CN105147677A - A group of 2-substituted formamido-3-alkyloxypyridine derivatives and pharmaceutical application thereof - Google Patents

Abstract

The invention relates to application of a compound as shown in formula I in preparing medicines for treating PPAR-mediated (peroxisome proliferator activated receptor) diseases and for improving or preventing PPAR-mediated symptoms. The invention is also applicable to a pharmaceutical composition of the compound for the purpose of treatment and/or prevention, wherein the pharmaceutical composition contains therapeutically effective amounts of selected active ingredients and optional pharmaceutically acceptable carriers.

Description

One group of 2-substituted formyl amido-3-alkoxy pyridines analog derivative and pharmaceutical applications thereof

Technical field

The present invention relates to 2-substituted formyl amido-3-alkoxy pyridines compounds, be applied to treating and/or preventing of type ii diabetes disease, belong to pharmaceutical field; Invent described compound active at activation nuclear receptors PPAR's γ, PPAR α or PPAR δ, reduce blood glucose, increase glucose uptake or utilization, improve glucose or insulin tolerance abnormal or increase insulin sensitivity; The invention still further relates to the pharmaceutical composition of described compound.

Background technology

Diabetes (DM, diabetesmellitus) be the endocrine metabolism disease being feature with hyperglycemia and glucose in urine because hypoinsulinism in body or insulin resistant cause, be a very important risk factor in metabolism syndrome, now become the disease of the third-largest harm humans health being only second to cardiovascular disease and cancer.In recent years, the sickness rate of global obesity rates and type ii diabetes (Type2DiabetesMellitus, T2DM) increases just rapidly.The number that World Health Organization (WHO) expects global type ii diabetes in 2035 will rise to 5.92 hundred million [1], at present only Chinese adult onset diabetes rate just up to 11.6%[2].Therefore in the urgent need to developing effective treatment and preventative strategies can.Type ii diabetes for principal character, simultaneously with fat, sugar and protein metabolism exception, is a kind of important risk factor of metabolism syndrome with hyperglycemia and insulin resistant.Except lifestyle modification, preventive medicine or the specific food supplement of application safety may contribute to the type ii diabetes [3] that treatment take insulin resistant as feature.

Nuclear receptor is the nuclear factor superfamily that a class has ligand activation, by participating in comprising the nearly all physiological process of the human bodies such as cell proliferation, differentiation, apoptosis, immunity, metabolism in body to the transcriptional control effect of gene.The major disease that the mankind are nearly all, as all relevant with its Abnormal regulation with the generation of cancer in cardiovascular disease, diabetes, obesity.Therefore nuclear receptor has not only become the focus of biomedical boundary study mechanism, and becomes the novel targets of kind new medicine research and development.International nearly all large drugmaker such as GlaxoWellcome, Roche, Lilly etc. all carry out the research and development of nuclear receptor medicine.

PPAR (PeroxisomeProliferator-ActivatedReceptors, PPARs), being also called peroxisome increment activated receptor, is one of nuclear receptor superfamily member, and adjustment glycolipid metabolism stable state plays an important role.PPAR has three hypotype: PPAR α, PPAR γ and PPAR β/δ.Have very high homology between three hypotypes, and mechanism of action being identical, is all when ligand activation, with retinoid X receptor (RetinoidXreceptor, RXR) form dimer, be combined transcribing [4,5] of downstream target gene with response element.But because three hypotypes exist tissue specificity, so the physiological function in body is also had nothing in common with each other.PPAR α mainly expresses in hepatocyte, has important regulating action at lipid metabolism and cholesterol metabolism, is the target spot of current hypolipidemic fibrate.PPAR γ expresses the highest in adipose cell, [6] are played a significant role in adjustment Adipocyte Differentiation, lipid metabolism, maintenance glucose homeostasis and insulin sensitivity, it is the action target spot of II phase diabetes insulin sensitivity enhancing medicine thiazolidinediones (Thiazolidinediones, TZDs) para-insulin sensitizer rosiglitazone (RGZ) and pioglitazone (PGZ) clinically.But RGZ and PGZ creates a series of less desirable side effect, as put on weight, edema, osteoporosis, tumor and cardiovascular risk [7,8].Then wide expression is in each tissue for PPAR β/δ, but the agonist of PPAR δ is in recent years tested proof in animal body and significantly can be suppressed fat, reduces insulin resistant.After PPARs is activated by endogenic ligand or pharmacological, promote the gene transcriptional activation or the Transcription inhibition that represent various physiological functions in a large number, and these gene expression products directly or indirectly relate to the metabolism of energy metabolism, lipid and carbohydrate.Thus, the medicine that to develop based on PPARs be target spot is used for the treatment of type ii diabetes and relevant disease is very attractive [9,10].

The typical structure schematic diagram of nuclear receptor, mainly comprises two domains, and one is DNA binding domain, and one is ligand binding domain.When part and receptors bind, DNA binding domain is transcribed in conjunction with controlling gene with response element.The mechanism of action of nuclear receptor: do not having under part, nuclear receptor can not be combined with the response element of receptor, so the target gene in downstream can not be transcribed.But after ligand activation, nuclear receptor occurred conformation changes, and is formed and has activated complex, be combined with response element, thus starts transcribing of downstream gene.

As mentioned above, because PPAR is the target that treatment comprises the metabolic disease of diabetes, diabetic complication or similar conditions, so have many exploitation PPAR parts so far for activating the trial of PPAR.United States Patent (USP) 6,939,875 disclose the disease and the compositions that uses that are used for the treatment of PPAR mediation, and described compositions is effective in cure to non-insulin-dependent diabetes mellitus, obesity, eating disorder (eatingdisorders), appetite low (suppressingappetite), leptin level modulation (leptinlevelmodulation), metabolic syndrome (metabolicsyndrome).In addition, United States Patent (USP) 6,967,212 disclose about having the effect of PPAR agonist and containing the compositions of minaline derivant replaced, and described compositions is impaired to insulin resistant, hyperglycemia, Hyperinsulinism, hyperlipemia, obesity, X syndrome, Developmental and Metabolic Disorder syndrome, inflammation, diabetic complication (diabeticcomplications), sugared stable state, impaired glucose tolerance, HTC (hypertriglyceridemia) etc. have curative effect.In addition, the material of the various diseases of known other treatment PPAR mediation (United States Patent (USP) 6,930,120, United States Patent (USP) 7,041,691 and United States Patent (USP) 7,037,914).

But, because PPAR part known at present causes side effect such as the toxicity of body weight increase, edema, osteoporosis, liver, be necessary to develop the side effect such as toxicity little, the part that can activate PPAR.

Country of Inst. of Medicinal Biological Technology, Chinese Academy of Medical Sciences new drug (microorganism) screening experiment room utilizes the single acrobatics art of mammal to set up PPARs activator screening system (specifically introducing as embodiment 1).The PPARs activator screening system set up, we find patent (number of patent application: 201310247772.0 in early stage, date of application: on June 21st, 2013) compound (i.e. 2-amide groups-3-alkoxy pyridines compounds) in [11], have in vitro and significantly activate PPAR γ, PPAR α, PPAR δ activity, there is the new role for the treatment of type Ⅱdiabetes mellitus.Particular content comprises, 2-substituted formyl amido-3-alkoxy pyridines class (10 μ g/ml) in vitro can activation PPAR γ, PPAR α in various degree, PPAR δ (table 1), wherein, LX-1, LX-3, LX-5, LX-7, LX-11, LS-14, LS-16, LS-20, LS-35 etc. are strong to the activation of PPAR γ, PPAR δ, and the activation to PPAR α such as LX-1, LX-3, LX-5, LX-7, LX-11, LS-4, LS-14, LS-16 is strong.Molecule and cellular level confirm, the inducing action to Adipocyte Differentiation such as LX-1, LX-5, LS-14, LS-35 more weak (Fig. 2).Hepatocyte metabolizable glucose (Fig. 3) can be promoted.Compared with rosiglitazone, compound L X-1, LX-5, LS-14, LS-35 are not only suppressed to bone cell differentiation, can also promote osteoblast differentiation (Fig. 4).At spontaneous type 2 diabetes mellitus model KKA ^yin Mice Body, gavage LX-1 (50mg/kg) obviously can reduce fasting glucose, improves glucose and insulin tolerance exception, increases insulin sensitivity; To reduce in blood the levels (Fig. 9) such as TC, LDL, reduce TC, CE equal size (Figure 10) in liver; But obvious body weight can not be induced to increase.Embodiment illustrates compound (i.e. 2-amide groups-3-alkoxy pyridines compounds), really has in vitro and significantly activates PPAR γ, PPAR α, PPAR δ activity, has treatment I type, type ii diabetes, osteoporotic new role.

In a word, 2-substituted formyl amido-3-alkoxy pyridines compounds there are no bibliographical information, is Late Cambrian of the present invention in type ii diabetes effect.2-substituted formyl amido-3-alkoxy pyridines compounds can increase cell to the picked-up of glucose and utilization, differentiation-inducing more weak to adipose cell, animal blood glucose can be reduced, improve glucose or insulin tolerance exception, improve insulin sensitivity, thus become the medicine of type ii diabetes.Therefore, this group 2-substituted formyl amido-3-alkoxy pyridines compounds has significant blood sugar reducing function, in the treatment of type ii diabetes, have wide development prospect.

Summary of the invention

2-substituted formyl amido-3-alkoxy pyridines class of the present invention, its structure is as logical formula I compound, the present invention is at patent (number of patent application: 201310247772.0 in early stage, date of application: on June 21st, 2013) basis on, synthesize new particular compound, and find 2-amide groups-3-alkoxy pyridines compounds external have significantly activate PPAR γ, PPAR α, PPAR δ is active, has the effect for the treatment of type ii diabetes.

Present invention also offers the pharmaceutical composition of described compound, it is characterized in that it with the above-claimed cpd containing treatment effective dose for active component, and containing one or more pharmaceutically acceptable carriers.

Compound of the present invention and the application of compositions in preparation treatment type ii diabetes medicine.

The various dosage forms of medicine of the present invention or compositions, can prepare according to the conventional production process of pharmaceutical field, such as, make active component mix with one or more carriers, be then made into required dosage form.

The present inventor utilizes PPAR γ, PPAR α, the activity of PPAR δ screening model to patents is evaluated, 0.1%DMSO is selected to be negative control, rosiglitazone is positive control, measures the rate of change of all compounds under 10 μ g/ml concentration in model (PPAR γ, PPAR α, PPAR δ).Repeatedly determination of activity the results show, the compounds of this invention all has agonism on PPAR γ, PPAR α, PPAR δ model, and activity and the measurement result of part of compounds are as shown in table 1.Table 1 is only and helps those skilled in the art to understand the present invention better, but does not limit the present invention in any way.

The present invention finds that general formula (I) LX-1, LX-3, LX-5, LX-7, LX-9, LX-11, LX-18, LX-22, LS-13, LS-14, LS-16, LS-20, LS-35 etc. have the effect of exciting PPAR γ, PPAR δ, and the activation to PPAR α such as LX-1, LX-3, LX-5, LX-7, LX-11, LX-20, LS-4, LS-14, LS-15, LS-16 is strong.Wherein, especially LX-1, LX-3, LX-5, LX-7, LX-11, LS-14, LS-16, LS-20, LS-35 demonstrate the activity of good exciting PPAR γ, the activation to PPAR α such as LX-1, LX-3, LX-5, LX-7, LX-11, LS-4, LS-14, LS-16 is strong, and provide them treating the application in type-II diabetes, lay a good foundation for this analog derivative develops into the novel anti-type-II diabetes medicine of a class.LX-1 under 0.016,0.08,0.4,2 and 10 μM of five concentration, on PPAR γ, PPAR α, PPAR δ tri-models all can dose-dependent activation PPAR γ, PPAR α, PPAR δ (Fig. 1).Prove thus, compound of the present invention or compositions can as the medicines of exciting or increase PPAR γ, PPAR α, PPAR δ activity.

In the present invention, the described induction of differentiation to adipose cell refers to, utilize 3T3-L1 PECTORAL LIMB SKELETON to induce 48h, then cultivate seven days in expansion culture medium, whether investigation compound breaks up the adipose cell becoming ripe process to 3T3-L1 PECTORAL LIMB SKELETON has inducing action.3T3-L1 PECTORAL LIMB SKELETON Induction experiments result shows, 3T3-L1 cell after induction 48h only cultivates seven days in containing the expansion culture medium of 10 μMs of RGZ, just can break up completely and become ripe adipose cell, fat drips number and almost reaches 100%, can be observed fat under mirror after oil red dyeing and drip interior abundant accumulation of lipid, illustrate that RGZ can accelerate the accumulation that the differentiation and maturation of 3T3-L1 cell and fat drip interior triglyceride.And under kindred circumstances, LX-1, LX-5, LS-14, LS-35 of 10 μMs then show more weak lipogenesis effect (Fig. 2), the pastille cultivation fat of seven days drips quantity and is no more than 50%, shows compound retards and reduces 3T3-L1 cell differentiation and lipogenetic inducing action.Prove thus, compound on fatty cell of the present invention differentiation-inducing more weak.

TZDs can increase the sensitivity of insulin sensitive tissues to insulin, and an important performance is the picked-up and the disposal that promote glucose.Therefore, we have investigated the impact that compound absorbs hepatic glucose.1,5-anhydroglucitol (2-deoxyglucose, 2-DG) is the derivant of natural D type glucose, is equally to be transported in cell by Glut1 to carry out metabolism.2-NBDG is the fluorescent analog of 1,5-anhydroglucitol (2-deoxyglucose, 2-DG), namely the oxygen atom of 2 ' position replace by fluorophor NBD.Therefore we utilize 2-NBDG to carry out the amount of indicator cells ingestion of glucose.LX-1, LX-5, LS-14, LS-35 of 10 μMs process 24h respectively all significantly can increase L02 hepatocyte (Fig. 3) to the picked-up of 2-NBDG.These results point out compound of the present invention having the glycometabolic effect of good improvement.

In the present invention, described osteoblast differentiation effect to be referred to, utilize pre-osteoblast MC3T3-E1 (Subclone14) vitro differentiation to be osteoblast, utilize the external investigation compound of Alizarin red staining on the impact of osteoblast differentiation.Result shows, after induction 21d, MC3T3-E1 (Subclone14) cell defines calcium scoring, after 10 μMs of compound L X-1 (Fig. 4 c), LX-5 (Fig. 4 d), LS-14 (Fig. 4 e), LS-35 (Fig. 4 f) intervene, (Fig. 4 a) obviously increases the more non-intervention group of area of calcification; And the more non-intervention group of area of the RGZ processed group of 10 μMs (Fig. 4 b) calcification (Fig. 4 a) obviously reduces.Prove that compound of the present invention can promote osteoblast differentiation thus, can not osteoporotic side effect caused by the medicine of TZDs together.Prove, compound of the present invention is useful to treatment osteoporosis simultaneously, can as the osteoporotic medicine for the treatment of.

Results of animal shows, and within the 0th, 3,7,14,21 day of oral administration gavage administration LX-1 (50mg/kg), measures the fasting glucose of mice.Relative to matched group, the LX-1 of 50mg/kg significantly can reduce the fasting glucose (Fig. 6 A) of mice.For the steady-state model insulin resistance index (HOMA-IR) calculating through fasting glucose and insulin and come, LX-1 then can reduce HOMA-IR (Fig. 6 B).Oral glucose tolerance test (OGTT) result shows, compared with matched group, LX-1 significantly can to slow down after glucose load 30,60, the rising (Fig. 7 A) of blood sugar level after 120min, reduce Area under the curve of blood glucose (Fig. 7 B) in OGTT.ITT result shows, and compared with matched group, LX-1 (50mg/kg) administration 21 days, significantly can reverse KKA ^yin mice fasting glucose and ITT test 40,90, the blood glucose (Fig. 8 A) of 120min, significantly reduce AUC (Fig. 8 B).Therefore, LX-1 has clear and definite improvement diabetes KKA ^ythe opposing of mouse islets element, increases the effect of its insulin sensitivity.Administration is after 21 days, and relative to body weight before administration, RGZ group Mouse Weight adds 24% (Fig. 5).To administration the 21st day, KKA ^ymouse Weight only increases by 13% relative to before administration, significantly lower than matched group 20%, therefore, shows the effect that compound can not cause body weight and increases.Administration 21 days, LX-1 significantly can lower serum total cholesterol (TC), triglyceride (TG) level (Fig. 9), reduce the accumulation (Figure 10) of T-CHOL (TC) in liver, cholesteryl ester (CE), illustrates that compound has and improve blood fat, reduce lipid accumulation, the effects such as obesity can not be caused.Prove thus, compound of the present invention or compositions can reduce blood glucose in animal body, improve glucose or insulin tolerance exception, can as treatment type ii diabetes medicine.

One aspect of the invention provides the purposes of a kind of formula I in the disease of preparation treatment PPAR mediation and the medicine of improvement or prevention PPAR mediation disease, and wherein, formula I is as follows,

R ₁represent the alkyl of H atom and C1-C4

R ₂, R ₃, R ₄independently represent the alkyl of C1-C3, the oxyl of C1-C3, halogen atom and H atom

R ₅for-(CH ₂) _n-R ₆or-(CH ₂) _n-O-R ₆or-(CH ₂) _n-S-R ₆, wherein, the integer of n=0-4;

R ₆independently representative includes H, C1-C4 alkyl, C1-C3 alcoxyl (sulfur) base, halogen atom, the substituent groups such as 1,3-dioxy (sulfur) Pentamethylene. base are at the cycloalkane, 1 of interior phenyl ring, pyridine ring, pyrimidine ring, piperidine ring, piperazine ring, C3-C6,2,3-triazole, 1,2,4-triazole, pyrrole ring, furan nucleus, oxazole ring, imidazole ring, thiphene ring, thiazole ring and benzimidazole, benzothiazole, benzoxazole, benzofuran, benzothiophene and indole ring.

Further, the disease of wherein said PPAR mediation and improvement or prevention PPAR mediate disease is the disease that mediates of PPAR γ, PPAR α or PPAR δ or disease.

Further, the disease of wherein said PPAR mediation is that diabetes, hyperinsulinemia, obesity, hyperglycemia, hyperlipemia, impaired fasting glucose (IFG), X syndrome, hypercholesterolemia, hyperlipoproteinemia, insulin resistant, dysmetabolic syndrome, diabetic complication, sugared stable state are impaired, impaired glucose tolerance, hypertriglyceridemia, osteoporosis, preferably, described diabetes are type i diabetes or type ii diabetes.

Further, wherein said improvement or prevention PPAR mediation disease are:

-increase glucose uptake or utilization;

-improve glucose-tolerant;

-improve insulin resistant or increase insulin secretion;

-improve and/or maintain glycemic control and/or reduce fasting glucose, post-prandial glycemia, blood glucose and/or glycosylated hemoglobin HbA1c after absorption;

-prevent, slow down, to postpone or reverting diabetes early stage, glucose tolerance reduce (IGT), impaired fasting glucose (IFG), insulin resistant and/or metabolism syndrome progress for type ii diabetes;

-prevent diabetes complication, reduce its risk, slow down its progress, postpone or treat this diabetic complication, as: blood capillary and macrovascular diseases, as nephropathy, trace or High-grade Proteinuria, albuminuria, retinopathy, cataract, neuropathy, learn or remember impaired, neurodegenerative or cognitive illnesses, cardiovascular or cerebrovascular disease, tissue ischemia, diabetic foot or ulcer, hypertension, Endothelial Dysfunction, myocardial infarction, acute coronary, unstable angina pectoris, stable angina pectoris, Peripheral arterial occlusive disease, cardiomyopathy, heart failure, heart rhythm disorders, vascular restenosis and/or apoplexy,

-to reduce in body weight and/or body fat and/or liver fat and/or myocyte fat in fat or prevention body weight and/or body fat and/or liver fat and/or myocyte and increase or promote that in body weight and/or body fat and/or liver fat and/or myocyte, fat reduces;

-prevent, slow down, postpone or treat the degeneration of pancreatic beta cell and/or going down and/or improving, keep and/or recover the function of pancreatic beta cell and/or the function of stimulation and/or the secretion of recovery pancreatic insulin of Pancreatic beta cells function;

-prevent, slow down, postpone or treat non-alcoholic fatty liver disease (NAFLD), it comprises fatty degeneration of liver, non-alcoholic stellato-hepatitis (NASH) and/or hepatic fibrosis, such as, prevent, slow down progress, postpone, weaken, treat or reverse fatty degeneration of liver, (liver) inflammation and/or the abnormal accumulation of liver fat;

-prevention, to postpone or treatment is single to conventional anti-diabetic or type ii diabetes that combined therapy is invalid, slow down its progress;

-reach the dosage reduced in order to the conventional antidiabetic medicine needed for abundant therapeutic effect;

The risk of the untoward reaction that-reduction is relevant with conventional antidiabetic medicine, such as hypoglycemia or body weight increase; And/or

-keep and/or improve insulin sensitivity and/or be used for the treatment of or prevent hyperinsulinemia and/or insulin resistant.

Further, in its Chinese style (I) compound, R ₁for H, alkyl is methyl or the tert-butyl group, and alcoxyl (sulfur) base is trifluoromethoxy or trifluoromethylthio, R ₆representative ring system is phenyl ring, furan nucleus, pyrrole ring, imidazole ring or benzimidazole ring.

Further, shown in its Chinese style (I), compound is as shown in the table,

The present invention another aspect provides the purposes of a kind of pharmaceutical composition in the disease of preparation treatment PPAR mediation and the medicine of improvement or prevention PPAR mediation disease, it is characterized in that described pharmaceutical composition contains formula (I) compound for the treatment of effective dose as active component, and one or more pharmaceutically acceptable carriers.

Further, described pharmaceutical composition contains the active component that weight ratio is 0.01%-99.5%, and preferably containing weight ratio is the active component of 0.5%-99.5%.

Further, the disease of described PPAR mediation and improvement or prevention PPAR mediate disease is the disease that mediates of PPAR γ, PPAR α or PPAR δ or disease.

Further, the disease of described PPAR mediation is that diabetes, hyperinsulinemia, obesity, hyperglycemia, hyperlipemia, impaired fasting glucose (IFG), X syndrome, hypercholesterolemia, hyperlipoproteinemia, insulin resistant, dysmetabolic syndrome, diabetic complication, sugared stable state are impaired, impaired glucose tolerance, hypertriglyceridemia, osteoporosis, preferably, described diabetes are type i diabetes or type ii diabetes.

The beneficial effect of the invention

1) having set forth 2-substituted formyl amido-3-alkoxy pyridines compounds has PPAR γ, PPAR α or PPAR δ activation for the first time in the present invention, and the purposes scope of this compounds has been expanded in this application greatly.

2) the present invention has set forth the definite effect of 2-substituted formyl amido-3-alkoxy pyridines compounds in type-II diabetes treatment first time; The compound that the present invention finds has good hypoglycemic effect in animal body, for development of new type-II diabetes medicine provides good lead compound, has broad application prospects.

3) the present invention sets forth the molecular mechanism of 2-substituted formyl amido-3-alkoxy pyridines compounds for treating type-II diabetes effect first, this at home and abroad belongs to first, and this anti-type ii diabetes medicine having independent intellectual property right for China's exploitation has great importance.

4) the present invention has set forth the effect that 2-substituted formyl amido-3-alkoxy pyridines compounds promotes osteoblast differentiation first, for development of new medicine for treating osteoporosis provides good lead compound, has broad application prospects.

Prepare the universal method of the employing amides compound of described compound.Key step is, 2-amino-3-alkoxy pyridines compounds (II) of the carboxylic acid (III) replaced by different ring system and replacement is as raw material, using EDCI as condensing agent, under suitable solvent and temperature conditions, final synthesis obtains corresponding compound in general formula (I), and general formula synthetic route is as follows:

According to the above route and method, can stablize, repeatable synthesis obtain all compounds of the present invention.

Compound of the present invention also can adopt number of patent application: the method for 201310247772.0 is prepared.

List of references

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[3]BergerJ,MollerDE.ThemechanismsofactionofPPARs[J].AnnuRevMed,2002,53:409-435.

[4]LembergerT,DesvergneB,WahliW.Peroxisomeproliferator-activatedreceptors:anuclearreceptorsignalingpathwayinlipidphysiology[J].AnnuRevCellDevBiol,1996,12:335-363.

[5]GelmanL,FeigeJN,DesvergneB.MolecularbasisofselectivePPARgammamodulationforthetreatmentofType2diabetes[J].BiochimBiophysActa,2007,1771(8):1094-1107.

[6]B.B.KahnandMcGrawT.E.,Rosiglitazone,PPARgamma,andtype2diabetes.NEnglJMed,2010.363(27):p.2667-9.

[7]WillsonTM,LambertMH,KliewerSA.Peroxisomeproliferator-activatedreceptorgammaandmetabolicdisease[J].AnnuRevBiochem,2001,70:341-367.

[8]E.Mannucci,MonamiM.,DiBariM.,etal.,Cardiacsafetyprofileofrosiglitazone:acomprehensivemeta-analysisofrandomizedclinicaltrials.IntJCardiol,2010.143(2):p.135-40.

[9]A.YessoufouandWahliW.,Multifacetedrolesofperoxisomeproliferator-activatedreceptors(PPARs)atthecellularandwholeorganismlevels.SwissMedWkly,2010.140:p.w13071.

[10]J.D.BrownandPlutzkyJ.,Peroxisomeproliferator-activatedreceptorsastranscriptionalnodalpointsandtherapeutictargets.Circulation,2007.115(4):p.518-33.

[11] official documents writer firm, Hong Bin, Xu Yanni, Jia Xiaojian, Li Yongzhen, Liu Chang, Wang Li, Liu Qi. the pyridine compounds and their that one group of 2-amide groups-3-alkoxyl replaces and novelty teabag thereof, China, on June 21st, 201310247772.0,2013.

Accompanying drawing explanation

Figure 1A, Figure 1B and Fig. 1 C be LX-1 under variable concentrations to the activation in PPAR γ, PPAR α or PPAR δ screening model;

Fig. 2 is the differentiation-inducing experiment of LX-1 murine preadipocyte 3T3-L1 cell, wherein a: blank; B: contrast RGZ; C-f is followed successively by: LX-1, LX-5, LS-14, LS-35 (compound concentration is 10 μMs);

Fig. 3 is the impact of compound on L02 grape cell Sugar intake, wherein a: blank; B: positive control; C-f is followed successively by: LX-1, LX-5, LS-14, LS-35 (compound concentration is 10 μMs);

Fig. 4 is the impact of Alizarin red staining detection compound on external osteoblast calcification, wherein LX-1, LX-3, LX-7 (compound concentration is 10 μMs);

Fig. 5 is the impact of compound L X-1 on body weight;

Fig. 6 A and Fig. 6 B is that compound L X-1 is at spontaneous type type ii diabetes mice KKA ^yin body, fasting glucose and HOMA-IR measure;

Fig. 7 A and Fig. 7 B is the impact of compound L X-1 on impaired glucose tolerance;

Fig. 8 A and Fig. 8 B is the impact of compound L X-1 on insulin tolerance exception;

Fig. 9 is the impact of compound L X-1 on blood lipid level;

Figure 10 is the impact of compound L X-1 on lipid in liver.

Detailed description of the invention

The concrete same patent (number of patent application: 201310247772.0 of the preparation embodiment of compound in the present invention,), it can make professional and technical personnel more fully understand the present invention, but does not limit the present invention in any way, and the structure of all compounds all determined through 1HNMR and MS.

Embodiment 1 the compounds of this invention is to PPAR γ, PPAR α or PPAR δ activation

Nuclear receptors PPAR's γ/α/δ has two main domains: ligand binding domains (LBD) and DNA binding structural domain (DBD), they have independently function separately.Utilize the single acrobatics art of mammal to set up PPARs activator screening system, build PPAR γ/α/δ-LBD/GAL4 cotransfection model inspection compound to the effect of PPAR γ/α/δ.This mechanism make use of two main domains total in the molecular structure of nuclear receptor: ligand binding domains (LBD) and DNA binding structural domain (DBD), and yeast cells transcription factor GAL4 has the feature of the analog structure of nuclear receptor.The ligand binding domains (LBD) of nuclear receptors PPAR's γ/α/δ and the DNA binding structural domain (DBD) of yeast cells transcription factor GAL4 are fused into chimeric protein expression plasmid pBIND-PPAR γ-LBD, pBIND-PPAR α-LBD, pBIND-PPAR δ-LBD.The respective element of synthetic 5 × GAL4, inserts reporter gene upstream and builds reporter plasmid pG5-promotor-GAL4 (being called for short GAL4).Chimeric protein expression plasmid and the reporter plasmid cotransfection containing the special corresponding original paper of GAL4 will be fused into, by measuring the expression of reporter gene thus evaluating the agonist activity of nuclear receptor ligands.

Concrete operation step is as follows: the recombiant plasmid pBIND-PPAR γ/α/δ-LBD of GAL4 reporter plasmid and structure mediates cotransfection HepG2 cell through LipofectamineTM2000 (Invitrogen), 5 × 104 cells/well (96 orifice plate), 200ng plasmid DNA (ratio of reporter plasmid GAL4 and expression plasmid pBIND-PPAR γ/α/δ-LBD is 10:1), 0.5 μ l liposome/hole, the MEM culture medium of serum-free is changed to after transfection 6h, continue to hatch 18h after adding certain density testing sample simultaneously, blank adds containing the MEM culture medium with testing sample same concentrations DMSO.Measure uciferase activity in the HepG2 cell of transient transfection.The determinand that testing sample adds in cell the indirect reaction of uciferase activity rate of change to the transcriptional activation activity of PPAR γ/α/δ, with following Equation for Calculating testing sample to the rate of change of the uciferase activity of cell:

Rate of change (%)=A/B × 100

Wherein, A is cell fluorescence element enzymatic activity (RLU) measured after adding testing sample, cell fluorescence element enzymatic activity (RLU) that B measures afterwards for adding blank control sample (DMSO).

Experimental result is as follows:

Table 1: part of compounds in 10 μ g/ml concentration to the activation of PPAR γ, PPAR α, PPAR δ, relative to the multiple (table 1) of blank.Wherein, LX-1, LX-3, LX-5, LX-7, LX-11, S-14, S-16, S-20, S-35 etc. are strong to the activation of PPAR γ, PPAR δ, and the activation to PPAR α such as LX-1, LX-3, LX-5, LX-7, LX-11, S-4, S-14, S-16 is strong.Tentatively can be drawn the structure activity relationship of compound by the data of table 1, the aromatic rings be directly connected with acyl group is larger to activity influence.If when aromatic rings is phenyl ring, furan nucleus, pyrrole ring, better active.If aromatic rings is phenyl ring, there is electron withdraw group and there is its activity favourable in para-position.If replace containing sulphur atom in the side chain be connected with parent nucleus, then activity reduces greatly.

Table 1

The differentiation-inducing experiment of embodiment 2 murine preadipocyte 3T3-L1 cell

1) induction of 3T3-L1 cell and differentiation

When 3T3-L1 cell (country of Inst. of Medicinal Biological Technology, Chinese Academy of Medical Sciences preserves new drug (microorganism) screening experiment room) grows to 70-80% degree of converging, by certain cell density kind (6 orifice plates: 8 × 10 in culture plate after digestion ⁴/ hole; 96 orifice plates: 2 × 10 ³/ hole; 24 orifice plates: 2 × 10 ⁴/ hole).Cultivation is continued after 3 days in expansion culture medium, grow to 100% degree of converging, change liquid, continue to cultivate 48h in Post-confluence after, change the division culture medium of appropriate volume into, inducing culture is 48h at least, division culture medium is abandoned in careful suction, and change into and maintain or expansion culture medium, now 3T3-L1 cell is induced, entering the idiophase, is DifferentiatingStage.If add maintain base after induction, within every 2 days, change liquid once, maintain after 10-14 days, 3T3-L1 PECTORAL LIMB SKELETON is then divided into ripe adipose cell completely, fat drips obviously, form the crowded cells layer of class loading block, the adipose cell now for having broken up, is DifferentiatedStage or MatureAdipocytes.

3T3-L1 Cell expansions culture medium (ExpansionMedium): add final concentration 10% (V/V) calf serum in DMEM-HG culture medium (Gibco), cultivate for 3T3-L1 passage.

3T3-L1 cell maintain base (MaintenanceMedium): add 280 μ L recombinant human insulin (100U/mL) injection and 10mL hyclone (FBS) in 90mLDMEM-HG culture medium, be mixed with the DMEM-HG culture medium containing 10 μ g/mL insulins and 10%FBS, i.e. maintain base, 4 DEG C of preservations.

2) the Oil-RedO dyeing of compound impact that the 3T3-L1 cytolipin of differential period is heaped and 3T3-L1 cell

The 3T3-L1 cell of just having induced in 24 orifice plates, abandons division culture medium, changes the fresh expansion culture medium containing prescribed concentration compound into, continue cultivation 7 days, within every two days, change once fresh pastille culture medium, treat the 7th day, carry out Oil-RedO dyeing, staining procedure is as follows:

A. elder generation is carefully light and slow siphons away culture fluid;

B. the light and slow rinsing cell of PBS 1 time is used;

C. add 4% paraformaldehyde and fix 15-30min, fixing is cell membrane;

D. the light and slow rinsing cell of PBS 1 time is used;

E. dilute Oil-RedO storage liquid, Oil-RedO: deionized water=3:2, filter paper filtering, room temperature places 10min;

F. dye about 15-30min, and the volume covering added is lived at the bottom of plate, and as 6 orifice plates add 1.5ml, 24 orifice plates add 0.5-1ml, and for avoiding Oil-Red to separate out, dyeing time is unsuitable long;

G. decolour, by 75% ethanol or 60% isopropyl alcohol, remove unnecessary dyestuff;

H. the light and slow rinsing cell of PBS 1 time is used;

I. basis of microscopic observation taking pictures.

Experimental result

We utilize mice 3T3-L1 PECTORAL LIMB SKELETON to evaluate the impact of reactive compound on Adipose Differentiation and generative capacity, and experimental result is see accompanying drawing 2, wherein a: blank; B: positive control; C-f is followed successively by: LX-1, LX-5, LS-14, LS-35 (compound concentration is 10 μMs).

As shown in Figure 2,3T3-L1 cell after induction 48h only cultivates seven days in containing the expansion culture medium of 10 μMs of RGZ, just can break up completely and become ripe adipose cell, fat drips number and almost reaches 100%, cell multilamellar and fat drips intensive, can be observed fat under mirror after oil red dyeing and drip interior abundant accumulation of lipid, illustrate that RGZ can accelerate the accumulation that the differentiation and maturation of 3T3-L1 cell and fat drip interior triglyceride.And under kindred circumstances, LX-1, LX-5, LS-14, LS-35 of 10 μMs then show more weak lipogenesis effect, the pastille cultivation fat of seven days drips quantity and is no more than 30%, and show relative to RGZ, reactive compound delays and reduces 3T3-L1 cell differentiation and lipogenetic inducing action.Prove thus, compound on fatty cell of the present invention differentiation-inducing more weak, reduce lipid and generate, may be used for the treatments such as hyperlipemia, hypercholesterolemia, dysmetabolic syndrome, hypertriglyceridemia.

The different cell 2-NBDG of embodiment 3 absorbs experiment and sugar utilizes experiment

1) compound is on the impact of L02 grape cell Sugar intake

When L02 cell (country of Inst. of Medicinal Biological Technology, Chinese Academy of Medical Sciences preserves new drug (microorganism) screening experiment room) grows to 80% degree of converging, digestion, by 2 × 10 ⁴/ hole is laid in 96 hole luciferase targets.After continuing to cultivate 24h, for experiment, process is as follows.

A. in 96 hole luciferase targets, 3T3-L1 PECTORAL LIMB SKELETON is induced to be divided into mature fat cell;

B. change the fresh expansion culture medium containing prescribed concentration medicine into, cultivate 24h;

C. exhaust each Kong Peiji, washes 2 times with PBS, and the PBS and the cell that slowly add 200 μ l/ holes hatch 30min altogether;

D. add with the 2-NBDG Incubating Solution (50 μ l/ hole) of the 0.4mM of PBS preparation, setting does not only add the blank control wells of 2-NBDG as background containing cell, gently shakes after mixing and puts into incubator, hatch 60min for 37 DEG C;

E. exhaust every hole liquid, slowly rinses cell 3 times in (200 μ l/ hole) with PBS, as far as possible soft to reduce cell loss;

F. exhaust every hole liquid completely, adds the neutrophil cell lysate (100 μ l/ hole) of pre-cooling, 4 DEG C of standing 10min, fully concussion cracking 2min, and carry out protein quantification by BCA method;

G. with multi-functional microplate reader record excitation wavelength for 485nm, fluorescence intensity when emission wavelength is 528nm, result is calibrated with total protein concentration.

Experimental result

TZDs can increase the sensitivity of insulin sensitive tissues to insulin, and an important performance is the picked-up and the disposal that promote glucose.Therefore, we utilize 2-NBDG to carry out the amount of indicator cells ingestion of glucose, thus have investigated the ability of compound to hepatocyte ingestion of glucose, and experimental result is see accompanying drawing 3.After LX-1, LX-5, LS-14, LS-35 of 10 μMs process L02 cell 24h respectively, all significantly can increase L02 hepatocyte (Fig. 3) to the picked-up of 2-NBDG.Prove that compound has the glycometabolic effect of good improvement, may be used for the treatment of dysmetabolic syndrome, diabetes etc. thus.

The differentiation-inducing experiment of embodiment 4 Development of Mouse Embryos osteoblast MC3T3-E1 cell

1) induction of MC3T3-E1 cell and differentiation

6 holes or 12 orifice plates are inoculated MC3T3-E1 cell (country of Inst. of Medicinal Biological Technology, Chinese Academy of Medical Sciences preserves new drug (microorganism) screening experiment room), cover with after monolayer until cell, culture medium is changed into the compound of the variable concentrations with the Osteoblast Differentiation inducing culture dilution containing 10mM β-phosphoglycerol and 50 μ g/mlL-ascorbic acid.37 DEG C, 5%CO ₂cultivate 21 days under condition, within every 3 days, change once fresh pastille culture medium.

2) Alizarin red staining detection compound is on the impact of external osteoblast calcification

A. after differentiation-inducing cultivation terminates, culture fluid is discarded, with PBS rinsing cell 2 times.

B.95% ethanol fixed cell 10min, distilled water rinsing 3 times.

C., under room temperature, to dye 10min with the alizarin red S solution (pH4.2) of 40mM.

Distilled water rinsing 3 times, takes pictures.

Experimental result

MC3T3-E1 cell is isolated pre-osteoblast from mice skull tissue, and its sub-clone 14 (Subcoloe14) can be divided into osteoblast in vitro, is a kind of model studied osteoblast differentiation in vitro and commonly use.A side effect of TZDs causes osteoporosis.Therefore we utilize Alizarin red staining detection compound on the impact of external osteoblast differentiation, and result as shown in Figure 4.We utilize β-phosphoglycerol and L-AA induction MC3T3-E1 (Subclone14) cell to osteoblast differentiation, Induction Process is intervened with compound L X-1, LX-5, LS-14, LS-35, RGZ and blank are set simultaneously, after 21d, carry out Alizarin red staining.As shown in Figure 4, after induction 21d, MC3T3-E1 (Subclone14) cell defines calcium scoring, after 10 μMs of compound L X-1 (Fig. 4 c), LX-5 (Fig. 4 d), LS-14 (Fig. 4 e), LS-35 (Fig. 4 f) intervene, (Fig. 4 a) obviously increases the more non-intervention group of area of calcification; And the more non-intervention group of area of the RGZ processed group of 10 μMs (Fig. 4 b) calcification (Fig. 4 a) obviously reduces.Prove that compound can promote osteoblast differentiation thus, useful to treatment osteoporosis, can as the medicine for the treatment of osteoporosis, diabetic complication.

Pharmacodynamic evaluation in embodiment 5 chemical combination object

1) laboratory animal grouping and experimental design

The spontaneous type 2 diabetes mellitus KKA of 4 month female ^ymice, first measures blood glucose and divides into groups.According to fasting glucose, body weight, be divided into 3 groups at random: matched group (Con), rosiglitazone group (RGZ, 5mg/kg), LX-1 group (50mg/kg), often organize 12.Press 0.05mL/10gBW gastric infusion every day 1 time, matched group gives normal saline, every two days record drinking water amounts and body weight, administration 21 days, measured fasting glucose in the 0th, 3,7,14,21 day, carried out oral glucose tolerance test in the 14th day, within the 21st day, carry out insulin tolerance test.

2) fasting plasma glucose

Equal more than the fasting 4h of animal (freely drinking water) before getting blood, gets tail point blood, measures fasting glucose with blood glucose meter and supporting reagent paper.

3) oral glucose tolerance test (Oralglucosetolerancetest, OGTT)

After animal fasting 6h, get blood as 0min, oral administration gavage gives glucose solution (2g/kg), after glucose load, 30min, 60min, 90min, 120min tail point gets blood, measure the blood glucose of 0min, 30min, 60min, 90min, 120min, calculate each group of Area under the curve of blood glucose (AUC).AUC (mg/dlh)=(BG ₀+ BG ₃₀) × 30/60+ (BG ₃₀+ BG ₆₀) × 30/60+ (BG ₆₀+ BG ₉₀) × 30/60+ (BG ₉₀+ BG ₁₂₀) × 30/60, wherein BG ₀, BG ₃₀, BG ₆₀and BG ₁₂₀representative to give after glucose load 0,30,60, the blood glucose of 120min.

4) insulin tolerance test (Insulintolerancetest, ITT)

After animal fasting 4h, get 0min blood, subcutaneous injection regular insulin solution (1U/kg), after injection, 40min, 90min, 120min tail point gets blood, measure the blood glucose of fasting glucose and 40min, 90min, 120min, calculate each group of Area under the curve of blood glucose (AUC).AUC (mg/dlh)=(BG ₀+ BG ₄₀) × 40/60+ (BG ₄₀+ BG ₉₀) × 50/60+ (BG ₉₀+ BG ₁₂₀) × 30/60, wherein BG ₀, BG ₄₀, BG ₉₀and BG ₁₂₀to represent after injection of insulin 0,40,90, the blood glucose of 120min.

5) level determination such as TC, TG in serum

Administration the 23rd day, after animal fasting 6h, plucks eyeball and gets blood, puts to death.After getting blood, centrifugal immediately, get serum.Utilize TC, TG, LDL to measure (enzyme process) test kit to complete.

6) TC, CE level determination

23 days, after dehematizing, fixing mice, cut off skin until abdominal part with operating scissors along the median line of cervical region, cuts off abdominal cavity, first leave and take fresh liver organization, divides 3 parts and puts into cryopreservation tube, put into liquid nitrogen immediately.

Take the liver organization 50mg of liquid nitrogen cryopreservation, utilize tissue T C, CE to measure test kit, measure after breaking into homogenate with refiner.Utilize BCA method to measure protein concentration to correct.

Experimental result

Experimental result is see accompanying drawing 5-8.Administration is after 21 days, and relative to body weight before administration, RGZ group Mouse Weight adds 23.8% (Fig. 5).To administration the 21st day, KKA ^ymouse Weight, relative to only increasing by 13.6% (Fig. 5) before administration, significantly lower than matched group, therefore, shows the effect that compound can not cause body weight and increases.

Results of animal shows, and within the 0th, 3,7,14,21 day of oral administration gavage administration LX-1 (50mg/kg), measures the fasting glucose of mice.Relative to matched group, the LX-1 of 50mg/kg significantly can reduce the fasting glucose (Fig. 6 A) of mice.Administration 21 days, for the steady-state model insulin resistance index (HOMA-IR) calculating through fasting glucose and insulin and come, LX-1 then can reduce HOMA-IR (Fig. 6 B).This illustrates that compound L X-1 significantly can reduce blood sugar level.

Oral glucose tolerance test (OGTT) result shows, compared with matched group, LX-1 significantly can to slow down after glucose load 30,60, the rising (Fig. 7 A) of blood sugar level after 120min, reduce Area under the curve of blood glucose (Fig. 7 B) in OGTT.Illustrate that LX-1 has the effect improving impaired glucose tolerance.

ITT result shows, compared with matched group, LX-1 administration 21 days, significantly can to reverse in KKAy mice fasting glucose and ITT test 40,90, the blood glucose (Fig. 8 A) of 120min, significantly reduce AUC (Fig. 8 B).Illustrate that LX-1 has the effect improving insulin tolerance exception, LX-1 has clear and definite improvement diabetic KKAY mice insulin resistant, increases the effect of its insulin sensitivity.

Administration 21 days, LX-1 significantly can lower serum total cholesterol (TC), triglyceride (TG) level (Fig. 9), reduce the accumulation (Figure 10) of T-CHOL (TC) in liver, cholesteryl ester (CE), illustrates that compound has and improve blood fat, reduce lipid accumulation, the effects such as obesity can not be caused.

Prove thus, compound of the present invention or compositions can reduce blood glucose in animal body, improve glucose or insulin tolerance is abnormal, can, impaired glucose tolerance, insulin tolerance exception, hypertriglyceridemia etc. impaired as treatment I type or type ii diabetes, hyperinsulinemia, obesity, hyperglycemia, hyperlipemia, impaired fasting glucose (IFG), X syndrome, hypercholesterolemia, hyperlipoproteinemia, insulin resistant, dysmetabolic syndrome, diabetic complication, sugared stable state.

The preparation of embodiment 6 compound L S-14 and LS-35

2-amino-3-alkoxy pyridines the compounds of the carboxylic acid replaced using corresponding ring system and replacement is as raw material, and using EDCI as condensing agent, under suitable solvent and temperature conditions, final synthesis obtains corresponding compound in LS-14 and LS-35.

LS-14：MS(ESI，m/z)：327.18[M+1]+

LS-35：MS(ESI，m/z)：650.98[2M+1]+

The mass spectrometric data of embodiment 7 part of compounds

Numbering	MS(ESI，m/z)	Numbering	MS(ESI，m/z)
				LS-6	283.09[M+1]+	LS-20	277.27[M+1]+
LS-7	329.05[M+1]+	LS-2	357.25[M+1]+
				LX-5	341.02[M+1]+	LS-36	263.23[M+1]+
LS-8	403.01[M+1]+	LS-29	322.19[M+1]+
				LX-7	313.17[M-1]+	LS-33	344.24[M+1]+
LX-11	301.27[M+1]+	LS-15	375.05[M+1]+
				LX-1	317.17[M-1]+	LS-1	355.20[M+1]+
LX-20	368.15[M+1]+	LS-5	355.18[M+1]+
				LS-17	376.29[M+1]+	LS-22	336.97[M+1]+
LS-12	410.29[M+1]+	LS-27	352.14[M+1]+
				LS-18	384.29[M+1]+	LS-19	304.19[M+1]+
LS-11	319.01[M+1]+	LS-25	362.12[M+1]+
				LS-13	352.15[M+1]+	LS-34	330.16[M+1]+
LS-24	205.11[M+1]+	LS-35	650.98[2M+1]+
				LS-14	327.18[M+1]+	LS-21	327.12[M+1]+
LX-9	219.11[M+1]+	LS-30	353.23[M+1]+
				LS-16	329.18[M+1]+	LS-32	261.15[M+1]+
LS-26	231.17[M+1]+

Claims (12)

1. the purposes of formula I in the disease of preparation treatment PPAR mediation and the medicine of improvement or prevention PPAR mediation disease, wherein, formula I is as follows,

R ₁represent the alkyl of H atom and C1-C4

R ₂, R ₃, R ₄independently represent the alkyl of C1-C3, the oxyl of C1-C3, halogen atom and H atom

R ₅for-(CH ₂) _n-R ₆or-(CH ₂) _n-O-R ₆or-(CH ₂) _n-S-R ₆, wherein, the integer of n=0-4;

R ₆independently representative includes H, C1-C4 alkyl, C1-C3 alcoxyl (sulfur) base, halogen atom, the substituent groups such as 1,3-dioxy (sulfur) Pentamethylene. base are at the cycloalkane, 1 of interior phenyl ring, pyridine ring, pyrimidine ring, piperidine ring, piperazine ring, C3-C6,2,3-triazole, 1,2,4-triazole, pyrrole ring, furan nucleus, oxazole ring, imidazole ring, thiphene ring, thiazole ring and benzimidazole, benzothiazole, benzoxazole, benzofuran, benzothiophene and indole ring.

2. purposes according to claim 1, it is the disease that mediates of PPAR γ, PPAR α or PPAR δ or disease that the disease of wherein said PPAR mediation and improvement or prevention PPAR mediate disease.

3. purposes according to claim 1, the disease of wherein said PPAR mediation is that diabetes, hyperinsulinemia, obesity, hyperglycemia, hyperlipemia, impaired fasting glucose (IFG), X syndrome, hypercholesterolemia, hyperlipoproteinemia, insulin resistant, dysmetabolic syndrome, diabetic complication, sugared stable state are impaired, impaired glucose tolerance, insulin tolerance exception, hypertriglyceridemia, osteoporosis, preferably, described diabetes are type i diabetes or type ii diabetes.

4. purposes according to claim 1, wherein said improvement or prevention PPAR mediate disease and are:

-increase glucose uptake or utilization;

-improve impaired glucose tolerance;

-improve insulin tolerance exception or increase insulin secretion;

-improve and/or maintain glycemic control and/or reduce fasting glucose, post-prandial glycemia, blood glucose and/or glycosylated hemoglobin HbA1c after absorption;

-prevent, slow down, to postpone or reverting diabetes early stage, glucose tolerance reduce (IGT), impaired fasting glucose (IFG), insulin resistant and/or metabolism syndrome progress for type ii diabetes;

-prevent diabetes complication, reduce its risk, slow down its progress, postpone or treat this diabetic complication, as: blood capillary and macrovascular diseases, as nephropathy, trace or High-grade Proteinuria, albuminuria, retinopathy, cataract, neuropathy, learn or remember impaired, neurodegenerative or cognitive illnesses, cardiovascular or cerebrovascular disease, tissue ischemia, diabetic foot or ulcer, hypertension, Endothelial Dysfunction, myocardial infarction, acute coronary, unstable angina pectoris, stable angina pectoris, Peripheral arterial occlusive disease, cardiomyopathy, heart failure, heart rhythm disorders, vascular restenosis and/or apoplexy,

-to reduce in body weight and/or body fat and/or liver fat and/or myocyte fat in fat or prevention body weight and/or body fat and/or liver fat and/or myocyte and increase or promote that in body weight and/or body fat and/or liver fat and/or myocyte, fat reduces;

-prevent, slow down, postpone or treat the degeneration of pancreatic beta cell and/or going down and/or improving, keep and/or recover the function of pancreatic beta cell and/or the function of stimulation and/or the secretion of recovery pancreatic insulin of Pancreatic beta cells function;

-prevent, slow down, postpone or treat non-alcoholic fatty liver disease (NAFLD), it comprises fatty degeneration of liver, non-alcoholic stellato-hepatitis (NASH) and/or hepatic fibrosis, such as, prevent, slow down progress, postpone, weaken, treat or reverse fatty degeneration of liver, (liver) inflammation and/or the abnormal accumulation of liver fat;

-prevention, to postpone or treatment is single to conventional anti-diabetic or type ii diabetes that combined therapy is invalid, slow down its progress;

-reach the dosage reduced in order to the conventional antidiabetic medicine needed for abundant therapeutic effect;

The risk of the untoward reaction that-reduction is relevant with conventional antidiabetic medicine, such as hypoglycemia or body weight increase; And/or

-keep and/or improve insulin sensitivity and/or be used for the treatment of or prevent hyperinsulinemia and/or insulin resistant.

5. the purposes according to any one of claim 1-4, in its Chinese style (I) compound, R ₁be preferably H, alkyl is preferably methyl or the tert-butyl group, and alcoxyl (sulfur) base is trifluoromethoxy or trifluoromethylthio, R ₆representative ring system is preferably phenyl ring, furan nucleus, pyrrole ring, imidazole ring or benzimidazole ring.

6. the purposes according to any one of claim 1-4, shown in its Chinese style (I), compound is as shown in the table,

。

7. the purposes of a pharmaceutical composition in the disease of preparation treatment PPAR mediation and the medicine of improvement or prevention PPAR mediation disease, it is characterized in that described pharmaceutical composition contains formula (I) compound for the treatment of effective dose as active component, and one or more pharmaceutically acceptable carriers.

8. the purposes of the pharmaceutical composition of claim 7, is characterized in that described pharmaceutical composition contains the active component that weight ratio is 0.01%-99.5%, and preferably containing weight ratio is the active component of 0.5%-99.5%.

9. the purposes of pharmaceutical composition as claimed in claim 7, is characterized in that the disease that described PPAR mediates and improvement or prevention PPAR mediate the disease or disease that disease is PPAR γ, PPAR α or PPAR δ mediation.

10. the purposes of pharmaceutical composition as claimed in claim 7, it is characterized in that disease that described PPAR mediates is that diabetes, hyperinsulinemia, obesity, hyperglycemia, hyperlipemia, impaired fasting glucose (IFG), X syndrome, hypercholesterolemia, hyperlipoproteinemia, insulin resistant, dysmetabolic syndrome, diabetic complication, sugared stable state are impaired, impaired glucose tolerance, insulin tolerance exception, hypertriglyceridemia, osteoporosis, preferably, described diabetes are type i diabetes or type ii diabetes.

The formula I of the disease of 11. treatment PPAR mediations and improvement or prevention PPAR mediation disease, wherein, formula I is as follows,

R ₁represent the alkyl of H atom and C1-C4

R ₂, R ₃, R ₄independently represent the alkyl of C1-C3, the oxyl of C1-C3, halogen atom and H atom

R ₅for-(CH ₂) _n-R ₆or-(CH ₂) _n-O-R ₆or-(CH ₂) _n-S-R ₆, wherein, the integer of n=0-4;

R ₆independently representative includes H, C1-C4 alkyl, C1-C3 alcoxyl (sulfur) base, halogen atom, the substituent groups such as 1,3-dioxy (sulfur) Pentamethylene. base are at the cycloalkane, 1 of interior phenyl ring, pyridine ring, pyrimidine ring, piperidine ring, piperazine ring, C3-C6,2,3-triazole, 1,2,4-triazole, pyrrole ring, furan nucleus, oxazole ring, imidazole ring, thiphene ring, thiazole ring and benzimidazole, benzothiazole, benzoxazole, benzofuran, benzothiophene and indole ring.

12. formula I according to claim 11, shown in its Chinese style (I), compound is as shown in the table,

。