CN108703970A - Compound for activating AMPK and application thereof - Google Patents

Abstract

The present invention relates to a compound which activates AMPK and its use in the preparation of a medicament for the treatment of diseases or physiological conditions ameliorated by AMPK activators, the prevention or treatment of diseases including pre-diabetes, insulin resistance, type ii diabetes, syndrome X, metabolic syndrome and obesity. The compound provided by the invention can reduce the amount of plasma glucose by more than 30 wt%, reduce the amount of triglyceride by more than 35 wt% and reduce the weight by more than 15%.

Description

A compound for activating AMPK and its use

This application is a divisional application of the application with the application number 201310430034.X, the application date is September 18, 2013, and the application name is "a compound for activating AMPK and its use".

technical field

The present invention relates to compounds, especially a compound that activates AMPK (AMP-activated protein kinase) and its preparation for the treatment of diseases or physiological conditions that can be improved by AMPK activators to prevent or treat diseases, including pre-diabetes, insulin resistance, Use in medicine for type 2 diabetes, X-syndrome, metabolic syndrome and obesity.

Background technique

AMPK is clearly a sensor of cellular energy and a responder to energy demands. AMPK is a heteroternary body composed of catalytic α subunits, regulatory β and γ subunits, and all subunits are highly reserved in eukaryotes. AMPK is activated through its upstream kinases such as LKB1, calcium ion/calcitonin-dependent protein phosphokinase (Ca ²⁺ /Calmodulin dependent kinase) and TAK1 phosphorylation of the 172th threonine residue in the α-subunit body, which is determined by physiological Or high AMP/ATP ratio caused by pathological stress also activates AMPK. AMPK activation promotes catabolic pathways and inhibits anabolism, thereby restoring cellular energy balance by reducing ATP consumption and promoting ATP production.

As a regulator of energy metabolism balance, AMPK is considered to be a potential drug target for metabolic syndrome, including type 2 diabetes, cardiovascular disease, fatty liver, etc. Many metabolic syndromes are associated with insulin resistance. Insulin resistance is a pathological condition in which cells fail to respond to insulin so excess glucose in the blood cannot be removed to skeletal muscle or adipose tissue. In muscle cells, AMPK activation increases the expression of glucose transport protein (GLUT4) through transcriptional regulation in a non-insulin-dependent manner, and induces the transfer of GLUT4 to the cell membrane, resulting in an increase in the rate of glucose uptake by cells. AMPK activation also inhibits the synthesis of fatty acids and cholesterol by inhibiting acetyl-CoA carboxylase and HMG-CoA reductase, respectively. Furthermore, AMPK activation leads to the repression of multiple transcription factors, including sterol regulatory element binding protein (SREBP-1c), carbohydrate response element binding protein transcription factor (ChREBP) and hepatic nuclear factor 4a (HNF-4a), and down-regulates Protein expression of enzymes involved in fatty acid synthesis and gluconeogenesis. The research findings mentioned above all support AMPK as a therapeutic target for metabolic syndrome, especially diabetes.

AMP is a natural AMPK activator, however, AMP is an unstable compound, and extracellular administration of AMP will trigger the transmission of messages from purinergic receptors (may lead to cell apoptosis, etc.). Therefore, many researchers have devoted themselves to the development of AMPK activators. It is currently known that high concentrations of 5-aminoimidazole-4-carboxamide-1-β-D-ribofuranoside (AICAR) and metformin compounds can activate AMPK in organisms. Among them, metformin has been used in the treatment of pre-diabetes, insulin resistance, X-syndrome, and type 2 diabetes. However, such AMPK activators have side effects including lactic acidosis, especially in patients with renal insufficiency. Therefore, there is an urgent need to develop novel AMPK activators with lower effective concentrations and fewer side effects.

Contents of the invention

In view of this, the object of the present invention is to provide a compound that activates AMPK and its preparation for the treatment of diseases or physiological conditions that can be improved by AMPK activators to prevent or treat diseases, including pre-diabetes, type II diabetes, X- Syndrome, metabolic syndrome and obesity medicine.

To achieve the above object, the present invention provides compounds of formula (1) and formula (2) and tautomers thereof:

and pharmaceutically acceptable salts thereof, wherein R2 represents a hydrogen atom, a halogen atom, a hydroxyl group, an amino group, an amino group with one or two substituents with up to ten carbons, a sulfhydryl group, a carboxyl group, a nitro group, a sulfo group, Alkyl, alkylamino, alkylthio, alkoxy, cycloalkyl, substituted alkyl, substituted alkenyl, substituted alkynyl, acyl, aryl, substituted aryl, aryloxy; R6 Represents hydroxyl, amine, mercapto, -NHR (monosubstituted amine) where R represents a halogen atom, hydroxy, amine, amine with one or two substituents with up to ten carbons, mercapto, carboxyl, Nitro, sulfo, alkyl, alkylamino, alkylthio, alkoxy, cycloalkyl, substituted alkyl, substituted alkenyl, substituted alkynyl, acyl, aryl, substituted aryl , aryloxy; R3 represents a hydrogen atom or an alkyl group with up to ten carbons; or a tautomer or a pharmaceutically acceptable salt thereof, and an AMPK activator includes the purine and pyrimidine and its derivatives or Pharmaceutically acceptable salts as an active compound.

The general substituents have the same meaning as defined, wherein

The hydroxyl group represents a hydroxyl group—OH;

The halogen atom represents fluorine, chlorine, bromine, iodine;

The amine group represents —NH ₂ ;

The mercapto group represents —SH;

The carboxy group represents —C(O)OR, wherein R is defined as a hydrogen atom, an alkyl group, a substituted alkyl group, an aromatic hydrocarbon group, or a substituted aromatic hydrocarbon group;

The nitro group represents —NO ₂ ;

The sulfo group represents —SO ₃ R; wherein R here is defined as a hydrogen atom, an alkyl group, or a substituted alkyl group;

The alkyl group means a straight or branched chain containing up to ten carbon atoms, wherein saturated or unsaturated groups include methyl, propyl, isopropyl, butyl, ethoxy, vinyl, ethynyl, propyne group, 2-alkynylhexyl group and other groups meeting the definition;

The substituted alkyl means that the alkyl group includes 1 to 7 substituents such as hydroxyl, mercapto, alkylamino, mercapto, halogen atom, alkoxy, acyloxy, amine, carboxyl, sulfo , acyl and others, so that these groups can be attached to any carbon of the alkyl moiety;

The alkylamino group represents —NRR', wherein R and R' independently represent a hydrogen atom, an alkyl group, a substituted alkyl group, an aromatic hydrocarbon group, and a substituted aromatic hydrocarbon group as defined herein;

The alkylthiol represents —SR, wherein R refers to an alkyl group, a substituted alkyl group, an aromatic hydrocarbon group, and a substituted aromatic hydrocarbon group as defined herein;

The alkoxy group represents —OR, wherein R refers to alkyl, substituted alkyl, aryl, substituted aryl as defined herein;

The cycloalkyl represents a single or multiple cycloalkyl groups containing 3-15 carbon atoms;

The acyl group represents —C(O)R, wherein R refers to a hydrogen atom, an alkyl group, a substituted alkyl group, an aromatic hydrocarbon group, a substituted aromatic hydrocarbon group as defined herein;

The aromatic hydrocarbon group represents an aromatic cyclized carbon group;

The substituted aromatic hydrocarbon group means that the aromatic hydrocarbon group contains substituents including halogen atoms, hydroxyl groups, amino groups, mercapto groups, alkoxy groups, sulfo groups, carboxyl groups, and alkyl groups as defined herein;

The aryloxy group represents —OAr, wherein Ar refers to an aromatic hydrocarbon group, and the substituted aromatic hydrocarbon group is as defined herein;

Therefore preferred AMPK activating groups include: adenine, 2-amino-6-methylaminopurine, 2-amino-6-ethylaminopurine, 2-amino-6-isobutylaminopurine, 2 -Amino-6-propylaminopurine, 2-amino-6-isoamylaminopurine, 2-amino-6-hexylaminopurine, 2-amino-6-cyclopropylaminopurine, 2-amine Base-6-cyclobutylaminopurine, 2-amino-6-cyclopentylaminopurine, 2-amino-6-cyclohexylaminopurine, 2-amino-6-anilinylpurine, 2-amine Base-6-(2-chloroanilino)purine, 2-amino-6-(3-chloroanilino)purine, 2-amino-6-(4-chloroanilino)purine, 2-amino- 6-(2-bromoanilino)purine, 2-amino-6-(3-bromoanilino)purine, 2-amino-6-(4-bromoanilino)purine, 2-amino-6- (2-fluoroanilino)purine, 2-amino-6-(3-fluorobromoanilino)purine, 2-amino-6-(4-fluoroanilino)purine, 2-amino-6-benzene Methylaminopurine, 2-amino-6-(2-methylbenzylamino)purine, 2-amino-6-(3-methylbenzylamino)purine, 2-amino-6- (4-methylbenzylamino)purine, 2-amino-6-(2-chlorobenzylamino)purine, 2-amino-6-(3-chlorobenzylamino)purine, 2- Amino-6-(4-chlorobenzylamino)purine, 2-amino-6-(2-fluorobenzylamino)purine, 2-amino-6-(3-fluorobenzylamino) Purine, 2-amino-6-(4-fluorobenzylamino)purine, 2-amino-6-(3-iodobenzylamino)purine, 2-amino-6-(4-hydroxybenzene Methylamino) purine, 2-amino-6-(2,3-dihydroxybenzylamino)purine, 2-amino-6-(3,4-dihydroxybenzylamino)purine, 2- Amino-6-(2,4-dihydroxybenzylamino)purine, 2-amino-6-(2-methoxybenzylamino)purine, 2-amino-6-(2,3 -dimethoxybenzylamino)purine, 2-amino-6-(3,5-dimethoxybenzylamino)purine, 2-amino-6-(2,4,5-trimethyl oxybenzylamino)purine, 2-amino-6-(3,4,5-trimethoxybenzylamino)purine, 6-methylaminopurine, 6-ethylaminopurine, 6-propylamine Base purine, 6-isobutylamino purine, 6-isopentylamino purine, 6-hexylamino purine, 6-cyclopropylamino purine, 6-cyclobutylamino purine, 6-cyclopentylamino purine, 6 -cyclohexylaminopurine, 6-anilinopurine, 6-(2-chloroanilino)purine, 6-(3-chloroanilino)purine, 6-(4-chloroanilino)purine, 6-(2 -Bromoanilino)purine, 6-(3-bromoanilino)purine, 6-(4-bromoanilino)purine, 6-(2-fluoroanilino)purine, 6-(3-fluoroanilino)purine , 6-(4-fluoroanilino)purine, 6-benzylaminopurine, 6-(2-methylbenzylamino)purine, 6-(3-methylbenzylamino)purine, 6- (4-methylbenzylamino)purine, 6-(2-chlorobenzylamino)purine, 6-(3-chlorobenzylamino)purine, 6-(4-chlorobenzylamino)purine , 6-(2-fluorobenzylamino)purine, 6-(3-fluorobenzylamino)purine, 6-(4-fluorobenzylamino)purine, 6-(3-iodobenzylamino)purine ) purine, 6-(4-hydroxybenzylamino)purine, 6-(2,3-dihydroxybenzylamino)purine, 6-(3,4-dihydroxybenzylamino)purine, 6- (2,4-dihydroxybenzylamino)purine, 6-(2-methoxybenzylamino)purine, 6-(2,3-dimethoxybenzylamino)purine, 6-( 3,5-dimethoxybenzylamino)purine, 6-(2,4,5-trimethoxybenzylamino)purine, 6-(3,4,5-trimethoxybenzylamino)purine ) purine, 2-hydroxy-6-methylaminopurine, 2-hydroxy-6-ethylaminopurine, 2-hydroxy-6-isobutylaminopurine, 2-hydroxy-6-propylaminopurine, 2-hydroxy -6-isopentylaminopurine, 2-hydroxy-6-hexylaminopurine, 2-hydroxy-6-cyclopropylaminopurine, 2-hydroxy-6-cyclobutylaminopurine, 2-hydroxy-6-cyclo Amylaminopurine, 2-hydroxy-6-cyclohexylaminopurine, 2-hydroxy-6-anilinylpurine, 2-hydroxy-6-(2-chloroanilino)purine, 2-hydroxy-6-(3 -Chloroanilino)purine, 2-hydroxy-6-(4-chloroanilino)purine, 2-hydroxy-6-(2-bromoanilino)purine, 2-hydroxy-6-(3-bromoanilino) Purine, 2-hydroxy-6-(4-bromoanilino)purine, 2-hydroxy-6-(2-fluoroanilino)purine, 2-hydroxy-6-(3-fluorobromoanilino)purine, 2- Hydroxy-6-(4-fluoroanilino)purine, 2-hydroxy-6-benzylaminopurine, 2-hydroxy-6-(2-methylbenzylamino)purine, 2-hydroxy-6-( 3-methylbenzylamino)purine, 2-hydroxy-6-(4-methylbenzylamino)purine, 2-hydroxy-6-(2-chlorobenzylamino)purine, 2-hydroxy- 6-(3-chlorobenzylamino)purine, 2-hydroxy-6-(4-chlorobenzylamino)purine, 2-hydroxy-6-(2-fluorobenzylamino)purine, 2-hydroxy -6-(3-fluorobenzylamino)purine, 2-hydroxy-6-(4-fluorobenzylamino)purine, 2-hydroxy-6-(3-iodobenzylamino)purine, 2- Hydroxy-6-(4-hydroxybenzylamino)purine, 2-hydroxy-6-(2,3-dihydroxybenzylamino)purine, 2-hydroxy-6-(3,4-dihydroxybenzyl Amino)purine, 2-hydroxy-6-(2,4-dihydroxybenzylamino)purine, 2-hydroxy-6-(2-methoxybenzylamino)purine, 2-hydroxy-6- (2,3-dimethoxybenzylamino)purine, 2-hydroxy-6-(3,5-dimethoxybenzylamino)purine, 2-hydroxy-6-(2,4,5 -trimethoxybenzylamino)purine, 2-hydroxy-6-(3,4,5-trimethoxybenzylamine base) purine, 2-methyl-6-methylaminopurine, 2-methyl-6-ethylaminopurine, 2-methyl-6-isobutylaminopurine, 2-methyl-6-propylaminopurine Purine, 2-methyl-6-isoamylaminopurine, 2-methyl-6-hexylaminopurine, 2-methyl-6-cyclopropylaminopurine, 2-methyl-6-cyclobutylaminopurine Purine, 2-methyl-6-cyclopentylaminopurine, 2-methyl-6-cyclohexylaminopurine, 2-methyl-6-anilinylpurine, 2-methyl-6-(2-chloro Anilino)purine, 2-methyl-6-(3-chloroanilino)purine, 2-methyl-6-(4-chloroanilino)purine, 2-methyl-6-(2-bromoanilino) ) purine, 2-methyl-6-(3-bromoanilino)purine, 2-methyl-6-(4-bromoanilino)purine, 2-methyl-6-(2-fluoroanilino)purine , 2-methyl-6-(3-fluorobromoanilino)purine, 2-methyl-6-(4-fluoroanilino)purine, 2-methyl-6-benzylaminopurine, 2-methyl Base-6-(2-methylbenzylamino)purine, 2-methyl-6-(3-methylbenzylamino)purine, 2-methyl-6-(4-methylbenzylamine) base) purine, 2-methyl-6-(2-chlorobenzylamino)purine, 2-methyl-6-(3-chlorobenzylamino)purine, 2-methyl-6-(4- Chlorobenzylamino)purine, 2-methyl-6-(2-fluorobenzylamino)purine, 2-methyl-6-(3-fluorobenzylamino)purine, 2-methyl-6 -(4-fluorobenzylamino)purine, 2-methyl-6-(3-iodobenzylamino)purine, 2-methyl-6-(4-hydroxybenzylamino)purine, 2- Methyl-6-(2,3-dihydroxybenzylamino)purine, 2-methyl-6-(3,4-dihydroxybenzylamino)purine, 2-methyl-6-(2, 4-dihydroxybenzylamino)purine, 2-methyl-6-(2-methoxybenzylamino)purine, 2-methyl-6-(2,3-dimethoxybenzylamine) base) purine, 2-methyl-6-(3,5-dimethoxybenzylamino)purine, 2-methyl-6-(2,4,5-trimethoxybenzylamino)purine , 2-methyl-6-(3,4,5-trimethoxybenzylamino)purine, 2-methylamino-6-aminopurine, 2-ethylamino-6-aminopurine, 2 -Isobutylamino-6-aminopurine, 2-propylamino-6-aminopurine, 2-isoamylamino-6-aminopurine, 2-hexylamino-6-aminopurine, 2- Cyclopropylamino-6-aminopurine, 2-cyclobutylamino-6-aminopurine, 2-cyclopentylamino-6-aminopurine, 2-cyclohexylamino-6-aminopurine, 2 -anilino-6-aminopurine, 2-(2-chloroanilino)-6-aminopurine, 2-(3-chloroanilino)-6-aminopurine, 2-(4-chloroanilino )-6-aminopurine, 2-(2-fluoroanilino)-6-aminopurine, 2-(3-fluoroanilino)-6-aminopurine, 2-(4-fluoroanilino)- 6-aminopurine, 2-(2-bromoanilino )-6-aminopurine, 2-(3-bromoanilino)-6-aminopurine, 2-(4-bromoanilino)-6-aminopurine, 2-benzylamino-6-amine Base purine, 2-(2-methylbenzylamino)-6-aminopurine, 2-(3-methylbenzylamino)-6-aminopurine, 2-(4-methylbenzylamino) Amino)-6-aminopurine, 2-(2-chlorobenzylamino)-6-aminopurine, 2-(3-chlorobenzylamino)-6-aminopurine, 2-(4 -Chlorobenzylamino)-6-aminopurine, 2-(2-fluorobenzylamino)-6-aminopurine, 2-(3-fluorobenzylamino)-6-aminopurine, 2-(4-fluorobenzylamino)-6-aminopurine, 2-(3-iodobenzylamino)-6-aminopurine, 2-(4-hydroxybenzylamino)-6- Aminopurine, 2-(2,3-dihydroxybenzylamino)-6-aminopurine, 2-(2,4-dihydroxybenzylamino)-6-aminopurine, 2-(3 ,4-dihydroxybenzylamino)-6-aminopurine, 2-(2,3-dimethoxybenzylamino)-6-aminopurine, 2-(2,3-dimethoxy Benzylamino)-6-aminopurine, 2-(3,5-dimethoxybenzylamino)-6-aminopurine, 2-(2,4,5-trimethoxybenzyl Amino)-6-aminopurine, 2-(3,4,5-trimethoxybenzylamino)-6-aminopurine, 2-methylamino-6-hydroxypurine, 2-ethylamino -6-hydroxypurine, 2-isobutylamino-6-hydroxypurine, 2-propylamino-6-hydroxypurine, 2-isoamylamino-6-hydroxypurine, 2-hexylamino-6-hydroxypurine , 2-cyclopropylamino-6-hydroxypurine, 2-cyclobutylamino-6-hydroxypurine, 2-cyclopentylamino-6-hydroxypurine, 2-cyclohexylamino-6-hydroxypurine, 2- Anilino-6-hydroxypurine, 2-(2-chloroanilino)-6-hydroxypurine, 2-(3-chloroanilino)-6-hydroxypurine, 2-(4-chloroanilino)-6- Hydroxypurine, 2-(2-fluoroanilino)-6-hydroxypurine, 2-(3-fluoroanilino)-6-hydroxypurine, 2-(4-fluoroanilino)-6-hydroxypurine, 2- (2-bromoanilino)-6-hydroxypurine, 2-(3-bromoanilino)-6-hydroxypurine, 2-(4-bromoanilino)-6-hydroxypurine, 2-benzylamino- 6-hydroxypurine, 2-(2-methylbenzylamino)-6-hydroxypurine, 2-(3-methylbenzylamino)-6-hydroxypurine, 2-(4-methylbenzylamino) Amino)-6-hydroxypurine, 2-(2-chlorobenzylamino)-6-hydroxypurine, 2-(3-chlorobenzylamino)-6-hydroxypurine, 2-(4-chlorobenzene Methylamino)-6-hydroxypurine, 2-(2-fluorobenzylamino)-6-hydroxypurine, 2-(3-fluorobenzylamino)-6-hydroxypurine, 2-(4-fluoro Benzylamino)-6-hydroxypurine, 2-(3-iodobenzylamino)-6-hydroxypurine, 2-(4-hydroxybenzylamino)-6-hydroxy Purine, 2-(2,3-dihydroxybenzylamino)-6-hydroxypurine, 2-(2,4-dihydroxybenzylamino)-6-hydroxypurine, 2-(3,4-di Hydroxybenzylamino)-6-hydroxypurine, 2-(2,3-dimethoxybenzylamino)-6-hydroxypurine, 2-(2,3-dimethoxybenzylamino) -6-hydroxypurine, 2-(3,5-dimethoxybenzylamino)-6-hydroxypurine, 2-(2,4,5-trimethoxybenzylamino)-6-hydroxypurine , 2-(3,4,5-trimethoxybenzylamino)-6-hydroxypurine, 6-hydroxypurine, 6-hydrothiopurine, N ⁶ -methyladenine, 2-aminoadenine , 2-hydroxyadenine, 2-methyladenine, 2-amino-6-hydroxypurine, 2,6-dihydroxypurine, 2-methyl-6-hydroxypurine, 2-amino-6-hydrosulfide base purine, 2-hydroxy-6-hydrothiopurine, 2-methyl-6-hydrothiopurine, 2-hydrogenthioadenine, 2-hydrogen-6-hydroxypurine, 2,6-di Thiopurine, 2-mercapto-6-methylpurine, 2-mercapto-6-ethylpurine, 2-mercapto-6-propylpurine, 2-ethyladenine, 2- Ethyl-6-hydroxypurine, 2-ethyl-6-hydrothiopurine, 2-ethyl-6-methylaminopurine, 2-ethyl-6-ethylaminopurine, 2-ethyl-6 -Propylaminopurine, 2-propyladenine, 2-propyl-6-hydroxypurine, 2-propyl-6-hydrothiopurine, 2-propyl-6-methylaminopurine, 2-propyl -6-ethylaminopurine, 2-propyl-6-propylaminopurine, 2-amino-6-benzylaminopurine, 2-ethyl-6-benzylaminopurine, 2-propyl- 6-benzylamino purine, 2-ethyl-6-anilino purine, 2-propyl-6-anilino purine, 2-benzyl adenine, 2-benzyl-6-hydroxy purine, 2 -Benzyl-6-mercaptopurine, 2-benzyl-6-methylaminopurine, 2-benzyl-6-ethylaminopurine, 2-benzyl-6-propylaminopurine, 2-phenyladenine, 2-phenyl-6-hydroxypurine, 2-phenyl-6-hydrothiopurine, 2-phenyl-6-methylaminopurine, 2-phenyl-6-ethylamine Base purine, 2-phenyl-6-propylamino purine, 2-mercapto-N ⁶ -2-prenyl adenine, 2-ethyl-N ⁶ -2-prenyl adenine, 2 -Propyl-N ⁶ -2-prenyl adenine, 2-benzyl-N ⁶ -2-prenyl adenine, 2-phenyl-N ⁶ -2-prenyl adenine , 2-chloroadenine, 2-chloro-6-hydroxypurine, 2-chloro-6-hydrothiopurine, 2-chloro-6-methylaminopurine, 2-chloro-6-ethylaminopurine, 2 -Chloro-6-propylaminopurine, 2-chloro-6-benzylaminopurine, 2-chloro- ⁶ -anilininopurine, 2-chloro-N6-2-prenyladenine, 2-fluoro Adenine, 2-fluoro-6-hydroxypurine, 2-fluoro-6-hydrothiopurine, 2-fluoro-6- Methylaminopurine, 2-fluoro-6-ethylaminopurine, 2-fluoro-6-propylaminopurine, 2-fluoro-6-benzylaminopurine, 2-fluoro-6-anilininopurine, 2- Fluoro-N ⁶ -2-prenyl adenine, 2-bromoadenine, 2-bromo-6-hydroxypurine, 2-bromo-6-hydrothiopurine, 2-bromo-6-methylaminopurine , 2-bromo-6-ethylaminopurine, 2-bromo-6-propylaminopurine, 2-bromo-6-benzylaminopurine, 2-bromo-6-anilinylpurine, 2-bromo-N ⁶ -2-Prenyl adenine, 2-iodoadenine, 2-iodo-6-hydroxypurine, 2-iodo-6-hydrothiopurine, 2-iodo-6-methylaminopurine, 2-iodo -6-ethylaminopurine, 2-iodo-6-propylaminopurine, 2-iodo-6-benzylaminopurine, 2-iodo-6-anilinylpurine, 2-iodo-N ⁶ -2-iso Pentenyl adenine, 2-mercapto-N ⁶ -cyclohexyl adenine, 2-propyl-N ⁶ -cyclohexyl adenine, 2-ethyl-N ⁶ -cyclohexyl adenine, 2-benzyl Base-N ⁶ -cyclohexyladenine, 2-phenyl-N ⁶ -cyclohexyladenine, 2-chloro-N ⁶ -cyclohexyladenine, 2-fluoro-N ⁶ -cyclohexyladenine, 2-bromo -N ⁶ -cyclohexyladenine, 2-iodo-N ⁶ -cyclohexyladenine, 2-amino-6-(4-hydroxy-3-methyl-but2enylamino)purine, 2-hydroxy- 6-(4-hydroxy-3-methyl-but2enylamino)purine, 2-mercapto-6-(4-hydroxy-3-methyl-but2enylamino)purine, 2-methyl -6-(4-hydroxy-3-methyl-but2enylamino)purine, 2-ethyl-6-(4-hydroxy-3-methyl-but2enylamino)purine, 2-propyl -6-(4-hydroxy-3-methyl-but2enylamino)purine, 2-benzyl-6-(4-hydroxy-3-methyl-but2enylamino)purine, 2-benzene Base-6-(4-hydroxy-3-methyl-but2enylamino)purine, 2-chloro-6-(4-hydroxy-3-methyl-but2enylamino)purine, 2-fluoro -6-(4-hydroxy-3-methyl-but2enylamino)purine, 2-bromo-6-(4-hydroxy-3-methyl-but2enylamino)purine, 2-iodo-6 -(4-hydroxy-3-methyl-but2enylamino)purine, 2-cyclohexyl-6-(4-hydroxy-3-methyl-but2enylamino)purine, 2-amino-6 -Furfuryl alcohol aminopurine, 2-hydroxy-6-furfuryl alcohol aminopurine, 2-mercapto-6-furfuryl alcohol aminopurine, 2-methyl-6-furfuryl alcohol aminopurine, 2-ethyl-6-furfuryl alcohol Aminopurine, 2-propyl-6-furfuryl alcohol aminopurine, 2-benzyl-6-furfuryl alcohol aminopurine, 2-phenyl-6-furfuryl alcohol aminopurine, 2-chloro-6-furfuryl alcohol aminopurine Purine, 2-fluoro-6-furfuryl alcohol amino purine, 2-bromo-6-furfuryl alcohol amino purine, 2-iodo-6-furfuryl alcohol amino purine, 2-cyclohexyl-6-furfuryl alcohol amino purine, 2-amine Base-6-acetamidopurine, 2-hydroxy-6-acetyl Aminopurine, 2-mercapto-6-acetamidopurine, 2-methyl-6-acetamidopurine, 2-ethyl-6-acetamidopurine, 2-propyl-6-acetamide Base purine, 2-benzyl-6-acetamido purine, 2-phenyl-6-acetamido purine, 2-chloro-6-acetamido purine, 2-bromo-6-acetamido purine, 2-iodo-6-acetamidopurine, 2-cyclohexyl-6-acetamidopurine, 2-dimethylamino-6-hydroxypurine, guanine, xanthine, hypoxanthine, 6-thio Hydrogen guanine, 5-methylcytosine, 5,6-dihydrouridine, thymine, cytosine, uracil, 5-fluorouracil, fluorouridine, 6-azauracil each of which may be Exists as a salt, hydrate, as a prodrug, or as a metabolite.

Further, the compound of formula (1) is an AMPK activator.

Further, the compound of formula (2) is an AMPK activator.

The present invention also provides an application of an AMPK-activating compound in the preparation of a medicament for treating diseases or physiological conditions that can be improved by AMPK activators, wherein the AMPK-activating compound is at least one effective dose of the compound selected from Claim 1 or 2 and/or pharmaceutically and nutritionally acceptable salts thereof, for administration to mammals in need of such treatment.

The present invention also provides the use of a compound that activates AMPK in the preparation of a medicament for preventing or treating a physiological condition or disease selected from the group consisting of: pre-diabetes, insulin resistance, type 2 diabetes, X-syndrome, metabolic syndrome purposes, wherein the compound for activating AMPK is at least one effective dose of compound selected from compounds of formula (1) and/or pharmaceutically and nutritionally acceptable salts thereof, to be administered to mammals in need of such treatment , thereby increasing cellular glucose.

The present invention also provides the use of a compound that activates AMPK in the preparation of a medicament for preventing or treating a physiological condition or disease selected from the group consisting of: pre-diabetes, insulin resistance, type 2 diabetes, X-syndrome, metabolic syndrome purposes, wherein the compound is at least one effective dosage of the compound selected from the compound of formula (2) and/or its pharmaceutically and nutritionally acceptable salts, to be given to the mammals needing this treatment, thereby increasing Cellular Glucose.

The present invention also provides an application of an AMPK-activating compound in the preparation of a medicament for preventing or treating obesity, wherein the AMPK-activating compound is at least one effective dosage compound selected from compounds of formula (1) and/or Or its pharmaceutically and nutritionally acceptable salts are administered to mammals in need of such treatment, thereby reducing plasma triglycerides and reducing body weight of mammals.

The present invention also provides an application of an AMPK-activating compound in the preparation of a medicament for preventing or treating obesity, wherein the AMPK-activating compound is at least one compound with an effective dosage, which is selected from compounds of formula (2) and/or Or its pharmaceutically and nutritionally acceptable salts are administered to mammals in need of such treatment, thereby reducing plasma triglycerides and reducing body weight of mammals.

The present invention further provides a medicine, which comprises an effective dose of the compound of formula (1) in combination with a pharmaceutically suitable carrier.

The present invention further provides a medicine, which comprises an effective dose of the compound of formula (2) in combination with a pharmaceutically suitable carrier.

The medicine for treating pre-diabetes, insulin resistance, type 2 diabetes, X-syndrome and metabolic syndrome of the present invention may contain the second medicine. Suitable second agents include various biguanides, thiazolediones, thienopyridones, and other AMPK activators.

As described herein, the present invention includes administering to a mammal a pharmaceutically effective dose of any compound of formula (1) and formula (2), and salts and prodrugs thereof. More suitably, the present invention also includes administering a pharmaceutically effective dose of any compound of formula (1) and formula (2) to humans, especially humans in need of treatment for any of the diseases described herein.

The term "AMPK" as used herein refers to adenosine monophosphate-activated protein kinase.

The term "prediabetes" as used herein refers to a physiological condition characterized by fasting blood glucose above 100 mg/dL but below 140 mg/dL.

The term "insulin resistance" as used herein refers to a physiological condition in which the whole body or tissues including liver, skeletal muscle, adipose tissue fail to respond to insulin.

The term "type 2 diabetes" as used herein also refers to non-insulin-dependent diabetes or adult-onset diabetes; refers to insufficient insulin production or insulin resistance caused by metabolic disorders, usually characterized by fasting blood glucose above 140 mg/dl.

The term "Syndrome X" as used herein refers to a physiological condition characterized by at least two of the following symptoms: fasting hyperglycemia (non-insulin dependent diabetes mellitus), hypertension, high triglycerides, low HDL cholesterol.

The compound provided by the invention can reduce the amount of plasma glucose by more than 30wt%, the amount of triglyceride by more than 35wt%, and the body weight by more than 15%.

Detailed ways

Example 1

2-Amino-6-(3-chlorobenzylamino)purine

After dissolving 4 mmol of 2-amino-6-chloropurine in 20 ml of butanol, 5 mmol of 3-chlorobenzylamine and 6 mmol of triethylamine were added. The mixture was reacted at 90°C for 4 hours. After cooling, the product was obtained by filtration, washed with water and butanol and crystallized from dimethylformamide or ethanol. HPLC: more than 98% purity. Yield 95%.

Table (1) compound manufactured by the method of embodiment 1

Example 2

6-(3-Chlorobenzylamino)purine

After dissolving 4 mmol of 6-chloropurine in 20 ml of butanol, 5 mmol of 3-chlorobenzylamine and 6 mmol of triethylamine were added. The mixture was reacted at 90°C for 4 hours. After cooling, the product was obtained by filtration, washed with water and butanol and crystallized from dimethylformamide or ethanol. HPLC: greater than 97% purity. Yield 94%.

Table (2) compound manufactured by the method of embodiment 2

Example 3

2-Hydroxy-6-chloropurine

After dissolving 4 mmol of 2-amino-6-chloropurine in 35 ml of 50 wt% sulfuric acid, 5 mmol of sodium nitrate was added. The mixture was reacted at -10°C for 2 hours and then at 50°C for 1 hour. After cooling, the product was obtained by filtration, washed with water and butanol and crystallized from dimethylformamide or ethanol. HPLC: more than 98% purity. Yield 86%. MS (ESI) m/e 170.88 (M+H ⁺ ); 1H NMR (DMSO-d6): 8.01 (s, 1H, =CH-N), 13.26 (s, 2H, OH and NH).

Example 4

2-Hydroxy-6-(3-chlorobenzylamino)purine

After dissolving 3 mmol of 2-hydroxy-6-chloropurine obtained in Example 3 in 20 ml of butanol, 4 mmol of 3-chlorobenzylamine and 6 mmol of triethylamine were added. The mixture was reacted at 90°C for 4 hours. After cooling, the product was obtained by filtration, washed with water and butanol and crystallized from dimethylformamide or ethanol. HPLC: greater than 97% purity. Yield 93%.

Table (3) compound manufactured by the method of embodiment 4

Example 5

2-methyl-6-chloropurine

After dissolving 3 mmol of 2-iodo-6-chloropurine in 20 ml of butanol, 4 mmol of methyl manganese chloride was added. The mixture was reacted at 80° C. for 24 hours under palladium catalysis. After cooling, the product was obtained by filtration, washed with water and butanol and crystallized from dimethylformamide or ethanol. HPLC: more than 97% purity. Yield 91%.

Example 6

2-Methyl-6-(3-chlorobenzylamino)purine

After dissolving 3 mmol of 2-methyl-6-chloropurine obtained in Example 5 in 20 ml of butanol, 4 mmol of 3-chlorobenzylamine and 6 mmol of triethylamine were added. The mixture was reacted at 90°C for 4 hours. After cooling, the product was obtained by filtration, washed with water and butanol and crystallized from dimethylformamide or ethanol. HPLC: more than 98% purity. Yield 93%.

Table (4) is manufactured with the compound of embodiment 6 method

Example 7

2-(3-Chlorobenzylamino)-6-aminopurine

After dissolving 4 mmol of 2-chloro-6-aminopurine in 20 ml of butanol, 5 mmol of 3-chlorobenzylamine and 6 mmol of triethylamine were added. The mixture was reacted at 90°C for 4 hours. After cooling, the product was obtained by filtration, washed with water and butanol and crystallized from dimethylformamide or ethanol. HPLC: more than 95% purity. Yield 92%.

Table (5) is manufactured with the compound of embodiment 7 methods

Example 8

2-(3-Chlorobenzylamino)-6-hydroxypurine

After dissolving 4 mmol of 2-chloro-6-hydroxypurine in 20 ml of butanol, 5 mmol of 3-chlorobenzylamine and 6 mmol of triethylamine were added. The mixture was reacted at 90°C for 4 hours. After cooling, the product was obtained by filtration, washed with water and butanol and crystallized from dimethylformamide or ethanol. HPLC: greater than 91% purity. Yield 88%.

Table (6) is manufactured with the compound of embodiment 8 method

AMPK activity analysis

The effects of target compounds on AMPK activation were analyzed in mouse muscle cells C2C12, mouse fibroblasts 3T3-L1 and human liver cancer cells Hep G2. Cells were cultured in high-glucose DMEM containing 10wt% fetal bovine serum (FBS), 4mM L-glutamine, 2mM sodium pyruvate, and 1wt% penicillin/streptomycin. Streptomycin) (Invitrogen) were cultured at 37°C and 5% (v/v) CO ₂ . 3×10 ⁵ cells were seeded in 6-well plates, and after 24 hours, the cells were treated with the indicated compounds for 30 minutes, and then the cells were lysed and analyzed by Western blot. Equal amounts of proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis followed by transfer to polyvinylidene fluoride membranes. The transferred polyvinylidene fluoride membrane was soaked in 3wt% bovine serum albumin dissolved in PBS buffer for 60 minutes, and anti-phosphorylated AMPK (Thr172) antibody (1:2000 (v/v), Cellsignaling ), anti-AMPK antibody (1:2000 (v/v), Cell signaling), anti-glucose transport protein-4 antibody (1:1000, Millipore) or anti-motor protein antibody (1:5000 (v/v) ); Merck) at 4 ° C. After 16 hours, the corresponding secondary antibody was added and reacted for 1 hour at room temperature. The immunoreactive bands were detected with luminescent substrates, and the signals were recorded with film. The obtained signal was scanned and analyzed with image analysis and quantification software (TotalLab Quant) (TotalLab).

The effects of various compounds on AMPK activation are summarized in Table 7. Most of the tested compounds significantly activated AMPK in mouse muscle cells C2C12, mouse fibroblasts 3T3-L1 and human hepatoma cells Hep G2 cells.

Table (7)

Glucose Uptake - In Vitro Assay

Fluorescent glucose analogs (2-NBDG, Molecular Probes) were used to analyze the effects of representative compounds on glucose uptake in muscle cells C2C12. After C2C12 cells were treated with the selected new AMPK activator at 37°C for 30 minutes, 500 μM fluorescent glucose analogue was added, and after incubation at room temperature for 5 minutes, the cells were washed three times with phosphate buffer solution and washed with 70% (v/ v) Ethanol fixation. Fluorescence of intracellular glucose analogues was detected with a fluorometer.

The impact of some selected compounds on glucose uptake is shown in Table 8. Most of the compounds tested significantly enhanced glucose uptake in C2C12 cells. Data are expressed as mean ± standard deviation of three independent experiments.

Table (8)

in vivo testing

In order to further evaluate the effects of representative compounds on the regulation of plasma glucose levels, mice were fed with high-fat diet as an animal model of type 2 diabetes. C57BL/6J mice were housed at 22°C on a 12-hour day/night cycle and fed either a high-fat diet (60% kcal % fat) or normal diet in an ad libitum manner. 0.1-50 mg/kg of selected compounds were administered intraperitoneally to 24-week-old mice, and blood glucose levels were measured 1 and 3 hours after injection. The mice were fed intraperitoneally with high-fat feed twice a day for 6 days. One hour after the last administration, the plasma was collected and the plasma glucose and triglyceride levels were measured.

Selected compounds were found to reduce plasma glucose by more than 30 wt%, lower triglycerides by more than 35 wt%, and reduce body weight by more than 15% compared to high-fat diet-fed mice administered saline.

The above descriptions are only preferred embodiments of the present invention, and are not intended to limit the scope of patent protection of the present invention. Other equivalent changes made by using the spirit of the patent of the present invention should also fall within the scope of patent protection of the present invention.

Claims (6)

1. a kind of composition of activation AMPK, which is characterized in that the composition of the activation AMPK includes at least one effective agent The compound of amount, its tautomer, racemic modification, optical isomer and/or it is acceptable pharmaceutically or in nutrition Salt and pharmaceutical carrier, to give the mammal that need to activate AMPK, wherein the compound is 2- anilino- -6- amido purine.

2. composition as described in claim 1, wherein the composition is used to increase the intake of grape cell sugar, to treat Physiological status selected from the group being made up of：Pre-diabetes, the second patients with type Ⅰ DM, X- syndromes and metabolic syndrome.

3. composition as described in claim 1, wherein the composition is used to reduce the blood plasma triglyceride of mammal And weight is reduced, to treat obesity.

4. the compound of activation AMPK a kind of and/or its pharmaceutically or in nutrition acceptable salt preparing for treat can Purposes in the drug of the disease or physiological status that are improved by AMPK activators, which is characterized in that the purposes is consisting of the following： By the compound of the activation AMPK and/or its acceptable salt pharmaceutically or in nutrition, with 0.1 mg/kg to 50 Mg/kg dosage give need this treat mammal, wherein detecting the grape cell Sugar intake of the mammal Increase and blood plasma triglyceride reduction, wherein the compound is selected from at least one of group being made up of：Gland Purine, 2- anilino- -6- amidos purine, the chloro- hypoxanthines of 2-, 2- bromines adenine, Guanine, hypoxanthine, 5- methyl Cytimidine, thymidine, cytimidine, the close pyridine of urine, 5,6- dihydros urinate close pyridine.

5. purposes as claimed in claim 4, wherein giving at least one compound of the mammal and/or its medicine Acceptable salt increases the intake of the grape cell sugar of the mammal on or in nutrition, to which treatment is selected from by following The physiological status of the group of composition：Pre-diabetes, the second patients with type Ⅰ DM, X- syndromes and metabolic syndrome.

6. purposes as claimed in claim 4, wherein giving at least one compound of the mammal and/or its medicine Acceptable salt reduces the blood plasma triglyceride of the mammal and reduces the mammal on or in nutrition Weight, to treat obesity.