阅读说明：本技术 作为组蛋白去乙酰化酶抑制剂的巯基化合物及其用途 (Sulfhydryl compound as histone deacetylase inhibitor and application thereof ) 是由 李建其 *** 郭政 浦强 曾景 钱浩 于 2018-09-10 设计创作，主要内容包括：本发明公开了一种作为组蛋白去乙酰化酶抑制剂的巯基化合物及其用途,药理实验结果表明,本发明化合物对多种肿瘤细胞显示较高抗增殖活性,在细胞模型上对谷氨酸诱发的神经元细胞损伤显示保护作用,对正常细胞毒性低,潜在心脏毒性小,且药代特征理想,具有较高的血脑屏障通透性,具有作为高效低毒抗肿瘤或神经退行性疾病治疗剂的开发前景。所述的巯基化合物,为结构通式如式(I)所示的化合物或其药学上可接受的盐、酯或前药：<Image he="253" wi="700" file="DDA0001795009620000011.GIF" imgContent="drawing" imgFormat="GIF" orientation="portrait" inline="no"></Image>(Pharmacological experiment results show that the compound has higher antiproliferative activity on various tumor cells, has a protective effect on glutamic acid induced neuronal cell injury on a cell model, has low toxicity on normal cells, low potential cardiotoxicity and ideal pharmacokinetic characteristics, has higher blood brain barrier permeability, and has a development prospect as a high-efficiency low-toxicity anti-tumor or neurodegenerative disease therapeutic agent. The sulfhydryl compound is a compound with a structural general formula shown in a formula (I) or a pharmaceutically acceptable salt, ester or prodrug thereof:)

1. The sulfhydryl compound is characterized by being a compound with a structural general formula shown as a formula (I) or a pharmaceutically acceptable salt, ester or prodrug thereof:

wherein:

r is hydrogen, halogen, alkyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl, cyano, trifluoromethyl, NH₂NH (alkyl), N (alkyl), NH-aryl, NH-heteroaryl, N (aryl), N (aryl) (heteroaryl), N (aryl) (acyl), N (aryl) (sulfonyl), or N (heteroaryl);

the alkyl is C1-C5;

x is CH or N;

m is 0, 1, 2 or 3;

n is 0 or 1.

2. Mercapto-compound according to claim 1, characterized in that said halogen is fluorine or chlorine.

3. The mercapto compound according to claim 1, wherein the alkyl group is a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group or an isobutyl group.

4. The mercapto compound according to claim 1, wherein the cycloalkyl group is a cyclopropyl group, a cyclopentyl group or a cyclohexyl group.

5. The mercapto compound of claim 1, wherein the heterocycloalkyl group is pyrrolyl, morpholinyl, piperidinyl, tetrahydroquinolinyl, tetrahydrotriazolopyrazinyl, diazepanyl, or piperazinyl.

6. The mercapto compound of claim 1, wherein the aryl or heteroaryl is phenyl, naphthyl, anthracenyl, pyridinyl, pyrimidinyl, pyrazinyl, indolyl, imidazolyl, (benzo) oxazolyl, (benzo) furanyl, (benzo) thienyl, (benzo) thiazolyl, triazolyl, isoxazolyl, quinolinyl, pyrrolyl, pyrazolyl, or 5, 6,7, 8-tetrahydroisoquinoline.

7. The mercapto compound according to claim 1, wherein the acyl group is an acetyl group, a propionyl group, an isobutyryl group or an arylacyl group.

8. The thiol compound according to claim 1, wherein the sulfonyl group is a methylsulfonyl group or an arylsulfonyl group.

9. The compound containing a sulfydryl structure is a compound shown as a formula (II) or a pharmaceutically acceptable salt, ester or prodrug thereof,

wherein:

R₁is hydrogen, alkyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl, acyl or sulfonyl.

The alkyl is C1-C5;

y is CH or N;

p is 0, 1, 2 or 3;

q is 0 or 1.

10. The mercapto compound of claim 9, wherein the alkyl group is a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, or an isobutyl group.

11. The mercapto compound of claim 9, wherein the cycloalkyl is cyclopropyl, cyclopentyl or cyclohexyl.

12. The mercapto compound of claim 9, wherein the heterocycloalkyl group is pyrrolyl, morpholinyl, piperidinyl, tetrahydroquinolinyl, tetrahydrotriazolopyrazinyl, diazepanyl, or piperazinyl.

13. The mercapto compound of claim 9, wherein the aryl or heteroaryl is phenyl, naphthyl, anthracenyl, pyridinyl, pyrimidinyl, pyrazinyl, indolyl, imidazolyl, (benzo) oxazolyl, (benzo) furanyl, (benzo) thienyl, (benzo) thiazolyl, triazolyl, isoxazolyl, quinolinyl, pyrrolyl, pyrazolyl, or 5, 6,7, 8-tetrahydroisoquinoline.

14. The mercapto compound of claim 9, wherein said acyl group is an acetyl, propionyl, isobutyryl, or aryloyl group.

15. The thiol compound according to claim 9, wherein the sulfonyl group is a methanesulfonyl group or an arylsulfonyl group.

16. A sulfhydryl compound, characterized in that it is selected from the following compounds or a pharmaceutically acceptable salt, ester or prodrug thereof:

i-12- (diphenylamino) -N- (5- (2-mercaptoacetylamino) pentyl) pyrimidine-5-carboxamide,

I-22- (diphenylamino) -N- (6-mercaptohexyl) pyrimidine-5-carboxamide,

I-34- (diphenylamino) -N- (5- (2-mercaptoacetylamino) pentyl) benzamide,

I-44- (diphenylamino) -N- (6-mercaptohexyl) benzamide,

I-54- (di (pyrimidin-2-yl) amino) -N- (5- (2-mercaptoacetylamino) pentyl) benzamide,

I-64- (di (pyrimidin-2-yl) amino) -N- (6-mercaptohexyl) benzamide,

I-72- ((2, 6-dichlorophenyl) (methyl) amino) -N- (5- (2-mercaptoacetylamino) pentyl) pyrimidine-5-carboxamide,

I-82- ((2, 6-dichlorophenyl) (methyl) amino) -N- (6-mercaptohexyl) pyrimidine-5-carboxamide,

I-9N- (5- (2-mercaptoacetylamino) pentyl) -2- (N-phenylacetylamino) pyrimidine-5-carboxamide,

I-10N- (6-mercaptohexyl) -2- (N-phenylacetylamino) pyrimidine-5-carboxamide,

I-112- (benzo [ d ] oxazol-2-ylamino) -N- (5- (2-mercaptoacetylamino) pentyl) pyrimidine-5-carboxamide,

I-122- (benzo [ d ] oxazol-2-ylamino) -N- (6-mercaptohexyl) pyrimidine-5-carboxamide,

I-13N- (4- ((5- (2-mercaptoacetylamino) pentyl) carbamoyl) phenyl) -N-phenylpyridinecarboxamide,

I-14N- (4- ((6-mercaptohexyl) carbamoyl) phenyl) -N-phenylpyridine-carboxamide,

I-15N- (5- (2-mercaptoacetylamino) pentyl) -2- (N-phenylphenylsulphonamido) pyrimidine-5-carboxamide,

I-16N- (6-mercaptohexyl) -2- (N-phenylphenylsulfonamide) pyrimidine-5-carboxamide,

I-17N- (5- (2-mercaptoacetylamino) pentyl) -2- (piperidin-1-yl) pyrimidine-5-carboxamide,

I-18N- (6-mercaptohexyl) -2- (piperidin-1-yl) pyrimidine-5-carboxamide,

I-19N- (5- (2-mercaptoacetylamino) pentyl) -2- (4-phenylpiperazin-1-yl) pyrimidine-5-carboxamide,

I-20N- (6-mercaptohexyl) -2- (4-phenylpiperazin-1-yl) -pyrimidine-5-carboxamide,

I-212- (6, 7-dimethoxy-3, 4-dihydroisoquinolin-2 (1H) -yl) -N- (5- (2-mercaptoacetylamino) pentyl) pyrimidine-5-carboxamide or

I-222- (6, 7-dimethoxy-3, 4-dihydroisoquinolin-2 (1H) -yl) -N- (6-mercaptohexyl) pyrimidine-5-carboxamide.

17. The sulfhydryl compound according to any of claims 1 to 16, wherein the pharmaceutically acceptable salt is a pharmaceutically acceptable anion salt.

18. A pharmaceutical composition comprising a therapeutically effective amount of a sulfhydryl compound as claimed in any one of claims 1 to 17 and a pharmaceutically acceptable carrier.

19. Use of the sulfhydryl compound as claimed in any of claims 1-17 in the preparation of a medicament for the treatment of a disease caused by abnormal gene expression.

20. The use of claim 19, wherein the disease caused by abnormal gene expression comprises a tumor, an endocrine disorder, an immune system disease, a genetic disease or a neurological disease.

Technical Field

The invention relates to a histone deacetylase inhibitor containing a sulfhydryl structure and application thereof in treating tumors or neurodegenerative diseases.

Background

Histone Deacetylases (HDACs) are capable of catalyzing the deacetylation process of histone and non-histone proteins, and regulating intracellular acetylation levels with Histone Acetyltransferases (HATs), thereby regulating gene expression. Currently, there are 18 subtypes of mammalian HDACs known, which fall into four categories: class I (HDAC1, HDAC2, HDAC3, HDAC 8); class II is further divided into two subfamilies of IIa (HDAC4, HDAC5, HDAC7, HDAC9) and IIb (HDAC6, HDAC 10); class III (Sirt 1-Sirt 7); class IV (HDAC 11).

There are 5 histone deacetylase inhibitors (HDACi) currently on the market, namely vorinostat (vorinostat), belinostat (belinostat), panobinostat (panobinostat), romidepsin (romidepsin) and chidamide (chidamide), which are broad-spectrum inhibitors and selectively act on class I subtypes. Vorinostat and romidepsin are used for treating cutaneous T-cell lymphoma (CTCL), belinostat and cideraniline are used for treating relapsed and refractory peripheral T-cell lymphoma (PTCL), panobinostat is used in combination with bortezomib and dexamethasone for treating Multiple Myeloma (MM).

Although the above HDAC inhibitors have achieved good clinical efficacy, the following disadvantages are common to broad-spectrum HDAC inhibitors:

(1) strong toxic and side effects, such as nausea, vomiting, bone marrow suppression and the like;

(2) genotoxicity;

(3) poor pharmacokinetic properties, low bioavailability, short half-life and the like.

The above disadvantages both cause inconvenience to tumor patients and hinder the application of broad-spectrum HDAC inhibitors in fields other than tumor therapy.

Hdac subtype selective inhibitors are currently the focus of research in this field. Of these, inhibitors of the HDAC6 subtype have received much attention. HDAC6 is involved in a wide range of diseases, including neurodegenerative diseases, inflammation, autoimmune responses, tumors, and bacterial infections.

HDAC6 is closely related to tumor development, invasion and metastasis. HDAC6 promotes tumor growth by upregulating protein kinase B in the cytoplasm and the phosphorylation levels of extracellular regulated kinases, thereby activating more oncogenic Ras signaling pathways and tumor cell survival signaling pathways. High expression of HDAC6 was also associated with accelerated metastasis and increased invasiveness of cancer cells.

HDAC6 is closely related to neurodegenerative diseases, neurodegenerative diseases are senile diseases in which nerve cells undergo progressive damage and apoptosis, and the pathological mechanism of the senile diseases comprises mitochondrial dysfunction, oxidative stress theory, protein misfolding aggregation, inflammation, immunodeficiency, gene mutation and the like, the diseases mainly comprise Alzheimer disease, amyotrophic lateral sclerosis, ataxia telangiectasia, Creutzfeldt-Jakob disease, Huntington disease, multiple sclerosis, Parkinson disease and the like, with the recent intensive research on the neurodegenerative diseases, a selective HDAC6 inhibitor has the effects of protecting nerve cell growth and promoting axon regeneration, in the neurodegenerative diseases such as Alzheimer disease, Parkinson disease and Huntington disease, inhibition of HDAC6 can enhance the transport of brain-derived neurotrophic factor, is favorable for cell protection and survival, a selective HDAC6 inhibitor tubastatin A TA can possibly inhibit HDAC6 de-activation in cerebral infarction, lead microtubule 2-acetylated protein 387 level and fibroblast growth factor-21 to be regulated, thus the increase the activity of rat tau-derived from tau structure, and further lead to increase of rat secondary mouse tau embolism, thus the effect of rat secondary mouse is probably caused by inhibition of microtubule 6 de-acetylated protein activity.

The HDAC6 inhibitor reported in the literature at present mainly comprises three parts, including a surface recognition region, a connection region and a zinc ion binding region, wherein the zinc ion binding region mostly contains a hydroxamic acid structure, and the structure has the defects of genotoxicity, poor pharmacokinetic property and the like.

Aiming at the defect analysis of the prior art, the invention adopts a drug design and synthesis means to obtain a series of compounds containing a sulfhydryl structure, and pharmacological experiments show that the compounds can selectively inhibit HDAC6, show higher antiproliferative activity on various tumor cells, have low toxicity on normal cells, small potential cardiotoxicity, ideal pharmacokinetic characteristics and higher blood brain barrier permeability, and have development prospects as high-efficiency low-toxicity anti-tumor or neurodegenerative disease therapeutic agents.

Disclosure of Invention

The invention aims to disclose a sulfhydryl compound as a histone deacetylase inhibitor and application thereof, so as to meet the requirement of clinical application. The compound has HDAC6 subtype selectivity, has the characteristics of high efficiency, low toxicity, ideal pharmacokinetic property and the like, and can meet the clinical requirement when being used as a high-efficiency low-toxicity anti-tumor or neurodegenerative disease therapeutic agent.

The sulfhydryl compound is a compound with a structural general formula shown in a formula (I) or a pharmaceutically acceptable salt, ester or prodrug thereof:

wherein:

r is hydrogen, halogen, alkyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl, cyano, trifluoromethyl, NH₂NH (alkyl), N (alkyl), NH-aryl, NH-heteroaryl, N (aryl), N (aryl) (heteroaryl), N (aryl) (acyl), N (aryl) (sulfonyl) or N (heteroaryl).

The alkyl is C1-C5;

x is CH or N;

m is 0, 1, 2 or 3;

n is 0 or 1.

Preferably, the halogen is fluorine or chlorine.

Preferably, said alkyl is methyl, ethyl, propyl, isopropyl, butyl or isobutyl, which may be optionally substituted;

the cycloalkyl is cyclopropyl, cyclopentyl or cyclohexyl, and the cycloalkyl can be optionally substituted;

the heterocycloalkyl is pyrrolyl, morpholinyl, piperidinyl, tetrahydroquinolinyl, tetrahydrotriazolopyrazinyl, diazepanyl or piperazinyl, and the heterocycloalkyl can be optionally substituted;

said aryl or heteroaryl is phenyl, naphthyl, anthracenyl, pyridinyl, pyrimidinyl, pyrazinyl, indolyl, imidazolyl, (benzo) oxazolyl, (benzo) furanyl, (benzo) thienyl, (benzo) thiazolyl, triazolyl, isoxazolyl, quinolinyl, pyrrolyl, pyrazolyl, or 5, 6,7, 8-tetrahydroisoquinoline; the aryl or heteroaryl group may be optionally substituted;

the acyl is acetyl, propionyl, isobutyryl or arylacyl;

the sulfonyl is methylsulfonyl or arylsulfonyl;

the compound containing the sulfhydryl structure is a compound shown in a formula (II) or a pharmaceutically acceptable salt, ester or prodrug thereof:

wherein:

R₁is hydrogen, alkyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl, acyl or sulfonyl;

the alkyl is C1-C5;

y is CH or N;

p is 0, 1, 2 or 3;

q is 0 or 1.

The alkyl is methyl, ethyl, propyl, isopropyl, butyl or isobutyl, and the alkyl can be optionally substituted;

the cycloalkyl is cyclopropyl, cyclopentyl or cyclohexyl, and the cycloalkyl can be optionally substituted;

the heterocycloalkyl is pyrrolyl, morpholinyl, piperidinyl, tetrahydroquinolinyl, tetrahydrotriazolopyrazinyl, diazepanyl or piperazinyl, and the heterocycloalkyl can be optionally substituted;

said aryl or heteroaryl is phenyl, naphthyl, anthracenyl, pyridinyl, pyrimidinyl, pyrazinyl, indolyl, imidazolyl, (benzo) oxazolyl, (benzo) furanyl, (benzo) thienyl, (benzo) thiazolyl, triazolyl, isoxazolyl, quinolinyl, pyrrolyl, pyrazolyl, or 5, 6,7, 8-tetrahydroisoquinoline; the aryl or heteroaryl group may be optionally substituted;

the acyl is acetyl, propionyl, isobutyryl or arylacyl.

The sulfonyl is methylsulfonyl or arylsulfonyl.

The compound containing a sulfhydryl structure is selected from the following compounds:

1-12- (diphenylamino) -N- (5- (2-mercaptoacetylamino) pentyl) pyrimidine-5-carboxamide;

1-22- (diphenylamino) -N- (6-mercaptohexyl) pyrimidine-5-carboxamide;

i-34- (diphenylamino) -N- (5- (2-mercaptoacetylamino) pentyl) benzamide;

i-44- (diphenylamino) -N- (6-mercaptohexyl) benzamide;

i-54- (di (pyrimidin-2-yl) amino) -N- (5- (2-mercaptoacetylamino) pentyl) benzamide;

i-64- (di (pyrimidin-2-yl) amino) -N- (6-mercaptohexyl) benzamide;

i-72- ((2, 6-dichlorophenyl) (methyl) amino) -N- (5- (2-mercaptoacetylamino) pentyl) pyrimidine-5-carboxamide;

1-82- ((2, 6-dichlorophenyl) (methyl) amino) -N- (6-mercaptohexyl) pyrimidine-5-carboxamide;

1-9N- (5- (2-mercaptoacetylamino) pentyl) -2- (N-phenylacetamido) pyrimidine-5-carboxamide;

1-10N- (6-mercaptohexyl) -2- (N-phenylacetamido) pyrimidine-5-carboxamide;

i-112- (benzo [ d ] oxazol-2-ylamino) -N- (5- (2-mercaptoacetylamino) pentyl) pyrimidine-5-carboxamide;

i-122- (benzo [ d ] oxazol-2-ylamino) -N- (6-mercaptohexyl) pyrimidine-5-carboxamide;

1-13N- (4- ((5- (2-mercaptoacetylamino) pentyl) carbamoyl) phenyl) -N-phenylpyridinecarboxamide;

1-14N- (4- ((6-mercaptohexyl) carbamoyl) phenyl) -N-phenylpyridine carboxamide;

1-15N- (5- (2-mercaptoacetylamino) pentyl) -2- (N-phenylphenylsulfonamide) pyrimidine-5-carboxamide;

I-16N- (6-mercaptohexyl) -2- (N-phenylphenylsulfonamide) pyrimidine-5-carboxamide;

1-17N- (5- (2-mercaptoacetylamino) pentyl) -2- (piperidin-1-yl) pyrimidine-5-carboxamide;

1-18N- (6-mercaptohexyl) -2- (piperidin-1-yl) pyrimidine-5-carboxamide;

1-19N- (5- (2-mercaptoacetylamino) pentyl) -2- (4-phenylpiperazin-1-yl) pyrimidine-5-carboxamide;

I-20N- (6-mercaptohexyl) -2- (4-phenylpiperazin-1-yl) -pyrimidine-5-carboxamide

I-212- (6, 7-dimethoxy-3, 4-dihydroisoquinolin-2 (1H) -yl) -N- (5- (2-mercaptoacetylamino) pentyl) pyrimidine-5-carboxamide or

I-222- (6, 7-dimethoxy-3, 4-dihydroisoquinolin-2 (1H) -yl) -N- (6-mercaptohexyl) pyrimidine-5-carboxamide.

The structural formula of the compound is shown in the following table:

the compound of formula (I) and the salt of any compound of I-1 to I-22 are pharmaceutically acceptable anion salts, preferably hydrochloride, hydrobromide, sulfate, acetate, trifluoroacetate, citrate, tartrate, maleate, fumarate, methanesulfonate, malate, p-toluenesulfonate or oxalate.

The compounds of the invention can be prepared by the following general synthetic method:

1) when n is 1, the synthesis is performed according to synthesis general method 1, as follows:

2) when n is 0, the synthesis was performed according to synthesis general method 2, as follows:

the compound with the structure of the formula (I) is obtained by the synthesis general method 1 or 2, and can be further reacted with inorganic acid and organic acid in a solvent, and the salt of the corresponding compound with the structure of the formula (I) is precipitated by cooling.

The raw materials, compounds and reagents used in the above-mentioned production methods can be purchased from commercial sources.

Pharmacological experiments show that the compound has the following beneficial effects:

1) the compounds of the invention have high inhibitory activity (nM level) on HDAC6, and show certain selective inhibitory activity (relative to HDAC 1).

2) Compared with the existing HDAC6 inhibitor, the compound of the invention has weak inhibition effect on normal cells while effectively inhibiting multiple tumor cells, and shows better selective inhibition activity.

3) The compound shows a protective effect on the glutamic acid induced neuronal cell injury on a cell model, and a certain neuroprotective effect is suggested.

4) The compound of the invention has less potential cardiotoxicity and low acute toxicity.

5) The compound of the invention has ideal pharmacokinetic characteristics and higher blood brain barrier permeability.

In conclusion, when the compound is applied as an anti-tumor medicament, the compound has higher blood brain barrier permeability and has a protective effect on neuronal cells, and the compound can be used as a high-efficiency low-toxicity therapeutic agent for treating tumors or neurodegenerative diseases.

The compound of the invention has novel structure, and the efficacy and safety of the compound have creative and substantial scientific progress.

The compound of the invention can be applied to mammals (including human beings) needing tumor treatment in the form of composition by oral administration, injection and the like; especially, oral administration is preferred. The dosage is 0.0001 mg/kg-200 mg/kg body weight per day. The optimum dose depends on the individual, and usually the dose is initially smaller and then gradually increased.

The composition comprises a therapeutically effective amount of a compound of formula (I) and a pharmaceutically acceptable carrier;

the carrier refers to a carrier which is conventional in the pharmaceutical field, such as: diluents, excipients such as water, etc.; binders such as cellulose derivatives, gelatin, polyvinylpyrrolidone, etc.; fillers such as starch and the like; disintegrating agents such as calcium carbonate, sodium bicarbonate; in addition, other adjuvants such as flavoring agents and sweeteners may also be added to the composition.

The composition can be prepared into conventional solid preparations, such as tablets, capsules and the like, and is used for oral administration; it can also be made into injection.

The compound has selective inhibition activity on HDAC6 tumor cell signal transduction pathway, has low toxicity on normal cells, low potential cardiotoxicity and high blood brain barrier permeability, has protective effect on neuron cells, and is suitable for being used as a high-efficiency low-toxicity therapeutic agent for treating tumors or neurodegenerative diseases.

The invention has the advantages that the compound and the medicinal preparation thereof can be used for treating diseases caused by abnormal gene expression, such as: endocrine disorders, immune system diseases, genetic diseases and neurological diseases may also have better therapeutic effects.

Detailed Description

The content of the invention is further elucidated with reference to examples, without the scope of protection of the invention being limited to these examples.