CN109879790B - Amide micromolecule organic compound with indole or indole analogue as mother nucleus structure, application and preparation method thereof - Google Patents
Amide micromolecule organic compound with indole or indole analogue as mother nucleus structure, application and preparation method thereof Download PDFInfo
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- CN109879790B CN109879790B CN201711276112.XA CN201711276112A CN109879790B CN 109879790 B CN109879790 B CN 109879790B CN 201711276112 A CN201711276112 A CN 201711276112A CN 109879790 B CN109879790 B CN 109879790B
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Abstract
The invention discloses amide micromolecule organic compounds which are shown in the formulas (I) and (II) and take indole or indole analogs as parent nucleus structures, or hydrates or pharmaceutically acceptable salts thereof. The invention also discloses application of the compound and a pharmaceutical composition containing the compound or hydrate or pharmaceutically acceptable salt thereof in preventing and/or treating diseases such as various tumors and the like, and application of the compound and the pharmaceutical composition as a histone deacetylase inhibitor in preventing and treating diseases such as autoimmune diseases, allergy and inflammation. The invention also provides a preparation method of the compound and the derivative thereof.
Description
Technical Field
The invention belongs to the field of drug synthesis, and relates to amide micromolecule organic compounds based on taking indole or indole analogs as parent nucleus structures, and a synthesis method and application thereof.
Background
Epigenetic modifications are of great significance for the development, diagnosis and treatment of tumors. Mainly comprises methylation of DNA, histone modification, chromatin remodeling and non-coding RNA regulation. Aberrant epigenetic modifications can cause misexpression of genes, leading to metabolic disorders and disease and even tumorigenesis.
Histone modification mainly includes histone methylation, acetylation, phosphorylation, histone ubiquitination and the like. The most intensive of these is the acetylation of histones. Under normal conditions, acetylation and deacetylation of histones are in a dynamic equilibrium, regulated by both Histone Acetylases (HAT) and Histone Deacetylases (HDAC). HAT can transfer acetyl of acetyl coenzyme A to specific lysine residue at the N terminal of histone, neutralize positive charge of histone, make chromatin present loose structure and promote transcription. HDAC can hydrolyze acetylated lysine, deacetylate it and inhibit gene transcription.
Research shows that most of histones show low acetylation in tumor cells, and chromatin is densely coiled, so that some cancer suppressor genes cannot be normally expressed. HDAC inhibitor can inhibit activity of high expression HDACs, restore dynamic equilibrium state of HATs and HDACs, and inhibit tumor cell growth. With the intensive study of HDACs in tumors, more and more HDAC inhibitors are being applied to tumor studies and exhibit multiple anti-tumor mechanisms of action, such as: inhibiting the proliferation of tumor cells, promoting the differentiation and apoptosis of tumor cells, retarding the progression of the cell cycle of tumor cells, inhibiting the angiogenesis of tumor cells, reducing the motility of tumor cells, etc. There are four HDAC inhibitors currently approved by the FDA for marketing: saha (merck) and FK228(Celgene) for cutaneous T-cell lymphoma, PXD101(Spectrum) for the treatment of peripheral T-cell lymphoma, and 2015 LBH589(Novartis) by FDA accelerated approval procedure, this drug could be used in combination with dexamethasone and bortezomib to treat multiple myeloma. In addition, more than 20 compounds are in different clinical stages. Therefore, the HDAC inhibitor is a very popular anti-tumor target and has wide market prospect.
In addition, (HDACs) inhibitors have been used extensively in the prevention and treatment of autoimmune, allergic and inflammatory diseases. Research on a plurality of animal models of inflammatory diseases shows that HDACI has an anti-inflammatory effect. HDACi, in addition to causing the expression of inflammatory cytokines by regulating the acetylation status of histones, can also inhibit or activate the expression of inflammatory factors by regulating the acetylation status of non-histones including transcription factors, most of which is achieved by the inhibition of nuclear transcription factor KB. However, different HDACs have different acetylation patterns, regulating different genes. Such as systemic lupus erythematosus MRL-Ipr/1pr mouse model, TSA and SAHA can down-regulate the level of inflammatory cytokines IL-12 and IL-6mRNA and protein; in a mouse model of asthma, TSA can reduce the concentrations of IL-4, IL-5, and lgE in bronchoalveolar lavage fluid, thereby slowing the progression of asthma. There are reports that HDACi can promote the expression of inflammatory cytokines. Suuronen et al reported that TSA could enhance the production of NO and IL-6 induced by lipopolysaccharide on microglia N9 cells. Thus, the regulation of inflammatory responses by HDACi may be gene transcription activators or inhibitors. Systemic sclerosis is an autoimmune disease characterized primarily by widespread vascular disease and progressive fibrosis of the skin and internal organs. In the study of the pathogenesis of systemic sclerosis, a variety of pro-inflammatory factors are involved, including TGF-. beta.1, IL-4, PDGF, etc., and HDACs activity is an essential factor in the pathogenesis and disease progression of systemic sclerosis. The studies of Huber et al indicate that TSA can weaken the expression of skin fibroblast type I collagen and fibronectin and reduce the accumulation of skin fibroblast total collagen and extracellular matrix by inhibiting the activity of HDACs in animal models of bleomycin-induced skin fibrosis.
The invention is based on SAHA structure, uses indole and its analogues as mother nucleus, and designs and synthesizes a series of antitumor drugs with novel structure, higher activity and stronger specificity. The compound of the invention is a broad-spectrum HDAC inhibitor, not only shows stronger inhibition activity on enzyme level, but also can inhibit the proliferation of various tumor cells,
disclosure of Invention
The invention provides an amide micromolecule organic compound or hydrate or pharmaceutically acceptable salt which has a novel structure, can be used as an HDAC inhibitor and has an anti-tumor effect and takes indole or indole analogues as a parent nucleus structure, and the structure of the amide micromolecule organic compound or hydrate is shown as the formula (I):
wherein X is N or C;
m, n, p is 0-8; preferably, m, n is 0, 1, 2; p is 1,4, 5, 6, 7;
u is CR 1 Or N;
v is CR 2 Or N;
w is CR 3 Or N;
z is CR 4 Or N;
wherein R is 1 ，R 2 ，R 3 ，R 4 Independently selected from any one of the following groups, including hydrogen, C 1-8 Alkyl groups and isomers thereof, halogen,nitro, amino, cyano, difluoromethyl, trifluoromethyl, methoxy, ethoxy, isopropoxy, difluoromethoxy, trifluoromethoxy, hydroxy.
Preferably, R 1 ，R 2 ，R 3 ，R 4 Independently selected from any one of the following groups, including hydrogen, C 1-8 Alkyl and isomers thereof, wherein alkyl is selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-n-butyl, isobutyl, sec-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, isohexyl, n-octyl, n-heptyl; halogen (F, Cl, Br, I), nitro, amino, cyano, difluoromethyl, trifluoromethyl, methoxy, ethoxy, isopropoxy, difluoromethoxy, trifluoromethoxy, hydroxy.
A is selected from one or none of the following groups: -O-; -SO-; -SO 2 -; -CONR-; -NRCO-; -NR-; -CO-, wherein R is hydrogen; c 1-8 Alkyl and isomers thereof; c 3-8 Cycloalkyl of (2).
Preferably, a is selected from one or none of the following groups: -O-; -SO-; -SO 2 -; -CONR-; -NRCO-; -NR-; -CO-, wherein R is hydrogen; c 1-8 Alkyl and isomers thereof, wherein alkyl is selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-n-butyl, isobutyl, sec-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, isohexyl, n-octyl, n-heptyl; c 3-8 Including monocyclic alicyclic rings (including cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and isomers thereof) and fused cyclic rings, wherein the cycloalkyl group contains one or more heteroatoms O, N, S or is substituted with one or more halogens or hydroxyl groups, and wherein the fused cyclic rings include 1-adamantyl, 2-adamantyl, bicyclo [2.2.0]Pentane, bicyclo [4.4.0]Decane, etc.
R 5 Selected from any one of the following groups: c 1-8 Alkyl and isomers thereof; c 3-8 Cycloalkyl groups of (a); a phenyl group; a heterocyclic aromatic group; substituted phenyl or substituted heterocyclic aromatic group; a benzoheterocyclyl group.
Preferably, R 5 Selected from any one of the following groups:C 1-8 alkyl and isomers thereof, wherein alkyl is selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-n-butyl, isobutyl, sec-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, isohexyl, n-octyl, n-heptyl; c 3-8 Including monocyclic alicyclic rings (including cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and isomers thereof) and fused cyclic rings, wherein the cycloalkyl group contains one or more heteroatoms O, N, S or is substituted with one or more halogens or hydroxyl groups, and wherein the fused cyclic rings include 1-adamantyl, 2-adamantyl, bicyclo [2.2.0]Pentane, bicyclo [4.4.0]Decane, etc.; a phenyl group; heterocyclic aromatic groups (including: isoxazolyl, isothiazolyl, pyrazolyl, pyridyl, oxazolyl, thiazole, imidazole, triazole, tetrazole, furyl, triazinyl, thiophene, pyrimidine, pyridazine, pyrazine); substituted phenyl or substituted heterocyclic aromatic group (the substituted phenyl and the substituted five-membered or six-membered heterocyclic aromatic group have one or more substituents comprising F, Cl, Br, I, hydroxyl, methyl, ethyl, isopropyl, methoxy, trifluoromethyl, difluoromethoxy, trifluoromethoxy, nitro, -CN, amino); benzoheterocyclyl, the heterocycle including C 3-6 Wherein the cycloalkyl group contains 1 or more heteroatoms O, N, S or is substituted with one or more halogens or hydroxyl groups.
Preferably, the first and second liquid crystal display panels are,
selected from any one of the following groups: a phenyl group; heterocyclic aromatic groups (including: isoxazolyl, isothiazolyl, pyrazolyl, pyridyl, oxazolyl, thiazole, imidazole, triazole, tetrazole, furyl, triazinyl, thiophene, pyrimidine, pyridazine, pyrazine); substituted phenyl or substituted heterocyclic aromatic radical (the substituted phenyl and the substituted five-membered or six-membered heterocyclic aromatic radical have one or more substituentsA substituent group comprising: F. cl, Br, I, hydroxy, methyl, ethyl, isopropyl, methoxy, trifluoromethyl, difluoromethoxy, trifluoromethoxy, nitro, -CN, amino).
C is selected from any one of the following groups: - (CH) 2 ) q-, wherein q is 0-3;
preferably, C is
ZBG is a zinc chelating group selected from any one of the following groups:
wherein R is 6 Selected from any one of the following groups: comprising hydrogen, C 1-8 Alkyl and its isomers, halogen, nitro, amino, cyano, difluoromethyl, trifluoromethyl, methoxy, ethoxy, isopropoxy, difluoromethoxy, trifluoromethoxy, hydroxy; r 7 Selected from any one of the following groups: comprising C 1-8 Alkyl and its isomer, C 3-8 The cycloalkyl, phenyl, heterocyclic aryl, substituted phenyl or substituted heterocyclic aryl of (a); r 8 And R 9 Independently selected from any one of the following groups: comprising hydrogen, C 1-8 Alkyl and isomers thereof, or R 8 And R 9 Linked to form a cycloalkyl group of 3 to 7 chain lengths.
Preferably, ZBG is a zinc chelating group selected from any one of the following groups:
wherein R is 6 Selected from any one of the following groups: comprising hydrogen, C 1-8 Alkyl and isomers thereof, wherein alkyl is selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-n-butyl, isobutyl, sec-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, isohexyl, n-octyl, n-heptyl, halo, nitro, amino, cyano, difluoromethyl, trifluoromethyl, methoxy, ethoxy, isopropoxy, difluoromethoxy, trifluoromethoxy, hydroxy; r 7 Selected from any one of the following groups: comprising C 1-8 Alkyl and isomers thereof, wherein alkyl is selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-n-butyl, isobutyl, sec-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, isohexyl, n-octyl, n-heptyl, C 3-8 Cycloalkyl groups of (a) include monocyclic alicyclic (including cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and isomers thereof) and fused-ring alicyclic wherein the cycloalkyl group contains one or more heteroatoms O, N, S or is substituted with one or more halogens or hydroxyl groups, phenyl, heterocyclic aromatic (including isoxazolyl, isothiazolyl, pyrazolyl, pyridyl, oxazolyl, thiazole, imidazole, triazole, tetrazole, furyl, triazinyl, thiophene, pyrimidine, pyridazine, pyrazine), substituted phenyl or substituted heterocyclic aromatic (the substituted phenyl and substituted five-or six-membered heterocyclic aromatic group having one or more substituents including F, Cl, Br, I, hydroxyl, methyl, ethyl, isopropyl, methoxy, trifluoromethyl, difluoromethoxy, trifluoromethoxy, nitro, -CN, amino). R 8 And R 9 Independently selected from any one of the following groups: comprising hydrogen, C 1-8 Alkyl and isomers thereof, wherein alkyl is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-n-butyl, isobutyl, sec-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, isohexyl, n-octyl, n-heptyl, or R 8 And R 9 Linked to form a cycloalkyl group of 3 to 7 chain length, wherein the cycloalkyl group contains one or more heteroatoms O, N, S or is substituted with one or more halogens or hydroxyl groups.
The invention also provides an amide micromolecule organic compound which has a novel structure, can be used as an HDAC inhibitor and has an anti-tumor effect and takes indole or indole analogues as a parent nucleus structure, and the structure of the amide micromolecule organic compound is shown as the formula (II):
wherein Y is N or C; m, n, p is 0-8; preferably, m, n is 0, 1, 2; p is 1,4, 5, 6, 7;
u is CR 1 Or N;
v is CR 2 Or N;
w is CR 3 Or N;
z is CR 4 Or N;
wherein R is 1 ，R 2 ，R 3 ，R 4 Independently selected from any one of the following groups, including hydrogen, C 1-8 Alkyl and its isomers, halogen, nitro, amino, cyano, difluoromethyl, trifluoromethyl, methoxy, ethoxy, isopropoxy, difluoromethoxy, trifluoromethoxy, hydroxy.
Preferably, R 1 ，R 2 ，R 3 ，R 4 Independently selected from any one of the following groups, including hydrogen, C 1-8 Alkyl and isomers thereof, wherein alkyl is selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-n-butyl, isobutyl, sec-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, isohexyl, n-octyl, n-heptyl; halogen (F, Cl, Br, I), nitro, amino, cyano, difluoromethyl, trifluoromethyl, methoxy, ethoxy, isopropoxy, difluoromethoxy, trifluoromethoxy, hydroxy.
A is selected from one or none of the following groups: -O-; -SO-; -SO 2 -; -CONR-; -NRCO-; -NR-; -CO-, wherein R is hydrogen; c 1-8 Alkyl and isomers thereof; c 3-8 A cycloalkyl group of (a).
Preferably, a is selected from one or none of the following groups: -O-; -SO-; -SO 2 -; -CONR-; -NRCO-; -NR-; -CO-, wherein R is hydrogen; c 1-8 An alkyl group and isomers thereof, wherein,wherein alkyl is selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-n-butyl, isobutyl, sec-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, isohexyl, n-octyl, n-heptyl; c 3-8 Cycloalkyl groups of (a) include monocyclic alicyclic rings (including: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and isomers thereof) and fused cyclic alicyclic rings wherein the cycloalkyl group contains one or more heteroatoms O, N, S or is substituted with one or more halogens or hydroxyl groups, and wherein the fused cyclic alicyclic rings include 1-adamantyl, 2-adamantyl, bicyclo [2.2.0 ] and]pentane, bicyclo [4.4.0]Decane, etc.
R 5 Selected from any one of the following groups: c 1-8 Alkyl and isomers thereof; c 3-8 Cycloalkyl groups of (a); a phenyl group; a heterocyclic aromatic group; substituted phenyl or substituted heterocyclic aromatic group; a benzoheterocyclyl group.
Preferably, R 5 Selected from any one of the following groups: c 1-8 Alkyl and isomers thereof, wherein alkyl is selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-n-butyl, isobutyl, sec-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, isohexyl, n-octyl, n-heptyl; c 3-8 Including monocyclic alicyclic rings (including cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and isomers thereof) and fused cyclic rings, wherein the cycloalkyl group contains one or more heteroatoms O, N, S or is substituted with one or more halogens or hydroxyl groups, and wherein the fused cyclic rings include 1-adamantyl, 2-adamantyl, bicyclo [2.2.0]Pentane, bicyclo [4.4.0]Decane, etc.; a phenyl group; heterocyclic aromatic groups (including: isoxazolyl, isothiazolyl, pyrazolyl, pyridyl, oxazolyl, thiazole, imidazole, triazole, tetrazole, furyl, triazinyl, thiophene, pyrimidine, pyridazine, pyrazine); substituted phenyl or substituted heterocyclic aromatic group (the substituted phenyl and the substituted five-membered or six-membered heterocyclic aromatic group have one or more substituents comprising F, Cl, Br, I, hydroxyl, methyl, ethyl, isopropyl, methoxy, trifluoromethyl, difluoromethoxy, trifluoromethoxy, nitro, -CN, amino); benzoheterocyclyl, the heterocycle including C 3-6 Wherein the cycloalkyl group contains1 or more heteroatoms O, N, S or one or more halogen or hydroxyl substitutions.
Preferably, the first and second electrodes are formed of a metal,
selected from any one of the following groups: a phenyl group; heterocyclic aromatic groups (including: isoxazolyl, isothiazolyl, pyrazolyl, pyridyl, oxazolyl, thiazole, imidazole, triazole, tetrazole, furyl, triazinyl, thiophene, pyrimidine, pyridazine, pyrazine); substituted phenyl or substituted heterocyclic aromatic group (the substituted phenyl and substituted five-membered or six-membered heterocyclic aromatic group have one or more substituents, including F, Cl, Br, I, hydroxyl, methyl, ethyl, isopropyl, methoxy, trifluoromethyl, difluoromethoxy, trifluoromethoxy, nitro, -CN, amino).
C is selected from any one of the following groups: - (CH) 2 ) q-, wherein q is 0-3;
preferably, C is
ZBG is a zinc chelating group selected from any one of the following groups:
wherein R is 6 Selected from any one of the following groups: comprising hydrogen, C 1-8 Alkyl and its isomers, halogen, nitro, amino, cyano, difluoromethyl, trifluoromethyl, methoxy, ethoxy,isopropoxy, difluoromethoxy, trifluoromethoxy, hydroxy; r 7 Selected from any one of the following groups: comprising C 1-8 Alkyl and isomers thereof, C 3-8 Cycloalkyl, phenyl, heterocycloaryl, substituted phenyl or substituted heterocycloaryl of (a); r 8 And R 9 Independently selected from any one of the following groups: comprising hydrogen, C 1-8 Alkyl and isomers thereof, or R 8 And R 9 Linked to form a cycloalkyl group of 3 to 7 chain lengths.
Preferably, ZBG is a zinc chelating group selected from any one of the following groups:
wherein R is 6 Selected from any one of the following groups: comprising hydrogen, C 1-8 Alkyl and isomers thereof, wherein alkyl is selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-n-butyl, isobutyl, sec-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, isohexyl, n-octyl, n-heptyl, halogen, nitro, amino, cyano, difluoromethyl, trifluoromethyl, methoxy, ethoxy, isopropoxy, difluoromethoxy, trifluoromethoxy, hydroxy; r 7 Selected from any one of the following groups: comprising C 1-8 Alkyl and isomers thereof, wherein alkyl is selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-n-butyl, isobutyl, sec-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, isohexyl, n-octyl, n-heptyl, C 3-8 Cycloalkyl groups include monocyclic alicyclic rings (including cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and isomers thereof) and fused cyclic alicyclic rings in which the cycloalkyl group contains one or more heteroatoms O, N, S or is substituted with one or more halogens or hydroxy groups, phenyl groups, heterocyclic aromatic groups (including isoxazolyl, isothiazolyl, pyrazolyl, pyridyl, oxazolyl, thiazole, imidazole, triazole, tetrazole, furyl, triazinyl, thiophene, pyrimidine, pyridazine, pyrazine), substituted phenyl groups or substituted phenyl groupsSubstituted heterocyclic aromatic group (the substituted phenyl and the substituted five-membered or six-membered heterocyclic aromatic group have one or more substituents comprising F, Cl, Br, I, hydroxyl, methyl, ethyl, isopropyl, methoxy, trifluoromethyl, difluoromethoxy, trifluoromethoxy, nitro, -CN and amino). R 8 And R 9 Independently selected from any one of the following groups: comprising hydrogen, C 1-8 Alkyl and isomers thereof, wherein alkyl is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-n-butyl, isobutyl, sec-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, isohexyl, n-octyl, n-heptyl, or R 8 And R 9 Linked to form a cycloalkyl group of 3 to 7 chain length, wherein the cycloalkyl group contains one or more heteroatoms O, N, S or is substituted with one or more halogens or hydroxyl groups.
The invention also provides pharmaceutically acceptable salts of amide micromolecule organic compounds which are shown in formulas (I) to (II) and take indole or indole analogues as parent structures or hydrates thereof, including but not limited to acid addition salts formed by the amide micromolecule compounds or the hydrates thereof and the following acids, wherein the acids comprise hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, acetic acid, tartaric acid, salicylic acid, citric acid, methanesulfonic acid, p-toluenesulfonic acid, lactic acid, pyruvic acid, maleic acid and succinic acid.
In the invention, the amide small molecule compound or the hydrate thereof can be combined with radioactivity, a fluorescent group or biotin.
The invention relates to amide micromolecule organic compounds which are shown in formulas (I) to (II) and take indole or indole analogs as parent nucleus structures, or hydrates or pharmaceutically acceptable salts thereof, which comprise:
(7- (3- ((1-adamantanamine) formyl)) -indolyl) heptanoylhydroxylamine
(7- (3- ((1-adamantanamine) formyl)) -indolyl) heptanoic acid methyl ester
(6- (3- ((1-adamantanamine) formyl)) -indolyl) hexanoylhydroxylamine
Methyl (6- (3- ((1-adamantanamine) formyl)) -indolyl) hexanoate
(7- (3- (cyclooctylcarbonyl)) -indolyl) heptanoylhydroxylamine
(7- (3- (Cyclooctylformyl)) -indolyl) heptanoic acid methyl ester
(7- (2- ((1-adamantanamine) formyl)) -indolyl) heptanoylhydroxylamine
(7- (2- ((1-adamantanamine) formyl)) -indolyl) heptanoic acid methyl ester
(7- (3- ((1-adamantanamine) formyl) -5-fluoro) -indolyl) heptanoylhydroxylamine
(7- (3- ((1-adamantanamine) formyl) -5-fluoro) -indolyl) heptanoic acid methyl ester
(7- (3- ((4-chlorobenzyl) formyl)) -indolyl) heptanoylhydroxylamine
(7- (3- ((4-chlorobenzyl) formyl)) -indolyl) heptanoic acid methyl ester
(7- (2- ((2-adamantanamine) formyl) -6-bromo) -indolyl) heptanoylhydroxylamine
(7- (2- ((2-adamantanamine) formyl) -6-bromo) -indolyl) heptanoic acid methyl ester
(7- (2- ((2-adamantanamine) formyl) -5-chloro) -indolyl) heptanoylhydroxylamine
(7- (2- ((2-adamantanamine) formyl) -5-chloro) -indolyl) heptanoic acid methyl ester
(7- (2- ((2-adamantanamine) formyl)) -indolyl) heptanoylhydroxylamine
(7- (2- ((2-adamantanamine) formyl)) -indolyl) heptanoic acid methyl ester
(7- (2- ((2-adamantanamine) formyl) -5-methoxy) -indolyl) heptanoylhydroxylamine
(7- (2- ((2-Adamantadine) formyl) -5-methoxy) -indolyl) heptanoic acid methyl ester
(7- (2- ((4-fluorophenyl) formyl)) -indolyl) heptanoylhydroxylamine
(7- (2- ((4-fluorophenyl) formyl)) -indolyl) heptanoic acid methyl ester
(7- (3- (cyclohexanecarboxoyl)) -indolyl) heptanoylhydroxylamine
Methyl (7- (3- (cyclohexanecarbonyl)) -indolyl) heptanoate
(7- (3- ((1-adamantanamine) formyl)) -7-azaindolyl) heptanoylhydroxylamine
(7- (3- ((1-adamantanamine) formyl)) -7-azaindolyl) heptanoic acid methyl ester
(7- (3- ((2-adamantanamine) formyl)) -indolyl) heptanoylhydroxylamine
(7- (3- ((2-adamantanamine) formyl)) -indolyl) heptanoic acid methyl ester
(5- (3- ((2-adamantanamine) formyl)) -indolyl) pentanoylhydroxylamine
(5- (3- ((2-adamantanamine) formyl)) -indolyl) pentanoic acid methyl ester
(7- (3- (cycloheptylcarbonyl)) -indolyl) heptanoylhydroxylamine
(7- (3- (cycloheptylcarbonyl)) -indolyl) heptanoic acid methyl ester
(5- (3- ((1-adamantanamine) formyl)) -indolyl) pentanoylhydroxylamine
(5- (3- ((1-adamantanamine) formyl)) -indolyl) pentanoic acid methyl ester
(7- (3- ((1-adamantanamine) formyl) -5-chloro) -indolyl) heptanoylhydroxylamine
(7- (3- ((1-adamantanamine) formyl) -5-chloro) -indolyl) heptanoic acid methyl ester
(7- (3- ((2, 4-dichlorophenyl) formyl)) -indolyl) heptanoylhydroxylamine
(7- (3- ((2, 4-dichlorophenyl) formyl)) -indolyl) heptanoic acid methyl ester
(7- (3- ((1-adamantanamine) formyl) -5-methoxy) -indolyl) heptanoylhydroxylamine
(7- (3- ((1-adamantanamine) formyl) -5-methoxy) -indolyl) heptanoic acid methyl ester
(7- (3- ((1-adamantanamine) formyl)) -indolyl) heptacetylmercaptoester
(6- (3- ((2-adamantanamine) formyl)) -indolyl) hexyl (acetyl) mercapto ester
(6- (3- ((1-adamantanamine) formyl)) -indolyl) hexanethiol
(7- (3- ((2-adamantanamine) formyl)) -indolyl) heptacetylmercaptoester
(6- (3- ((1-adamantanamine) formyl)) -indolyl) hexyl (acetyl) mercapto ester
(7- (3- ((2-adamantanamine) formyl)) -indolyl) heptanethiol
(7- (2- ((1-adamantanamine) formyl)) -3-azaindolyl) heptanoylhydroxylamine
N- (2-adamantane) -1- (4-hydroxycarbamoylbenzyl) indolecarboxamides
The invention also provides a pharmaceutical composition which contains the amide micromolecule organic compounds shown in formulas (I) to (II) or hydrates or pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable carrier. The pharmaceutical composition is formulated as an injectable fluid, aerosol, cream, gel, pill, capsule, syrup, transdermal patch, or excipient. The pharmaceutical composition of the present invention can be used alone or in combination with other drugs.
The invention also provides an amide micromolecule organic compound shown in formulas (I) to (II) or a hydrate or pharmaceutically acceptable salt thereof, or an application of the pharmaceutical composition in preparing Histone Deacetylase (HDACs) inhibitors; preferably, for the preparation of HDAC1, HDAC6 inhibitors.
The invention also provides an amide micromolecule organic compound shown in formulas (I) to (II), or a hydrate or a pharmaceutically acceptable salt thereof, or an application of the pharmaceutical composition in preparing a preparation for promoting deacetylation of cell histones H3 and H4 and alpha-Tubulin.
The invention also provides an amide micromolecule organic compound shown in formulas (I) to (II) or a hydrate or a pharmaceutically acceptable salt thereof, or an application of the pharmaceutical composition in preparing a medicament for preventing and/or treating tumors; wherein the compound or hydrate or pharmaceutically acceptable salt thereof, or the pharmaceutical composition is used for inhibiting proliferation, growth, infiltration and migration, clone formation of tumor cells, promoting apoptosis of tumor cells, wherein the compound or hydrate or pharmaceutically acceptable salt thereof, or the pharmaceutical composition inhibits proliferation of tumor cells by retarding cell cycle. The invention also provides an application of the amide micromolecule organic compounds shown in formulas (I) to (II) or hydrates or pharmaceutically acceptable salts thereof or the pharmaceutical composition in preparing anti-tumor therapeutic drugs, which are used for anti-tumor therapy after chemotherapy failure caused by acquired drug resistance. Wherein, the tumor includes but is not limited to lung cancer, breast cancer, epidermoid cancer, rectal/colon cancer, liver cancer, stomach cancer, prostate cancer, pancreatic cancer, leukemia, ovarian cancer, bladder cancer, kidney cancer, oral cancer, melanoma, head and neck cancer, uterine cancer, brain tumor, esophageal cancer, lymph cancer, etc.; preferably, non-small cell lung cancer, breast cancer, gastric cancer, colon cancer and promyelocytic leukemia; further preferably, the tumor cells are a549 cells (non-small cell lung cancer), MDA-MB-231 cells (breast cancer), SGC7901 (gastric cancer), HCT116 (colon cancer) and HL60 cells (promyelocytic leukemia).
The invention also provides an application of the amide micromolecule organic compounds shown in formulas (I) to (II) or hydrates or pharmaceutically acceptable salts thereof, or the pharmaceutical composition as a histone deacetylase inhibitor in preparation of medicines for preventing and/or treating thioredoxin-mediated diseases, wherein the diseases comprise but are not limited to autoimmune diseases, allergy, inflammation, neurodegenerative diseases, Alzheimer's disease, peroneal muscular atrophy, traumatic brain injury, hypertension, stroke and the like; preferably, autoimmune diseases, allergy or inflammation.
The invention also provides an application of the amide micromolecule organic compound, hydrate or pharmaceutically acceptable salt with the indole or the indole analogue as a parent nucleus structure or the pharmaceutical composition in preparing a medicament for treating malignant tumors, wherein the compound, the hydrate or pharmaceutically acceptable salt is used for inhibiting the growth, the metastasis and the recurrence of the malignant tumors; wherein, the malignant tumor includes but is not limited to liver cancer, lung cancer, prostate cancer, skin cancer, rectal/colon cancer, pancreatic cancer, breast cancer, leukemia, ovarian cancer, stomach cancer, bladder cancer, renal cancer, oral cancer, melanoma, head and neck cancer, uterine cancer, brain tumor, esophageal cancer, lymph cancer, etc.; preferably, non-small cell lung cancer, breast cancer, gastric cancer, colon cancer and promyelocytic leukemia; further preferably, the tumor cells are a549 cells (non-small cell lung cancer), MDA-MB-231 cells (breast cancer), SGC7901 (gastric cancer), HCT116 (colon cancer) and HL60 cells (promyelocytic leukemia).
In one embodiment, the tumor cells include, but are not limited to, lung cancer cells, breast cancer cells, epidermal cancer cells, colon cancer cells, liver cancer cells, stomach cancer cells, prostate cancer cells, pancreatic cancer cells, leukemia cells, ovarian cancer cells, bladder cancer cells, kidney cancer cells, oral cancer cells.
In a particular embodiment, the composition is formulated as an injectable fluid, aerosol, cream, gel, pill, capsule, syrup, transdermal patch, or excipient. In another embodiment, wherein said compound is radiolabeled, fluorophore or Biotin (Biotin).
The invention also provides a prodrug and a drug metabolite of the compounds shown in the formulas (I) and (II) or hydrates or pharmaceutically acceptable salts thereof, wherein the prodrug is a compound which is obtained by modifying a chemical structure of a drug, has no or little activity in vitro, releases an active drug through enzymatic or non-enzymatic conversion in vivo and exerts the drug effect; the drug metabolite refers to a substance having a physiological or pharmacological activity.
The invention also provides a preparation method of the amide micromolecule compound which takes indole or indole analogue as a parent nucleus structure and is shown in the formula (I) and/or (II), suitable reaction conditions for the reaction routes are known in the field, and appropriate changes of the reaction conditions, including replacement of solvents, reagents and the like, are within the capability of a person skilled in the art, and the preparation method has guiding value for small tests, pilot tests and amplification.
The first method is shown as a reaction formula (i):
the method comprises the steps of firstly carrying out condensation reaction on carboxylic acid of various indoles or indole analogues and different amines to form corresponding amide compounds, then reacting with sodium hydride in dimethylformamide, and adding bromide under the ice bath condition to form corresponding intermediates. The intermediate is coupled with ZBG in a proper solvent such as dichloromethane, methanol and dimethylformamide to generate a target compound, the target compound is generally quenched by ice water after the reaction is finished, extracted by solvents such as ethyl acetate or dichloromethane and the like, washed by water and saturated saline solution respectively, dried by anhydrous sodium sulfate, decompressed and distilled to remove the solvent, and the final product is obtained by column chromatography. The structure and purity of the compound are proved by methods such as nuclear magnetic resonance, mass spectrum, high performance liquid chromatography and the like.
Method two, as shown in equation (ii):
firstly, various indole or indole analogue carboxylic acids and different amines are subjected to condensation reaction to form corresponding amide compounds, and then the corresponding amide compounds and terminal olefin-containing bromization reaction are subjected to reaction to form corresponding intermediates. The intermediate generates corresponding acetyl mercapto ester through free radical reaction, and then mercaptan is obtained through further hydrolysis. Quenching with ice water after reaction, extracting with ethyl acetate or dichloromethane, washing with water and saturated sodium chloride, drying with anhydrous sodium sulfate, distilling under reduced pressure to remove solvent, and performing column chromatography to obtain the final product. The structure and purity of the compound are proved by methods such as nuclear magnetic resonance, mass spectrum, high performance liquid chromatography and the like.
In recent years, the incidence of tumors is increasing year by year, and the research and development market of anti-tumor drugs is very wide. With the rapid development of molecular oncology and molecular pharmacology, the drug therapy of malignant tumors is no longer limited to common chemotherapeutic drugs, and the current research focus is on the development from traditional cytotoxic drugs to novel anti-tumor drugs with multi-link action on the mechanism. The invention takes histone deacetylase as a target, is based on the structural characteristics and the anti-tumor effect of SAHA, reserves partial key structural skeleton and functional group, and creatively obtains a novel HDAC inhibitor. Experiments show that the compounds not only have stronger activity on the enzyme level, but also show good inhibitory activity in various tumor cells.
The invention is mainly characterized in the following aspects:
(1) the invention provides an HDAC inhibitor with a novel structure, which comprises pharmaceutically acceptable salts, solvent compounds (such as hydrates), metabolites, isomers, prodrugs and the like, and can be used as antitumor drug lead compounds and clinical drug candidate compounds.
(2) The compound of the invention has the advantages of simple synthetic route, cheap raw materials and high reaction yield.
(3) The compounds of the invention are broad-spectrum HDAC inhibitors, have stronger activity improvement on the enzyme level compared with SAHA, and in addition, the compounds of the invention all show good activity in a plurality of experiments of in vitro anti-tumor proliferation.
(4) The compound BE018 has an effect of promoting apoptosis of non-small cell lung cancer A549 and can inhibit cloning of tumor cells.
(5) The compound BE018 can block the cell cycle of the non-small cell lung cancer A549, so that the proliferation of tumor cells is inhibited.
Drawings
FIG. 1 is a graph showing the deacetylation inhibition effect of a portion of the compounds of the present invention on histones H3 and H4 and alpha-Tubulin.
FIG. 2 is a graph showing the effect of certain compounds of the present invention on apoptosis of non-small cell type lung cancer A549. Wherein 2A is the apoptosis of non-small cell lung cancer, and 2B is the cleavage experiment picture of poly (adenosine diphosphate-ribose) polymerase (PARP).
FIG. 3 is a graph showing the effect of some compounds of the present invention on the cell cycle of non-small cell lung cancer A549. Fig. 3A is a schematic diagram of a cell cycle, and fig. 3B is a statistical calculation diagram.
FIG. 4 is a graph showing the effect of the compounds of the present invention on the clonality of human non-small cell lung cancer cell A549 at various concentrations. Fig. 4A is an effect graph, and fig. 4B is a statistical result graph.
Detailed Description
The present invention will be described in further detail with reference to the following specific examples and drawings, but the present invention is not limited to the following examples. Variations and advantages that may occur to those skilled in the art may be incorporated into the invention without departing from the spirit and scope of the inventive concept, and the scope of the appended claims is intended to be protected.
1 H-NMR was measured using a Bruker model 500MHz instrument; MS is measured by a Bruker MicroTOF-Q LCMS model instrument, and is in an ESI mode except for indication; all solvents are redistilled before use, and the used anhydrous solvents are obtained by drying treatment according to a standard method; all reactions were carried out under argon protection and followed by TLC except for the indication, and the post-treatment was carried out by washing with saturated saline and drying with anhydrous magnesium sulfate; purification of the product except for the indication silica gel (200 and 300 mesh) column chromatography was used; the silica gel used comprises 200-300 mesh and GF 254 Is produced by Qingdao oceanic chemical plants or Nicotin Bo silica gel company. Example 1-1 preparation of the Compound (7- (3- ((1-adamantanamine) formyl)) -indolyl) heptanoylhydroxylamine (BE001)
3-indolecarboxylic acid (806mg,5.0mmol) was taken and dissolved in dimethylformamide (10ml), and after cooling the reaction system in an ice-water bath, carbonyldiimidazole (970mg,6.0mmol) was added, and after reacting at room temperature for 45min, 1-amantadine (1513mg,10.0mmol) was added, and the mixture was refluxed at 120 ℃ overnight. And cooling the reaction system at room temperature, extracting with ethyl acetate and water respectively, taking the organic phase, evaporating to dryness, and passing through a silica gel column to obtain an intermediate N- (1-amantadine) -3-indolecarboxamide.
N- (1-Adamantadine) -3-indolecarboxamide (294mg,1.0mmol) was dissolved in dimethylformamide (5ml), sodium hydride (80mg,2,0mmol) was slowly added under ice bath conditions until no bubbles were generated, and after methyl 7-bromoheptanoate (275mg,1.2mmol) was slowly added, and reaction was carried out at room temperature for 30 min. Extracting with ethyl acetate and water, evaporating organic phase, and passing through silica gel column to obtain intermediate compound (7- (3- ((1-adamantine amine) formyl)) -indolyl) methyl heptanoate.
KOH (2.06g,36.7mmol) was added to a solution of hydroxylamine hydrochloride (2.55g,36.7mmol) in methanol (10ml) at 40 ℃ to react for 10min, the reaction was placed in an ice bath, the filtrate was filtered after 30min, the above ester (80mg,0.18mmol) was added to the filtrate and additional KOH (206mg,3.67mmol) was added to the system,the reaction was carried out at room temperature for 2 hours. The solvent was evaporated to dryness by distillation under the reduced pressure, and the residue was extracted with ethyl acetate and saturated ammonium chloride and then subjected to column chromatography to give BE001(34mg, 43%). 1 HNMR(500MHz,DMSO)δ10.32(s,1H),8.64(s,1H),8.11(d,J＝8.0Hz,1H),8.09(s,1H),7.50(d,J＝8.0Hz,1H),7.17(dd,J＝8.0Hz,7.5Hz,1H),7.10(dd,J＝8.0Hz,7.5Hz,1H),7.00(s,1H),4.15(t,J＝7.0Hz,2H),2.10–2.07(m,9H),1.92(t,J＝7.0Hz,2H),1.78–1.68(m,8H),1.48–1.46(m,2H),1.28–1.24(m,4H).
EXAMPLES 1-2 TO 1-9, PREPARATION OF BE SERIES COMPOUNDS TABLE 1 (see references hereinafter for details)
TABLE 1
Examples 1 to 10 preparation of the Compound (7- (3- ((2-adamantanamine) formyl)) -indolyl) heptanoylhydroxylamine (BE010)
3-indolecarboxylic acid (323mg,2.0mmol), 2-amantadine hydrochloride (376mg,2.0mmol), EDC.HCl (384mg,2.0mmol) and HOBt (271mg,2.0mmol) were taken in dimethylformamide (5 ml). Reacting for 4h to obtain an intermediate.
Intermediate (135mg,0.46mmol) was taken in dimethylformamide (5ml), sodium hydride (37mg,0.92mmol) was slowly added under ice-water bath conditions until no bubbles were produced, and methyl 7-bromoheptanoate (123mg,0.55mmol) was added. Obtaining intermediate ester after 30 min.
KOH (2.83g,50.7mmol) was added to a solution of hydroxylamine hydrochloride (3.33g,50.7mmol) in methanol (10ml) at 40 ℃, the reaction was left in an ice bath after 10min of reaction, the filtrate was filtered after 30min, the above ester (118mg,0.27mmol) was added to the filtrate and KOH (333mg,5.0 mmol) was added to the system7mmol) were reacted at room temperature for 2 hours and then extracted through the column to give BE010(65mg, 55%). 1 H NMR(500MHz,DMSO):δ10.32(s,1H),8.64(s,1H),8.27(s,1H),8.11(d,J＝8.0Hz,1H),7.52(d,J＝8.5Hz,1H),7.28(d,J＝7.0Hz,1H),7.19(dd,J＝7.0Hz,7.5Hz,1H),7.12(dd,J＝7.5Hz,7.5Hz,1H),4.18(t,J＝7.0Hz,2H),4.11-4.07(m,1H),2.14-2.12.(m,2H),1.97-1.74(m,14H),1.57-1.54(m,2H),1.50-1.42(m,2H),1.35-1.25(m,4H).
EXAMPLES 1-10 TO 1-24, PREPARATION OF BE SERIES COMPOUNDS TABLE 2 (see references hereinafter for details)
TABLE 2
Examples 1-25 preparation of (7- (3- ((1-adamantanamine) formyl)) -indolyl) heptacetylmercaptoester (BE025)
N- (1-adamantane) -3-indolecarboxamide (322mg,1.10mmol) was taken in dimethylformamide (5ml), sodium hydride (88mg,2.20mmol) was slowly added under ice-water bath conditions until no bubbles were produced, and 7-bromo-1-heptene (244mg,1.30mmol) was added. After 30min reaction, intermediate (7- (3- ((1-adamantanamine) formyl)) -indolyl) heptene (187mg, 44%) was obtained.
This intermediate (187mg,0.48mmol) was placed in 1, 4-dioxane (10ml), AcSH (365mg,4,80mmol) and AIBN (79mg,0.48mmol) were added and reacted at 75 ℃ for 1 h. The reaction system was cooled under ice bath conditions, excess cyclohexene was added, the reaction solution was evaporated to dryness and column chromatography was performed to obtain the product (7- (3- ((1-amantadine) formyl)) -indolyl) heptacetylmercaptoester (140mg, 63%). 1 H NMR(500MHz,CDCl 3 )δ7.89(d,J＝7.55Hz,1H),7.66(s,1H),7.37(d,J＝7.5Hz,1H),7.27-7.23(m,2H),5.71(s,1H),4.12(t,J＝7.0Hz,2H),2.85(t,J＝7.0Hz,2H),2.33(s,3H),2.25-7.20(m,6H),2.20-2.13(m,3H),1.90-1.82(m,2H),1.80-1.72(m,6H),1.58-1.50(m,2H),1.40-1.30(m,6H).
EXAMPLES 1-25 TO 1-30, PREPARATION OF BE SERIES COMPOUNDS TABLE 3 (see references hereinafter for details)
TABLE 3
Example 2 inhibition of HDAC1 and HDAC6 enzymatic Activity by Compounds of the invention
MDA-MB231 cells are inoculated in a 6 cm cell culture dish, when the cells grow to 80% density, the compound of the invention and SAHA are respectively added as a control group, after the cells are treated for 24 hours, the culture medium is removed, precooled PBS is added, then PBS is sucked up, cell lysate is added, the cells are scraped to a centrifuge tube with the volume of 1.5 ml, the centrifuge tube is placed on ice, the centrifuge tube is vortexed once every five minutes for 5 times, after the cells are vortexed, the centrifuge tube is centrifugated for 15 minutes at 12000 ℃ at 4 ℃, supernatant is sucked up to a new precooled centrifuge tube, the BCA method is used for measuring the protein concentration, loading buffer solution containing 40 micrograms of protein is used for carrying out gel running on 8% -12% SDS protein separation gel, after bromophenol blue runs out of the gel plate, the gel plate is rotated for 1 hour at 100 volts, primary antibody corresponding to HDAC1 and HDAC6 is incubated on a shaker at 4 ℃ overnight, TBST is used for washing for three times each 10 minutes, then secondary antibody corresponding to 1 hour, strips of each mesh were exposed using the ECL kit, washed three times in TBST for 10 minutes each time.
As shown in table 4, most compounds had inhibitory effects on HDAC1 and HDAC6 enzyme activities at a certain concentration, which was comparable to or better than the inhibitory effect of SAHA, a positive control. Among them, compound BE023 has a certain selectivity to HDAC6 subtype.
TABLE 4
Example 3 preliminary screening for the cellular Activity of Compounds of the invention
The partial compound with better primary screening activity and the positive compound (SAHA) were tested for cytotoxic activity on different cell types, 3000-10000 cells per 100. mu.L of medium, and incubated overnight in a 96-well plate (Falcon, CA). Dividing the cells to be detected into 10 parts, treating the 10 parts by using medicines with gradient concentration, culturing at least 3 multiple wells with each concentration at 37 ℃ for 72h, and detecting by adopting an MTS method, namely adding 20 mu L of CellTiter 96Aqueous One Solution into each well for incubation for 0.5-3h, and measuring the OD value at 490nm of an enzyme-labeling instrument. The 50% drug concentration at which cell growth inhibition is to be detected is IC 50 IC of drug was calculated using GraphPadprism 5.0 software 50 。
As a result, as shown in Table 5, Compound BE018 showed strong activities in A549 cells (non-small cell lung cancer), MDA-MB-231 cells (breast cancer), SGC7901 (gastric cancer), HCT116 (colon cancer) and HL60 cells (promyelocytic leukemia). Compared with the positive medicine SAHA, the activity of the compound is obviously improved.
TABLE 5
a A549 cell (non-small cell lung cancer)
b MDA-MB-231 cells (Breast cancer)
c SGC7901 (stomach cancer)
d HCT116 (Colon cancer)
e HL60 cell (promyelocytic leukemia)
Example 4: the compounds of the invention promote the deacetylation of histones H3 and H4 and alpha-Tubulin
The test method is the same as example 2, A549 cells are inoculated in a 6 cm cell culture dish, when the cells grow to 80% density, the compounds BE018, SAHA and DMSO of the invention are respectively added as control groups, after the cells are treated for 24 hours, the culture medium is removed, precooled PBS is added, then PBS is absorbed, cell lysate is added, the cells are scraped to a centrifuge tube with the volume of 1.5 ml, the centrifuge tube is placed on ice, the centrifuge tube is vortexed once every five minutes, after 5 times of vortexing, centrifugation is carried out for 15 minutes at a 4 ℃ centrifuge 12000, supernatant is absorbed to a new precooled centrifuge tube, the protein concentration is measured by a BCA method, loading buffer solution containing 40 micrograms of protein is run on 8% -12% SDS protein separation gel, after bromophenol blue runs out of the gel plate, the centrifugation is stopped at 100V for 1 hour, H3 and H4 acetylated antibody is incubated at the temperature of 4 ℃ shaking table for overnight, and washed three times at 10 minutes by TBST, the corresponding secondary antibodies were then incubated for 1 hour, washed three times with TBST for 10 minutes each, and exposed to light using the ECL kit to reveal bands of each.
As a result, as shown in fig. 1, the compound BE018 according to the present invention can effectively promote acetylation of intracellular histones in cancer cells, and the level of acetylation promotion is gradient-dependent, since α -tubulin is a specific substrate of HDAC6, and BE018 not only contributes to acetylation of H3 and H4, but also acts on α -tubulin, indicating that the compound BE018 according to the present invention has an inhibitory effect on both HDAC1 and HDAC 6.
Example 5: effect of some Compounds of the invention on apoptosis of non-Small cell Lung cancer A549
DMSO, SAHA (3. mu.M, 8. mu.M) and the present compound BE018 (0.3. mu.M, 1. mu.M, 3. mu.M and 10. mu.M) were added to the non-small cell lung cancer cell A549, respectively. Cells were harvested after 48h and analyzed using the Annexin V-FITC Apoptosis Detection Kit (eBioscience/Bender MedSystems GmbH, Vienna, Austria). Annexin V-FITC and propidium iodide are used for staining, a flow cytometer is used for detecting the apoptosis condition, and a western blot method is used for detecting the PARP lysis in A549 cells without judging the apoptosis condition.
The results are shown in fig. 2A, with the increase of the concentration of the compound BE018 in a549 cells, the pro-apoptosis effect is gradually enhanced, and the effect is obviously better than that of SAHA. In addition, the results of the polyadenylation-ribose polymerase (PARP) cleavage assay (FIG. 2B) show that PARP lysates were detected in A549 cells when BE018 concentrations reached 3. mu.M and 10. mu.M, further demonstrating the pro-apoptotic effect of the compounds on cancer cells.
Example 6: effect of some Compounds of the invention on the cell cycle of non-Small cell Lung cancer A549
A549 cells were treated with DMSO, SAHA (8 μ M) or BE018(0.3 μ M,1 μ M,3 μ M) for 24 hours, stained with Propidium Iodide (PI), and analyzed by cell cycle detection using a BD flow cytometer in the United states.
The results are shown in FIG. 3. From the figure, we can find that when SAHA (8 μ M) acts on A549 cells, cell cycle block is generated, G2/M phase is prolonged, and S phase is shortened. From the experimental results, it is also clear that compound BE018 exhibited a concentration gradient dependence on cell cycle arrest and was more effective than SAHA. Therefore, BE018 has been experimentally confirmed to inhibit the proliferation of tumor cells by arresting the cell cycle.
Example 7: influence of the inventive Compounds on the clonogenic potency of human non-Small cell type Lung cancer cell A559 at different concentrations
Solid tumors are finally formed by tumor cells dividing and proliferating themselves. Therefore, the compound of the present invention can be tested for inhibition of the clonogenic capacity of tumor cells in an in vitro experimental model to evaluate the anticancer effect of the compound.
Non-small cell lung cancer cell A549 at 1x10 3 The density of each dish is inoculated in a 35mm culture dish, the mixture is divided into groups after being cultured for 24h, the compounds to be tested with different concentrations are respectively added to make the final concentrations respectively be0 mu mol/L, 0.1 mu mol/L, 0.3 mu mol/L and 1.0 mu mol/L, and the same amount of DMSO is added to a control group. The culture was continued, and the medium and the compound at the corresponding concentration were changed every 3 days. After 7 days, the medium was discarded, washed 3 times with PBS, fixed with 4% paraformaldehyde at room temperature for 10min, stained with 1% crystal violet for 10min, and rinsed with tap water. The number of cell clones formed was counted by taking pictures under a microscope.
The results of the experiment are shown in FIG. 4. The compound BE018 can effectively inhibit the formation of A549 cell populations, and the inhibition effect is concentration gradient dependent. The effect of inhibiting cell colony formation was quite significant when BE018 was used at a concentration of only 0.1. mu.M, compared to the control group treated with DMSO only and the control group treated with 0.3. mu.M of SAHA. When the used concentration of BE018 was increased to 1. mu.M, the clonogenic activity of A549 cells was almost completely inhibited.
Example 1-2 preparation of Compound (6- (3- ((1-Adamantadine) formyl)) -indolyl) hexanoylhydroxylamine (BE002)
The compound (BE001) was prepared by substituting methyl 7-bromoheptanoate with methyl 6-bromohexanoate. 1 H NMR(500MHz,DMSO):δ10.35(s,1H),8.67(s,1H),8.11(d,J＝8.0Hz,1H),8.08(s,1H),7.51(d,J＝8.0Hz,1H),7.17(dd,J＝7.0Hz,7.0Hz,1H),7.10(dd,J＝7.5Hz,7.0Hz,1H),7.00(s,1H),4.17(t,J＝7.0Hz,2H),2.14-204(m,9H),2.00-1.95(m,2H),1.80-1.72(m,4H),1.71-1.55(m,6H),1.51-1.46(m,2H).
Examples 1-3 preparation of the Compound (7- (2- ((1-adamantanamine) formyl)) -indolyl) heptanoylhydroxylamine (BE003)
The 3-indolecarboxylic acid was replaced by 2-indolecarboxylic acid and prepared accordingly by the method for preparing the compound BE 001. 1 H NMR(500MHz,DMSO):δ10.23(s,1H),8.53(s,1H),7.62(s,1H),7.59(d,J＝8.0Hz,1H),7.49(d,J＝8.0Hz,1H),7.22(dd,J＝7.5Hz,8Hz,1H),7.06(dd,J＝7.5Hz,7.5Hz,1H),6.94(s,1H),4.46(t,J＝7.0Hz,2H),2.15–1.99(m,9H),1.91(t,J＝7.0Hz,2H),1.74–1.58(m,8H),1.47–1.40(m,2H),1.25–1.19(m,4H).
Examples 1-4 preparation of the Compound (7- (3- ((1-Adamantadine) formyl) -5-fluoro) -indolyl) heptanoylhydroxylamine (BE004)
The 3-indolecarboxylic acid is replaced with 5-fluoro-2-indolecarboxylic acid, and the compound BE001 is prepared accordingly. 1 H NMR(500MHz,DMSO):δ10.23(s,1H),8.54(s,1H),7.70(s,1H),7.54(d,J＝9.0Hz,1H),7.36(d,J＝9.5Hz,1H),7.07(d,J＝9.0Hz,1H),6.92(s,1H),4.46(t,J＝7.0Hz,2H),2.21-2.01(m,9H),1.91(t,J＝7.0Hz,2H),1.75-1.54(m,8H),1.49-1.39(m,2H),1.26-1.17(m,4H).
Examples 1 to 5 preparation of the Compound (7- (3- ((1-adamantanamine) formyl)) -7-azaindolyl) heptanoylhydroxylamine (BE005)
The 3-indolecarboxylic acid was replaced by 7-aza-3-indolecarboxylic acid and prepared accordingly by the method for preparing the compound BE 001. 1 H NMR(500MHz,DMSO):δ10.32(s,1H),8.64(s,1H),8.41(d,J＝9.0Hz,1H),8.30-8.27(m,2H),7.19-7.16(m,2H),4.25(t,J＝7.0Hz,2H),2.10-2.07(m,9H),1.92(t,J＝7.5Hz,2H),1.85-1.80(m,2H),1.70-1.65(s,6H),1.49-1.44(m,2H),1.30-1.21(m,4H).
Examples 1-6 preparation of Compound (5- (3- ((1-adamantanamine) formyl)) -indolyl) pentanoylhydroxylamine (BE006)
The compound BE001 was prepared by substituting methyl 7-bromoheptanoate with methyl 5-bromovalerate. 1 H NMR(500MHz,DMSO):δ10.35(s,1H),8.67(s,1H),8.11(d,J＝8.0Hz,1H),8.08(s,1H),7.51(d,J＝8.0Hz,1H),7.17(dd,J＝7.0Hz,7.0Hz,1H),7.10(dd,J＝7.5Hz,7.0Hz,1H),7.00(s,1H),4.17(t,J＝7.0Hz,2H),2.14-204(m,9H),2.00-1.95(m,2H),1.80-1.72(m,2H),1.71-1,55(m,6H),1.51-1.46(m,2H).
Examples 1 to 7 preparation of Compound (7- (3- ((1-adamantanamine) formyl) -5-chloro) -indolyl) heptanoylhydroxylamine (BE007)
The compound BE001 can BE produced by substituting 3-indolecarboxylic acid with 5-chloro-3-indolecarboxylic acid. 1 H NMR(500MHz,DMSO):δ10.32(s,1H),8.64(s,1H),8.19(s,1H),8.13(s,1H),7.56(d,J＝8.5Hz,1H),7.19(d,J＝8.5Hz,1H),7.12(s,1H),4.16(t,J＝7.0Hz,2H),2.15-2.00(m,9H),1.92(t,J＝7.5Hz,2H),1.82-1.72(m,2H),1.72-1.60(m,6H),1.50-1.40(s,2H),1.31-1.20(m,4H).
Examples 1 to 8 preparation of the Compound (7- (3- ((1-adamantanamine) formyl) -5-methoxy) -indolyl) heptanoylhydroxylamine (BE008)
The 3-indolecarboxylic acid is substituted with 5-methoxy-3-indolecarboxylic acid, and the compound BE001 is produced according to the method. 1 H NMR(500MHz,DMSO):δ10.32(s,1H),8.64(s,1H),8.06(s,1H),7.65(s,1H),7.39(d,J＝9.0Hz,1H),6.96(s,1H),6.80(d,J＝9.0Hz,1H),4.10(t,J＝6.0Hz,2H),3.77(s,3H),2.15-2.05(m,9H),1.92(t,J＝6.5Hz,2H),1.80-1.73(m,2H),1.70-1.50(m,6H),1.50-1.42(m,2H),1.30-1.20(m,4H).
Examples 1 to 9 preparation of Compound (7- (2- ((1-adamantanamine) formyl)) -3-azaindolyl) heptanoylhydroxylamine (BE009)
The 3-indolecarboxylic acid is substituted with 3-aza-2-indolecarboxylic acid, and the compound BE001 is produced according to the method for producing the compound. 1 H NMR(500MHz,DMSO):δ10.23(s,1H),8.53(s,1H),7.59(d,J＝8.0Hz,1H),7.49(d,J＝8.0Hz,1H),7.22(dd,J＝7.5Hz,8Hz,1H),7.06(dd,J＝7.5Hz,7.5Hz,1H),6.94(s,1H),4.46(t,J＝7.0Hz,2H),2.15-1.99(m,9H),1.91(t,J＝7.0Hz,2H),1.74-1.58(m,8H),1.47-1.40(m,2H),1.25-1.19(m,4H).
Examples 1 to 10 preparation of the Compound (7- (3- ((2-adamantanamine) formyl)) -indolyl) heptanoylhydroxylamine (BE010)
3-indolecarboxylic acid (323mg,2.0mmol), 2-amantadine hydrochloride (376mg,2.0mmol), EDC & HCl (384mg,2.0mmol), HOBt & HCl (271mg,2.0mmol) was taken in dimethylformamide (5 ml). The reaction was stirred at room temperature for 4h, extracted with ethyl acetate and water, the organic phase evaporated to dryness and chromatographed to give intermediate N- (2-adamantane) -3-indolecarboxamide (302mg, 51%).
N- (2-adamantane) -3-indolecarboxamide (135mg,0.46mmol) was taken in dimethylformamide (5ml), and sodium hydride (37mg,0.92mmol) was slowly added under ice-water bath conditions until no bubbles were produced, and methyl 7-bromoheptanoate (123mg,0.55mmol) was added. After 30min, extraction was performed with water and ethyl acetate, the organic phase was evaporated to dryness and then subjected to column chromatography to give intermediate (7- (3- ((2-amantadine) formyl)) -indolyl) heptanoylmethyl ester (118mg, 55%).
KOH (2.83g,50.7mmol) was added to a solution of hydroxylamine hydrochloride (3.33g,50.7mmol) in methanol (10ml) at 40 ℃ and after 10min of reaction the reaction was placed in an ice bath and after 30min suction filtered to give a filtrate, and the above ester (118mg,0.27mmol) was added to the filtrate and KOH (333mg,5.07mmol) was added to the system and allowed to react for 2 hours at room temperature. The solvent was evaporated to dryness by distillation under the reduced pressure, extracted with ethyl acetate and saturated ammonium chloride, and then passed through a column to give product BE010(65mg, 55%). 1 H NMR(500MHz,DMSO):δ10.32(s,1H),8.64(s,1H),8.27(s,1H),8.11(d,J＝8.0Hz,1H),7.52(d,J＝8.5Hz,1H),7.28(d,J＝7.0Hz,1H),7.19(dd,J＝7.0Hz,7.5Hz,1H),7.12(dd,J＝7.5Hz,7.5Hz,1H),4.18(t,J＝7.0Hz,2H),4.11-4.07(m,1H),2.14-2.12.(m,2H),1.97-1.74(m,14H),1.57-1.54(m,2H),1.50-1.42(m,2H),1.35-1.25(m,4H).
Examples 1-11 preparation of the Compound (7- (2- ((2-adamantanamine) formyl)) -indolyl) heptanoylhydroxylamine (BE011)
The 3-indolecarboxylic acid was substituted with 2-indolecarboxylic acid, and the compound BE010 was prepared. 1 H NMR(500MHz,DMSO):δ10.30(s,1H),8.64(s,1H),8.25(s,1H),8.10(s,1H),7.49(s,1H),7.26(s,1H),7.20-7.05(m,2H),4.26-4.05(m,3H),2.20-2.05(m,2H),1.99-1.70(m,14H),1.38-1.17(m,8H).
Examples 1 to 12 preparation of the Compound (7- (3- (cyclooctylcarbonyl)) -indolyl) heptanoylhydroxylamine (BE012)
The 2-adamantanamine hydrochloride was substituted for cyclooctylamine, and the preparation was carried out according to the method for preparing compound BE 010. 1 H NMR(500MHz,DMSO):δ10.26(s,1H),8.55(s,1H),8.14(d,J＝8.0Hz,1H),8.06(s,1H),7.52(d,J＝8.5Hz,1H),7.50(s,1H),7.18(did,J＝7.0Hz,8.0Hz,1H),7.11(dd,J＝7.5Hz,7.0Hz,1H),4.17(t,J＝7.0Hz,2H),4.07-4.02(m,1H),1.94(t,J＝7.5Hz,2H),1.78-1.72(m,4H),1.70-1.60(m,4H),1.61-1.45(m,8H),1.31-1.25(m,6H).K
Examples 1 to 13, preparation of Compound (7- (3- ((2-adamantylamino) formyl)) -indazolyl) heptanoylhydroxylamine (BE013)
The compound BE010 can BE produced by substituting 3-indazolecarboxylic acid with 3-indolecarboxylic acid. 1 HNMR(500MHz,DMSO)δ10.32(s,1H),8.66(s,1H),8.14(d,J＝8.2Hz,1H),7.78(d,J＝8.6Hz,1H),7.56(d,J＝7.6Hz,1H),7.46(dd,J＝7.4Hz,8.2Hz,1H),7.28(dd,J＝7.5Hz,8.6Hz,1H),4.48(t,J＝7.0Hz,2H),2.04–1.73(m,16H),1.69–1.56(m,2H),1.52–1.39(m,2H),1.37–1.17(m,5H).
Examples 1 to 14, preparation of Compound (7- (2- ((2-adamantylamino) formyl)) -3-azaindolyl) heptanoylhydroxylamine (BE014)
The 3-indole carboxylic acid was substituted with 3-aza-2-indole carboxylic acid, and the compound BE010 was prepared. 1 HNMR(400MHz,DMSO)δ10.32(s,1H),8.65(s,1H),8.32(s,1H),7.89–7.57(m,2H),7.49–7.21(m,2H),4.74–4.53(m,2H),4.18–3.99(m,1H),2.09–1.37(m,24H).
Examples 1 to 15 preparation of the Compound (7- (3- ((4-chlorobenzyl) formyl)) -indolyl) heptanoylhydroxylamine (BE015)
The 2-adamantanamine hydrochloride was substituted for p-chlorobenzylamine, and prepared according to the method for preparing compound BE 010. 1 H NMR(500MHz,DMSO):δ10.26(s,1H),8.55(s,1H),8.38(t,J＝6.0Hz,1H),8.16(d,J＝8.0Hz,1H),8.07(s,1H),7.53(d,J＝8.0Hz,1H),7.37(m,4H),7.21(dd,J＝7.5Hz,8.0Hz,1H),7.14(dd,J＝7.5Hz,7.0Hz,1H),4.47(d,J＝6.0Hz,2H),4.19(t,J＝7.0Hz,2H),1.93(t,J＝7.0Hz,2H),1.85-1.74(m,2H),1.53-1.43(m,2H),1.35-1.25(m,4H).
Examples 1-16 preparation of the Compound (7- (2- ((2-adamantanamine) formyl) -6-bromo) -indolyl) heptanoylhydroxylamine (BE016)
The 3-indolecarboxylic acid was substituted with 6-bromo-2-indolecarboxylic acid, and the compound BE010 was prepared. 1 H NMR(500MHz,DMSO):δ10.30(s,1H),8.62(s,1H),8.15(d,J＝7.0Hz,1H),7.82(s,1H),7.59(d,J＝8.5Hz,1H),7.21(d,J＝8.5Hz,1H),7.09(s,1H),4.46(t,J＝7.0Hz,2H),4.05-4.03(m,1H),2.13-2.11(m,2H),2.01-1.98(m,2H),1.92-1.79(m,7H),1.74-1.71(m,2H),1.66-1.37(m,6H),1.29-1.11(m,5H).
Examples 1-17 preparation of the Compound (7- (2- ((2-adamantanamine) formyl) -5-chloro) -indolyl) heptanoylhydroxylamine (BE017)
The 3-indolecarboxylic acid is substituted with 5-chloro-2-indolecarboxylic acid, and the compound BE010 is produced by the method. 1 H NMR(500MHz,DMSO):δ10.30(s,1H),8.63(s,1H),8.19(d,J＝7.0Hz,1H),7.70(s,1H),7.58(d,J＝8.5Hz,1H),7.24(d,J＝8.5Hz,1H),7.06(s,1H),4.47(t,J＝7.0Hz,2H),4.07-4.00(m,1H),2.14-2.09(m,2H),2.05-1.98(m,2H),1.90-1.80(m,7H),1.73-1.70(m,2H),1.62-1.58(m,2H),1.54-1.52(m,2H),1.45-1.38(m,2H),1.25-1.16(m,5H).
Examples 1-18 preparation of the Compound (7- (2- ((2-adamantanamine) formyl) -5-methoxy) -indolyl) heptanoylhydroxylamine (BE018)
The preparation of the compound BE010 is carried out by replacing 3-indolecarboxylic acid with 5-methoxy-2-indolecarboxylic acid. 1 H NMR(500MHz,DMSO):δ10.30(s,1H),8.63(s,1H),7.99(s,1H),7.45(s,1H),7.14-6.82(m,3H),4.51-4.35(m,2H),4.10-3.98(m,1H),3.77(s,3H),2.14-1.23(m,24H).
Examples 1 to 19 preparation of the compound (7- (2- ((4-fluorophenyl) formyl)) -indolyl) heptanoylhydroxylamine (BE019)
The 2-adamantanamine hydrochloride was substituted for p-chloroaniline, and prepared according to the method for preparing the compound BE 010. 1 HNMR(500MHz,DMSO):δ10.40(s,1H),10.30(s,1H),8.63(s,1H),7.81-7.78(m,2H),7.70(d,J＝8.0Hz,1H),7.59(d,J＝8.5Hz,1H),7.31(d,J＝8.0Hz,1H),7.29(s,1H),7.21(m,2H),7.13(dd,J＝8.0Hz,7.0Hz,1H),4.55(t,J＝7.0Hz,2H),1.90(t,J＝7.5Hz,2H),1.74-1.65(m,2H),1.47-1.39(m,2H),1.31-1.17(m,4H).
Examples 1-20 preparation of the Compound (7- (3- (cyclohexanecarboxoyl)) -indolyl) heptanoylhydroxylamine (BE020)
By replacing 2-adamantanamine hydrochloride with cyclohexylamine, according to a process for preparing the compound BE010And (4) preparation. 1 HNMR(500MHz,DMSO):δ10.32(s,1H),8.64(s,1H),8.14(d,J＝8.0Hz,1H),8.06(s,1H),7.59(d,J＝8.0Hz,1H),7.51(d,J＝8.5Hz,1H),7.18(dd,J＝7.5Hz,7.5Hz 1H),7.11(dd,J＝8.0Hz,7.0Hz 1H),4.17(t,J＝7.0Hz,2H),3.80-3.74(m,1H),1.92(t,J＝7.5Hz,2H),1.86-1.84(m,2H),1.79-1.74(m,4H),1.68-1.59(m,2H),1.50-1.43(m,2H),1.33-1.24(m,8H).
Examples 1 to 21 preparation of Compound (5- (3- ((2-adamantylamino) formyl)) -indolyl) pentanoylhydroxylamine (BE021)
The compound was prepared by substituting methyl 7-bromoheptanoate with methyl 5-bromovalerate according to the method for preparing the compound BE 010. 1 HNMR(500MHz,DMSO):δ10.35(s,1H),8.67(s,1H),8.26(s,1H),8.10(d,J＝7.5Hz,1H),7.53(d,J＝8.0Hz,1H),7.29(d,J＝7.0Hz,1H),7.19(dd,J＝7.0Hz,7.0Hz,1H),7.13(dd,J＝7.5Hz,7.0Hz,1H),4.20(t,J＝7.0Hz,2H),4.09-4.01(m,1H),2.14-2.12(m,2H),2.00-1.97(m,5H),1.89-1.73(m,9H),1.58-1.47(m,4H).
Examples 1-22 preparation of Compound (7- (3- (cycloheptylcarbonyl)) -indolyl) heptanoylhydroxylamine (BE022)
The 2-adamantanamine hydrochloride was replaced with cycloheptylamine, and the compound BE010 was prepared. 1 HNMR(500MHz,DMSO)δ10.32(s,1H),8.64(s,1H),8.14(d,J＝8.0Hz,1H),8.07(s,1H),7.62(d,J＝8.0Hz,1H),7.51(d,J＝8.0Hz,1H),7.18(dd,J＝7.5Hz,7.5Hz,1H),7.11(dd,J＝7.5Hz,7.0Hz,1H),4.17(t,J＝7.0Hz,2H),4.02-3.93(m,1H),1.97-1.83(m,4H),1.82-1.74(m,2H),1.70-1.42(m,11H),1.30-1.24(m,5H).
Examples 1-23 preparation of the Compound N- (2-adamantane) -1- (4-Hydroxycarbamoylbenzyl) indole carboxamide (BE023)
The compound BE010 can BE produced by substituting methyl 7-bromoheptanoate with methyl 4-bromomethylbenzoate. 1 HNMR(500MHz,DMSO)δ11.15(s,1H),9.01(s,1H),8.39(s,1H),8.12(d,J＝8.5Hz,1H),7.70(d,J＝8.0Hz,2H),7.47(d,J＝8.0Hz,1H),7.38(d,J＝7.0Hz,1H),7.30(d,J＝8.0Hz,2H),7.18-7.10(m,2H),5.52(s,2H),4.13-4.05(m,1H),2.17-2.10(m,2H),2.01-1.96(m,2H),1.90-1.77(m,6H),1.77-1.71(m,2H),1.60-1.53(m,2H).
Examples 1 to 24, preparation of compound (7- (3- ((2, 4-dichlorophenyl) formyl)) -indolyl) heptanoylhydroxylamine (BE024)
The 2-adamantanamine hydrochloride was replaced with 2, 4-dichloroaniline and prepared according to the method for preparing compound BE 010. 1 HNMR(500MHz,DMSO)δ10.32(s,1H),9.42(s,1H),8.64(s,1H),8.35(s,1H),8.16(d,J＝7.5Hz,1H),7.80(d,J＝8.5Hz,1H),7.71(s,1H),7.60(d,J＝8.0Hz,1H),7.46(d,J＝8.0Hz,1H),7.27-7.18(m,2H),4.25(m,2H),1.93(t,J＝6.5Hz,2H),1.84-1.78(m,2H),1.50-1.48(m,2H),1.30-1.24(m,4H).
Examples 1 to 25 preparation of the Compound (7- (3- ((1-adamantanamine) formyl)) -indolyl) heptacetylmercaptoester (BE025)
N- (1-adamantane) -3-indolecarboxamide (322mg,1.10mmol) was taken in dimethylformamide (5ml), sodium hydride (88mg,2.20mmol) was slowly added under ice-water bath conditions until no bubbles were produced, and 7-bromo-1-heptene (244mg,1.30mmol) was added. After 30min, extraction was performed with water and ethyl acetate, the organic phase was evaporated to dryness and then subjected to column chromatography to give intermediate (7- (3- ((1-adamantanamine) formyl)) -indolyl) heptene (187mg, 44%).
Taking (7- (3- ((1-amantadine) formyl)) -indolyl) heptene (187mg,0.48mmol) inTo 1, 4-dioxane (10ml) were added AcSH (365mg,4,80mmol) and AIBN (79mg,0.48mmol) and reacted at 75 ℃ for 1 h. Cooling the reaction system under the ice bath condition, adding excessive cyclohexene, extracting with water and ethyl acetate, taking an organic phase, evaporating to dryness, and performing column chromatography to obtain a product (7- (3- ((1-amantadine) formyl)) -indolyl) heptaacetyl mercapto ester (140mg, 63%). 1 H NMR(500MHz,CDCl3)δ7.89(d,J＝7.55Hz,1H),7.66(s,1H),7.37(d,J＝7.5Hz,1H),7.27-7.23(m,2H),5.71(s,1H),4.12(t,J＝7.0Hz,2H),2.85(t,J＝7.0Hz,2H),2.33(s,3H),2.25-7.20(m,6H),2.20-2.13(m,3H),1.90-1.82(m,2H),1.80-1.72(m,6H),1.58-1.50(m,2H),1.40-1.30(m,6H).
Examples 1 to 26 preparation of Compound (6- (3- ((2-adamantanamine) formyl)) -indolyl) Hexyacetyl mercapto ester (BE026)
The 7-bromo-1-heptene is substituted with 6-bromo-1-hexene, by the method for preparing the compound BE 025. 1 H NMR(500MHz,CDCl3)δ7.89(d,J＝5.8Hz,1H),7.74(s,1H),7.39(d,J＝6.0Hz,1H),7.28-7.30(m,2H),6.34(s,1H),4.37-4.35(m,1H),4.14(t,J＝7.0Hz,2H),2.84(t,J＝7.0Hz,2H),2.32(s,3H),2.12-2.09(m,2H),1.95-1.73(m,14H),1.58-1.50(m,2H),1.41-1.33(m,4H).
Examples 1 to 27 preparation of Compound (7- (3- ((2-adamantylamino) formyl)) -indolyl) heptacetylmercapto ester (BE027)
The compound BE025 was prepared by substituting 1-adamantanamine with 2-adamantanamine. 1 H NMR(500MHz,DMSO)δ8.27(s,1H),8.10(d,J＝8.0Hz,1H),7.51(d,J＝8.0Hz,1H),7.28(d,J＝7.0Hz,1H),7.18(dd,J＝7.5Hz,7.0Hz,1H),7.11(dd,J＝7.5Hz,7.5Hz,1H),4.17(t,J＝7.0Hz,2H),4.12-4.05(s,1H),2.79(t,J＝7.0Hz,2H),2.30(s,3H),2.15-2.10(m,2H),2.00-1.94(m,2H),1.90-1.75(m,7H),1.75-1.70(m,2H),1.60-1.50(s,2H),1.49-1.43(m,2H),1.35-1.15(m,7H).
Examples 1-28 preparation of the Compound (6- (3- ((1-adamantanamine) formyl)) -indolyl) hexyl acetyl mercapto ester (BE028)
The 7-bromo-1-heptene is substituted with 6-bromo-1-hexene, by the method for preparing the compound BE 025. 1 H NMR(500MHz,DMSO)δ8.13-8.08(m,2H),7.50(d,J＝8.0Hz,1H),7.17(dd,J＝7.5Hz,7.5Hz,1H),7.10(dd,J＝7.5Hz,7.0Hz,1H),7.00(s,1H),4.15(t,J＝7.0Hz,2H),2.81(t,J＝7.0Hz,2H),2.31(s,3H),1.92-1.73(m,2H),1.72-1.65(m,6H),1.52-1.45(m,2H),1.38-1.32(m,2H),1.30-1.22(m,2H).
Examples 1-29 preparation of Compound (6- (3- ((2-adamantanamine) formyl)) -indolyl) Hexyacetyl mercapto alcohol (BE029)
(6- (3- ((2-amantadine) formyl)) -indolyl) hexanethiol mercaptoester (428mg, 0.95mmol) was taken in EtOH (10ml), sodium hydroxide (380mg, 9.50mmol) was added, and after reacting overnight, the product was evaporated to dryness and passed through a column to give the product (268mg, 66%). 1 H NMR(500MHz,CDCl3)δ7.91(d,J＝8.0Hz,1H),7.77(s,1H),7.42(d,J＝6.0Hz,1H),7.32-7.29(m,2H),6.36(s,1H),4.37(d,J＝8.0Hz,1H),4.17(t,J＝7.0Hz,2H),2.52(q,J＝7.5Hz,2H),2.16-2.15(m,2H),2.00-1.77(m,14H),1.64-1.60(m,2H),1.48-1.42(m,2H),1.36-1.32(m,2H).
Examples 1-30 preparation of the Compound (7- (3- ((2-adamantanamine) formyl)) -indolyl) heptacetylmercaptol (BE030)
The 6-bromo-1-hexene is substituted with 7-bromo-1-heptene, by the method for producing the compound BE 029. 1 HNMR(500MHz,DMSO)δ8.28(s,1H),8.11(d,J＝8.0Hz,1H),7.53(d,J＝8.0Hz,1H),7.30(d,J＝7.0Hz,1H),7.19(dd,J＝7.5Hz,7.5Hz,1H),7.12(dd,J＝7.5Hz,7.5Hz,1H),4.19(t,J＝7.0Hz,2H),4.10-4.07(m,1H),2.46-2.41(m,2H),2.14-2.11(m,2H),1.86-1.83(m,2H),1.80-1.73(m,8H),1.79-1.73(m,2H),1.57-1.48(m,4H),1.34-1.30(m,6H).
The present invention is not limited to the above embodiments. Variations and advantages that may occur to those skilled in the art may be incorporated into the invention without departing from the spirit and scope of the inventive concept, and the scope of the appended claims is intended to be protected.
Claims (15)
1. An amide micromolecule organic compound or pharmaceutically acceptable salt with indole or indole analogue as a parent nucleus structure is characterized by being represented by the following structural formula (I):
wherein X is N or C;
a is selected from-CONR-; wherein R is hydrogen;
m, n = 0 and p = 5-8;
u is CR 1 ；
V is CR 2 ；
W is CR 3 ；
Z is CR 4 Or N;
wherein X and Z are not N at the same time;
wherein R is 1 ，R 3 ，R 4 Independently selected from hydrogen;
R 2 selected from any one of the following groups: including hydrogen, fluoro, chloro, methoxy;
R 5 selected from any one of the following groups: 1-adamantyl; 2-adamantyl; c 6-8 Cycloalkyl groups of (a); chloro-substituted phenyl;
wherein q = 0-3;
ZBG is a zinc chelating group selected from
。
2. An amide micromolecule organic compound or pharmaceutically acceptable salt with indole or indole analogue as a parent nucleus structure is characterized by being represented by the following structural formula (II):
wherein Y is N or C;
a is selected from-CONR-; wherein R is hydrogen; m, n = 0 and p = 5-8;
u is CR 1 ；
V is CR 2 ；
W is CR 3 ；
Z is CR 4 ；
Wherein R is 1 ，R 4 Independently selected from hydrogen;
R 2 any one selected from the group consisting of hydrogen, chlorine, methoxy;
R 3 any one selected from the group consisting of hydrogen, bromine;
R 5 selected from any one of the following groups: 1-adamantyl; 2-adamantyl; fluoro-substituted phenyl;
wherein q = 0-3;
ZBG is a zinc chelating group selected from
。
3. The amide small-molecule organic compound or pharmaceutically acceptable salt with indole or indole analog as a parent nucleus structure according to claim 1 or 2, which comprises:
(7- (3- ((1-adamantanamine) formyl)) -indolyl) heptanoylhydroxylamine
(6- (3- ((1-adamantanamine) formyl)) -indolyl) hexanoylhydroxylamine
(7- (3- (cyclooctylcarbonyl)) -indolyl) heptanoylhydroxylamine
(7- (2- ((1-adamantanamine) formyl)) -indolyl) heptanoylhydroxylamine
(7- (3- ((1-adamantanamine) formyl) -5-fluoro) -indolyl) heptanoylhydroxylamine
(7- (3- ((4-chlorobenzyl) formyl)) -indolyl) heptanoylhydroxylamine
(7- (2- ((2-adamantanamine) formyl) -6-bromo) -indolyl) heptanoylhydroxylamine
(7- (2- ((2-adamantanamine) formyl) -5-chloro) -indolyl) heptanoylhydroxylamine
(7- (2- ((2-adamantanamine) formyl)) -indolyl) heptanoylhydroxylamine
(7- (2- ((2-adamantanamine) formyl) -5-methoxy) -indolyl) heptanoylhydroxylamine
(7- (2- ((4-fluorophenyl) formyl)) -indolyl) heptanoylhydroxylamine
(7- (3- (cyclohexanecarboxoyl)) -indolyl) heptanoylhydroxylamine
(7- (3- ((1-adamantanamine) formyl)) -7-azaindolyl) heptanoylhydroxylamine
(7- (3- ((2-adamantanamine) formyl)) -indolyl) heptanoylhydroxylamine
(5- (3- ((2-adamantanamine) formyl)) -indolyl) pentanoylhydroxylamine
(7- (3- (cycloheptylcarbonyl)) -indolyl) heptanoylhydroxylamine
(7- (3- ((1-adamantanamine) formyl) -5-chloro) -indolyl) heptanoylhydroxylamine
(7- (3- ((2, 4-dichlorophenyl) formyl)) -indolyl) heptanoylhydroxylamine
(7- (3- ((1-adamantanamine) formyl) -5-methoxy) -indolyl) heptanoylhydroxylamine
(7- (2- ((1-adamantanamine) formyl)) -3-azaindolyl) heptanoylhydroxylamine
N- (2-adamantane) -1- (4-hydroxycarbamoylbenzyl) indolecarboxamide.
4. A pharmaceutical composition, which comprises the amide small-molecule organic compound or pharmaceutically acceptable salt with the indole or indole analog as the parent nucleus structure according to claim 1 or 2, and a pharmaceutically acceptable carrier.
5. The use of the amide small-molecule organic compound or the pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein the amide small-molecule organic compound or the pharmaceutically acceptable salt thereof has a parent nucleus structure of indole or indole analogues, for preparing histone deacetylase inhibitors.
6. Use of the amide small-molecule organic compound or the pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein the amide small-molecule organic compound or the pharmaceutically acceptable salt thereof has a core structure of indole or indole analogues, or the pharmaceutical composition according to claim 4, in preparation of a preparation for promoting deacetylation of cell histones H3 and H4 and alpha-Tubulin.
7. The use of the amide small-molecule organic compound or the pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein the amide small-molecule organic compound or the pharmaceutically acceptable salt thereof has an indole or an indole analog as a parent nucleus, or the pharmaceutical composition according to claim 4, for preparing a medicament for preventing and/or treating tumors.
8. Use of the amide small-molecule organic compound or the pharmaceutically acceptable salt thereof according to claim 1 or 2, or the pharmaceutical composition according to claim 4, wherein the amide small-molecule organic compound or the pharmaceutically acceptable salt thereof has a parent nucleus structure of indole or indole analogues, for the preparation of a medicament for the antitumor treatment after chemotherapy failure induced by acquired drug resistance.
9. Use according to claim 7 or 8, wherein the tumour comprises lung cancer, breast cancer, epidermoid cancer, colon cancer, rectal cancer, liver cancer, stomach cancer, prostate cancer, pancreatic cancer, leukaemia, ovarian cancer, bladder cancer, kidney cancer, oral cancer, melanoma, head and neck cancer, uterine cancer, brain tumour, oesophageal cancer, lymphatic cancer.
10. The use according to claim 7 or 8, wherein the compound or pharmaceutically acceptable salt is used for inhibiting proliferation, growth, migration, infiltration and clonality of tumor cells, and promoting apoptosis of tumor cells.
11. The use of the amide small-molecule organic compound or the pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein the amide small-molecule organic compound or the pharmaceutically acceptable salt thereof has an indole or an indole analog as a parent nucleus structure, or the pharmaceutical composition according to claim 4, for the preparation of a medicament for preventing and/or treating thioredoxin-mediated diseases.
12. The use according to claim 11, wherein the disease comprises autoimmune diseases, allergy, inflammation, neurodegenerative diseases, alzheimer's disease, peroneal muscular atrophy, traumatic brain injury, hypertension and stroke.
13. The use of the amide small-molecule organic compound or the pharmaceutically acceptable salt thereof according to claim 1 or 2, or the pharmaceutical composition according to claim 4, wherein the amide small-molecule organic compound or the pharmaceutically acceptable salt thereof has a parent-nucleus structure of indole or indole analogues, for the preparation of a medicament for the treatment of malignant tumors.
14. The use of claim 13, wherein the malignancy comprises liver cancer, lung cancer, prostate cancer, skin cancer, colon cancer, rectal cancer, pancreatic cancer, breast cancer, leukaemia, ovarian cancer, stomach cancer, bladder cancer, kidney cancer, oral cancer, melanoma, head and neck cancer, uterine cancer, brain tumour, oesophageal cancer, lymphoma.
15. The use of claim 13, wherein the compound or pharmaceutically acceptable salt is used to inhibit the growth, metastasis and recurrence of a malignant tumor.
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Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102046240A (en) * | 2008-03-25 | 2011-05-04 | 阿费克蒂斯药品公司 | Novel P2X7R antagonists and their use |
| CN103467359A (en) * | 2013-09-27 | 2013-12-25 | 山东大学 | Cinnamon amides histone deacetylase inhibitor with benzpyrole and preparation method and application of same |
| CN105102446A (en) * | 2013-02-11 | 2015-11-25 | 星座制药公司 | Modulators of methyl modifying enzymes, compositions and uses thereof |
| CN105198789A (en) * | 2015-10-26 | 2015-12-30 | 山东大学 | Substituted3-indole Bcl-2 protein inhibitor and preparation method and application thereof |
| CN105367479A (en) * | 2015-11-16 | 2016-03-02 | 青岛大学 | Histone deacetylase inhibitor 2-(1-(4-chlorobenzoyl)-5-methoxy-2-methyl-1 hydro-indole-3-yl-N-(2-(hydroxyamino)-2-ketoacyl) acetamide and preparation method and application thereof |
| CN105820104A (en) * | 2016-03-10 | 2016-08-03 | 温州医科大学 | Indole-2-acetamide compound with anti-inflammatory effect, and application thereof in preparation of anti-inflammatory drugs |
| CN106478606A (en) * | 2016-09-21 | 2017-03-08 | 沈阳药科大学 | N substituted indole analog derivative and its application |
| CN106957315A (en) * | 2016-01-08 | 2017-07-18 | 中国人民解放军第二军医大学 | N- replaces benzenesulfonyl-azaindole oxybenzamide class compound and its prepares the purposes of medicine |
| CN107141244A (en) * | 2017-05-08 | 2017-09-08 | 潍坊博创国际生物医药研究院 | Indolebutyric acid class inhibitors of histone deacetylase and its preparation method and application |
| CN107417592A (en) * | 2017-04-24 | 2017-12-01 | 浙江师范大学行知学院 | A kind of oxoaGetamide derivative of 1H indoles 2 and preparation method and application |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007079239A2 (en) * | 2005-12-30 | 2007-07-12 | Acadia Pharmaceuticals Inc. | Bicyclic nitrogen compounds as modulators of ghrelin receptor and uses thereof |
-
2017
- 2017-12-06 CN CN201711276112.XA patent/CN109879790B/en active Active
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102046240A (en) * | 2008-03-25 | 2011-05-04 | 阿费克蒂斯药品公司 | Novel P2X7R antagonists and their use |
| CN105102446A (en) * | 2013-02-11 | 2015-11-25 | 星座制药公司 | Modulators of methyl modifying enzymes, compositions and uses thereof |
| CN103467359A (en) * | 2013-09-27 | 2013-12-25 | 山东大学 | Cinnamon amides histone deacetylase inhibitor with benzpyrole and preparation method and application of same |
| CN105198789A (en) * | 2015-10-26 | 2015-12-30 | 山东大学 | Substituted3-indole Bcl-2 protein inhibitor and preparation method and application thereof |
| CN105367479A (en) * | 2015-11-16 | 2016-03-02 | 青岛大学 | Histone deacetylase inhibitor 2-(1-(4-chlorobenzoyl)-5-methoxy-2-methyl-1 hydro-indole-3-yl-N-(2-(hydroxyamino)-2-ketoacyl) acetamide and preparation method and application thereof |
| CN106957315A (en) * | 2016-01-08 | 2017-07-18 | 中国人民解放军第二军医大学 | N- replaces benzenesulfonyl-azaindole oxybenzamide class compound and its prepares the purposes of medicine |
| CN105820104A (en) * | 2016-03-10 | 2016-08-03 | 温州医科大学 | Indole-2-acetamide compound with anti-inflammatory effect, and application thereof in preparation of anti-inflammatory drugs |
| CN106478606A (en) * | 2016-09-21 | 2017-03-08 | 沈阳药科大学 | N substituted indole analog derivative and its application |
| CN107417592A (en) * | 2017-04-24 | 2017-12-01 | 浙江师范大学行知学院 | A kind of oxoaGetamide derivative of 1H indoles 2 and preparation method and application |
| CN107141244A (en) * | 2017-05-08 | 2017-09-08 | 潍坊博创国际生物医药研究院 | Indolebutyric acid class inhibitors of histone deacetylase and its preparation method and application |
Non-Patent Citations (2)
| Title |
|---|
| 一类含吲哚酰胺结构的新型组蛋白去乙酰化酶抑制剂的设计、合成及其抗肿瘤活性的研究;刘志韬;《中国优秀博硕士学位论文全文数据库（硕士）医药卫生科技辑》;20200515;E079-64 * |
| 作用于多靶点的组蛋白去乙酰化酶抑制剂结构设计的研究进展;黄亚天，陆涛;《中国现代应用药学》;20131231;第30卷(第9期);第1037-1042页 * |
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