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

Abstract

The invention discloses the new pyrimidine thiazole amines derivative of a class and its preparation method and pharmaceutical composition and purposes.Specifically, it is related to the pyrimidine thiazole amines derivative shown in formula I, its officinal salt, and preparation method thereof, composition containing one or more this compounds, and such compound is in the purposes of the treatment disease relevant with the glucokinase and peroxisome proliferation activated receptor-α such as aspect such as diabetes, obesity.

Description

The pyrimidine thiazole amines derivative of GK and PPAR double excitation activity

Invention field

The present invention relates to the pyrimidine thiazole amines derivative shown in formula I and II, its officinal salt, its same biological function Precursor or derivative, and preparation method thereof, the composition containing one or more this compounds, and such compound treatment The purposes of the disease relevant with the glucokinase and peroxisome proliferation activated receptor-α such as aspect such as diabetes, obesity.

Background of invention

Sugar is the important energy of organism and carbon source.Sugar decomposition produces energy, can supply the need of organism vital movement Will, the intermediate product of glycometabolism can be transformed into other carbon compounds, such as amino acid, aliphatic acid, nucleosides again.Glycometabolism The decomposition of sugar and the aspect of synthesis two of sugar can be divided into.Catabolism of carbohydrate includes glycolysis, i.e., sugared common decomposition approach；With three Carboxylic acid recycle, i.e., sugared last oxidative pathway.Glycolysis is enzyme by glucose degradation into pyruvic acid and along with the mistake of generation ATP Journey.It is the common metabolic approach of breakdown of glucose generation energy in animal, plant, microbial cell.In oxygen consumption organism, The pyruvic acid of glycolysis generation enters mitochondria, through tricarboxylic acid cycle by exhaustive oxidation into CO₂And water；The NADH of glycolysis generation is through exhaling Inhale chain oxidation and produce ATP and water.So, glycolysis is the prelude of oxidative phosphorylation and tricarboxylic acid cycle.Glucokinase (GK) It is one of four kinds of hexokinases of discovery corridor Cofowick, S.P.The Enzymes, Vol.9 with mammal .1-48 pages of (P.Boyer ' ed.) Academic Press, New York, N bifurcation, 1973], it is first in glycolytic pathway Key enzyme, it glucose phosphorylation can be turned into G6P, into the generation in downstream under conditions of ATP presence Apologize for having done sth. wrong journey.Therefore, this step is first rate-limiting step of whole glycometabolism process, and GK is first speed limit during glycometabolism Enzyme, plays important biological action.

The cell distribution of GK is limited, is mainly seen in pancreas beta cell and hepatic tissue cell.In addition, GK control blood glucose balance and Key player is play in metabolism, on the one hand the metabolism of regulation glycogen, when empty stomach or blood sugar are low, GK hypoactivities, glycogen output Increase, to ensure the energy supply of vitals；After the meal or when blood sugar is high, GK increased activities, so as to promote hepatic glycogen to synthesize, press down Liver gluconeogenesis processed, to maintain glycaemic homeostasis.On the other hand the secretion of insulin is regulated and controled as the receptor of glucose.When internal When blood sugar concentration is higher than normal value, the glucose in blood is transported into beta Cell of islet by glucose transporter 2 (GLUT2), The lower phosphorylation generation G6P of GK effects.The glycolysis of glucose, oxidative metabolism increase ATP/ADP ratios, K+ ions Passage is closed, membrane depolarization, voltage sensitivity Ca2+ channel openers, stream in Ca2+, insulin storage vesica and plasma membrane fusion, is made Insulin releasing enters blood, and enters the regulation that liver, fat and muscle cell participate in blood sugar with blood circulation.In liver Glucose phosphoric acid in the presence of GK turns to -6 glucose 1-phosphate1-s, then synthesizes hepatic glycogen under insulin action.Fat and Muscle cell, insulin triggers glucose transporter 4 (GLUT4) and is migrated to plasma membrane from cell memory storage vesica and promotes glucose Intake and metabolism.GK is metabolized double action mechanism reduction blood sugar by adjusting insulin releasing and glycogen, is maintaining glycaemic homeostasis During play a significant role [Al-Hasani H；Tschopl M H.Mol Interv, 2003,3 (7), 367-370]. [Chipkin, S.R., Kelly, K.L. and Ruderrnam, N.B. see Joslin ' Diabetes (C.R.Khan and G.C.Wier is compiled), Lea and 97-115 pages of Febiger, Philadelphia, PA, the, 1994].

Young type early onset diabetes (MODY) are to be lost by function after GK gene mutations and caused, and show GK in human body Play glucose sensor Liang, Y, Kesavan, P., Wang, L. et al., Biochem.J.309,167-173, 1995).In addition to MODY, also generally existing GK activity decreases in general diabetic individual.Pathogenesis to diabetes B enters one Walk research to find, liver GK activity is reduced and may participate in insulin resistance mechanism, cause blood glucose rise, islet function is damaged and pancreas Insulin resistance is aggravated.So finding the medicine that can increase GK activity to prevent or delay advancing of disease, it is possible to be early stage Prevention and treatment diabetes open up a new way.

Recognize in recent years：Pancreatic beta type glucokinase is confined to the brain of rat, is wherein particularly feeding center (hypothalamus Ventromedial nucleus, VMH) middle expression.The nerve cell of the about twenty percent in VMH is referred to as glucose response nerve unit (glucose- Responsive neutrons), it was considered as being played a significant role in terms of control body weight in the past.Portugal is given to the intracerebral of rat Grape sugar, then food ration reduction, and if intracerebral gives the aminoglucose of analog one of glucose to suppress glucose metabolism, then occur Many foods.Electrophysiology experiment shows：Glucose response nerve unit is corresponding with physiological concentration of glucose change (5-20mM) Be activated, but by aminoglucose etc. suppress glucose metabolism, then its activity be suppressed.The glucose that VHM can be estimated is dense It is via the mechanism of glucokinase that degree sensory perceptual system is same with the insulin secretion of pancreatic beta cell.Therefore, except liver, pancreatic beta are thin Outside born of the same parents, activating the material of the glucokinase activity activity of VHM not only has the effect of regulation blood sugar, it is also possible to adjust many 2 type sugar The worried obesity of urine patient.From above-mentioned record, the compound with Glucokinase Activation effect can be used as diabetes Curative and/or preventive medicine, or the sugar such as retinopathy, ephrosis, neurosis, ischemic heart disease, artery sclerosis Treatment and/or the preventive medicine of the chronic complicating diseases of disease are urinated, further can be used as treatment and/or the preventive medicine of obesity.

It has been found that many GK activation of small molecule agent, with different architectural features：Such as the phenylacetamides of substitution (WO0058293WO0185706WO0208209WO0185707WO0183465WO0246173WO2004072066WO0246173) , substituted hydantoins (WO0183478), substituted azole class (WO2006112549), substituent indole (US0067939WO031179), the Propionamides (WO0248106) of isoindoline substitution, substituted is anthranilamide-based (WO03080585), substituted α phenylacryloyls amine (WO0214312) etc., wherein phenylacetamides are that many one are in research Class.Although very big contribution has been made in above-mentioned GK activators research to this area, lived to improve compound structure and GK activators Property, this area still is continuing to study.

Peroxisome proliferation activated receptor-α (PPARs) belong to nuclear hormone receptor superfamily, have three kinds of hypotypes (PPAR α, PPARγ、PPARδ).Three kinds of acceptors exercise the function of lipid sensor, the expression of coordinated regulation several genes sequence, regulation weight The organism metabolism wanted.PPARs has various biological effects, and Adipocyte Differentiation and fat can be promoted to generate, and enhancing body is to pancreas The sensitiveness of island element, sugar balance in mediation body is effectively treatment metabolic disease (such as type ii diabetes and atherosclerosis) Drug targets.And suppress inflammatory factor generation and inflammatory reaction, and tumour growth is influenceed, protection effect is produced to angiocarpy Should.Recent studies indicate that, PPARs also has neuroprotection, can mitigate alzheimers disease, Parkinson's, brain and lack The damage of nerve cell in the nerve degenerative diseases such as blood and multiple sclerosis.

PPAR γ are different from the nuclear receptor of classics, after it combines activation by sepcific ligands, are still unable to Direct Recognition and combine spy Different DNA regulating and controlling sequences, it is necessary to form heterodimer by with vitamin A acid x acceptors (retinoid X receptor, RXR), In the presence of many auxiliary adjustment factors (cofactors), special genetic transcription is adjusted.Early stage research turns activated gene The PPAR γ parts of record are referred to as PPAR gamma agonists, and suppressor transcription is referred to as inhibitor.Latest Progress shows, The particular conformation that PPAR γ are combined to form with different ligands, determines the RXR heterodimers of PPAR γ mono- and special confactor Selective binding effect, further determine the alternative transcription adjustment effect to different genes.Just because of this structure With the selectivity for interacting, make different PPAR γ parts by selected gene transcriptional control, produce different biology Effect.For the complexity of gene regulation, the activation that transcription of the same PPAR γ parts to target gene has, some suppression, because This, increasing scholar replaces the unilateral definition of " activator/inhibitor " with " conditioning agent ", and then the known selection of research Property acts on some PPAR γ parts referred to as " selective PPARγ modulator " (selective of specific gene transcriptional control PPAR γ modulator, SPPARM).If any micromolecular compound activate PPAR γ, it is then mutual from different confactors Effect, can separate Adipose Differentiation process with glycolipid metabolism process in gene regulation level, i.e., some promote glycolipid generation The PPAR gamma agonists thanked not induced lipolysis differentiation.

Research to PPAR gamma modulators at present is concentrated mainly on two problems：One is that we need the PPAR γ parts to have Great Activation Activity, to reach appropriateness activation PPAR γ, the purpose of regulation target gene transcription；Two be whether can be in gene regulation Level makes Adipose Differentiation be separated with insulin-sensitizing effectAlign compound under study for action or material answers further investigated its PPAR The molecular mechanism of γ regulations, screen existing insulin-sensitizing effect does not influence Adipose Differentiation even to promote the PPAR of fat metabolism again Gamma modulators.

Because PPAR α/δ participates in regulation lipid metabolism, improve hyperlipidemia, PPAR γ activation is mitigated to a certain extent and is lured The Adipose Differentiation led；PPAR δ are also more extensive than PPAR α in Tissue distribution, and the research to PPAR δ in recent years also attracts attention, Many researchers are devoted to developing PPAR alpha/gamma double agonists or the activators of α/δ/γ tri-, expect that such medicine can reach drop blood Sugar plays the effect of regulation blood fat, immunotherapy targeted autoantibody obesity, insulin resistance, metabolic syndrome simultaneously.Animal and facing at present The result of study prompting of bed experiment, PPAR alpha/gamma double agonists or the activators of α/δ/γ tri- can really improve insulin resistance, improve Dyslipidemia, but due to still lacking drug safety and toxicity test foundation, the application prospect of this kind of compound need to be seen Examine.

Diabetes are a kind of whole body chronic metabolic diseases, and its pathological characters is mainly hyperglycaemia.It has been generally acknowledged that causing height (also referred to as three is big for the imbalance of reduction of the main pathological change of blood sugar including insulin secretion, the decrease of effect and glycogen metabolism Pathological change).The activation energy of glucokinase (GK) promotes insulin secretion and glycogen metabolism simultaneously；Peroxidase paraphyte The activation energy of activated receptor (PPAR) increases sensitiveness of the body cell to insulin.Therefore, for the three of diabetes hyperglycaemia Big pathological change, structure and pharmacological action based on known GK and PPAR small molecule agonists build and synthesis GK and PPAR Double target spot ligand compounds, research and development one kind can simultaneously improve insulin secretion, glycogen metabolism and promote peripheral tissues pair ' one medicine Mutiple Targets of the sensitiveness of insulin ' medicine, the new way for finding universal effective treatment Glucovance will be turned into.

The content of the invention

Glucokinase activators are can be used as the invention provides one kind to be activated but also as peroxidase paraphyte The pyrimidine thiazole amines derivative of the general structure I of receptor stimulating agent：

Spread out it is an object of the invention to provide a kind of new pyrimidine thiazole amines with GK and PPAR double excitation activity Biology, its officinal salt.

Another object of the present invention is to provide a kind of method for preparing new pyrimidine thiazole amines derivative.

It is still another object of the present invention to provide a kind of pharmaceutical composition containing one or more this compounds.

A further object of the present invention is, based on GK and PPAR biological functions, to propose Mutiple Targets combinational drug therapy diabetes New ideas.

Yet another object of the invention is to provide the double ligand compounds of GK and PPAR, and such compound in treatment and GK and PPAR is about the purposes in disease medicament.

The present invention is to be related to the pyrimidine thiazole amines derivative with general formula I：

In formula

R₁Selected from hydrogen, halogen, amino, hydroxyl, COOC1-8 alkyl, carboxyl, aminoacyl, C1-8 alkyl replaces aminoacyl, C1-8 alkyl, carboxy-methyl mercapto-group, methylol, carboxyethyl；

Preferred R₁Selected from hydrogen, fluorine, chlorine, bromine, amino, COOC1-4 alkyl, carboxyl, aminoacyl, C1-4 alkyl substitution aminoacyl Base, C1-4 alkyl, carboxy-methyl mercapto-group, methylol, carboxyethyl；

Preferred R₁Selected from hydrogen, fluorine, chlorine, bromine, amino, carbomethoxy, ethoxycarbonyl, isopropyl ester group, carbomethoxy, carboxyl, ammonia Acyl group, methylamine acyl group, isopropylamine acyl group, ring the third methylamine acyl group, methyl, ethyl, carboxy-methyl mercapto-group, methylol, carboxyethyl；

Most preferred R₁Selected from hydrogen, fluorine, chlorine, ethoxycarbonyl, isopropyl ester group, carbomethoxy, carboxyl, aminoacyl, methylamine acyl group is different Propylamine acyl group, ring the third methylamine acyl group, methyl, carboxy-methyl mercapto-group, methylol.

R₂Selected from hydrogen, halogen, amino, hydroxyl, COOC1-8 alkyl, carboxyl, aminoacyl, C1-8 alkyl replaces aminoacyl, C1-8 alkyl, carboxy-methyl mercapto-group, methylol, carboxyethyl；

Preferred R₂Selected from hydrogen, fluorine, chlorine, bromine, amino, COOC1-4 alkyl, carboxyl, aminoacyl, C1-4 alkyl substitution aminoacyl Base, C1-4 alkyl, carboxy-methyl mercapto-group, methylol, carboxyethyl；

Preferred R₂Selected from hydrogen, fluorine, chlorine, bromine, amino, carbomethoxy, ethoxycarbonyl, isopropyl ester group, carbomethoxy, carboxyl, ammonia Acyl group, methylamine acyl group, isopropylamine acyl group, ring the third methylamine acyl group, methyl, ethyl, carboxy-methyl mercapto-group, methylol, carboxyethyl；

Most preferred R₂Selected from hydrogen, fluorine, chlorine, ethoxycarbonyl, isopropyl ester group, carbomethoxy, carboxyl, aminoacyl, methylamine acyl group is different Propylamine acyl group, ring the third methylamine acyl group, methyl, carboxy-methyl mercapto-group, methylol.

R₃Selected from hydrogen, halogen, C1-8 alkylaminos, fragrant amino, C1-8 alkoxies, aryloxy group, C1-8 alkylthio groups, arylthio, Nitro, cyano group,

Preferred R₃Selected from hydrogen, chlorine, C1-6 alkylaminos, fragrant amino, C1-6 alkoxies, aryloxy group, C1-6 alkylthio groups, benzene sulphur Base, pyridine thio, nitro, cyano group,

Preferred R₃Selected from hydrogen, chlorine, methylamino, ethylamino, isopropylamino, the methylamino of ring third, phenylamino, pyridine ammonia Base, methoxyl group, ethyoxyl, butoxy, phenoxy group, pyridomethoxy, methyl mercapto, thiophenyl, pyridine thio, nitro, cyano group,

Most preferred R₃Selected from ethylamino, isopropylamino, the methylamino of ring third, phenylamino, pridylamino, ethyoxyl, fourth oxygen Base, pyridomethoxy,

R₄Selected from hydrogen, methyl, trifluoromethyl, fluorine or chlorine, methoxyl group, methyl mercapto, methylamino.

Preferred R₄Selected from hydrogen, methyl, trifluoromethyl, fluorine or chlorine, methoxyl group, methyl mercapto.

Preferred R₄Selected from hydrogen, methyl, trifluoromethyl, fluorine or chlorine, methoxyl group.

Most preferred R₄Selected from hydrogen, methyl, trifluoromethyl.

Preferred aryl is selected from phenyl, pyridine radicals.

In order to prepare the compound described in formula I of the present invention, according to the structure of formula I, the present invention prepares compound of Formula I It is classified into two kinds of routes.

In first route, with thiazole amine derivative 1 as raw material, reacted in the basic conditions with dichloro pyrimidine derivative 2 Generation midbody compound 3, then with R₃Reaction obtains product 4.

In Article 2 route, reacted with pyrimidinamine derivatives 6 with halo thiazole derivative 5 as raw material, under alkaline environment and given birth to Into target compound 7.

In addition, the initiation material and intermediate in above-mentioned reaction are readily obtained, each step reaction can be according to the document reported Or can be easy to synthesis with the conventional method in organic synthesis to those skilled in the art.Pharmacy described in formula I Upper acceptable salt includes different acid-addition salts, such as acid-addition salts of following inorganic acid or organic acid：Hydrochloric acid, hydrobromic acid, phosphoric acid, Sulfuric acid, methanesulfonic acid, p-methyl benzenesulfonic acid, trifluoroacetic acid, matrimony vine acid, maleic acid, tartaric acid, fumaric acid, citric acid, lactic acid.Formula I The pharmaceutically acceptable salt also includes different alkali metal salts (lithium, sodium, sylvite), alkali salt (calcium, magnesium salts) and ammonium salt, With the salt of the organic base that can provide physiologically acceptable cation, such as methylamine, dimethylamine, trimethylamine, piperidines, morpholine and three The salt of (2- ethoxys) amine.All these salt within the scope of the present invention can all be prepared using conventional method.

The invention further relates to the pharmaceutical composition using the compounds of this invention as active ingredient.The pharmaceutical composition can basis It is prepared by method well known in the art.Can be by by the compounds of this invention and one or more pharmaceutically acceptable solid or liquid Excipient and/or assistant agent are combined, and are made and are suitable to any formulation that human or animal uses.The compounds of this invention is in its pharmaceutical composition In content be usually 0.1-95 weight %.

The compounds of this invention or the pharmaceutical composition containing it can be administered in a unit, and method of administration can be enteron aisle Or non-bowel, such as oral, intravenous injection, intramuscular injection, hypodermic injection, nasal cavity, oral mucosa, eye, lung and respiratory tract, skin, Vagina, rectum etc..

Form of administration can be liquid dosage form, solid dosage forms or semisolid dosage form.Liquid dosage form can be solution (including True solution and colloidal solution), emulsion (including o/w types, w/o types and emulsion), supensoid agent, injection (including liquid drugs injection, powder-injection And transfusion), eye drops, nasal drop, lotion and liniment etc.；Solid dosage forms can be tablet (including ordinary tablet, enteric coatel tablets, lozenge, Dispersible tablet, chewable tablets, effervescent tablet, oral disnitegration tablet), capsule (including hard shell capsules, soft capsule, capsulae enterosolubilis), granule, dissipate Agent, micropill, dripping pill, suppository, film, paster, the agent of gas (powder) mist, spray etc.；Semisolid dosage form can be ointment, gel Agent, paste etc..

It is sustained release preparation, controlled release preparation, targeting preparation and various that the compounds of this invention can be made ordinary preparation, also be made Particulate delivery system.

In order to the compounds of this invention is made into tablet, various excipient well known in the art can be widely used, including it is dilute Release agent, binder, wetting agent, disintegrant, lubricant, glidant.Diluent can be starch, dextrin, sucrose, glucose, breast Sugar, mannitol, sorbierite, xylitol, microcrystalline cellulose, calcium sulfate, calcium monohydrogen phosphate, calcium carbonate etc.；Wetting agent can be water, second Alcohol, isopropanol etc.；Adhesive can be starch slurry, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, Arabic gum Slurry, gelatine size, sodium carboxymethylcellulose, methylcellulose, hydroxypropyl methyl cellulose, ethyl cellulose, acrylic resin, card Ripple nurse, polyvinylpyrrolidone, polyethylene glycol etc.；Disintegrant can be dried starch, microcrystalline cellulose, low substituted hydroxy-propyl fiber Element, PVPP, Ac-Di-Sol, sodium carboxymethyl starch, sodium acid carbonate and citric acid, polyoxy second Alkene sorbitan fatty acid ester, dodecyl sodium sulfate etc.；Lubricant and glidant can be talcum powder, silica, tristearin Hydrochlorate, tartaric acid, atoleine, polyethylene glycol etc..

Tablet can also be further made coating tablet, such as sugar coated tablet, thin membrane coated tablet, ECT, or it is double Synusia and multilayer tablet.

In order to administration unit is made into capsule, active ingredient the compounds of this invention can be mixed with diluent, glidant Close, during mixture is placed directly within into hard shell capsules or soft capsule.Also can be by active ingredient the compounds of this invention elder generation and diluent, bonding Agent, disintegrant are made particle or micropill, then are placed in hard shell capsules or soft capsule.For preparing each dilute of the compounds of this invention tablet Release the capsule that agent, binder, wetting agent, disintegrant, glidant kind can also be used for preparing the compounds of this invention.

It is that the compounds of this invention is made injection, water, ethanol, isopropanol, propane diols or their mixture can be used Make solvent and add appropriate solubilizer commonly used in the art, cosolvent, pH adjustments agent, osmotic pressure regulator.Solubilizer or hydrotropy Agent can be poloxamer, lecithin, HP-β-CD etc.；PH adjustment agent can be phosphate, acetate, hydrochloric acid, hydrogen Sodium oxide molybdena etc.；Osmotic pressure regulator can be sodium chloride, mannitol, glucose, phosphate, acetate etc..Such as prepare freeze-dried powder Injection, can also add mannitol, glucose etc. as proppant.

Additionally, if desired, can also be to addition colouring agent, preservative, spices, flavouring or other additions in pharmaceutical preparation Agent.

To reach medication purpose, strengthen therapeutic effect, medicine of the invention or pharmaceutical composition known can be given with any Prescription method is administered.

The dosage of the compounds of this invention pharmaceutical composition is according to the property to be prevented or treated disease and serious journey The individual instances of degree, patient or animal, method of administration and formulation etc. can have change on a large scale.In general, the present inventionization The daily Suitable dosage ranges of compound are 0.001-150mg/Kg body weight, preferably 0.01-100mg/Kg body weight.Above-mentioned dosage With a dosage unit or several dosage unit administrations can be divided into, this depends on the clinical experience of doctor and including using other The dosage regimen for the treatment of means.

Compound of the invention or composition can individually be taken, or merge with other treatment medicine or symptomatic drugs and use. When compound of the invention and other medicines have synergy, its dosage should be adjusted according to actual conditions.

The compounds of this invention is the dual activators of GK and PPAR or its precursor, can be by adjusting insulin releasing, and it is right to organize Sensitiveness and glycogen the metabolism multiple action mechanism reduction blood sugar of insulin, can be used to preventing and treating 1 type or 2 types especially 2 The complication of patients with type Ⅰ DM and correlation, or the Other diseases relevant with GK and PPAR.

Specific embodiment

Invention is described further below with reference to embodiment, but is not limit the scope of the invention.

Determining instrument：The NMR spectrum type NMRs of Vaariaan Mercury 300.Mass spectrum ZAD-2F With VG300 mass spectrographs.

Embodiment 1：2- (4- (2- methyl -6- (4- methylthiazol -2- amino) pyrimidine -4- amino) phenoxy group) ethyl acetate

a：The preparation of N- (the chloro- 2 methylpyrimidine -4- bases of 6-) -4- methylthiazol -2- amine：

2- methyl -4,6- dichloro pyrimidines (16.3g, 0.1mol), 2- amino -4- methylthiazols (11.4g, 0.1mol) are added Enter in DMF (250mL), under ice bath stirring, 60%NaH (8g, 0.2mol) is dividedly in some parts in reaction solution, room temperature reaction 6h, adds glacial acetic acid (15mL) terminating reaction after reaction completely.Reaction solution is poured into 1L saturated sodium bicarbonate ice-water baths, there is big Amount solid is separated out, and filtering, washing, dry cake is recrystallized to give N- (the chloro- 2 methylpyrimidine -4- bases of 6-) -4- methylthiazols -2- 16.5 grams of amine.

b：The system of 2- (4- (2- methyl -6- (4- methylthiazol -2- amino) pyrimidine -4- amino) phenoxy group) ethyl acetate It is standby：

By N- (the chloro- 2 methylpyrimidine -4- bases of 6-) -4- methylthiazol -2- amine (2.4g, 10mmol), 2- (4- aminobenzene oxygen Base)-ethyl acetate (1.75g, 10mmol), NaI (50mg) is added in DMF (20mL), and 150 DEG C of reaction 36h, reaction is complete, Room temperature is down to, evaporated under reduced pressure solvent adds ethyl acetate (200mL), washes with water successively, and saturated sodium-chloride is washed, anhydrous sodium sulfate Dry, column chromatography obtains 1.32 grams of white solid after concentration.¹H NMR(DMSO-d₆, 300MHz), 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 ,-CH₂O-), 4.27 (q, 2H, Me-CH₂-), 2.60 (s, 3H ,-CH₃), 2.32 (s, 3H ,-CH₃), 1.32 (t, 3H ,-CH₃).MS(FAB)：400(M+1)

Embodiment 2：2- (4- (2- methyl -6- (4- methylthiazol -2- amino) pyrimidine -4- amino) phenoxy group) acetic acid

By 2- (4- (2- methyl -6- (4- methylthiazol -2- amino) pyrimidine -4- amino) phenoxy group) ethyl acetate (300mg) is added in methyl alcohol (10mL) and 2.5N NaOH (15mL), 60 DEG C of stirring 2h, is down to room temperature, adds water (30mL), is used Ether (30mL) is extracted, and water layer concentrated hydrochloric acid adjusts pH to 2, has a large amount of white native lands to separate out, and filtering, dry cake obtains white Solid 255mg.¹H NMR(DMSO-d₆, 300MHz), 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 ,-CH₃), 2.30 (s, 3H ,-CH₃).MS(FAB)：372(M+1).

Embodiment 3：2- (3- (2- methyl -6- (4- methylthiazol -2- amino) pyrimidine -4- amino) phenoxy group) ethyl acetate

Operating procedure obtains white solid (585mg, 29%) with embodiment 1.¹H NMR(DMSO-d₆, 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 ,-CH₂O-), 4.17 (q, 2H, Me-CH₂-), 2.43 (s, 3H ,- CH₃), 2.22 (s, 3H ,-CH₃), 1.20 (t, 3H ,-CH₃).MS(FAB)：400(M+1)

Embodiment 4：2- (3- (2- methyl -6- (4- methylthiazol -2- amino) pyrimidine -4- amino) phenoxy group) acetic acid

Operating procedure obtains white solid (260mg, 95%) with embodiment 2.¹H NMR(DMSO-d₆, 300MHz), 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 ,-CH₂O-), 2.42 (s, 3H ,-CH₃), 2.17 (s, 3H ,-CH₃).MS(FAB)：372(M+1).

Embodiment 5：2- methyl -2- (4- (2- methyl -6- (4- methylthiazol -2- amino) pyrimidine -4- amino) phenoxy group) Ethyl propionate

Operating procedure obtains white solid 585mg with embodiment 1.¹H NMR(DMSO-d₆, 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 ,-CH₃), 1.38 (s, 6H ,-CH₃×2).MS(FAB)： 414(M+1).

Embodiment 6：2- methyl -2- (4- (2- methyl -6- (5- ethoxycarbonyl thiazole -2- amino) pyrimidine -4- amino) benzene Epoxide) ethyl propionate

Operating procedure obtains white solid 556mg with embodiment 1.¹H NMR(DMSO-d₆, 300MHz), 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-CH₂- × 2), 2.45 (s, 3H ,-CH₃), 1.45 (s, 6H ,-CH₃ × 2), 1.28 (t, 3H ,-CH₃), 1.19 (t, 3H ,-CH₃).MS(FAB)：486(M+1).

Embodiment 7：2- (4- (2- methyl -6- (4- methylthiazol -2- amino) pyrimidine-4-yl) piperazine -1- bases) ethanol

a：The preparation of N- (the chloro- 2 methylpyrimidine -4- bases of 6-) -4- methylthiazol -2- amine is with embodiment 1.

b：The preparation of 2- (4- (2- methyl -6- (4- methylthiazol -2- amino) pyrimidine-4-yl) piperazine -1- bases) ethanol：

By N- (the chloro- 2 methylpyrimidine -4- bases of 6-) -4- methylthiazol -2- amine (2.4g, 10mmol), hydroxyethyl piperazine (1.73g, 10mmol), DIPEA (1.94g, 15mmol), DMSO (5mL) is added in dioxane (60mL), 110 DEG C of reactions 10h, reaction is complete.Reaction solution is poured into water (200mL), 1h, filtering is stirred.Dry cake, re-crystallizing in ethyl acetate is obtained White solid (1.7g, 50%)¹H NMR(DMSO-d₆, 300MHz), δ (ppm)：10.98 (s, 1H ,-NH-), 6.55 (s, 1H, ArH), 5.98 (s, 1H, ArH), 3.53-3.47 (m, 6H ,-CH₂× 3), 2.47-2.39 (m, 6H, CH₂× 3), 2.34 (s, 3H ,- CH₃), 2.21 (s, 3H ,-CH₃).MS(FAB)：335(M+1).

Embodiment 8：2- (4- (2- methyl -6- (5- ethoxycarbonyl thiazole -2- amino) pyrimidine-4-yl) piperazine -1- bases) Ethanol

a：The preparation of 2- (the chloro- 2 methylpyrimidine -4- amidos of 6-) thiazole-5-carboxylic acid ethyl ester：

By 2- methyl -4,6- dichloro pyrimidines (16.3g, 0.1mol), thiazolamine -5- Ethyl formates (17.2g, 0.1mol) it is added in DMF (250mL), under ice bath stirring, 60%NaH (8g, 0.2mol) is dividedly in some parts in reaction solution, Room temperature reaction 6h, reaction is complete.Add glacial acetic acid (15mL) terminating reaction.Reaction solution is poured into 1L saturated sodium bicarbonate ice-water baths In, there are a large amount of solids to separate out, filtering, washing, dry cake obtains 2- (6- chloro-2-methyl pyrimidine -4- amidos) thiazole -5- first Acetoacetic ester (18.5g, 65%) crude product.

b：2- (4- (2- methyl -6- (5- ethoxycarbonyl thiazole -2- amino) pyrimidine-4-yl) piperazine -1- bases) ethanol Prepare：

By 2- (6- chloro-2-methyl pyrimidine -4- amidos) thiazole -5- Ethyl formates (149mg, 5mmol), hydroxyethyl piperazine (865mg, 5mmol), DIPEA (970mg, 7.5mmol), DMSO (5mL) is added in dioxane (60mL), 110 DEG C of reactions 10h, reaction is complete.Reaction solution is poured into water (200mL), 1h, filtering is stirred.Dry cake, re-crystallizing in ethyl acetate is obtained 2- (4- (2- methyl -6- (5- ethoxycarbonyl thiazole -2- amino) pyrimidine-4-yl) piperazine -1- bases) ethanol white solid¹H NMR(DMSO-d₆, 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 ,-CH₂), 3.51 (m, 6H, CH₂× 3), 3.32 (s, 4H ,-CH₂), 2.43 (m, 2H ,-CH₂), 2.41 (s, 3H ,-CH₃), 1.28 (t, 3H ,-CH₃).MS(FAB)：393(M+1).

Embodiment 9：2- (1- (2- methyl -6- (5- ethoxycarbonyl thiazole -2- amino) pyrimidine-4-yl) piperidin-4-yl) Ethanol

Operating procedure obtains white solid 256mg with embodiment 8.¹H NMR(DMSO-d₆, 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 ,-CH₂ × 2), 3.53-3.50 (m, 1H ,-CH), 3.25 (t, 2H ,-CH₂), 2.84 (t, 2H ,-CH₂), 2.49-2.45 (m, 2H ,-CH₂), 2.39 (s, 3H ,-CH₃), 1.73-1.70 (m, 2H ,-CH₂), 1.27 (t, 3H ,-CH₃), 1.11-1.02 (m, 2H ,-CH₂).MS (FAB)：392(M+1).

Embodiment 10：2- (1- (2- methyl -6- (5- ethoxycarbonyl thiazole -2- amino) pyrimidine-4-yl) piperidin-4-yl) Acetic acid

Operating procedure obtains white solid 356mg with embodiment 8.¹H NMR(DMSO-d₆, 400MHz), δ (ppm)： 11.53 (s, 1H ,-NH-), 8.02 (s, 1H, ArH), 6.06 (s, 1H, ArH), 4.25 (q, 2H ,-CH₂), 4.13 (m, 2H ,- CH₂), 3.53-3.50 (m, 1H ,-CH), 3.03-2.97 (m, 2H ,-CH₂), 2.55-2.45 (m, 2H ,-CH₂), 2.40 (s, 3H ,- CH₃), 1.90-1.86 (m, 2H ,-CH₂), 1.53-1.43 (m, 2H ,-CH₂), 1.28 (t, 3H ,-CH₃).MS(FAB)：406(M+ 1).

Embodiment 11：2- ((1- (2- methyl -6- (5- ethoxycarbonyl thiazole -2- amino) pyrimidine-4-yl) piperidines -4- Base) methoxyl group) acetic acid

Operating procedure obtains white solid 342mg with embodiment 8.¹H NMR(DMSO-d₆, 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 ,-CH₂), 3.58 (s, 2H ,-CH₂), 2.92 (d, 2H ,-CH₂), 2.33 (s, 3H ,-CH₃), 2.10 (m, 4H ,-CH₂), 1.46 (m, 1H ,-CH), 1.37-1.33 (m, 2H ,-CH₂), 0.89 (t, 3H ,-CH₃), 0.74-0.71 (m, 2H ,-CH₂).MS(FAB)： 436(M+1)

Embodiment 12：2- (4- (6- (5- isopropyl oxygen formyl thiazole -2- amino) pyrimidine-4-yl) piperazine -1- bases) ethanol

Operating procedure obtains white solid 342mg with embodiment 8.¹H NMR(DMSO-d₆, 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 ,-CH₂-), 3.51 (m, 6H, CH₂× 3), 2.48 (m, 4H, CH₂× 2), 2.42 (m, 2H ,- CH₂), 1.28 (d, 6H ,-CH₃×2).MS(FAB)：393(M+1)

Embodiment 13：2- (4- (6- (5- ethoxycarbonyl thiazole -2- amino) pyrimidine-4-yl) piperazine -1- bases) ethanol

Operating procedure obtains white solid 242mg with embodiment 8.¹H NMR(DMSO-d₆, 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 ,-CH₂), 3.53-3.52 (m, 6H, CH₂× 3), 2.50 (m, 4H ,-CH₂), 2.44-2.40 (m, 2H ,- CH₂), 2.42 (t, 2H ,-CH₃).MS(FAB)：379(M+1)

Embodiment 14：2- methyl -2- (4- (2- methyl -6- (5- carboxyl thiazole -2- amino) pyrimidine -4- amino) phenoxy group) Propionic acid

Operating procedure obtains white solid 223mg with embodiment 1 and embodiment 2.¹H NMR(DMSO-d₆, 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 ,-CH₃), 1.48 (s, 6H ,-CH₃× 2).MS(FAB)：430(M+1).

Pharmacological activity

External activity is evaluated：

To the activation of glucokinase

1. reaction principle：

2. reaction system is constituted：

Reaction system includes 5mmol/l ATP, 0.2U/ml G6PDH, 0.2mmol/l NADP, 5mmol/lMgCl₂, 1mmol/l DTT, 25mmol/l KCl, 100mmol/l Tris-HCl, different glucose, 1%DMSO, various concentrations are received Examination compound and recombination human source liver GK protein liquids.

3. operating process：

Prepare reaction mixture (ATP, G6PDH, NADP, MgCl₂, DTT, KCl, glucose, Tris-HCl) → add and receive Examination compound → addition restructuring GK protein liquids → room temperature measuring 340nm absorbances, and it is designated as initial value (0min) → 37 DEG C temperature Educate, every 10 minutes 340nm readings once to 60min → result of calculation.

4. computational methods：

Activation multiple=(OD_t-OD₀)_{Sample cell}/(OD_t-OD₀)_{Reaction tube}

Note：To add test-compound in system, reaction tube is the reaction control for being not added with test-compound to sample cell.Activation Multiple ＞ 1.5 is considered as the positive.

Section Example Activity Results

To the activation of peroxisome proliferation activated receptor-α

1. principle：

Activated after PPAR γ and its ligand binding, in indexing to nucleus, heterodimeric is formed with another nuclear receptor RXR Body, can regulate and control the expression of series of genes with specific recognition DNA sequence dna PPRE (PPAR responsive element), the latter. The luciferase reporter gene of PPRE regulation and control is set up, the expression of reporter gene is reacted the level that PPAR is activated.

2. method：

1) plasmid vector of construction expression PPAR γ, RXR, and the response element PPRE regulation and control of PPAR γ, RXR fluorescence Plain enzyme reporter gene plasmid vector.

2) with the method (Lipofectamine2000, invitrogen) of liposome transfection, cotransfection PPAR, RXR, PPRE-luciferase expression plasmids enter mammal cell line 293E cells.

3) 293E cell transfectings are digested after 24 hours with pancreatin, and several pieces are divided into after counting cell, respectively with addition sample The culture medium mixing of product, cultivates 24 hours in suitable culture plate, and this step must set up negative control (plus DMSO), positive control (such as Rosiglitazone), each sample sets up parallel group, and sample concentration can set some gradients, such as 10 as needed^-9 ~10^-5M。

4) after dosing 24-48 hours, with the abundant cell lysis of cell pyrolysis liquid, each hole cell cracking in culture plate is collected Liquid, adds luciferase reaction substrate (Luciferase Assay System, Promega), with chemiluminescence detector immediately Measurement fluorescence reading.

3. compound PPAR screening active ingredients：

With the luciferase reporter gene method of above-mentioned foundation, the PPAR γ Activation Activities of comparative compound.Calculating sifting The relative activity of compound：By the fluorescent value reading of compound compared with the fluorescent value reading of positive control Rosiglitazone, if sieve The activity of lattice row ketone is 100%, and the activity of other compounds is expressed as relative activity, i.e.,：

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

Section Example Activity Results

Claims (14)

1. the pyrimidine thiazole amines compound and its officinal salt as shown in formula I,

In formula

R₁Selected from hydrogen, fluorine, chlorine, bromine, C1-4 alkyl, methylol；

R₂Selected from hydrogen, fluorine, chlorine, bromine, COOC1-4 alkyl, carboxyl, aminoacyl, C1-4 alkyl substitution aminoacyl, C1-4 alkyl, hydroxyl first Base；

R₃It is selected from

R₄Selected from hydrogen, methyl, trifluoromethyl, fluorine or chlorine.

2. compound according to claim 1 and its officinal salt, it is characterised in that

R₁Selected from hydrogen, fluorine, chlorine, bromine, methyl, ethyl；

R₂Selected from hydrogen, fluorine, chlorine, bromine, methyl formate base, group-4 ethyl formate, isopropyl formate base, carboxyl, aminoacyl, methylamine acyl group, Isopropylamine acyl group, methyl, ethyl, methylol；

R₃It is selected from

R₄Selected from hydrogen, methyl, trifluoromethyl, fluorine or chlorine.

3. compound according to claim 2 and its officinal salt, it is characterised in that

R₁Selected from hydrogen, fluorine, chlorine, methyl；

R₂Selected from hydrogen, fluorine, chlorine, group-4 ethyl formate, isopropyl formate base, methyl formate base, carboxyl, aminoacyl, methylamine acyl group is different Propylamine acyl group, methyl, methylol；

R₃It is selected from

R₄Selected from hydrogen, methyl, trifluoromethyl.

4., according to the compound and its officinal salt of claim any one of 1-3, described compound is selected from：

2- (1- (2- methyl -6- (5- ethoxycarbonyl thiazole -2- amino) pyrimidine-4-yl) piperidin-4-yl) ethanol

2- (1- (2- methyl -6- (5- ethoxycarbonyl thiazole -2- amino) pyrimidine-4-yl) piperidin-4-yl) acetic acid

2- ((1- (2- methyl -6- (5- ethoxycarbonyl thiazole -2- amino) pyrimidine-4-yl) piperidin-4-yl) methoxyl group) acetic acid

。

5. according to the compound and its officinal salt of claim any one of 1-3, it is characterised in that described medicinal salt includes With inorganic acid, Organic Acid and Base metal ion, alkaline-earth metal ions or can provide the organic base knot of physiologically acceptable cation Close the salt for being formed.

6. compound according to claim 4 and its officinal salt, it is characterised in that described medicinal salt include with it is inorganic Acid, Organic Acid and Base metal ion, alkaline-earth metal ions can provide the organic base of physiologically acceptable cation and combine to form Salt.

7. compound according to claim 5 and its officinal salt, it is characterised in that described inorganic acid is selected from hydrochloric acid, hydrogen bromine Acid, phosphoric acid or sulfuric acid；Described organic acid be selected from methanesulfonic acid, p-methyl benzenesulfonic acid, trifluoroacetic acid, matrimony vine 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 from Attached bag includes calcium ion, magnesium ion；The described organic base that physiologically acceptable cation can be provided be selected from methylamine, dimethylamine, Trimethylamine, piperidines, morpholine or three (2- ethoxys) amine.

8. compound according to claim 6 and its officinal salt, it is characterised in that described inorganic acid is selected from hydrochloric acid, hydrogen bromine Acid, phosphoric acid or sulfuric acid；Described organic acid be selected from methanesulfonic acid, p-methyl benzenesulfonic acid, trifluoroacetic acid, matrimony vine 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 from Attached bag includes calcium ion, magnesium ion；The described organic base that physiologically acceptable cation can be provided be selected from methylamine, dimethylamine, Trimethylamine, piperidines, morpholine or three (2- ethoxys) amine.

9. the method for preparing compound described in claim any one of 1-8, comprises the following steps：

With thiazole amine derivative 1 as raw material, generation midbody compound 3 is reacted in the basic conditions with dichloro pyrimidine derivative 2, Again with R₃H reactions obtain product 4；Wherein R₁-R₄Definition it is identical with any one of claim 1-8；

10. the method for preparing compound described in claim any one of 1-8, comprises the following steps：

With halo thiazole derivative 5 as raw material, with the reaction generation target compound 7 of pyrimidinamine derivatives 6 under alkaline environment；Wherein R₁-R₄Definition it is identical with any one of claim 1-8；X is halogen；

A kind of 11. pharmaceutical compositions, including as active ingredient claim any one of 1-8 described in compound or its can medicine The carrier commonly used on salt and pharmaceutics.

Compound and its officinal salt described in 12. claim any one of 1-8 prepare prevention and or treatment and glucokinase Application in the medicine of the enzyme disease relevant with peroxisome proliferation activated receptor-α.

13. applications according to claim 12, it is characterised in that described swashs with glucokinase and peroxidase paraphyte The receptor related disease of work is selected from diabetes, the chronic complicating diseases of diabetes and obesity.

14. applications according to claim 13, it is characterised in that described diabetes are selected from type 1 diabetes or diabetes B；Institute It is hard that the chronic complicating diseases of the diabetes stated are selected from retinopathy, ephrosis, neurosis, ischemic heart disease or artery Change.