CN102786517B - The pyrimidine thiazole amines derivative of GK and PPAR double excitation activity - Google Patents

Abstract

The invention discloses a new class of pyrimidinethiazole amine derivatives, its preparation method, pharmaceutical composition and application. Specifically, it relates to pyrimidinethiazole amine derivatives shown in general formula I, pharmaceutically acceptable salts thereof, and preparation methods thereof, compositions containing one or more of these compounds, and such compounds in the treatment of glucokinase and The use of the peroxisome proliferator-activated receptor-related diseases such as diabetes and obesity.

Description

Pyrimidine Thiazolamine Derivatives with Dual Agonistic Activities of GK and PPAR

field of invention

The present invention relates to pyrimidine thiazole amine derivatives represented by general formulas I and II, their pharmaceutically acceptable salts, their precursors or derivatives with the same biological function, their preparation methods, and compositions containing one or more of these compounds , and the use of the compounds in treating diseases related to glucokinase and peroxisome proliferator-activated receptors, such as diabetes and obesity.

Background of the invention

Sugar is an important energy and carbon source for organisms. The decomposition of sugar produces energy, which can supply the needs of the organism's life activities, and the intermediate products of sugar metabolism can be converted into other carbon-containing compounds, such as amino acids, fatty acids, and nucleosides. Sugar metabolism can be divided into sugar decomposition and sugar synthesis. Sugar catabolism includes glycolysis, the common breakdown pathway for sugars; and the tricarboxylic acid cycle, the final oxidation pathway for sugars. Glycolysis is the enzymatic degradation of glucose to pyruvate with concomitant generation of ATP. It is a common metabolic pathway for the decomposition of glucose in animal, plant, and microbial cells to generate energy. In aerobic organisms, pyruvate produced by glycolysis enters the mitochondria and is completely oxidized into CO ₂ and water through the tricarboxylic acid cycle; NADH produced by glycolysis is oxidized by the respiratory chain to produce ATP and water. Therefore, glycolysis is the prelude to oxidative phosphorylation and the Krebs cycle. Glucokinase (GK) is one of four hexokinases found in mammals. Gallery Cofowick, SPThe Enzymes, Vol.9 (P.Boyer'ed.) Academic Press, New York, N Y .1-48 pages, 1973], is the first key enzyme in the glycolysis pathway, it can phosphorylate glucose into glucose 6-phosphate in the presence of ATP, and enter the downstream metabolic process. Therefore, this step is the first rate-limiting step in the whole glucose metabolism process, and GK is the first rate-limiting enzyme in the process of glucose metabolism, which plays an important biological role.

The cellular distribution of GK is limited, mainly found in pancreatic β-cells and liver tissue cells. In addition, GK plays an important role in controlling blood sugar balance and metabolism. On the one hand, it regulates glycogen metabolism. When fasting or low blood sugar, GK activity is low, and glycogen output increases to ensure the energy supply of vital organs; When it is high, GK activity is enhanced, thereby promoting liver glycogen synthesis, inhibiting liver gluconeogenesis, and maintaining blood sugar homeostasis. On the other hand, as a sensor of glucose, it regulates the secretion of insulin. When the blood glucose concentration in the body is higher than the normal value, the glucose in the blood is transported by the glucose transporter 2 (GLUT2) into the pancreatic β-cell, and phosphorylated under the action of GK to generate glucose-6-phosphate. The glycolysis and oxidative metabolism of glucose increase the ratio of ATP/ADP, the K+ ion channel is closed, the membrane is depolarized, the voltage-sensitive Ca2+ channel is opened, Ca2+ flows in, the insulin storage vesicle fuses with the plasma membrane, and insulin is released into the blood. And enter the liver, fat and muscle cells with the blood circulation to participate in the regulation of blood sugar. Glucose in the liver is phosphorylated into -6-phosphate glucose under the action of GK, and then synthesizes liver glycogen under the action of insulin. In adipose and muscle cells, insulin triggers the migration of glucose transporter 4 (GLUT4) from intracellular storage vesicles to the plasma membrane to facilitate glucose uptake and metabolism. GK lowers blood sugar by regulating insulin release and liver glucose metabolism, and plays an important role in maintaining blood sugar homeostasis [Al-Hasani H; Tschopl M H. Mol Interv, 2003, 3(7), 367-370]. [Chipkin, S.R., Kelly, K.L. and Ruderrnam, N.B. In Joslin' Diabetes (eds. C.R. Khan and G.C. Wier), Lea and Febiger, Philadelphia, PA, pp. 97-115, 1994].

Diabetes of youth onset diabetes (MODY) is caused by loss-of-function mutations in the GK gene, suggesting that GK also functions as a glucose sensor in humans Liang, Y, Kesavan, P., Wang, L. et al., Biochem.J. 309, 167-173, 1995). In addition to MODY, decreased GK activity is also common in individuals with diabetes in general. Further studies on the pathogenesis of type 2 diabetes have found that decreased liver GK activity may be involved in the mechanism of insulin resistance, resulting in elevated blood sugar, impaired islet function and increased insulin resistance. Therefore, looking for drugs that can increase the activity of GK to prevent or delay the development of the disease may open up a new way for early prevention and treatment of diabetes.

In recent years, it has been found that pancreatic β-glucokinase is expressed only in the rat brain, particularly in the feeding center (ventromedial nucleus of the hypothalamus, VMH). About 20% of the nerve cells in the VMH are called glucose-responsive neurons (glucose-responsive neutrons), which were previously thought to play an important role in weight control. Administration of glucose into the brain of rats reduces the food intake, and administration of glucosamine, an analog of glucose, to the brain of rats inhibits glucose metabolism, resulting in hyperphagia. Electrophysiological experiments have shown that glucose-responsive neurons are activated corresponding to changes in physiological glucose concentration (5-20 mM), but their activity is inhibited by inhibiting glucose metabolism by glucosamine or the like. It is presumed that the glucose concentration sensing system of the VHM is through the same mechanism as the insulin secretion of pancreatic β cells via glucokinase. Therefore, in addition to liver and pancreatic β cells, substances that activate the glucokinase activity of VHM not only have the effect of regulating blood sugar, but may also regulate the obesity that many patients with type 2 diabetes suffer from. As can be seen from the above description, compounds having glucokinase activation can be used as therapeutic and/or preventive drugs for diabetes, or chronic complications of diabetes such as retinopathy, nephropathy, neurosis, ischemic heart disease, and arteriosclerosis. Therapeutic and/or preventive medicine for obesity, and further can be used as therapeutic and/or preventive medicine for obesity.

Many GK small molecule activators have been found, with different structural features: such as substituted phenylacetamides (WO0058293WO0185706WO208209WO185707WO0183465WO246173WO2004072066WO0246173), substituted hydantoins (WO0183478), substituted pyrroles (WO209611254 US0067939WO031179), isoindoline substituted propionamides (WO0248106), substituted anthranilamides (WO03080585), substituted α-phenylacrylamides (WO0214312), etc. Among them, phenylacetamides are the most researched of a class. Although the research on GK activators mentioned above has made great contributions to this field, in order to improve the structure of compounds and the activity of GK activators, this field is still continuing to study.

Peroxisome proliferator-activated receptors (PPARs) belong to the nuclear hormone receptor superfamily, and there are three subtypes (PPARα, PPARγ, PPARδ). The three receptors function as lipid sensors, coordinately regulate the expression of various gene sequences, and regulate important body metabolism. PPARs have a variety of biological effects, which can promote adipocyte differentiation and lipogenesis, enhance the body's sensitivity to insulin, and mediate glucose balance in the body. They are effective drug targets for the treatment of metabolic diseases (such as type II diabetes and atherosclerosis). Moreover, it also inhibits the production of inflammatory factors and inflammatory response, affects tumor growth, and has a protective effect on cardiovascular. Studies in recent years have shown that PPARs also have neuroprotective effects, which can reduce the damage of nerve cells in neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, cerebral ischemia and multiple sclerosis.

PPARγ is different from the classic nuclear receptors. After being activated by specific ligand binding, PPARγ cannot directly recognize the binding specific DNA regulatory sequence, and must form a heterodimer with retinoid X receptor (RXR). , under the action of many co-regulators (cofactors), regulate specific gene transcription. Early studies referred to PPARγ ligands that activate gene transcription as PPARγ agonists, and those that inhibit gene transcription as inhibitors. The latest research progress shows that the specific conformation formed by the combination of PPARγ and different ligands determines the selective binding of PPARγ-RXR heterodimer and specific cofactors, and further determines the selective transcriptional regulation of different genes. It is precisely because of the selectivity of this structure and interaction that different PPARγ ligands produce different biological effects through selective gene transcription regulation. In terms of the complexity of gene regulation, the same PPARγ ligand can activate or inhibit the transcription of target genes. Therefore, more and more scholars use "modulator" instead of the one-sided definition of "agonist/inhibitor". , and then some PPARγ ligands known to selectively act on the transcriptional regulation of specific genes are called "selective PPARγ modulators" (selective PPARγ modulator, SPPARM). If some small molecular compounds activate PPARγ, and then interact with different cofactors, the fat differentiation process can be separated from the glucose and lipid metabolism process at the gene regulation level, that is, some PPARγ agonists that promote glucose and lipid metabolism do not induce fat differentiation.

The current research on PPARγ regulators is mainly focused on two issues: one is how much activation activity we need PPARγ ligands to achieve the purpose of moderately activating PPARγ and regulating the transcription of target genes; Differentiation and insulin sensitization dissociated? For the compounds or substances under study, the molecular mechanism of PPARγ regulation should be deeply explored, and the PPARγ regulators that have insulin-sensitizing effects without affecting fat differentiation or even promoting fat metabolism should be screened.

Because PPARα/δ is involved in regulating lipid metabolism, improving hyperlipidemia, and to a certain extent, reducing fat differentiation induced by PPARγ activation; PPARδ is more extensive than PPARα in tissue distribution, and research on PPARδ has also attracted attention in recent years. Researchers are committed to the development of PPARα/γ dual agonists or α/δ/γ triple agonists, hoping that these drugs can lower blood sugar and regulate blood lipids, and target obesity, insulin resistance, and metabolic syndrome. The current research results in animals and clinical experiments suggest that PPARα/γ dual agonists or α/δ/γ triple agonists can indeed improve insulin resistance and dyslipidemia, but due to the lack of drug safety and toxicity experimental evidence, this type of The application prospects of the compound remain to be seen.

Diabetes mellitus is a systemic chronic metabolic disease, and its pathological feature is mainly hyperglycemia. It is generally believed that the main pathological changes leading to hyperglycemia include the reduction of insulin secretion, the weakening of its action and the disorder of hepatic glucose metabolism (also known as the three major pathological changes). The activation of glucokinase (GK) can simultaneously promote insulin secretion and liver glucose metabolism; the activation of peroxisome proliferator-activated receptor (PPAR) can increase the sensitivity of body cells to insulin. Therefore, aiming at the three major pathological changes of hyperglycemia in diabetes, based on the structure and pharmacological effects of known GK and PPAR small molecule agonists, construct and synthesize GK and PPAR dual-target ligand compounds, and research and develop a compound that can improve insulin resistance at the same time. Secretion, glycogen metabolism and the promotion of insulin sensitivity in peripheral tissues will become a new way to find universally effective new drugs for the treatment of diabetes.

Contents of the invention

The present invention provides a pyrimidine thiazole amine derivative of the general structural formula I that can be used as a glucokinase activator and as a peroxisome proliferator-activated receptor agonist:

The object of the present invention is to provide a novel pyrimidinethiazolamine derivative with dual agonistic activity of GK and PPAR, and a pharmaceutically acceptable salt thereof.

Another object of the present invention is to provide a method for preparing novel pyrimidinethiazole amine derivatives.

Another object of the present invention is to provide a pharmaceutical composition containing one or more of these compounds.

Another object of the present invention is to propose a new concept of multi-target drug combination for treating diabetes based on the biological functions of GK and PPAR.

Still another object of the present invention is to provide GK and PPAR double ligand compounds, and the use of such compounds in the treatment of diseases related to GK and PPAR.

The present invention relates to pyrimidine thiazole amine derivatives having the following general formula I:

In the formula

R1 is selected from hydrogen, halogen, amino, hydroxyl, _COOC1-8 alkyl, carboxyl, aminoacyl, C1-8 alkyl substituted aminoacyl, C1-8 alkyl, carboxymethylthio, hydroxymethyl, carboxyethyl ;

Preferred R is selected from hydrogen, fluorine, chlorine, bromine, amino, _COOC1-4 alkyl, carboxyl, aminoacyl, C1-4 alkyl substituted aminoacyl, C1-4 alkyl, carboxymethylthio, hydroxymethyl , carboxyethyl;

More preferred R is selected from the group consisting _of hydrogen, fluorine, chlorine, bromine, amino, carbocarboxylate, carbocarboxylate, carbocarboxylate, carboxyl, carboxyl, aminoacyl, carbamoyl, isopropanyl, cyclopropylmethyl Aminoacyl, methyl, ethyl, carboxymethylthio, hydroxymethyl, carboxyethyl;

Most preferred R is selected from the group consisting _of hydrogen, fluorine, chlorine, carbocarboxylate, carbocarboxylate, carbomethoxy, carboxyl, aminoacyl, carbamoyl, isopropanyl, cyclopropylcarboxamide, methyl, carboxymethyl Thio, hydroxymethyl.

R2 is selected from _hydrogen , halogen, amino, hydroxyl, COOC1-8 alkyl, carboxyl, aminoacyl, C1-8 alkyl substituted aminoacyl, C1-8 alkyl, carboxymethylthio, hydroxymethyl, carboxyethyl ;

Preferred R is selected from _hydrogen , fluorine, chlorine, bromine, amino, COOC1-4 alkyl, carboxyl, aminoacyl, C1-4 alkyl substituted aminoacyl, C1-4 alkyl, carboxymethylthio, hydroxymethyl , carboxyethyl;

More preferred R is selected from the group consisting of _hydrogen , fluorine, chlorine, bromine, amino, carbocarboxylate, carbocarboxylate, carbocarboxylate, carboxyl, carboxyl, aminoacyl, carbamoyl, isopropanyl, cyclopropylmethyl Aminoacyl, methyl, ethyl, carboxymethylthio, hydroxymethyl, carboxyethyl;

Most preferred R is selected from the group consisting of _hydrogen , fluorine, chlorine, carboethoxy, carbocarboxylate, carbomethoxy, carboxyl, aminoacyl, carbamoyl, isopropanyl, cyclopropylcarboxamide, methyl, carboxymethyl Thio, hydroxymethyl.

_R is selected from hydrogen, halogen, C1-8 alkylamino, arylamino, C1-8 alkoxy, aryloxy, C1-8 alkylthio, arylthio, nitro, cyano,

Preferred _R is selected from hydrogen, chlorine, C1-6 alkylamino, arylamino, C1-6 alkoxy, aryloxy, C1-6 alkylthio, phenylthio, pyridylthio, nitro, cyano ,

More preferred _R is selected from the group consisting of hydrogen, chlorine, methylamino, ethylamino, isopropylamino, cyclopropylmethylamino, aniline, pyridylamino, methoxy, ethoxy, butoxy, phenoxy, picolyl Oxygen, methylthio, phenylthio, pyridylthio, nitro, cyano,

Most preferred _R is selected from ethylamino, isopropylamino, cyclopropylmethylamino, aniline, pyridylamino, ethoxy, butoxy, pyridylmethoxy,

_R4 is selected from hydrogen, methyl, trifluoromethyl, fluorine or chlorine, methoxy, methylthio, methylamino.

Preferred _R4 is selected from hydrogen, methyl, trifluoromethyl, fluorine or chlorine, methoxy, methylthio.

More preferred _R4 is selected from hydrogen, methyl, trifluoromethyl, fluoro or chloro, methoxy.

Most preferred _R4 is selected from hydrogen, methyl, trifluoromethyl.

Preferred aryl groups are selected from phenyl, pyridyl.

In order to prepare the compound described in the general formula I of the present invention, according to the structure of the general formula I, the preparation of the compound of the general formula I in the present invention will be divided into two routes.

In the first route, the thiazolamine derivative 1 is used as a raw material, reacted with a dichloropyrimidine derivative 2 under basic conditions to generate an intermediate compound 3, and then reacted with _R3 to obtain a product 4.

In the second route, the halogenated thiazole derivative 5 was used as a raw material to react with the pyrimidinamine derivative 6 in an alkaline environment to generate the target compound 7.

In addition, the starting materials and intermediates in the above reactions are easy to obtain, and each step of the reaction can be easily synthesized according to the reported literature or by a person skilled in the art using conventional methods in organic synthesis. The pharmaceutically acceptable salts described in general formula I include different acid addition salts, such as the acid addition salts of the following inorganic acids or organic acids: hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, methanesulfonic acid, p-toluenesulfonic acid, Trifluoroacetic acid, Lycic acid, Maleic acid, Tartaric acid, Fumaric acid, Citric acid, Lactic acid. Pharmaceutically acceptable salts described in general formula I also include different alkali metal salts (lithium, sodium, potassium salts), alkaline earth metal salts (calcium, magnesium salts) and ammonium salts, and organic compounds that can provide physiologically acceptable cations. Salts of bases such as methylamine, dimethylamine, trimethylamine, piperidine, morpholine and tris(2-hydroxyethyl)amine. All such salts within the scope of this invention may be prepared by conventional methods.

The present invention also relates to pharmaceutical compositions containing the compound of the present invention as an active ingredient. The pharmaceutical composition can be prepared according to methods known in the art. Any dosage form suitable for human or animal use can be prepared by combining the compounds of the present invention with one or more pharmaceutically acceptable solid or liquid excipients and/or adjuvants. The content of the compound of the present invention in its pharmaceutical composition is usually 0.1-95% by weight.

The compound of the present invention or the pharmaceutical composition containing it can be administered in the form of a unit dosage, and the route of administration can be enteral or parenteral, such as oral, intravenous injection, intramuscular injection, subcutaneous injection, nasal cavity, oral mucosa, eye, lung and Respiratory tract, skin, vagina, rectum, etc.

The dosage form for administration may be a liquid dosage form, a solid dosage form or a semi-solid dosage form. Liquid dosage forms can be solutions (including true solutions and colloid solutions), emulsions (including o/w type, w/o type and double emulsion), suspensions, injections (including aqueous injections, powder injections and infusion solutions), eye drops Agents, nasal drops, lotions and liniments, etc.; solid dosage forms can be tablets (including ordinary tablets, enteric-coated tablets, buccal tablets, dispersible tablets, chewable tablets, effervescent tablets, orally disintegrating tablets), capsules ( Including hard capsules, soft capsules, enteric-coated capsules), granules, powders, pellets, dripping pills, suppositories, films, patches, gas (powder) aerosols, sprays, etc.; semi-solid dosage forms can be ointments, Gels, pastes, etc.

The compound of the present invention can be made into common preparations, sustained-release preparations, controlled-release preparations, targeted preparations and various microparticle drug delivery systems.

To form the compound of the present invention into tablets, various excipients known in the art can be widely used, including diluents, binders, wetting agents, disintegrants, lubricants, glidants. Diluents can be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate, calcium carbonate, etc.; wetting agents can be water, ethanol, iso Propanol, etc.; binders can be starch slurry, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, arabic mucilage, gelatin slurry, sodium carboxymethylcellulose, methylcellulose, hypromellose Base cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone, polyethylene glycol, etc.; disintegrants can be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, cross-linked poly Vinylpyrrolidone, croscarmellose sodium, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitan fatty acid ester, sodium dodecylsulfonate, etc.; lubricant and flow aid The agent can be talc, silicon dioxide, stearate, tartaric acid, liquid paraffin, polyethylene glycol and the like.

Tablets can also be further made into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or double-layer tablets and multi-layer tablets.

In order to make the administration unit into a capsule, the active ingredient compound of the present invention can be mixed with a diluent and a glidant, and the mixture can be directly placed in a hard capsule or a soft capsule. The active ingredient compound of the present invention can also be made into granules or pellets with diluents, binders, and disintegrants, and then placed in hard capsules or soft capsules. Various diluents, binders, wetting agents, disintegrants, and glidants used in the preparation of tablets of the compound of the present invention can also be used in the preparation of capsules of the compound of the present invention.

In order to make the compound of the present invention into injection, water, ethanol, isopropanol, propylene glycol or their mixtures can be used as solvent and an appropriate amount of commonly used solubilizers, cosolvents, pH regulators and osmotic pressure regulators in this field can be added. The solubilizer or co-solvent can be poloxamer, lecithin, hydroxypropyl-β-cyclodextrin, etc.; the pH regulator can be phosphate, acetate, hydrochloric acid, sodium hydroxide, etc.; the osmotic pressure regulator can be Sodium chloride, mannitol, glucose, phosphate, acetate, etc. For preparation of freeze-dried powder injection, mannitol, glucose, etc. can also be added as proppants.

In addition, coloring agents, preservatives, fragrances, flavoring agents or other additives can also be added to the pharmaceutical preparations, if necessary.

In order to achieve the purpose of medication and enhance the therapeutic effect, the medicine or pharmaceutical composition of the present invention can be administered by any known administration method.

The dosage of the pharmaceutical composition of the compound of the present invention can vary widely depending on the nature and severity of the disease to be prevented or treated, individual conditions of the patient or animal, administration route and dosage form, etc. Generally, the suitable daily dosage range of the compound of the present invention is 0.001-150 mg/Kg body weight, preferably 0.01-100 mg/Kg body weight. The above-mentioned dosage can be administered in one dosage unit or divided into several dosage units, depending on the clinical experience of the doctor and the dosage regimen including the use of other therapeutic means.

The compound or composition of the present invention can be taken alone, or used in combination with other therapeutic drugs or symptomatic drugs. When the compound of the present invention has a synergistic effect with other therapeutic drugs, its dose should be adjusted according to the actual situation.

The compound of the present invention is a dual activator of GK and PPAR or its precursor, which can lower blood sugar by regulating insulin release, tissue sensitivity to insulin and liver glucose metabolism, and can be used to prevent and treat type 1 or type 2, especially type 2 Type 2 diabetes and related complications, or other diseases related to GK and PPAR.

detailed description

The invention will be further described below in conjunction with the examples, but the scope of the invention is not limited.

Measuring instrument: Vaariaan Mercury 300 nuclear magnetic resonance instrument for nuclear magnetic resonance spectroscopy. Mass spectrometry uses ZAD-2F and VG300 mass spectrometers.

Example 1: 2-(4-(2-methyl-6-(4-methylthiazole-2-amino)pyrimidine-4-amino)phenoxy)ethyl acetate

a: Preparation of N-(6-chloro-2methylpyrimidin-4-yl)-4-methylthiazol-2-amine:

Add 2-methyl-4,6-dichloropyrimidine (16.3g, 0.1mol), 2-amino-4-methylthiazole (11.4g, 0.1mol) into DMF (250mL), stir in ice bath, 60% NaH (8 g, 0.2 mol) was added to the reaction solution in batches, and reacted at room temperature for 6 h. After the reaction was complete, glacial acetic acid (15 mL) was added to terminate the reaction. The reaction solution was poured into 1L saturated sodium bicarbonate ice-water bath, a large amount of solids precipitated, filtered, washed with water, and the filter cake was dried and recrystallized to obtain N-(6-chloro-2methylpyrimidin-4-yl)-4-methanol Thiazol-2-amine 16.5 g.

b: Preparation of ethyl 2-(4-(2-methyl-6-(4-methylthiazole-2-amino)pyrimidine-4-amino)phenoxy)acetate:

N-(6-chloro-2methylpyrimidin-4-yl)-4-methylthiazol-2-amine (2.4g, 10mmol), 2-(4-aminophenoxy)-ethyl acetate (1.75 g, 10mmol), NaI (50mg) was added to DMF (20mL), reacted at 150°C for 36h, the reaction was complete, cooled to room temperature, evaporated the solvent under reduced pressure, added ethyl acetate (200mL), washed with water successively, saturated chlorination Washed with sodium, dried over anhydrous sodium sulfate, concentrated, and column chromatographed to obtain 1.32 g of a white solid. ¹ H NMR (DMSO-d ₆ , 300 MHz), δ (ppm): 11.15 (s, 1H, -NH-), 9.20 (s, 1H, -NH-), 7.52 (d, 2H, ArH), 6.99 ( d, 2H, ArH), 6.67 (d, 1H, ArH), 6.23 (s, 1H, ArH), 4.83 (s, 2H, -CH ₂ O-), 4.27 (q, 2H, Me-CH ₂ -) , 2.60 (s, 3H, -CH ₃ ), 2.32 (s, 3H, -CH ₃ ), 1.32 (t, 3H, -CH ₃ ). MS (FAB): 400 (M+1)

Example 2: 2-(4-(2-methyl-6-(4-methylthiazole-2-amino)pyrimidine-4-amino)phenoxy)acetic acid

Add ethyl 2-(4-(2-methyl-6-(4-methylthiazole-2-amino)pyrimidine-4-amino)phenoxy)acetate (300 mg) to methanol (10 mL) and 2.5N NaOH (15mL), stirred at 60°C for 2h, cooled to room temperature, added water (30mL), extracted with diethyl ether (30mL), adjusted the pH of the aqueous layer to 2 with concentrated hydrochloric acid, a large amount of white soil was precipitated, filtered, and dried the filter cake to obtain White solid 255mg. ¹ H NMR (DMSO-d ₆ , 300 MHz), δ (ppm): 12.59 (s, 1H, COOH), 11.11 (s, 1H, -NH-), 9.17 (s, 1H, -NH-), 7.62 ( d, 2H, ArH), 6.98(d, 2H, ArH), 6.65(d, 1H, ArH), 6.23(s, 1H, ArH), 4.83(s, 2H, _-CH2O- ), 2.55(s , 3H, -CH ₃ ), 2.30 (s, 3H, -CH ₃ ). MS (FAB): 372 (M+1).

Example 3: 2-(3-(2-methyl-6-(4-methylthiazole-2-amino)pyrimidine-4-amino)phenoxy)ethyl acetate

The operation steps were the same as in Example 1 to obtain a white solid (585 mg, 29%). ¹ H NMR (DMSO-d ₆ , 300MHz), δ (ppm): 11.12 (s, 1H, -NH-), 9.33 (s, 1H, -NH-), 7.38 (s, 1H, ArH), 7.20- 7.09(m, 2H, ArH), 6.51(d, 1H, ArH), 6.27(s, 1H, ArH), 4.74(s, 2H, -CH ₂ O-), 4.17(q, 2H, Me-CH ₂ -), 2.43 (s, 3H, -CH ₃ ), 2.22 (s, 3H, -CH ₃ ), 1.20 (t, 3H, -CH ₃ ). MS (FAB): 400 (M+1)

Example 4: 2-(3-(2-methyl-6-(4-methylthiazole-2-amino)pyrimidine-4-amino)phenoxy)acetic acid

The operation steps were the same as in Example 2 to obtain a white solid (260 mg, 95%). ¹ H NMR (DMSO-d ₆ , 300 MHz), δ (ppm): 11.17 (s, 1H, -NH-), 9.31 (s, 1H, -NH-), 7.33 (s, 1H, ArH), 7.15 ( t, 1H, ArH), 7.07(d, 1H, ArH), 6.60(s, 1H, ArH), 6.49(d, 1H, ArH), 6.27(s, 1H, ArH), 4.55(s, 2H, - CH ₂ O-), 2.42(s, 3H, -CH ₃ ), 2.17(s, 3H, -CH ₃ ). MS(FAB): 372(M+1).

Example 5: 2-methyl-2-(4-(2-methyl-6-(4-methylthiazole-2-amino)pyrimidine-4-amino)phenoxy)ethyl propionate

The operation steps were the same as in Example 1 to obtain 585 mg of white solid. ¹ H NMR (DMSO-d ₆ , 300MHz), δ (ppm): 12.85 (s, 1H, COOH), 11.60 (s, 1H, -NH-), 9.20 (s, 1H, -NH-), 7.96 ( s, 1H, ArH), 7.31(m, 3H, ArH), 6.73(d, 2H, ArH), 6.10(s, 1H, ArH), 2.35(s, 3H, -CH ₃ ), 1.38(s, 6H , -CH ₃ ×2). MS (FAB): 414 (M+1).

Example 6: Ethyl 2-methyl-2-(4-(2-methyl-6-(5-ethoxyformylthiazole-2-amino)pyrimidine-4-amino)phenoxy)propionate

The operation steps were the same as in Example 1 to obtain 556 mg of white solid. ¹ H NMR (DMSO-d ₆ , 300 MHz), δ (ppm): 11.70 (s, 1H, -NH-), 9.30 (s, 1H, -NH-), 8.03 (s, 1H, ArH), 7.20 ( d, 2H, ArH), 6.79 (d, 2H, ArH), 6.20 (s, 1H, ArH), 4.33-4.16 (m, 4H, Me-CH ₂ -×2), 2.45 (s, 3H, -CH ₃ ), 1.45(s, 6H, -CH ₃ ×2), 1.28(t, 3H, -CH ₃ ), 1.19(t, 3H, -CH ₃ ). MS(FAB): 486(M+1).

Example 7: 2-(4-(2-methyl-6-(4-methylthiazol-2-amino)pyrimidin-4-yl)piperazin-1-yl)ethanol

a: The preparation of N-(6-chloro-2methylpyrimidin-4-yl)-4-methylthiazol-2-amine is the same as in Example 1.

b: Preparation of 2-(4-(2-methyl-6-(4-methylthiazol-2-amino)pyrimidin-4-yl)piperazin-1-yl)ethanol:

N-(6-chloro-2methylpyrimidin-4-yl)-4-methylthiazol-2-amine (2.4g, 10mmol), hydroxyethylpiperazine (1.73g, 10mmol), DIPEA (1.94g , 15mmol), DMSO (5mL) was added into dioxane (60mL), reacted at 110°C for 10h, and the reaction was complete. The reaction solution was poured into water (200 mL), stirred for 1 h, and filtered. The filter cake was dried and recrystallized from ethyl acetate to obtain a white solid (1.7g, 50%). ¹ H NMR (DMSO-d ₆ , 300MHz), δ (ppm): 10.98 (s, 1H, -NH-), 6.55( s, 1H, ArH), 5.98(s, 1H, ArH), 3.53-3.47(m, 6H, -CH ₂ ×3), 2.47-2.39(m, 6H, CH ₂ ×3), 2.34(s, 3H , -CH ₃ ), 2.21 (s, 3H, -CH ₃ ). MS (FAB): 335 (M+1).

Example 8: 2-(4-(2-Methyl-6-(5-ethoxyformylthiazole-2-amino)pyrimidin-4-yl)piperazin-1-yl)ethanol

a: Preparation of ethyl 2-(6-chloro-2-methylpyrimidin-4-amino)thiazole-5-carboxylate:

2-Methyl-4,6-dichloropyrimidine (16.3g, 0.1mol), ethyl 2-aminothiazole-5-carboxylate (17.2g, 0.1mol) were added to DMF (250mL), stirred in an ice bath , 60% NaH (8 g, 0.2 mol) was added to the reaction liquid in batches, and reacted at room temperature for 6 h, and the reaction was complete. Glacial acetic acid (15 mL) was added to quench the reaction. The reaction solution was poured into 1L saturated sodium bicarbonate ice-water bath, a large amount of solids were precipitated, filtered, washed with water, and the filter cake was dried to obtain 2-(6-chloro-2-methylpyrimidine-4-amino)thiazole-5- Ethyl formate (18.5 g, 65%) crude.

b: Preparation of 2-(4-(2-methyl-6-(5-ethoxyformylthiazol-2-amino)pyrimidin-4-yl)piperazin-1-yl)ethanol:

2-(6-Chloro-2-methylpyrimidine-4-amino)thiazole-5-carboxylic acid ethyl ester (149mg, 5mmol), hydroxyethylpiperazine (865mg, 5mmol), DIPEA (970mg, 7.5mmol) , DMSO (5 mL) was added into dioxane (60 mL), reacted at 110° C. for 10 h, and the reaction was complete. The reaction solution was poured into water (200 mL), stirred for 1 h, and filtered. The filter cake was dried and recrystallized from ethyl acetate to obtain 2-(4-(2-methyl-6-(5-ethoxyformylthiazole-2-amino)pyrimidin-4-yl)piperazin-1-yl)ethanol White solid. ¹ HNMR (DMSO-d ₆ , 300MHz), δ (ppm): 11.60 (s, 1H, -NH-), 8.03 (s, 1H, ArH), 7.04 (s, 1H, ArH), 4.43 ( m, 1H, OH), 4.29-4.22 (q, 2H, -CH ₂ ), 3.51 (m, 6H, CH ₂ ×3), 3.32 (s, 4H, -CH ₂ ), 2.43 (m, 2H, - CH ₂ ), 2.41 (s, 3H, -CH ₃ ), 1.28 (t, 3H, -CH ₃ ). MS (FAB): 393 (M+1).

Example 9: 2-(1-(2-Methyl-6-(5-ethoxyformylthiazole-2-amino)pyrimidin-4-yl)piperidin-4-yl)ethanol

The operation steps were the same as in Example 8 to obtain 256 mg of white solid. ¹ H NMR (DMSO-d ₆ , 400MHz), δ(ppm): 11.53(s, 1H, -NH-), 8.02(s, 1H, ArH), 6.04(s, 1H, ArH), 4.47(t, 1H, -OH), 4.25(m, 4H, -CH ₂ ×2), 3.53-3.50(m, 1H, -CH), 3.25(t, 2H, -CH ₂ ), 2.84(t, 2H, -CH ₂ ), 2.49-2.45 (m, 2H, -CH ₂ ), 2.39 (s, 3H, -CH ₃ ), 1.73-1.70 (m, 2H, -CH ₂ ), 1.27 (t, 3H, -CH ₃ ) , 1.11-1.02 (m, 2H, -CH ₂ ). MS (FAB): 392 (M+1).

Example 10: 2-(1-(2-Methyl-6-(5-ethoxyformylthiazole-2-amino)pyrimidin-4-yl)piperidin-4-yl)acetic acid

The operation steps were the same as in Example 8 to obtain 356 mg of white solid. ¹ H NMR (DMSO-d ₆ , 400MHz), δ(ppm): 11.53(s, 1H, -NH-), 8.02(s, 1H, ArH), 6.06(s, 1H, ArH), 4.25(q, 2H, -CH ₂ ), 4.13 (m, 2H, -CH ₂ ), 3.53-3.50 (m, 1H, -CH), 3.03-2.97 (m, 2H, -CH ₂ ), 2.55-2.45 (m, 2H , -CH ₂ ), 2.40(s, 3H, -CH ₃ ), 1.90-1.86(m, 2H, -CH ₂ ), 1.53-1.43(m, 2H, -CH ₂ ), 1.28(t, 3H, - CH ₃ ). MS (FAB): 406 (M+1).

Example 11: 2-((1-(2-Methyl-6-(5-ethoxyformylthiazole-2-amino)pyrimidin-4-yl)piperidin-4-yl)methoxy)acetic acid

The operation steps were the same as in Example 8 to obtain 342 mg of white solid. ¹ H NMR (DMSO-d ₆ , 300MHz), δ (ppm): 12.16 (s, 1H, COOH), 11.18 (s, 1H, -NH-), 7.63 (s, 1H, ArH), 5.66 (s, 1H, ArH), 3.88-3.83 (m, 2H, -CH ₂ ), 3.58 (s, 2H, -CH ₂ ), 2.92 (d, 2H, -CH ₂ ), 2.33 (s, 3H, -CH ₃ ) , 2.10 (m, 4H, -CH ₂ ), 1.46 (m, 1H, -CH), 1.37-1.33 (m, 2H, -CH ₂ ), 0.89 (t, 3H, -CH ₃ ), 0.74-0.71 ( m, 2H, -CH ₂ ). MS (FAB): 436 (M+1)

Example 12: 2-(4-(6-(5-isopropoxyformylthiazol-2-amino)pyrimidin-4-yl)piperazin-1-yl)ethanol

The operation steps were the same as in Example 8 to obtain 342 mg of white solid. ¹ H NMR (DMSO-d ₆ , 300MHz), δ(ppm): 11.66(s, 1H, -NH-), 8.41(s, 1H, ArH), 8.02(s, 1H, ArH), 6.20(s, 1H, ArH), 5.07(m, 1H, CH), 4.44(m, 1H, OH), 4.25(m, 2H, -CH ₂ -), 3.51(m, 6H, CH ₂ ×3), 2.48(m , 4H, CH ₂ ×2), 2.42 (m, 2H, -CH ₂ ), 1.28 (d, 6H, -CH ₃ ×2). MS (FAB): 393 (M+1)

Example 13: 2-(4-(6-(5-carboethoxythiazol-2-amino)pyrimidin-4-yl)piperazin-1-yl)ethanol

The operation steps were the same as in Example 8 to obtain 242 mg of white solid. ¹ H NMR (DMSO-d ₆ , 300MHz), δ(ppm): 11.68(s, 1H, -NH-), 8.41(s, 1H, ArH), 8.04(s, 1H, ArH), 6.21(s, 1H, ArH), 4.44 (m, 1H, OH), 4.28-4.22 (q, 2H, -CH ₂ ), 3.53-3.52 (m, 6H, CH ₂ × 3), 2.50 (m, 4H, -CH ₂ ), 2.44-2.40 (m, 2H, -CH ₂ ), 2.42 (t, 2H, -CH ₃ ). MS (FAB): 379 (M+1)

Example 14: 2-methyl-2-(4-(2-methyl-6-(5-carboxythiazole-2-amino)pyrimidine-4-amino)phenoxy)propanoic acid

The operation steps were the same as in Example 1 and Example 2 to obtain 223 mg of white solid. ¹ H NMR (DMSO-d ₆ , 300MHz), δ (ppm): 12.88 (s, 2H, COOH×2), 11.62 (s, 1H, -NH-), 9.27 (s, 1H, -NH-), 7.95(s, 1H, ArH), 7.40(d, 2H, ArH), 6.84(d, 1H, ArH), 6.19(s, 1H, ArH), 2.44(s, 3H, -CH ₃ ), 1.48(s , 6H, -CH ₃ ×2). MS (FAB): 430 (M+1).

pharmacological activity

Evaluation of in vitro activity:

Activation of glucokinase

1. Reaction principle:

2. Reaction system composition:

The reaction system includes 5mmol/l ATP, 0.2U/ml G6PDH, 0.2mmol/l NADP, 5mmol/lMgCl ₂ , 1mmol/l DTT, 25mmol/l KCl, 100mmol/l Tris-HCl, different concentrations of glucose, 1% DMSO , different concentrations of test compounds and recombinant human liver GK protein solution.

3. Operation process:

Prepare the reaction mixture (ATP, G6PDH, NADP, MgCl ₂ , DTT, KCl, glucose, Tris-HCl) → add the test compound → add the recombinant GK protein solution → measure the absorbance value at 340nm at room temperature, and record it as the initial value (0min) → Incubate at 37°C, read at 340nm every 10 minutes until 60 minutes → calculate the result.

4. Calculation method:

Activation multiple = (OD _t -OD ₀ ) _{sample tube} /(OD _t -OD ₀ ) _{reaction tube}

Note: The sample tube is the test compound added to the system, and the reaction tube is the reaction control without the test compound. Activation fold > 1.5 was considered positive.

Activity results of some examples

Activation of peroxisome proliferator-activated receptor

1. Principle:

PPARγ is activated after binding to its ligand, translocates into the nucleus, forms a heterodimer with another nuclear receptor RXR, and can specifically recognize the DNA sequence PPRE (PPAR responsive element), which regulates the expression of a series of genes. Establish a PPRE-regulated luciferase reporter gene, so that the expression level of the reporter gene reflects the level of PPAR activation.

2. Method:

1) Construct a plasmid vector expressing PPARγ, RXR, and a luciferase reporter gene plasmid vector regulated by the response element PPRE of PPARγ, RXR.

2) Using liposome transfection method (Lipofectamine2000, invitrogen), co-transfect PPAR, RXR, PPRE-luciferase expression plasmids into mammalian cell line 293E cells.

3) After 24 hours of transfection, 293E cells were digested with trypsin, counted cells were divided into several parts, mixed with the culture medium added to the sample, and cultured in a suitable culture plate for 24 hours. In this step, a negative control (plus DMSO), positive control (such as Rosiglitazone), set up parallel groups for each sample. Several gradients can be set according to the needs of the sample concentration, for example, 10 ^-9 ~ 10 ^-5 M.

4) 24-48 hours after adding the drug, fully lyse the cells with the cell lysate, collect the cell lysate from each well in the culture plate, add the luciferase reaction substrate (Luciferase Assay System, Promega), and measure immediately with a chemiluminescence detector Fluorescence readout.

3. Compound PPAR activity screening:

Compounds were compared for their PPARγ activating activity using the luciferase reporter gene method established above. Calculate the relative activity of the screening compound: compare the fluorescence value reading of the compound with the fluorescence value reading of the positive control rosiglitazone, set the activity of rosiglitazone as 100%, and the activity of other compounds is expressed as relative activity, namely:

Relative activity of screening compound = sample reading/positive control reading × 100%

Activity results of some examples

Claims (14)

1. pyrimidinethiazole amine compounds and pharmaceutically acceptable salts thereof as shown in general formula I, In the formula R is selected from hydrogen, fluorine, chlorine, bromine, C1-4 alkyl, hydroxymethyl _; R2 is selected from _hydrogen , fluorine, chlorine, bromine, COOC1-4 alkyl, carboxyl, aminoacyl, C1-4 alkyl substituted aminoacyl, C1-4 alkyl, hydroxymethyl; _{R3 is} selected from _R4 is selected from hydrogen, methyl, trifluoromethyl, fluorine or chlorine. 2. Compounds and pharmaceutically acceptable salts thereof according to claim 1, characterized in that, R is selected from hydrogen, fluorine, chlorine, bromine, methyl, ethyl _; R2 is selected from _hydrogen , fluorine, chlorine, bromine, methyl formate, ethyl formate, isopropyl formate, carboxyl, aminoacyl, carbamoyl, isopropanyl, methyl, ethyl, hydroxymethyl ; _{R3 is} selected from _R4 is selected from hydrogen, methyl, trifluoromethyl, fluorine or chlorine. 3. Compounds and pharmaceutically acceptable salts thereof according to claim 2, characterized in that, R is selected from hydrogen, fluorine, chlorine, methyl _; R is selected from _hydrogen , fluorine, chlorine, ethyl formate, isopropyl formate, methyl formate, carboxyl, aminoacyl, carbamoyl, isopropanyl, methyl, hydroxymethyl; _{R3 is} selected from _R4 is selected from hydrogen, methyl, trifluoromethyl. 4. according to the compound and pharmaceutically acceptable salt thereof any one of claim 1-3, described compound is selected from: 2-(1-(2-Methyl-6-(5-ethoxyformylthiazole-2-amino)pyrimidin-4-yl)piperidin-4-yl)ethanol 2-(1-(2-Methyl-6-(5-ethoxyformylthiazole-2-amino)pyrimidin-4-yl)piperidin-4-yl)acetic acid 2-((1-(2-Methyl-6-(5-ethoxyformylthiazole-2-amino)pyrimidin-4-yl)piperidin-4-yl)methoxy)acetic acid . 5. The compound according to any one of claims 1-3 and pharmaceutically acceptable salts thereof, characterized in that, said pharmaceutically acceptable salts include compounds with inorganic acids, organic acids, alkali metal ions, alkaline earth metal ions or can provide physiological Salts formed by combining organic bases with acceptable cations. 6. according to the compound of claim 4 and pharmaceutically acceptable salt thereof, it is characterized in that, described pharmaceutically acceptable salt comprises and inorganic acid, organic acid, alkali metal ion, alkaline earth metal ion or can provide physiologically acceptable cation Salts formed by combining organic bases. 7. according to the compound of claim 5 and pharmaceutically acceptable salt thereof, it is characterized in that, described inorganic acid is selected from hydrochloric acid, hydrobromic acid, phosphoric acid or sulfuric acid; Described organic acid is selected from methanesulfonic acid, p-toluenesulfonic acid Acid, trifluoroacetic acid, lycic acid, maleic acid tartaric acid, fumaric acid, citric acid or lactic acid; Described alkali metal ion is selected from lithium ion, sodium ion, potassium ion; Described alkaline earth metal ion comprises calcium ion, Magnesium ions; the organic base that can provide physiologically acceptable cations is selected from methylamine, dimethylamine, trimethylamine, piperidine, morpholine or tris(2-hydroxyethyl)amine. 8. according to the compound of claim 6 and pharmaceutically acceptable salt thereof, it is characterized in that, described inorganic acid is selected from hydrochloric acid, hydrobromic acid, phosphoric acid or sulfuric acid; Described organic acid is selected from methanesulfonic acid, p-toluenesulfonic acid Acid, trifluoroacetic acid, lycic acid, maleic acid tartaric acid, fumaric acid, citric acid or lactic acid; Described alkali metal ion is selected from lithium ion, sodium ion, potassium ion; Described alkaline earth metal ion comprises calcium ion, Magnesium ions; the organic base that can provide physiologically acceptable cations is selected from methylamine, dimethylamine, trimethylamine, piperidine, morpholine or tris(2-hydroxyethyl)amine. 9. The method for preparing the compound described in any one of claims 1-8, comprising the steps of: Using thiazolamine derivative 1 as raw material, reacting with dichloropyrimidine derivative 2 under basic conditions to generate intermediate compound ₃ , and then reacting with R3H to obtain product ₄ ; wherein the definition of R1 _- R4 is consistent with claim 1 Any one of -8 is the same; 10. The method for preparing the compound described in any one of claims 1-8, comprising the steps of: Using a halogenated thiazole derivative 5 as a raw material, reacting with a pyrimidinamine derivative 6 in an alkaline environment to generate the target compound 7; wherein the definition of R ₁ -R ₄ is the same as any one of claims 1-8; X is a halogen; 11. A pharmaceutical composition comprising, as an active ingredient, the compound of any one of claims 1-8 or a pharmaceutically acceptable salt thereof and a carrier commonly used in pharmacy. 12. The compound of any one of claims 1-8 and pharmaceutically acceptable salts thereof are used in the preparation of medicines for preventing and/or treating diseases related to glucokinase and peroxisome proliferator-activated receptors. 13. Use according to claim 12, characterized in that said diseases associated with glucokinase and peroxisome proliferator-activated receptors are selected from diabetes, chronic complications of diabetes and obesity. 14. according to the application of claim 13, it is characterized in that described diabetes is selected from type 1 diabetes or type 2 diabetes; The chronic complication of described diabetes is selected from retinopathy, nephropathy, neurosis, ischemic heart disease disease or arteriosclerosis.