitizer and a radiotherapy sensitizer, and has obvious convenient effect on tumor treatment.

The invention provides a compound which has good inhibitory activity to HDAC and PARP simultaneously, the compound has good inhibitory effect to HDAC and PARP, and can be used for preparing PARP/HDAC double-target inhibitor, preventing and/or treating diseases improved by PARP and/or HDAC activity inhibition, such as tumor, ischemic diseases, diabetes, inflammatory diseases and the like. Meanwhile, the compound has synthetic lethal effect, can be used as a chemo-therapeutic drug sensitizer and a radiotherapy sensitizer and is used together with other anti-tumor drugs, so that the treatment effect is improved, and the adverse reaction is reduced. In addition, the compound improves the compliance of patients, has more medicament economy advantages and has good application prospect.

Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.

The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.

Detailed Description

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

Example 1, 2- (4- ((2-fluoro-5- ((4-oxo-3, 4-dihydrophthalazin-1-yl) methyl) benzamide) methyl) piperidin-1-yl) -N-hydroxypyrimidine-5-carboxamide (Compound 1)

A: 4-Aminomethylpiperidin-1-yl-pyrimidine-5-carboxylic acid methyl ester

Into a 500ml three-necked flask, 200ml of N, N-dimethylacetamide, 34.3 g (0.3mol) of the compound 4-aminomethylpiperidine and 65.5 ml (0.375mol) of N, N-diisopropylethylamine were added, and 100ml of a solution of 4-chloropyrimidine-5-carboxylic acid methyl ester, 25.9 g (0.15mol), 100ml of N, N-dimethylacetamide, was added dropwise under stirring at room temperature, and after 3 hours, the reaction mixture was poured slowly into a stirred ice-water mixture, followed by extraction three times with 300 ml of ethyl acetate, and after the organic phases were combined, washed twice with an appropriate amount of saturated saline solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure, a pale yellow wax was obtained. Then recrystallizing with isopropyl ether, precipitating solid, filtering, drying to obtain light yellow powder target compound 25.3 g, ESI-MS: m/z 251.1.

B: 2- (4- ((2-fluoro-5- ((4-oxo-3, 4-dihydrophthalazin-1-yl) methyl) benzamide) methyl) piperidin-1-yl) -pyrimidine-5-carboxylic acid methyl ester

14.9 g (50mmol) of 2-fluoro-5- (4-oxo-3, 4-dihydrophthalazin-1-yl) methyl) benzoic acid and 200ml of N, N-dimethylformamide were charged into a 500ml three-necked flask, and 20.8 g (55mmol) of O-benzotriazol-tetramethyluronium Hexafluorophosphate (HBTU) was further added with stirring at room temperature, and after 2 hours, 12.5 g (50mmol) of 4-aminomethylpiperidin-1-yl-pyrimidine-5-carboxylic acid methyl ester was further added. Stirring overnight, the next day, slowly pouring the reaction solution into a stirred ice-water mixture, extracting with ethyl acetate, washing, drying, concentrating to obtain a yellow solid, and purifying by column chromatography to obtain 21 g of the target compound. ESI-MS: m/z 531.2.

C: 2- (4- ((2-fluoro-5- ((4-oxo-3, 4-dihydrophthalazin-1-yl) methyl) benzamide) methyl) piperidin-1-yl) -pyrimidine-5-carboxylic acid

21 g of methyl 2- (4- ((2-fluoro-5- ((4-oxo-3, 4-dihydrophthalazin-1-yl) methyl) benzamide) methyl) piperidin-1-yl) -pyrimidine-5-carboxylate was charged into a 500-ml single-neck flask, and 200ml of methanol was added thereto, and after stirring and dissolution, 50ml of water and 6.7 g (0.16mol) of lithium hydroxide monohydrate were further added thereto in an ice-water bath. After 4 hours, TLC detection was carried out, hydrolysis was complete, and 2M hydrochloric acid was added dropwise to neutrality. And (3) carrying out reduced pressure rotary evaporation, removing methanol, adjusting the pH value of the solution to about 5 with 2M hydrochloric acid, separating out a large amount of powdery solid, filtering, washing with water, and drying to obtain 15.8 g of light yellow powdery solid, wherein the light yellow powdery solid can be directly used for the next reaction without further purification.

D: 2- (4- ((2-fluoro-5- ((4-oxo-3, 4-dihydrophthalazin-1-yl) methyl) benzamide) methyl) piperidin-1-yl) -pyrimidine-5-carboxylic acid (tetrahydropyran-2-oxy) -amine

Adding the obtained 2- (4- ((2-fluoro-5- ((4-oxo-3, 4-dihydrophthalazin-1-yl) methyl) benzamide) methyl) piperidin-1-yl) -pyrimidine-5-carboxylic acid, tetrahydrofuran 100ml and N, N-dimethylformamide 30ml into a 250ml round bottom flask, stirring to dissolve, adding N, N-diisopropylethylamine 2.6ml and 2.46g O-benzotriazol-tetramethylurea hexafluorophosphate, stirring at room temperature for one hour, adding 0.89g O- (tetrahydro-2H-pyran-2-yl) hydroxylamine, monitoring the reaction progress by TLC, concentrating under reduced pressure after the reaction is completed to remove the tetrahydrofuran, extracting the residue with ethyl acetate, combining ethyl acetate phases, concentrating, and performing column chromatography to obtain 14.8g of target compound.

E: 2- (4- ((2-fluoro-5- ((4-oxo-3, 4-dihydrophthalazin-1-yl) methyl) benzamide) methyl) piperidin-1-yl) -N-hydroxypyrimidine-5-carboxamide (Compound 1)

2- (4- ((2-fluoro-5- ((4-oxo-3, 4-dihydrophthalazin-1-yl) methyl) benzamide) methyl) piperidin-1-yl) -pyrimidine-5-carboxylic acid (tetrahydropyran-2-oxy) -amine 2.0g and 20ml of hydrochloric acid gas solution of ethyl acetate are sequentially added into a 100ml single-neck bottle, the mixture is stirred and reacted at room temperature, the reaction process is monitored by TLC, after the reaction is completed, precipitated solid is filtered, and then methanol is used for recrystallization to obtain 1.1g of the product, wherein the yield is 70%. ESI-MS: m/z 532.21[ M + H ]]⁺；¹H NMR(600MHz，DMSO-d6)δ:1.10(dd,2H),1.76(d,2H),1.89(m,1H),2.94(t,2H),3.15(t,2H),4.33(s,2H),4.70(d,2H),7.20(t,1H),7.45(m,1H),7.55(d,1H),7.86(dt,2H),7.98(d,1H),8.27(d,1H),8.34(m,1H),8.66(s,2H),8.99(s,1H),11.04(s,1H),12.60(s,1H)。

F: 2- (4- ((2-fluoro-5- ((4-oxo-3, 4-dihydrophthalazin-1-yl) methyl) benzamide) methyl) piperidin-1-yl) -pyrimidine-5-carboxylic acid benzyloxyamine

2.6g of 2- (4- ((2-fluoro-5- ((4-oxo-3, 4-dihydrophthalazin-1-yl) methyl) benzamide) methyl) piperidin-1-yl) -pyrimidine-5-carboxylic acid, 100ml of tetrahydrofuran and 30ml of N, N-dimethylformamide are added into a 250ml round bottom flask, after stirring and dissolving, 2.6ml of N, N-diisopropylethylamine and 2.46g of O-benzotriazol-tetramethylurea hexafluorophosphate are added, stirring is carried out for one hour at room temperature, 1.2g of O-benzylhydroxylamine hydrochloride is added, the reaction progress is monitored by TLC, after the reaction is completed, the tetrahydrofuran is removed by decompression, residues are extracted by ethyl acetate, an ethyl acetate phase is combined, and the mixture is concentrated and subjected to column chromatography to obtain a pure product of 2.0 g.

G: 2- (4- ((2-fluoro-5- ((4-oxo-3, 4-dihydrophthalazin-1-yl) methyl) benzamide) methyl) piperidin-1-yl) -N-hydroxypyrimidine-5-carboxamide (Compound 1)

In a 100ml single-neck flask, 1.8g of 2- (4- ((2-fluoro-5- ((4-oxo-3, 4-dihydrophthalazin-1-yl) methyl) benzamide) methyl) piperidin-1-yl) -pyrimidine-5-carboxylic acid benzyloxyamine and 20ml of methanol were added, and after dissolution by stirring, 0.2g of palladium on carbon was added, hydrogen gas was introduced, and the progress of the reaction was monitored by TLC. After the reaction is completed, the reaction solution is filtered, concentrated and recrystallized by methanol to obtain 1.1g of the product with the yield of 74 percent. ESI-MS: m/z 532.21[ M + H ]]⁺；¹H NMR(600MHz，DMSO-d6)δ:1.10(dd,2H),1.76(d,2H),1.89(m,1H),2.94(t,2H),3.15(t,2H),4.33(s,2H),4.70(d,2H),7.20(t,1H),7.45(m,1H),7.55(d,1H),7.86(dt,2H),7.98(d,1H),8.27(d,1H),8.34(m,1H),8.66(s,2H),8.99(s,1H),11.04(s,1H),12.60(s,1H)。

Example 2, 2- (4- ((2H-indazole-7-carboxamide) methyl) piperidin-1-yl) -N-hydroxypyrimidine-5-carboxamide (Compound 2)

A: 2- (4- ((2H-indazole-7-carboxamide) methyl) piperidin-1-yl) -5-carboxylic acid methyl ester

To a 250ml single-neck flask, 8.1 g (50mmol) of the compound 2H-indazole-7-carboxylic acid and 100ml of N, N-dimethylformamide were added, and 8.1 g (50mmol) of N, N-carbonyldiimidazole was further added under stirring at room temperature, and after 2 hours, 12.5 g (50mmol) of methyl 4-aminomethylpiperidin-1-yl-pyrimidine-5-carboxylate was further added. Stirring overnight, the next day, slowly pouring the reaction solution into a stirred ice-water mixture, precipitating a light yellow solid, standing for 3 hours, filtering, washing with water, and drying to obtain a light yellow powdery product, namely 17.2g of the target compound, wherein the product is directly used for the next reaction without purification. ESI-MS: m/z 395.2.

B: 2- (4- ((2H-indazole-7-carboxamide) methyl) piperidin-1-yl) -5-carboxylic acid

Methyl 2- (4- ((2H-indazole-7-carboxamide) methyl) piperidin-1-yl) -5-carboxylate (17.2 g) was charged in a 500-ml single-neck flask, and then methanol (150 ml) was added thereto, followed by stirring and dissolution, and then under an ice-water bath, 50ml of water and 6.3 g (0.15mol) of lithium hydroxide monohydrate were added thereto. After 4 hours, TLC detection was carried out, hydrolysis was complete, and 2M hydrochloric acid was added dropwise to neutrality. And (3) carrying out reduced pressure rotary evaporation, removing methanol, adjusting the pH value of the solution to about 5 with 2M hydrochloric acid, separating out a large amount of powdery solid, filtering, washing with water, and drying to obtain 13.8 g of light yellow powdery solid, wherein the light yellow powdery solid is directly used for the next reaction without purification. ESI-MS: M/z381.1[ M + H ].

C: 2- (4- ((2H-indazole-7-carboxamide) methyl) piperidin-1-yl) -pyrimidine-5-carboxylic acid (tetrahydropyran-2-oxy) -amine

Adding 3.8g of 2- (4- ((2H-indazole-7-formamide) methyl) piperidine-1-yl) -5-formic acid, 100ml of tetrahydrofuran and 30ml of N, N-dimethylformamide into a 250ml round-bottom flask, stirring to dissolve, adding 5.3ml of N, N-diisopropylethylamine and 4.93g O-benzotriazole-tetramethylurea hexafluorophosphate, stirring for one hour at room temperature, adding 1.76g O- (tetrahydro-2H-pyran-2-yl) hydroxylamine, monitoring the reaction progress by TLC, concentrating under reduced pressure after the reaction is completed to remove tetrahydrofuran, extracting residues with ethyl acetate, combining ethyl acetate phases, concentrating, and performing column chromatography to obtain the target compound.

D: 2- (4- ((2H-indazole-7-carboxamide) methyl) piperidin-1-yl) -N-hydroxypyrimidine-5-carboxamide (Compound 2)

Adding 3.0g of 2- (4- ((2H-indazole-7-formamide) methyl) piperidin-1-yl) -pyrimidine-5-carboxylic acid (tetrahydropyran-2-oxy) -amine and 30ml of hydrochloric acid gas solution of ethyl acetate into a 100ml single-neck bottle, stirring at room temperature for reaction, monitoring the reaction process by TLC, filtering precipitated solid after the reaction is completed, and recrystallizing by using methanol to obtain the product with the yield of 75%. ESI-MS of M/z 418.16[ M + Na ]]⁺；¹H NMR(600MHz，DMSO-d6)δ:1.18(dd,2H),1.83(d,2H),1.97(m,1H),2.97(t,2H),3.28(t,2H),4.74(d,2H),7.19(t,1H),7.95(d,2H),8.16(s,1H),8.69(s,1H),8.81(t,1H)。

E: 2- (4- ((2H-indazole-7-carboxamide) methyl) piperidin-1-yl) -pyrimidine-5-carboxylic acid benzyloxyamine

3.04g of 2- (4- ((2H-indazole-7-formamide) methyl) piperidine-1-yl) -5-formic acid, 100ml of tetrahydrofuran and 30ml of N, N-dimethylformamide are added into a 250ml round-bottom flask, stirring is carried out to dissolve the mixture, 4.19ml of N, N-diisopropylethylamine and 3.94g O-benzotriazole-tetramethylurea hexafluorophosphate are added into the mixture, the mixture is stirred for one hour at room temperature, 1.9g O-benzylhydroxylamine hydrochloride is added into the mixture, the reaction progress is monitored by TLC, after the reaction is completed, the tetrahydrofuran is removed by concentration under reduced pressure, the residue is extracted by ethyl acetate, the ethyl acetate phases are combined, and the mixture is concentrated and subjected to column chromatography to obtain the target compound.

F: 2- (4- ((2H-indazole-7-carboxamide) methyl) piperidin-1-yl) -N-hydroxypyrimidine-5-carboxamide (Compound 2)

In a 100ml single-neck flask, 2.4g of 2- (4- ((2H-indazole-7-carboxamide) methyl) piperidin-1-yl) -pyrimidine-5-carboxylic acid benzyloxyamine was added, stirred to dissolve, and then 0.3g of palladium on carbon and 25ml of methanol were added, and hydrogen gas was introduced, followed by TLC to monitor the progress of the reaction. After the reaction is completed, the reaction solution is filtered, concentrated and recrystallized by methanol to obtain the product with the yield of 78 percent. ESI-MS of M/z 418.16[ M + Na ]]⁺；¹H NMR(600MHz，DMSO-d6)δ:1.18(dd,2H),1.83(d,2H),1.97(m,1H),2.97(t,2H),3.28(t,2H),4.74(d,2H),7.19(t,1H),7.95(d,2H),8.16(s,1H),8.69(s,1H),8.81(t,1H)。

Example 3, 2- ((1- ((2-fluoro-5- ((4-oxo-3, 4-dihydrophthalazin-1-yl) methyl) benzoyl) piperidin-4-yl) methylamino) -N-hydroxypyrimidine-5-carboxamide (Compound 3)

A: 2- (piperidin-4-ylmethylamino) pyrimidine-5-carboxylic acid methyl ester

4.0g of methyl 2- ((1- (tert-butoxycarbonyl) piperidin-4-yl) methylamino pyrimidine-5-carboxylate and 40ml of hydrochloric acid gas solution of ethyl acetate are added into a 100ml single-mouth bottle, the mixture is stirred at room temperature for reaction, the reaction process is monitored by TLC, and after the reaction is completed, the precipitated solid is filtered, so that 2.8g of the target product is obtained.

B: 2- ((1- (2-fluoro-5- ((4-oxo-3, 4-dihydrophthalazin-1-yl) methyl) benzoyl) piperidin-4-yl) methylamino) pyrimidine-5-carboxylic acid methyl ester

5.96 g of 2-fluoro-5- (4-oxo-3, 4-dihydrophthalazin-1-yl) methyl) benzoic acid and 100ml of N, N-dimethylformamide were added to a three-necked flask, and then O-benzotriazol-tetramethylurea Hexafluorophosphate (HB) was added thereto under stirring at room temperatureTU)8.3 g, and after 2 hours 5g of methyl 2- (piperidin-4-ylmethylamino) pyrimidine-5-carboxylate are added. Stirring overnight, the next day, slowly pouring the reaction solution into a stirred ice-water mixture, extracting with ethyl acetate, washing, drying, concentrating to obtain a yellow solid, and purifying by column chromatography to obtain 7.6 g of the target compound. ESI-MS of M/z 532.21[ M + H ]]⁺

C: 2- ((1- (2-fluoro-5- ((4-oxo-3, 4-dihydrophthalazin-1-yl) methyl) benzoyl) piperidin-4-yl) methylamino) pyrimidine-5-carboxylic acid

10 g of methyl 2- ((1- (2-fluoro-5- ((4-oxo-3, 4-dihydrophthalazin-1-yl) methyl) benzoyl) piperidin-4-yl) methylamino) pyrimidine-5-carboxylate was added to a flask, 100ml of methanol was added thereto, and after stirring and dissolution, 50ml of water and 4.2 g of lithium hydroxide monohydrate were added thereto in an ice-water bath. After 4 hours, TLC detection was carried out, hydrolysis was complete, and 2M hydrochloric acid was added dropwise to neutrality. And (3) carrying out reduced pressure rotary evaporation, removing methanol, adjusting the pH value of the solution to about 5 with 2M hydrochloric acid, separating out a large amount of powdery solid, filtering, washing with water, and drying to obtain 15.8 g of light yellow powdery solid, wherein the light yellow powdery solid can be directly used for the next reaction without further purification.

D: 2- ((1- ((2-fluoro-5- ((4-oxo-3, 4-dihydrophthalazin-1-yl) methyl) benzoyl) piperidin-4-yl) methylamino) -N-hydroxypyrimidine-5-carboxylic acid (tetrahydropyran-2-oxy) -amine

Adding 2.6g of 2- ((1- (2-fluoro-5- ((4-oxo-3, 4-dihydrophthalazin-1-yl) methyl) benzoyl) piperidin-4-yl) methylamino) pyrimidine-5-carboxylic acid, 100ml of tetrahydrofuran and 30ml of N, N-dimethylformamide into a round-bottom flask, stirring to dissolve the mixture, adding 2.6ml of N, N-diisopropylethylamine and 2.46g O-benzotriazol-tetramethylurea hexafluorophosphate, stirring at room temperature for one hour, adding 0.89g O- (tetrahydro-2H-pyran-2-yl) hydroxylamine, monitoring the progress of the reaction by TLC (thin layer chromatography), concentrating the mixture under reduced pressure after the reaction is completed to remove the tetrahydrofuran, extracting the residue with ethyl acetate, combining ethyl acetate phases, concentrating the mixture, adding a solvent, stirring, adding a mixture, and stirring to the mixture to obtain a solution, And (4) carrying out column chromatography to obtain the target compound.

E: 2- ((1- ((2-fluoro-5- ((4-oxo-3, 4-dihydrophthalazin-1-yl) methyl) benzoyl) piperidin-4-yl) methylamino) -N-hydroxypyrimidine-5-carboxamide (Compound 3)

In a 100ml single-mouth bottleSequentially adding 2- ((1- ((2-fluoro-5- ((4-oxo-3, 4-dihydrophthalazin-1-yl) methyl) benzoyl) piperidine-4-yl) methylamino) -N-hydroxypyrimidine-5-carboxylic acid (tetrahydropyran-2-oxy) -amine 2.0g and 20ml of hydrochloric acid gas solution of ethyl acetate, stirring for reaction at room temperature, monitoring the reaction process by TLC, filtering precipitated solid after the reaction is completed, recrystallizing by methanol to obtain 1.12g of a product, wherein the yield is 65% of ESI-MS (ESI-MS: M/z 532.21[ M + H ] M/z 532.21]⁺；¹H NMR(600MHz，DMSO-d6)δ:1.13(dd,2H),1.71(d,2H),1.85(m,1H),2.78(t,2H),3.08(t,2H),4.31(s,2H),4.58(d,2H),7.18(t,1H),7.43(m,1H),7.54(d,1H),7.87(dt,2H),7.88(d,1H),8.20(d,1H),8.31(m,1H),8.61(s,2H),8.92(s,1H),10.94(s,1H),12.56(s,1H)。

F: 2- ((1- ((2-fluoro-5- ((4-oxo-3, 4-dihydrophthalazin-1-yl) methyl) benzoyl) piperidin-4-yl) methylamino) -N-hydroxypyrimidine-5-carboxylic acid benzyloxyamine

Adding 2.6g of 2- ((1- (2-fluoro-5- ((4-oxo-3, 4-dihydrophthalazin-1-yl) methyl) benzoyl) piperidin-4-yl) methylamino) pyrimidine-5-carboxylic acid, 100ml of tetrahydrofuran and 30ml of N, N-dimethylformamide into a round-bottom flask, stirring to dissolve, adding 2.6ml of N, N-diisopropylethylamine and 2.46g of O-benzotriazol-tetramethylurea hexafluorophosphate, stirring for one hour at room temperature, adding 1.2g of O-benzylhydroxylamine hydrochloride, monitoring the reaction progress by TLC, concentrating under reduced pressure to remove the tetrahydrofuran after the reaction is completed, extracting residues with ethyl acetate, combining ethyl acetate phases, concentrating and carrying out column chromatography to obtain 1.8g of a pure product.

G: 2- ((1- ((2-fluoro-5- ((4-oxo-3, 4-dihydrophthalazin-1-yl) methyl) benzoyl) piperidin-4-yl) methylamino) -N-hydroxypyrimidine-5-carboxamide (Compound 3)

In a 100ml single-mouth bottle, 2- ((1- ((2-fluoro-5- ((4-oxo-3, 4-dihydrophthalazin-1-yl) methyl) benzoyl) piperidin-4-yl) methylamino) -N-hydroxypyrimidine-5-carboxylic acid benzyloxy amine 2g and methanol 25ml are stirred and dissolved, then 0.2g palladium carbon is added, hydrogen is introduced, TLC is used for monitoring the reaction process, after the reaction is completed, the reaction liquid is filtered and concentrated, and methanol is used for recrystallization, thus obtaining the product 1.3 g.ESI-MS: M/z 532.21[ M + H ] M/z 532.21]⁺；¹H NMR(600MHz，DMSO-d6)δ:1.13(dd,2H),1.71(d,2H),1.85(m,1H),2.78(t,2H),3.08(t,2H),4.31(s,2H),4.58(d,2H),7.18(t,1H),7.43(m,1H),7.54(d,1H),7.87(dt,2H),7.88(d,1H),8.20(d,1H),8.31(m,1H),8.61(s,2H),8.92(s,1H),10.94(s,1H),12.56(s,1H)。

Example 4, 2- ((1- (2H-indazole-7-formyl) piperidin-4-yl) methylamino) -N-hydroxypyrimidine-5-carboxamide (Compound 4)

A: 2- ((1- (2H-indazole-7-formyl) piperidin-4-yl) methylamino) pyrimidine-5-carboxylic acid methyl ester

To a 250ml single-neck flask, 8g of the compound 2H-indazole-7-carboxylic acid and 100ml of N, N-dimethylformamide were added, and 8g (50mmol) of N, N-carbonyldiimidazole was added under stirring at room temperature, and after 2 hours, 12.5 g of methyl 2- (piperidin-4-ylmethylamino) pyrimidine-5-carboxylate was added. Stirring overnight, and the next day, slowly pouring the reaction solution into a stirred ice-water mixture to precipitate a light yellow solid, standing for 3 hours, filtering, washing with water, and drying to obtain a light yellow powdery product, namely 16.8 g of the target compound, which is directly used for the next reaction without purification. ESI-MS: m/z 395.2.

B: 2- ((1- (2H-indazole-7-formyl) piperidin-4-yl) methylamino) pyrimidine-5-carboxylic acid

16g of methyl 2- ((1- (2H-indazole-7-formyl) piperidin-4-yl) methylamino) pyrimidine-5-carboxylate was charged into a 500-ml single-neck flask, 150 ml of methanol was further added, and after stirring and dissolution, 50ml of water and 6g of lithium hydroxide monohydrate were further added in an ice-water bath. After 4 hours, TLC detection was carried out, hydrolysis was complete, and 2M hydrochloric acid was added dropwise to neutrality. And (3) carrying out reduced pressure rotary evaporation, removing methanol, adjusting the pH value of the solution to about 5 with 2M hydrochloric acid, separating out a large amount of powdery solid, filtering, washing with water, and drying to obtain 12.5 g of light yellow powdery solid, wherein the light yellow powdery solid is directly used for the next reaction without purification. ESI-MS: M/z381.1[ M + H ].

C: 2- ((1- (2H-indazole-7-formyl) piperidin-4-yl) methylamino) pyrimidine-5-carboxylic acid (tetrahydropyran-2-oxy) -amine

3.5g of 2- ((1- (2H-indazole-7-formyl) piperidin-4-yl) methylamino) pyrimidine-5-carboxylic acid, 100ml of tetrahydrofuran and 30ml of N, N-dimethylformamide are added into a 250ml round bottom flask, stirring is carried out to dissolve the mixture, 5ml of N, N-diisopropylethylamine and 4.54g O-benzotriazole-tetramethylurea hexafluorophosphate are added into the mixture, the mixture is stirred for one hour at room temperature, 1.62g O- (tetrahydro-2H-pyran-2-yl) hydroxylamine is added into the mixture, the reaction progress is monitored by TLC, after the reaction is completed, the tetrahydrofuran is removed by concentration under reduced pressure, the residue is extracted by ethyl acetate, the ethyl acetate phases are combined, and the target compound is obtained by concentration and column chromatography, wherein the yield is 75 percent 3.3 g.

D: 2- ((1- (2H-indazole-7-formyl) piperidin-4-yl) methylamino) -N-hydroxypyrimidine-5-carboxamide (Compound 4)

Adding 5.0g of 2- ((1- (2H-indazole-7-formyl) piperidin-4-yl) methylamino) pyrimidine-5-carboxylic acid (tetrahydropyran-2-oxy) -amine and 50ml of hydrochloric acid gas solution of ethyl acetate into a three-necked bottle, stirring at room temperature for reaction, monitoring the reaction process by TLC, filtering precipitated solid after the reaction is completed, and recrystallizing by using methanol to obtain 2.89g of a product with the yield of 70%. ESI-MS of M/z 418.16[ M + Na ]]⁺；¹HNMR(600MHz，DMSO-d6)δ:1.24(dd,2H),1.78(d,2H),1.98(m,1H),3.07(t,2H),3.48(t,2H),4.54(d,2H),7.21(t,1H),7.88(d,2H),8.09(s,1H),8.65(s,1H),8.76(t,1H)。

E: 2- ((1- (2H-indazole-7-formyl) piperidin-4-yl) methylamino) -pyrimidine-5-carboxylic acid benzyloxyamine

2.8g of 2- ((1- (2H-indazole-7-formyl) piperidin-4-yl) methylamino) pyrimidine-5-carboxylic acid, 100ml of tetrahydrofuran and 30ml of N, N-dimethylformamide are added into a 250ml round bottom flask, after stirring and dissolving, 4.19ml of N, N-diisopropylethylamine and 3.88g O-benzotriazole-tetramethylurea hexafluorophosphate are added, stirring is carried out for one hour at room temperature, 1.9g O-benzylhydroxylamine hydrochloride is added, the reaction progress is monitored by TLC, after the reaction is completed, tetrahydrofuran is removed by concentration under reduced pressure, the residue is extracted by ethyl acetate, the ethyl acetate phase is combined, and the mixture is concentrated and subjected to column chromatography to obtain 2.78g of the target compound.

F: 2- ((1- (2H-indazole-7-formyl) piperidin-4-yl) methylamino) -N-hydroxypyrimidine-5-carboxamide (Compound 4)

In a 100ml single-necked bottle, 2- ((1- (2H-indazole-7-formyl) piperidin-4-yl) was added) Methylamino) -pyrimidine-5-carboxylic acid benzyloxy amine 2.4g, methanol 20ml, stirring and dissolving, then adding 0.3g palladium carbon, and introducing hydrogen, TLC monitoring reaction progress. After the reaction is completed, the reaction solution is filtered, concentrated and recrystallized by methanol to obtain 1.52g of a product with the yield of 78%. ESI-MS of M/z 418.16[ M + Na ]]⁺；¹HNMR(600MHz，DMSO-d6)δ:1.24(dd,2H),1.78(d,2H),1.98(m,1H),3.07(t,2H),3.48(t,2H),4.54(d,2H),7.21(t,1H),7.88(d,2H),8.09(s,1H),8.65(s,1H),8.76(t,1H)。

The compounds listed in Table 1 can all be prepared by methods similar to the examples described above. The reaction process is monitored by TLC, and the purification is carried out by adopting conventional column chromatography or recrystallization and other methods.

TABLE 1 numbering, Structure and characterization data for the remaining compounds of the invention

The advantageous effects of the present invention are demonstrated by specific test examples below.

Test example 1 HDAC/PARP1 bioactivity test

A. Experimental method and principle

Enzyme assay of PARP1

PARP is a cellular ribozyme that is present in eukaryotic cells to catalyze the saccharification of poly ADP nuclei. PARP plays an important role in DNA damage repair and maintenance of genomic stability. When DNA is damaged, PARP enzyme is activated, binds to DNA, catalyzes the poly (adenosine diphosphate) ribosylation, thereby initiating the DNA damage control and repair process.

PARP-1 is involved in recognition of DNA nicks, PARP-1 activated after recognition of the DNA nicks forms homodimers and catalyzes decomposition of NAD + into nicotinamide and ADP ribose, and the latter is used as a raw material to carry out nuclear saccharification on nuclear receptor protein poly ADP. HT Universal Chemicals PARP Assay Kit With Histone-coated Strip Wells (Cat # 4676-K096-K) was purchased from TREEVGEN and the activity of PARP1 was determined by measuring the amount of bioadenosine diphosphate bound to histones using a chemiluminescence Assay. AZD2281 is a positive drug.

HDAC enzymology experiment

Imbalance of histone acetylation state caused by Histone Deacetylase (HDAC) abnormality is closely related to the occurrence and development of tumor. The histone deacetylase inhibitor can cause the accumulation of acetylated nucleosome histone in vitro and in vivo experiments, improve the expression level of p21 gene, inhibit the proliferation of tumor cells, induce cell differentiation or apoptosis, and can be used for treating various malignant blood diseases and solid tumors.

HDAC can deacetylate Boc-lys (AC) -AMC under certain conditions, and then hydrolyze the substrate under the action of trypsin to generate AMC, which can emit 460nm fluorescence at 355nm excitation wavelength, so that the enzyme activity can be reflected by detecting the change of the product amount. In this experiment, HDAC enzyme was extracted from Hela cell nucleus. LBH-589 is a positive drug.

B. Experimental data

The results of the inhibition of HDAC and PARP1 by the compounds of the present invention are shown in table 2.

TABLE 2 inhibition of HDAC and PARP1 by the compounds of the present invention

Note: means not determined

The above test results show that: the compound of the invention has inhibitory activity on HDAC and PARP simultaneously, and both reach nanomolar level. Wherein, the inhibition rate of the compound 11 and the compound 13 on HDAC and PARP reaches more than 90 percent. The compound of the invention can be used for preparing HDAC/PARP dual-target inhibitors.

Test example 2 MTT method for cell growth inhibitory Activity

In vitro cell proliferation inhibition assays MMT was used with the following 9 cell lines: human breast cancer cell MDA-MB-436, human breast cancer cell HCC1937, human breast ductal carcinoma cell HCC1395, human breast cancer cell HCC1428, human breast cancer cell SUM-149PT, human breast cancer cell MDA-MB-231, human breast cancer MCF7, human cervical carcinoma Hela and non-mutation triple negative breast cancer MDA-MB-157.

A. Experimental method and principle

The detection principle is that succinate dehydrogenase in mitochondria of living cells can reduce exogenous MTT into water-insoluble blue-purple crystalline Formazan (Formazan) and deposit the blue-purple crystalline Formazan in the cells, while dead cells do not have the function. Dimethyl sulfoxide (DMSO) can dissolve formazan in cells, and the light absorption value is measured by an enzyme-labeling instrument at 490nm wavelength, and the MTT crystal formation amount is in direct proportion to the number of cells in a certain cell number range. The number of living cells is judged according to the measured absorbance value (OD value), and the larger the OD value is, the stronger the cell activity is (if the drug toxicity is measured, the lower the drug toxicity is).

B. Experimental data

The results of the inhibitory activity of the compounds of the present invention on some human tumor cells are shown in table 3:

TABLE 3 cytostatic Activity data for some of the compounds

Note: -indicates that the activity is low, not determined

Test results show that the compound has good in-vitro tumor cell proliferation inhibition activity and good application prospect.

In summary, the present invention provides a class of compounds having good inhibitory activity on both HDAC and PARP, which have good inhibitory effect on both HDAC and PARP, and can be used for preparing a PARP/HDAC dual-target inhibitor, and preventing and/or treating diseases, such as tumors, ischemic diseases, diabetes, inflammatory diseases, etc., which are improved by inhibiting the activity of PARP and/or HDAC. Meanwhile, the compound has synthetic lethal effect, can be used as a chemo-therapeutic drug sensitizer and a radiotherapy sensitizer and is used together with other anti-tumor drugs, so that the treatment effect is improved, and the adverse reaction is reduced. In addition, the compound improves the compliance of patients, has more medicament economy advantages and has good application prospect.

Claims

1. A compound represented by formula I, or a salt thereof, or a stereoisomer thereof, or a solvate thereof, or a hydrate thereof, or a prodrug thereof:

wherein the content of the first and second substances,

X₁selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

X₂Selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

Z₁Selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

Z₂Selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

Y₁、Y₂Each independently selected from N, CR₅；

the ring A is selected from 6-10 membered aryl and 5-10 membered heteroaryl;

2. The compound according to claim 1, or a salt thereof, or a stereoisomer thereof, or a solvate thereof, or a hydrate thereof, or a prodrug thereof, characterized in that:

X₁selected from none, O or NR₂；

X₂Selected from none or CR₃R₄；

Z₁Selected from none or CR₃R₄；

Z₂Selected from none, O or NR₂；

Y₁、Y₂Each independently selected from N, CR₅；

3. The compound according to claim 2, or a salt thereof, or a stereoisomer thereof, or a solvate thereof, or a hydrate thereof, or a prodrug thereof, characterized in that:

X₁selected from none, O or NR₂；

X₂Selected from none or CR₃R₄；

Z₁Selected from none or CR₃R₄；

Z₂Selected from none, O or NR₂；

And X₂And Z₁Cannot be simultaneously CR₃R₄Or simultaneously none;

Y₁、Y₂each independently selected from N, CR₅；

preferably, the first and second electrodes are formed of a metal,

X₁and X₂At the same time is noneOr not both;

Z₁and Z₂Simultaneously absent or not simultaneously absent;

Y₁and Y₂Are not identical.

4. The compound according to claim 2, or a salt thereof, or a stereoisomer thereof, or a solvate thereof, or a hydrate thereof, or a prodrug thereof, characterized in that:

n tableThe substituent R on the ring of A₁The number of (a) is an integer selected from 0, 1, 2 or 3, and when n is 0, it means that the ring A has no substituent;

preferably, the first and second electrodes are formed of a metal,

a ring is selected from

R₁' is selected from halogen.

5. The compound according to claim 1, or a salt thereof, or a stereoisomer thereof, or a solvate thereof, or a hydrate thereof, or a prodrug thereof, characterized in that: the compound is represented by formula II:

wherein the content of the first and second substances,

X₁selected from O or NR₂；

X₂Selected from the group consisting of CR₃R₄；

R₁is a substituent on the A ring and is respectively and independently selected from substituted or unsubstituted C₁～C₄Alkyl radical, C₁～C₄Alkoxy, halogen, substituted or unsubstituted 3-to 6-membered cycloalkylA substituted or unsubstituted 3-to 6-membered saturated heterocyclic group; the heteroatom in the heterocyclic group is N, O or S, and the number of the heteroatoms is 1, 2 or 3;

preferably, the first and second electrodes are formed of a metal,

substituents of said cycloalkyl, heterocyclylIs selected from C₁～C₄Alkyl radical, C₁～C₄Alkoxy, halogen, hydroxy, amino, carboxyl, nitro, cyano; or two substituents on the same carbon atom form a double bond with O;

more preferably still, the first and second liquid crystal compositions are,

a ring is selected from

R₁' is selected from halogen.

6. The compound according to claim 1, or a salt thereof, or a stereoisomer thereof, or a solvate thereof, or a hydrate thereof, or a prodrug thereof, characterized in that: the compound is shown as formula III:

wherein the content of the first and second substances,

X₁selected from O or NR₂；

preferably, the first and second electrodes are formed of a metal,

more preferably still, the first and second liquid crystal compositions are,

a ring is selected from

R₁' is selected from halogen.

7. The compound according to claim 1, or a salt thereof, or a stereoisomer thereof, or a solvate thereof, or a hydrate thereof, or a prodrug thereof, characterized in that: the compound is shown as formula IV:

wherein the content of the first and second substances,

X₁selected from O or NR₂；

R₂Selected from hydrogen, C₁～C₄An alkyl group;

preferably, the first and second electrodes are formed of a metal,

more preferably still, the first and second liquid crystal compositions are,

a ring is selected from

R₁' is selected from halogen.

8. The compound according to claim 1, or a salt thereof, or a stereoisomer thereof, or a solvate thereof, or a hydrate thereof, or a prodrug thereof, characterized in that: the compound is shown as a formula V:

wherein the content of the first and second substances,

Z₁selected from the group consisting of CR₃R₄；

Z₂Selected from O or NR₂；

preferably, the first and second electrodes are formed of a metal,

more preferably still, the first and second liquid crystal compositions are,

a ring is selected from

R₁' is selected from halogen.

9. The compound according to claim 1, or a salt thereof, or a stereoisomer thereof, or a solvate thereof, or a hydrate thereof, or a prodrug thereof, characterized in that: the compound is represented by formula VI:

wherein the content of the first and second substances,

Z₂selected from O or NR₂；

preferably, the first and second electrodes are formed of a metal,

more preferably still, the first and second liquid crystal compositions are,

a ring is selected from

R₁' is selected from halogen.

10. The compound according to claim 1, or a salt thereof, or a stereoisomer thereof, or a solvate thereof, or a hydrate thereof, or a prodrug thereof, characterized in that: the compound is represented by formula VII:

wherein the content of the first and second substances,

Z₂selected from O or NR₂；

R₂Selected from hydrogen, C₁～C₄An alkyl group;

the substituent of the heteroaryl is selected from C₁～C₄Alkyl radical, C₁～C₄Alkoxy, halogen, hydroxy, amino, carboxyl, nitro, cyano; or two on the same carbon atomThe substituents form a double bond to O;

preferably, the first and second electrodes are formed of a metal,

more preferably still, the first and second liquid crystal compositions are,

a ring is selected from

R₁' is selected from halogen.

11. The compound according to claim 1, or a salt thereof, or a stereoisomer thereof, or a solvate thereof, or a hydrate thereof, or a prodrug thereof, characterized in that: the compound is represented by formula VIII:

wherein the content of the first and second substances,

X₁selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

X₂Selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

Z₁Selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

Z₂Selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

Y₁、Y₂Each independently selected from N, CR₅；

preferably, the first and second electrodes are formed of a metal,

X₁selected from none, O or NR₂；

X₂Selected from none or CR₃R₄；

Z₁Selected from none or CR₃R₄；

Z₂Selected from none, O or NR₂；

And X₂And Z₁Cannot be simultaneously CR₃R₄Or simultaneously none;

Y₁、Y₂are respectively provided withIndependently selected from N, CR₅；

more preferably, the compound is of formula VIIIa:

wherein the content of the first and second substances,

X₁selected from O or NR₂；

R₂Selected from hydrogen, C₁～C₄An alkyl group;

alternatively, the compound is of formula VIIIb:

wherein the content of the first and second substances,

Z₂selected from O or NR₂；

R₂Selected from hydrogen, C₁～C₄An alkyl group.

12. The compound according to claim 1, or a salt thereof, or a stereoisomer thereof, or a solvate thereof, or a hydrate thereof, or a prodrug thereof, characterized in that: the compound is represented by formula IX:

wherein the content of the first and second substances,

X₁selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

X₂Selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

Z₁Selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

Z₂Selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

Y₁、Y₂Each independently selected from N, CR₅；

preferably, the first and second electrodes are formed of a metal,

X₁selected from none, O or NR₂；

X₂Selected from none or CR₃R₄；

Z₁Selected from none or CR₃R₄；

Z₂Selected from none, O or NR₂；

And X₂And Z₁Cannot be simultaneously CR₃R₄Or simultaneously none;

Y₁、Y₂each independently selected from N, CR₅；

more preferably, the compound is of formula IXa:

wherein the content of the first and second substances,

X₁selected from O or NR₂；

R₂Selected from hydrogen, C₁～C₄An alkyl group;

alternatively, the compound is of formula IXb:

wherein the content of the first and second substances,

Z₂selected from O or NR₂；

R₂Selected from hydrogen, C₁～C₄An alkyl group.

13. The compound according to claim 1, or a salt thereof, or a stereoisomer thereof, or a solvate thereof, or a hydrate thereof, or a prodrug thereof, characterized in that: the compound is shown as formula X:

wherein the content of the first and second substances,

X₁selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

X₂Selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

Z₁Selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

Z₂Selected from the group consisting of none, O, S, NR₂Or CR₃R₄；

Y₁、Y₂Each independently selected from N, CR₅；

R₁' is selected from halogen;

preferably, the first and second electrodes are formed of a metal,

X₁selected from none, O or NR₂；

X₂Selected from none or CR₃R₄；

Z₁Selected from none or CR₃R₄；

Z₂Selected from none, O or NR₂；

And X₂And Z₁Cannot be simultaneously CR₃R₄Or simultaneously none;

Y₁、Y₂each independently selected from N, CR₅；

R₁' is selected from halogen;

more preferably, the compound is of formula Xa:

wherein the content of the first and second substances,

X₁selected from O or NR₂；

R₂Selected from hydrogen, C₁～C₄An alkyl group;

R₁' is selected from halogen;

alternatively, the compound is of formula Xb:

wherein the content of the first and second substances,

Z₂selected from O or NR₂；

R₂Selected from hydrogen, C₁～C₄An alkyl group;

R₁' is selected from halogen.

14. The compound according to any one of claims 1 to 13, or a salt thereof, or a stereoisomer thereof, or a solvate thereof, or a hydrate thereof, or a prodrug thereof, characterized in that: the compound is one of the following compounds:

15. use of a compound according to any one of claims 1 to 14, or a salt thereof, or a stereoisomer thereof, or a solvate thereof, or a hydrate thereof, or a prodrug thereof, in the manufacture of a medicament for inhibiting PARP and/or HDAC activity.

16. Use according to claim 15, characterized in that: the medicament is a medicament for treating ischemic diseases, diabetes or inflammatory diseases.

17. Use according to claim 15, characterized in that: the medicine is a chemo-therapeutic medicine sensitizer, a radiotherapy sensitizer or a medicine for treating tumors;

preferably, the tumor is breast, ovarian, pancreatic or prostate cancer.

18. A pharmaceutical formulation characterized by: the compound is a preparation prepared by taking one or more of the compounds, or salts thereof, or stereoisomers thereof, or solvates thereof, or hydrates thereof, or prodrugs thereof according to any one of claims 1 to 14 as an active ingredient and adding pharmaceutically acceptable auxiliary materials or auxiliary ingredients.

19. A pharmaceutical composition characterized by: the pharmaceutical composition comprises a compound according to any one of claims 1 to 14, or a salt thereof, or a stereoisomer thereof, or a solvate thereof, or a hydrate thereof, or a prodrug thereof, and a chemotherapeutic agent or a radiotherapeutic agent.

20. Use of a compound according to any one of claims 1 to 14, or a salt thereof, or a stereoisomer thereof, or a solvate thereof, or a prodrug thereof, in combination with a chemotherapeutic agent or a radiotherapeutic agent for the manufacture of a medicament for use in combination.