CN101899047B

CN101899047B - Be used for the treatment of as depeptidyl peptidase inhibitors or the beta-amino tetrahydrochysene pyrazine of prevent diabetes, tetrahydropyrimidine and tetrahydropyridine

Info

Publication number: CN101899047B
Application number: CN200910085611.XA
Authority: CN
Inventors: 余强; 韦凤萍
Original assignee: Cgenetech Suzhou China Co Ltd
Current assignee: Shengshi Taike Biopharmaceutical Technology Suzhou Co ltd
Priority date: 2009-05-26
Filing date: 2009-05-26
Publication date: 2016-01-20
Anticipated expiration: 2029-05-26
Also published as: CN101899047A

Abstract

The present invention relates to a kind of being used for the treatment of as depeptidyl peptidase inhibitors or the amino tetrahydrochysene pyrazine of B of prevent diabetes, tetrahydropyrimidine and tetrahydropyridine, wherein each variable as in present specification define.Described compound is the inhibitor (" DP-IV inhibitor ") of dipeptidyl peptidase-IV enzyme, and it can be used for treating or prevent the disease relevant with dipeptidyl peptidase-IV enzyme as diabetes, particularly type ii diabetes.The invention still further relates to containing the pharmaceutical composition of these compounds and these compounds and composition in prevention or the purposes for the treatment of in these diseases relevant with dipeptidyl peptidase-IV enzyme.

Description

Be used for the treatment of as depeptidyl peptidase inhibitors or the beta-amino tetrahydrochysene pyrazine of prevent diabetes, tetrahydropyrimidine and tetrahydropyridine

Technical field

The present invention relates to compound, it is the inhibitor (" DP-IV inhibitor ") of dipeptidyl peptidase-IV enzyme and it is used for the treatment of or prevents the disease relevant with dipeptidyl peptidase-IV enzyme as diabetes, particularly type ii diabetes.The invention still further relates to containing the pharmaceutical composition of these compounds and these compounds and composition in prevention or the purposes for the treatment of in these diseases relevant with dipeptidyl peptidase-IV enzyme.

Background technology

Diabetes refer to a kind of lysis caused by multiple paathogenic factor, it is characterized in that, in the fasted state or after taking glucose during oral glucose tolerance test, plasma glucose levels raises or hyperglycemia.

To continue or uncontrolled property hyperglycemia is fallen ill with too early and mortality ratio improves relevant.Regular abnormal glucose homeostasis directly and indirect change with lipid, lipoprotein and Metabolism of Apolipoprotein and other metabolic relevant with hemodynamic disease.Therefore, type ii diabetes patient considerably increases the risk suffering from great vessels and microvascular complication, and described complication comprises coronary heart disease, apoplexy, peripheral vascular disease, hypertension, ephrosis, neuropathy and retinopathy.Therefore, in the Clinical Management of diabetes and the process for the treatment of, it is very important that glucose homeostasis, lipid metabolism and hypertensive therapeutic control.

Diabetes have been considered to two types usually.In type i diabetes or insulin-dependent diabetes (IDDM), patient produces or does not produce completely Regular Insulin (utilization of this hormone regulation glucose) hardly.In type ii diabetes or non-insulin-dependent diabetes mellitus (NIDDM) (NIDDM), compared with ND, patient has identical or higher plasma insulin level usually; But, the hormesis of the Regular Insulin of these patients in main insulin-sensitive tissues to glucose and lipid metabolism forms resistance, it is muscle, liver and fatty tissue, although and plasma insulin level rising, be still not enough to overcome obvious insulin resistance.

The minimizing of insulin resistance master except for insulin receptor number, but the rear insulin receptor binding deficient owing to not yet knowing.Cause intramuscular not enough to the cells activated by insulin of glucose uptake, oxidation and storage to this resistance of insulin replies, and to steatolysis in fatty tissue with to produce glucose in liver and that the Regular Insulin of secretion suppresses is inappropriate.

Substantially the limitation existing methods for the treatment of of type ii diabetes having been had realized that to it does not become for many years.Although the minimizing of physical activity and meals calorie intake significantly can improve the symptom of diabetes, but due to the mode of life be seated that fetters completely and excessive food consumption (especially containing the food of a large amount of saturated fatty), treatment is extremely poor like this.The blood plasma level of Regular Insulin is increased by giving sulfonylurea (such as tolbutamide and Glipizide) or meglitinide, its stimulating pancreas (3-cell is to secrete more Regular Insulin, and/or when sulfonylurea or meglitinide become invalid, by insulin injection, insulin concentration can be caused enough high to stimulate the tissue of very synalbumin.But by giving Regular Insulin or Insulin secretagogues (sulfonylurea or meglitinide) causes, and due to higher plasma insulin level, high-caliber insulin resistance can may be there is in dangerous low-level plasma glucose.Biguanides energy insulin sensitivity, thus hyperglycemia is relaxed to a certain extent.But two kinds of described biguanides, phenformin and N1,N1-Dimethylbiguanide, all may bring out lactic acidosis and/diarrhoea of feeling sick.N1,N1-Dimethylbiguanide is lower than phenformin side effect, therefore often as the prescription drugs for the treatment of type ii diabetes.

Glitazone (i.e. 5-substituted benzylthiazolidine-2,4-diketone) be recently disclosed in effectively improve a compounds of the multiple symptom of type ii diabetes.In several animal models of type ii diabetes, these medicines significantly improve the insulin sensitivity in muscle, liver and fatty tissue, thus make high plasma glucose level obtain correction partially or completely and not occur hypoglycemia.The glitazone of current sale is the agonist of the acceptor (PPAR) that peroxisome proliferators activates, mainly RPAR-γ hypotype.RPAR-γ agonism is considered to the reason of the insulin sensitizing agent effect causing improvement usually, and described insulin sensitizing agent effect is observed by glitazone compound.The newer RPAR agonist carrying out type ii diabetes treatment test is at present the agonist of α, γ or δ hypotype or these combinations, and is chemically different from glitazone (that is, they are not thiazolidinedioneses) under many circumstances.

There is severe side effect (such as liver poisoning) in some glitazones such as troglitazone.

Treat other method of this disease still under study for action.

Xinshenghua method nearest introducing or still under development comprises treats with alpha-glucosidase inhibitor (such as acarbose) and Protein Tyrosine Phosphatases-1B (PTP-1B) inhibitor.

As the compound of medicine, its be dipeptidyl peptidase-IV (" DP-IV " or " DPP-IV ") inhibitor also research among, they may be used for treat diabetes, particularly type ii diabetes.For example, see WO97/40832, WO98/19998, U.S. Patent number 5,939,560, Bioorg.Med.Chem.Lett., 6 (10), 1163-1166 (1996), and Bioorg.Med.Chem.Lett., 6 (22), 2745-2748 (1996), WO2004064778, Bioorganic & MedicinalChemistryLetters (2007), 17 (21), 5934-5939, WO2004032836, WO2003004498, JournalofMedicinalChemistry (2005), 48 (1), 141-151., Bioorganic & MedicinalChemistryLetters (2007), 17 (12), 3384-3387, CurrentMedicalResearchandOpinion (2008), 24 (2), 489-496.The purposes of DP-IV inhibitor in treatment type ii diabetes is the fact being easy to deactivation glucagon-like-peptide-1 (GLP-1) and gastrin inhibitory polypeptide (GIP) based on DP-IV in body.GLP-1 and GIP is incretin, and they produce when food consumption.Described incretin stimulates Regular Insulin to generate.The suppression of DP-IV causes the deactivation of incretin to decline, and this causes incretin stimulating by being improved in the effect of pancreas generation Regular Insulin conversely.

Therefore, DP-IV suppresses to cause serum insulin level to improve.

Advantageously, just produced by health when food consumption because incretin only has, therefore undesirably the unsuitable time as two meal between DP-IV suppress to improve insulin level, it may cause hypoglycemia (hypoglycemia).Therefore, expect that the suppression of DP-IV can increase Regular Insulin and not increase hypoglycemic risk, hypoglycemia is a kind of dangerous side-effects relevant with the use of Insulin secretagogues.

DP-IV inhibitor also has other therepic use described herein.Up to the present, also DP-IV inhibitor is not extensively studied, especially not yet the purposes except diabetes is studied.For the treatment of diabetes and potential Other diseases and illness, need new compound to find the DP-IV inhibitor improved.

Summary of the invention

Detailed description of the invention

The present invention relates to the compound of formula I:

Wherein: Ar is phenyl, it is unsubstituted or is replaced by 1-5 R5, wherein R5 independently selected from:

(1) halogen,

(2) C _1-6alkyl, it is linear or branch and be unsubstituted or by 1-5 halogen substiuted,

(3) OC _1-6alkyl, it is linear or branch and be unsubstituted or by 1-5 halogen substiuted, and

(4)CN；

X is selected from:

(1) N, and

(2)CR1；

Y is selected from:

(1) N, and

(2)CR2；

Z is selected from:

(1) N, and

(2)CR3；

R1, R2 and R3 independently selected from:

(1) hydrogen,

(2)CN，

(3) C1-10 alkyl, it is linear or branch and be unsubstituted or replaced by 1-5 halogen or phenyl, described phenyl is unsubstituted or is replaced by 1-5 substituting group, described substituting group independent selected from halo, CN, OH, R4, OR4, NHSO2R4, SO2R4, CO2H and CO2C _1-6alkyl, wherein said CO2C _1-6alkyl is linear or branch,

(4) phenyl, it is unsubstituted or is replaced by 1-5 substituting group, described substituting group independent selected from halo, CN, OH, R4, OR4, NHSO2R4, SO2R4, CO2H and CO2C _1-6alkyl, wherein said CO2C _1-6alkyl is linear or branch, and

(5) 5-or 6-unit heterocycle, it can be saturated or undersaturated and comprise 1-4 the heteroatoms independently selected from N, S and O, and described heterocycle is unsubstituted or individual independently selected from oxo, OH, halogen, C by 1-3 _1-6alkyl and OC _1-6the substituting group of alkyl replaces, wherein said C _1-6alkyl and OC _1-6alkyl is linear or branch and optionally by 1-5 halogen substiuted;

R4 is C _1-6alkyl, it is linear or branch and it is unsubstituted or by 1-5 independent selected from halo, CO2H and CO2C _1-6the group of alkyl replaces, wherein said CO2C _1-6alkyl is linear or branch;

And pharmacy acceptable salt and each diastereomer.

In the present invention, Ar is preferably phenyl, and it is unsubstituted or is replaced by 1-5 substituting group, described substituting group independently selected from:

(1) fluorine,

(2) bromine, and

(3)CF3。

In the present invention, Ar is more preferably selected from:

(1) phenyl,

(2) 2-fluorophenyl,

(3) 3,4-difluorophenyls,

(4) 2,5-difluorophenyls,

(5) 2,4,5-trifluorophenyls,

(6) the fluoro-4-of 2-(trifluoromethyl) phenyl, and

(7) bromo-2, the 5-difluorophenyls of 4-.

In the present invention, R1, R2 and R3 are preferably selected from:

(1) hydrogen, and

(2) C _1-6alkyl, its be linear or branch and its be unsubstituted or replaced by phenyl or 1-5 fluorine.

In the present invention, R2 and R3 is more preferably selected from:

(1) hydrogen,

(2) methyl,

(3) ethyl,

(4)CF3，

(5)CH2CF3，

(6)CF2CF3，

(7) phenyl, and

(8) benzyl.

In the present invention, R1 is more preferably hydrogen.

In the present invention, R2 and R3 is preferably selected from:

(1) hydrogen,

(2) C _1-6alkyl, it is linear or branch and be unsubstituted or replaced by 1-5 fluorine,

(3) phenyl, it is unsubstituted or is replaced by the individual substituting group independently selected from fluorine, OCH3 and OCF3 of 1-3.

In the present invention, R2 and R3 is more preferably selected from:

(1) hydrogen,

(2) methyl,

(3) ethyl,

(4)CF3，

(5)CH2CF3，

(6)CF2CF3，

(7) phenyl,

(8) (4-methoxyl group) phenyl,

(9) (4-trifluoromethoxy) phenyl,

(10) 4-fluorophenyl, and

(11) 3,4-difluorophenyls, and

(12) sec.-propyl.

In the present invention, R5 is preferably F, Br or CF3.

The present invention relates to the compound of formula II:

(1) halogen,

(4)CN；

X1 is selected from:

(1) N, and

(2)CR1；

Y1 is selected from:

(1) N, and

(2)CR2；

Z is selected from:

(1) N, and

(2)CR3；

R1, R2 and R3 independently selected from:

(1) hydrogen,

(2)CN，

(3) C1-10 alkyl, its be linear or branch and its be unsubstituted or replaced by 1-5 halogen or phenyl, described phenyl is unsubstituted or is replaced by 1-5 substituting group, described substituting group independent selected from halo, CN, OH, R4, OR4, NHSO2R4, SO2R4, CO2H and CO2C _1-6alkyl, wherein said CO2C _1-6alkyl is linear or branch,

And pharmacy acceptable salt and each diastereomer.

(1) fluorine,

(2) bromine, and

(3)CF3。

In the present invention, Ar is more preferably selected from:

(1) phenyl,

(2) 2-fluorophenyl,

(3) 3,4-difluorophenyls,

(4) 2,5-difluorophenyls,

(5) 2,4,5-trifluorophenyls,

(6) the fluoro-4-of 2-(trifluoromethyl) phenyl, and

(7) bromo-2, the 5-difluorophenyls of 4-.

In the present invention, R1, R2 and R3 are preferably selected from:

(1) hydrogen, and

In the present invention, R2 is more preferably selected from:

(1) hydrogen,

(2) methyl,

(3) ethyl,

(4)CF3，

(5)CH2CF3，

(6)CF2CF3，

(7) phenyl, and

(8) benzyl.

In the present invention, R5 is preferably F, Br or CF3.

The present invention relates to the compound of formula III:

(1) halogen,

(4)CN；

X2 is selected from:

(1) N, and

(2)CR1；

Y2 is selected from:

(1) N, and

(2)CR2；

Z2 is selected from:

(1) N, and

(2)CR3；

R1, R2 and R3 independently selected from:

(1) hydrogen,

(2)CN，

And pharmacy acceptable salt and each diastereomer.

(1) fluorine,

(2) bromine, and

(3)CF3。

In the present invention, Ar is more preferably selected from:

(1) phenyl,

(2) 2-fluorophenyl,

(3) 3,4-difluorophenyls,

(4) 2,5-difluorophenyls,

(5) 2,4,5-trifluorophenyls,

(6) the fluoro-4-of 2-(trifluoromethyl) phenyl, and

(7) bromo-2, the 5-difluorophenyls of 4-.

In the present invention, R1, R2 and R3 are preferably selected from:

(1) hydrogen, and

In the present invention, R5 is preferably F, Br or CF3.

Compound of the present invention can contain one or more asymmetric center, and therefore can occur with the form of racemic modification and racemic mixture, single enantiomer, non-enantiomer mixture and each diastereomer.Compound of the present invention has an asymmetric center at beta carbon place.

Depend on each substituent character in molecule, other asymmetric center can be there is.Each such asymmetric center will generate two kinds of optical isomers independently, the present invention includes all possible optical isomer and non-enantiomer mixture and pure or partial-purified compound.The present invention includes all these isomeric form of these compounds.

Compounds more described herein contain olefinic double bond, except as otherwise noted, the present invention includes E and Z two kinds of geometrical isomers.

Compounds more described herein can exist with the form of tautomer, and it has different hydrogen points of contact by moving with one or more double bond.Such as, ketone and enol form thereof are keto-enol tautomerism bodies.

Each tautomer and composition thereof is all included in compound of the present invention.

As known in the art by carrying out suitable improvement to method disclosed herein, independence synthesis or their chromatographic separation of these diastereomers can be realized.By the X-ray crystallographic of the crystalline intermediate (if necessary, deriving with the reagent of the asymmetric center containing known absolute configuration) of crystalline product or derivatize, their absolute stereochemical can be determined.

If necessary, the racemic mixture of described compound can be separated, to be separated each enantiomer.Can be separated by means commonly known in the art, the racemic mixture of such as compound and the coupling compounds of enantiomeric pure to form non-enantiomer mixture, then by standard method as fractionation crystallization or chromatography are separated each diastereomer.The acid of enantiomeric pure or the coupled reaction of alkali is used usually to generate salt.Then, by the cracking of chiral residue added, described non-enantiomer derivative is converted into pure enantiomer.The racemic mixture of described compound directly can also be separated by using the chromatography of chiral stationary phase, and the method is known in the art.

Or, use optically pure starting material or the reagent of configuration known, by means commonly known in the art, any enantiomorph of compound can be obtained by stereoselective syntheses.

Term " pharmacy acceptable salt " refers to by the obtained salt of pharmaceutically acceptable nontoxic alkali or acid (comprising inorganic or organic bases and inorganic or organic acid).The salt deriving from these mineral alkalis comprises the salt such as aluminium, ammonium, calcium, copper, iron, ferrous iron, lithium, magnesium, manganic salt, bivalent manganese, potassium, sodium, zinc.Particularly preferably be ammonium, calcium, magnesium, potassium and sodium salt.May be there is more than a kind of crystalline structure in the salt of solid-state form, and can with the form of hydrate.The salt deriving from pharmaceutically acceptable organic nontoxic alkali comprises the salt of following compound: primary, the second month in a season and tertiary amines, substitutional amine-group, comprise naturally occurring substitutional amine-group, cyclammonium and Zeo-karb, such as arginine, trimethyl-glycine, caffeine, choline, N, N '-dibenzyl-ethylenediamin, diethylamine, 2-diethylaminoethanol, DMAE, thanomin, quadrol, N-ethyl-morpholine, N-ethylpiperidine, glycosamine, glucosamine, Histidine, breathe out amine, Isopropylamine, Methionin, methylglucosamine, morpholine, piperazine, piperidines, polyamines resin, PROCAINE HCL, PHARMA GRADE, purine, Theobromine, triethylamine, Trimethylamine 99, tripropyl amine, tromethane etc.

When compound of the present invention is alkalescence, salt can be obtained by pharmaceutically acceptable non-toxic acid, comprises mineral acid and organic acid.These acid comprise, and acetic acid, Phenylsulfonic acid, phenylformic acid, camphorsulfonic acid, citric acid, ethyl sulfonic acid, fumaric acid, glyconic acid, L-glutamic acid, Hydrogen bromide, hydrochloric acid, hydroxyl second (base) sulfonic acid, lactic acid, toxilic acid, oxysuccinic acid, amygdalic acid, methylsulfonic acid, glactaric acid, nitric acid, flutter acid, pantothenic acid, phosphoric acid, succsinic acid, sulfuric acid, tartrate, tosic acid etc.Particularly preferably be citric acid, Hydrogen bromide, hydrochloric acid, toxilic acid, phosphoric acid, sulfuric acid, fumaric acid and tartrate.

When being appreciated that the compound as volume type I in this article, it refers to and also comprises its pharmacy acceptable salt.

Understandable as those skilled in the art, halo used herein or halogen are used to comprise fluorine, chlorine, bromine and iodine.Similarly, C1-8, when the group being defined in C1 alkyl to determine to have 1,2,3,4,5,6,7 or 8 carbon in linear or branch arrange, such C1 alkyl comprises methyl, ethyl, n-propyl, sec.-propyl, normal-butyl, isobutyl-, the tertiary butyl, amyl group, hexyl, heptyl and octyl group particularly.Equally, C0, is defined by there is direct covalent linkage in C0 alkyl.A kind of group, it is called as and is substituted with a substituent independently and can be replaced by these substituting groups multiple independently.Term used herein " heterocyclic radical " is used to be included in following institute: 5-or the 6-ring system benzimidazolyl-within the scope of row, Ben Bing oxane base, benzofuryl, benzopyrazoles base, diazosulfide base, benzotriazole base, benzothienyl, Ben Bing oxadiazolyl, benzoxazolyl, carbazyl, carbolinyl, chromanyl, cinnolines base, furyl, imidazolyl, indolinyl, indyl, indolazinyl, indazolyl, isobenzofuran-base, pseudoindoyl, isoquinolyl, isothiazolyl, isoxazolyl, naphthyridinyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridopyridine base, pyridazinyl, pyridyl, pyrimidyl, pyrryl, quinazolyl, quinolyl, quinoxalinyl, tetrazyl, thiadiazolyl group, thiazolidyl, thiazolyl, thienyl, triazolyl, azetidinyl, Isosorbide-5-Nitrae-alkyl dioxin, six hydrogen azepine bases, piperazinyl, piperidyl, pyrrolidyl, morpholinyl, thio-morpholinyl, dihydrobenzo imidazolyl, dihydro benzo furyl, dihydrobenzo thienyl, Er hydrogen benzoxazolyl, dihydrofuran base, glyoxalidine base, indolinyl, dihydro-isoxazole base, dihydro isothiazolyl, Er Qing oxadiazolyl, dihydro-oxazole base, dihydro pyrazinyl, pyrazoline base, dihydropyridine base, dihydro-pyrimidin base, pyrrolin base, dihydroquinoline base, dihydro tetrazyl, thiodiazoline base, dihydro-thiazolyl, dihydro-thiophene base, dihydro triazolyl, dihydro azetidinyl, methylenedioxyphenyl formyl radical, tetrahydrofuran base, imidazolidine base, tetrahydro isoquinolyl and tetrahydro-thienyl.

The present invention illustrates the purposes in embodiment and compound disclosed herein.

Particular compound of the present invention is included in compound disclosed in the following example and pharmacy acceptable salt thereof and each diastereomer.

Compound of the present invention is used for suppressing in the method for dipeptidyl peptidase-IV enzyme in needing the patient of this suppression as Mammals, comprises the described compound giving significant quantity.

The present invention relates to the purposes of compound disclosed herein as dipeptidyl peptidase-IV activity inhibitor.

Except primate is as except people, method of the present invention can be used for treating other Mammals multiple.Such as, treatable Mammals includes but not limited to ox, sheep, goat, horse, dog, cat, cavy, rat or other ox, sheep, horse, dog, cat family, rodent or murine species.But the method can also be used for other species as avian species (such as, chicken).

The invention still further relates to and to be prepared in humans and animals for suppressing the method for the medicine of dipeptidyl peptidase-IV enzymic activity, comprise compound of the present invention and pharmaceutical carrier or mixing diluents.

The treatment target of the inventive method normally needs the Mammals suppressing dipeptidyl peptidase-IV enzymic activity, preferred people, sex.Term " treatment significant quantity " refers to the quantity by causing tissue, system, the biology of animal or human or the compounds of this invention of medical response sought by researchist, animal doctor, attending doctor or other clinicist.

Term used herein " composition " refers to the product of the special component comprised containing specific quantity, and any directly or indirectly from the product of the combination of the special component of specific quantity.Term " pharmaceutical composition " refers to and comprises containing activeconstituents, the formation inert component of carrier and the product of any product, it is directly or indirectly by any two or more compositions mixing, complexing or gathering, or by one or more ingredient breakdown, or obtained by the reaction of other type of one or more compositions or interaction.

Therefore, pharmaceutical composition of the present invention comprises and mixes by compound of the present invention any composition obtained with pharmaceutically acceptable carrier.Term " pharmaceutically acceptable " refers to that carrier, thinner or vehicle must be compatible with other composition of preparation and do not poison experimenter.

Term " gives compound " and/or " administration of compound " is construed as the prodrug that the individuality for the treatment of to needs provides compound of the present invention or the compounds of this invention.

The compounds of this invention can be proved by methods known in the art as the practicality of dipeptidyl peptidase-IV activity inhibitor.Constant is suppressed to measure as follows.Use the continuous fluorescence assay method of band substrate Gly-Pro-AMC, it discharges fluorescent AMC leaving group by DP-IV cracking.The kinetic parameter describing this reaction is as follows: Km=50JAM; Kcat=75s-1; Kcat/Km=1.5x106M-ls-1.In 100l total reaction volume, type reaction comprises about 50pM enzyme, 50 μMs of Gly-Pro-AMC and damping fluid (100mMHEPES, pH7.5,0.1mg/mlBSA).Use the excitation wavelength of 360nm and the emission wavelength of 460nm, in 96 orifice plate photofluorometers, monitor the release of AMC continuously.Under these conditions, at 25 DEG C, in 30 minutes, generate about 0.8uMAMC.Solubility (cross-film district and tenuigenin expand except) the people's albumen generated in the baculovirus expression system (Bac-To-Bac, GibcoBRL) for the enzyme in these researchs.Find that the hydrolysis dynamics constant of Gly-Pro-AMC with GLP-1 conforms to the literature value of natural enzyme.In order to measure the dissociation constant of compound, in the reaction containing enzyme and substrate (final DMSO concentration is 1%), add the solution of inhibitor in DMSO.All experiments all at room temperature use above-mentioned standard reaction condition to carry out.In order to measure dissociation constant (Ki), by non-linear regression by speed of reaction matching in the Michaelis-Menton equation of competitive inhibition.The error that dissociation constant is reproduced typically is less than twice.

Especially, the compound of the following example has the activity suppressing dipeptidyl peptidase-IV enzyme in above-mentioned test, and usual IC50 is less than about 1uM.This result shows that described compound is used as the intrinsic activity of dipeptidyl peptidase-IV activity inhibitor.

Dipeptidyl peptidase-IV enzyme (DP-IV) relates to the cell surface protein of various biological function.It has tissue distribution (intestines, kidney, liver, pancreas, placenta, thymus gland, spleen, epithelial cell, blood vessel endothelium, lymph and medullary cell, serum) widely, and different tissues and cell-type expression levels.DP-IV is confirmed as t cell activation mark CD26, and it can cracking immunoregulatory, endocrine and neurologic peptide in a large number in vitro.This shows the latent effect in the various lysises of this peptase in human body or other species.

Therefore, compound of the present invention may be used in the prevention of following disease, obstacle and illness or the method for the treatment of.

Type ii diabetes and relative disease: now determine completely, incretin GLP-1 and GIP is in vivo by DP-IV rapid deactivation.DP-IV (~/~) defective mouse and initial clinical experience are shown: DP-IV suppresses to add the Css of GLP-1 and GIP, causes glucose tolerance to improve.Similar with GLP-1 and GIP, other relates to grape glycoregulatory hyperglycemic-glycogenolytic factor race peptide equally by DP-IV (such as, PACAP, hyperglycemic-glycogenolytic factor) inactivation.

These peptides can also be worked by the deactivation of DP-IV in glucose homeostasis.

Therefore, DP-IV inhibitor of the present invention can be used for treating type ii diabetes and be used for the treatment of and prevent type ii diabetes usual adjoint many illnesss, comprise metabolic syndrome X, reactive hypoglycemia and diabetic hyperlipemia.Obesity is hereinafter described the another kind of symptom often found with type ii diabetes, and it has response to the treatment of compound of the present invention.

Following disease, obstacle is relevant with type ii diabetes with illness, therefore can by treating with compounds for treating of the present invention, control or prevent in some cases: (1) hyperglycemia, (2) low dextrose tolerance, (3) insulin resistance, (4) obesity, (5) lipoid dyscrasias, (6) hyperlipemia, (7) hyperlipidaemia, (8) hypertriglyceridemia, (9) hypercholesterolemia, (10) low HDL levels, (11) high LDL level, (12) atherosclerosis and sequela thereof, (13) vascular restenosis, (14) irritable bowel syndrome, (15) enteritis comprises Crohn disease and ulcerative colitis, (16) other inflammation, (17) pancreatitis, (18) abdominal fatness, (19) neurodegenerative disease, (20) retinopathy, (21) ephrosis, (22) neuropathy, (23) X syndromes, (24) hyperandrogenism (polycystic ovary syndrome) and other there is the disease of insulin resistance.

Obesity: DP-IV inhibitor can be used for treatment of obesity.This is based on the restraining effect of GLP-1 and GLP-2 observed to food intake and stomach emptying.The exogenous GLP-1 of giving of human body can significantly reduce food intake and slow down stomach emptying (Am.J.Physiol.277, R910-R916 (1999)).ICV administration rat and mouse being carried out to GLP-1 has profound influence (NatureMedicine2,1254-1258 (1996)) to food intake equally.In GLP-1R mouse, do not observe this feed suppress, show that these are used as by brain GLP-1 regulation.Similar with GLP-1, GLP-2 also may regulate by DP-IV.

Be similar to the effect observed with GLP-1, the ICV administration of GLP-2 suppresses food intake (NatureMedicine6,802-807 (2000)) equally.

Growth hormone deficiency: the peptide discharged from prepituitary gland based on hypothesis and somatotropin releasing factor (GRF)-a kind of stimulating growth hormone-in vivo by DP-IV enzymatic lysis (WO00/56297), DP-IV suppresses to can be used for treating growth hormone deficiency.Following data provides the evidence that GRF is Endogenous Substrate: (1) GRF, in vitro by effective cracking, generates inactive products GRF [3-44] (BBA1122,147-153 (1992)); (2) GRF fast degradation in blood plasma becomes GRF [3-44]; This is stoped by DP-IV inhibitor diprotinA; And (3) find GRF [3-44] (J.Clin.Invest.83,1533-1540 (1989)) in the blood plasma of people GRF transgenic pig.Therefore, DP-IV inhibitor can be used for the indication that tethelin urgees to secrete the same range that agent is assert.

Damage of intestines: result of study shows glucagon-like-peptide-2 (GLP-2), the possible Endogenous Substrate of a kind of DP-IV, trophicity effect can be shown to enteric epithelium, this result of study illustrates and uses DP-IV inhibitor potentially can be used for the treatment of damage of intestines (RegulatoryPeptides90,27-32 (2000)).

The administration of GLP-2 causes rodent small intestine weight to increase, and reduces the damage of intestines in the rodent model of colitis and enteritis.

Immunosuppression: based on the effect about DP-IV inhibitor in the research of DP-IV enzyme in t cell activation and chemokine processing and disease In vivo model, DP-IV suppresses to can be used for immunity moderation response.

DP-IV identification in CD26, a kind of cell surface marker thing of immune cell activated.The expression of CD26 is regulated by the differentiation of immunocyte and state of activation.It is generally acknowledged that CD26 plays co stimulatory molecule in the external model of t cell activation.Many chemokines are containing proline(Pro) in position second from the bottom, and estimation may be prevent them from being degraded by non-specific aminopeptidases.Many proof in these is processed by DP-IV in vitro.In some situations (RANTES, LD78-beta, MDC, eotaxin, SDF-lalpha), cracking causes the change of chemotaxis and signal test active.In some cases (RANTES), receptor-selective might as well as if be changed.In vitro in cell culture system, identify many n terminal truncations form of some chemokines, comprise the prediction product of DP-IV hydrolysis.

In transplanting and arthritic animal model, DP-IV inhibitor has proved effective immunosuppressor.Prodipine (ProPro-diphenyl-phosphonate), the irreversible inhibitor of a kind of DP-IV, be proved to be the survival of rats 7 to 17 (Transplantation63,1495-1500 (1997)) making the dirty allograft of diplocardia.DP-IV inhibitor has carried out testing in the rat arthritis of collagen protein and alkyl diamine induction and the statistical significance showing rear solid end swelling in this model weakens (Int.J.Immunopharmacology19,15-24 (1997), Immunopharmacology40,21-26 (1998)).It is raise (ImmunologyToday20,367-375 (1999)) that DP-IV comprises in rheumatoid arthritis, multiple sclerosis, Exophthalmus goiter and struma lymphomatosa at some autoimmune diseases.

HIV: DP-IV suppresses to can be used for treatment or pre-preventing HIV infection or AIDS, because the many chemokines suppressing HIV cell to enter are potential substrates (ImmunologyToday20,367-375 (1999)) of DP-IV.When SDF-1 α, cracking decreases antiviral activity (PNAS95,6331-6 (1998)).

Therefore, wish by suppressing DP-IV to stablize SDF-1 α and reduce HIV.

Hemopoietic: because DP-IV may relate to hemopoietic, therefore DP-IV suppresses to can be used for treatment or prevention hemopoietic.In the mouse model that the neutrophilic leukocyte of Induced by Cyclophosphamide reduces, DP-IV inhibitor (Val-Boro-Pro) stimulates hemopoietic (WO99/56753).

Neuronal disease: because many peptides relevant with various neuronal process are in vitro by DP-IV cracking, therefore DP-IV suppresses can be used for treatment or prevent various neuronal disease or mental disorder.Therefore, DP-IV inhibitor has treatment benefit in treatment neuronal disease.

Endomorphin-2, beta-caseins morphine peptide and Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH2 are all proved to be the substrate of DP-IV in vitro.In all situations, external cracking is that efficiency is very high, and kcat/Km is about 106M-ls-1 or larger.In the analgesia electric shock jump test model of rat, DP-IV inhibitor shows significant effect, the existence irrelevant (BrainResearch, 815,278-286 (1999)) of this effect and exogenous endomorphin-2.

Tumour invades and transfer: DP-IV suppression can be used for treatment or prophylaxis of tumours invades and transfer, because at normal cell in malignant phenotype's conversion process, increase or minimizing (J.Exp.Med.190,301-305 (1999)) that several exopeptidase (comprising DP-IV) expresses are observed.The rise of these albumen or lower seemingly tissue and cell type-specific.Such as, t cell lymphoma, T cell acute lymphoblastic leukemia, cell-derived thyroid carcinoma, rodent cancer and mammary cancer are observed CD26/DP-IV and express increase.Therefore, DP-IV inhibitor can be used for treating these cancers.

Benign prostatauxe: DP-IV suppresses to can be used for treating benign prostatauxe, because observe DP-IV activity to increase (Eur.J.Clin.Chem.Clin.Biochem30,333-338 (1992)) in the prostata tissue of BPH.

Motility of sperm/male contraception: because in seminal fluid, it is active that the organoid important to motility of sperm that prostate gland, prostate gland derive has very high-caliber DP-IV, so DP-IV suppresses to can be used for changing motility of sperm and for male contraception (Eur.J.Clin.Chem.Clin.Biochem30,333-338 (1992)).

Gingivitis: because find that DP-IV is active in the research that gum seam fluid is relevant to periodontal disease severity with some, DP-IV suppresses to can be used for treatment gingivitis (Arch.OralBiol.37,167-173 (1992)).

Osteoporosis: because gip receptor is present in scleroblast, so DP-IV suppresses to can be used for treatment or preventing osteoporosis disease.

Compound of the present invention may be used for treatment or prevents one or more following illness or diseases: (1) hyperglycemia, (2) low dextrose tolerance, (3) insulin resistance, (4) obesity, (5) lipid disorder, (6) hyperlipemia, (7) hyperlipidaemia, (8) hypertriglyceridemia, (9) hypercholesterolemia, (10) low HDL levels, (11) high LDL level, (12) atherosclerosis and sequela thereof, (13) vascular restenosis, (14) irritable bowel syndrome, (15) inflammatory bowel, comprise Crohn disease and ulcerative colitis, (16) other inflammatory conditions, (17) pancreatitis, (18) abdominal fatness, (19) neurodegenerative disease, (20) retinopathy, (21) ephrosis, (22) neuropathy, (23) X syndromes, (24) ovarian hyperandrogenism (polycystic ovary syndrome), (25) type ii diabetes, (26) tethelin defect, (27) neutropenia, (28) neuronal disease, (29) metastases, (30) benign prostatauxe, (32) gingivitis, (33) hypertension, (34) osteoporosis, and other can carry out the illness for the treatment of by suppressing DP-IV.

Compound of the present invention can also be used from the prevention of above-mentioned disease, obstacle and illness or the method for the treatment of with other medicament one.

Compound of the present invention can combinationally use with one or more other medicines, one or more other medicines described are used for the treatment of, prevent, suppress or improve compound or other medicines possibility effectively disease or the illness of formula I, and the combination of wherein said medicine is more safer or more effective than drug alone.These other medicines can by the while of a kind of approach and its usual used quantity compound with formula I or administration successively.When the compound of formula I and one or more other medicines use simultaneously, the pharmaceutical composition of preferred unit formulation contains the compound of these other medicines and formula I.But described combination therapy can also comprise the therapy wherein by the compound of formula I and the administration in different overlapping schedules of one or more other medicines.Also consider when with one or more other active ingredient combination medications, the compound of the present invention than dosage lower during respective medication separately and other activeconstituents can be used.Therefore, except the compound of formula I, pharmaceutical composition of the present invention comprises containing those of one or more other activeconstituentss.

Can with the compound drug combination of formula I and respectively administration or in same medicinal compositions the example of other activeconstituents of administration include, but are not limited to: (a) other DPP IV (DP-IV) inhibitor, b () insulin sensitizer comprises (i) PPARy agonist such as glitazone (such as troglitazone, U-721017E, englitazone, MCC-555, rosiglitazone etc.) and other RPAR part, comprise PPARaJy dual agonists, such as KRP297 and PPARoc agonist such as fenofibric acid derivatives (gemfibrozil, clofibrate, fenofibrate and bezafibrate), (ii) biguanides such as N1,N1-Dimethylbiguanide and phenformin, and (iii) Protein Tyrosine Phosphatases-1B (PTP-1B) inhibitor, (c) Regular Insulin or insulin-mimickers, (d) sulfonylurea and other Insulin secretagogues such as tolbutamide and Glipizide, meglitinide, and related substance, (e) alpha-glucosidase inhibitor (such as acarbose), (f) glucagon receptor antagonist such as disclosed in WO98/04528, WO99/01423, WO00/39088 and WO00/69810 those, (g) GLP-1, GLP-1 stand-in and GLP-1 receptor stimulant such as disclosed in WO00/42026 and WO/59887 those, (h) GIP and GIP stand-in such as disclosed in WO00/58360 those, and gip receptor agonist, (i) PACAP, PACAP stand-in and PACAP acceptor 3 agonist such as disclosed in WO01/23420 those, (j) cholesterol decline reagent, such as (i) HMG-CoA reductase inhibitor (lovastatin, Simvastatin, Pravastatin, Fluvastatin, Zarator, stand and cut down statin, itavastatin, superstatin, and other statin), (ii) sequestrant (QUESTRAN, the dialkylaminoalkyl derivative of colestipol and cross linked dextran), (iii) nicotinic alcohol, nicotinic acid or its salt, (iv) PPAR alfa agonists such as fenofibric acid derivatives (gemfibrozil, clofibrate, fenofibrate and bezafibrate), (v) PPARon/ dual agonists, such as KRP-297, (vi) cholesterol absorption inhibitor, such as beta Sitosterol and ezetimibe, (vii) acyl CoA: chole-sterol acyltransferase inhibitor, such as avasimibe, (viii) antioxidant, such as probucol, (k) PPAR8 agonist, such as disclosed in WO97/28149 those, l () antiobesity compounds is Phenfluoramine, dexfenfluramine, phentermine, sibutramine, orlistat, neuropeptide Y 5 inhibitor and 33 adrenergic receptor agonists such as, (m) ileal bile acid transporter inhibitor, (n) medicine designed by inflammatory conditions is Asprin, nonsteroidal anti-inflammatory drug, glucocorticosteroid, sulfasalazine and COX-2 selective depressant such as.

Aforesaid combination comprises compound of the present invention and the combination of not only other active compound a kind of, also comprises the combination of compound of the present invention and two or more other active compounds.Non-limitative example comprises the combination of formula I and two or more active compounds, and two or more active compounds described are selected from biguanides, sulfonylurea, HMG-CoA reductase inhibitor, RPAR agonist, PTP-1B inhibitor, other DP-IV inhibitor and antiobesity compounds.

Equally, compound of the present invention can combine with other medicines, and described other medicines are used for the treatment of/prevent/suppress or improve the disease or illness that the compounds of this invention uses.These other medicines can by a kind of approach and its usual used quantity with the while of compound of the present invention or administration successively.When compound of the present invention and one or more other medicines use simultaneously, the pharmaceutical composition preferably in addition to the present compounds also containing these other medicines.Therefore, in addition to the present compounds, pharmaceutical composition of the present invention comprises also containing those of one or more other activeconstituentss.

The weight ratio of the compounds of this invention and the second active ingredient can change and will depend on the effective dose of each component.Usually, the effective dose of often kind of active principle will be used.

Therefore, such as, when compound of the present invention and other medicines conbined usage, the weight ratio of compound of the present invention and other medicines usually will in the scope of about 1000: 1-about 1: 1000, preferably in the scope of about 200: 1-about 1: 200.The drug combination of compound of the present invention and other activeconstituents usually also within above-mentioned scope, but in each case, should use the effective dose of often kind of activeconstituents.

In these drug combinations, compound of the present invention and other promoting agent can be administered separately or simultaneously administered.In addition, can by a kind of composition before giving other medicines, period or administration afterwards.

Compound of the present invention can by oral, parenteral (such as, intramuscular, intraperitoneal, intravenously, ICV, intracisternal injection or transfusion, subcutaneous injection or implantation), by sucking spraying, nasal cavity, vagina, rectum, sublingual or topical routes of administration, can be mixed with separately or be together appropriate to often kind of route of administration conventional non-toxic pharmaceutical on the suitable dosage unit preparation of acceptable carrier, adjuvant and vehicle.Except treatment warm-blooded animal is as except mouse, rat, horse, ox, sheep, dog, cat, monkey etc., compound of the present invention can effectively for human body.

Pharmaceutical composition for the compounds of this invention administration can exist with the form of unit dosage easily, and can be prepared by the known any method of pharmacy field.All methods all comprise the step mixed with the carrier forming one or more auxiliary agents by activeconstituents.Usually, by by activeconstituents and liquid vehicle or subdivided solids carrier or both are all even closely mixes, if then needed, make required type processed, described pharmaceutical composition can be prepared like this.In described pharmaceutical composition, the amount of described Active Target Compounds is enough to produce desired result to the course of disease of disease or illness.Term used herein " composition " refers to the product of the special component comprised containing concrete quantity, and any directly or indirectly from the product of the combination of the special component of concrete quantity.

Pharmaceutical composition containing described activeconstituents can be suitable for oral form, such as tablet, lozenge, lozenge, moisture or containing oil suspension, and dispersible powder or granula, emulsion, hard or soft capsule or syrup or elixir form exist.Oral compositions can be prepared according to any method of pharmaceutical compositions known in the art, and these compositions can be selected from reagent of sweeting agent, seasonings, tinting material and sanitas to provide pharmaceutically exquisite in good to eat preparation containing one or more.Tablet contains the activeconstituents with acceptable vehicle fusion on the non-toxic pharmaceutical being suitable for preparing tablet.These excipients are as can be that inert diluent is as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; Granulating agent and disintegrating agent are as W-Gum or alginic acid; Tackiness agent is as starch, gelatin or gum arabic; And lubricant is as Magnesium Stearate, stearic acid or talcum powder.These tablets can be non-dressings or carry out dressing to postpone disintegration in the gastrointestinal tract and absorption with known technology, and therefore within a long period, provide a kind of continuous action.Such as, time delay material such as glyceryl monostearate or distearin can be used.They can also use U.S. Patent number 4, and 256,108; 4,160,452 and 4,265, the technology described in 874 carries out dressing, to form the osmotic therapeutic tablets for Co ntrolled release.

Oral preparations can also be hard capsule, and wherein activeconstituents is with inert solid diluent as calcium carbonate, calcium phosphate or kaolin mix, or can be soft capsule, and wherein activeconstituents and water or oily medium are as peanut oil, whiteruss or mixed with olive oil.

Aqueous suspension contains the active substance together with being suitable for preparing the mixed with excipients of aqueous suspension.These vehicle are suspension agents, such as Xylo-Mucine, methylcellulose gum, Vltra tears, sodium alginate, Polyvinyl-pyrrolidone, tragacanth gum and gum arabic; Dispersion agent or wetting agent can be naturally occurring phosphatide, such as Yelkin TTS, or the condensation product of alkylene oxide and lipid acid, such as polyoxyethylene stearic acid ester, or the condensation product of oxyethane and long-chain fat race alcohol, such as heptadecaethylene oxycetanol, or oxyethane and derive from the condensation product of lipid acid and hexitol partial ester, such as polyoxyethylene 80 sorbitan monooleate, or oxyethane and derive from the condensation product of partial ester of lipid acid and hexitol anhydrides, such as polyoxyethylene sorbitol monoleate.Described aqueous suspension can also containing one or more sanitass if para hydroxybenzene acetoacetic ester or p-hydroxybenzoic acid n-propyl, one or more tinting materials, one or more seasoningss and one or more sweeting agents be as sucrose or asccharin.

Can be prepared by following containing oil suspension: activeconstituents is suspended in vegetables oil as in peanut oil, sweet oil, sesame oil or Oleum Cocois, or activeconstituents is suspended in mineral oil as in whiteruss.Can containing thickening material as beeswax, paraffinum durum or hexadecanol containing oil suspension.Sweeting agent as above and seasonings can be added, to provide a kind of good to eat oral preparations.These compositions can be undertaken anticorrosion by adding antioxidant such as xitix.

Be suitable for by adding water and prepare the dispersible powder of aqeous suspension and granula being prepared as follows: by activeconstituents and dispersion agent or wetting agent, suspension agent and one or more sanitas fusion.Suitable dispersion agent or wetting agent and suspension agent be above-mentioned illustrational those.Other vehicle can also be there is, such as sweeting agent, seasonings and tinting material.

Pharmaceutical composition of the present invention can also be the form of oil-water emulsifiers.Described oil phase can be vegetables oil if sweet oil or peanut oil or mineral oil are as the mixture of whiteruss or these oil.Suitable emulsifying agent can be naturally occurring natural gum, such as gum arabic or tragacanth gum, naturally occurring phosphatide, such as soybean, Yelkin TTS and derive from ester or the partial ester of lipid acid and hexitol anhydrides, such as sorbitan monooleate, and the condensation product of described partial ester and oxyethane is as polyoxyethylene sorbitan monoleate.

Described emulsion can also contain sweeting agent and seasonings.

Syrup and elixir can with sweeting agent as the preparations of glycerine, propylene glycol, Sorbitol Powder or sucrose.These preparations can also contain wetting agent, sanitas and seasonings and tinting material.

Described pharmaceutical composition can exist with sterile injectable aq suspension or containing the form of oil suspension.Use suitable dispersion agent or wetting agent and suspension agent according to known technology, this suspension can be prepared.Sterile injectable preparation can also be a kind of sterile injectable solution in the acceptable thinner of a kind of nontoxic parenteral or solvent or suspension, such as, solution in 1,3 butylene glycol.In the middle of acceptable vehicle and solvent, operable is water, Ringer's solution and isotonic sodium chlorrde solution.In addition, aseptic expressed oil is typically used as solvent or suspension medium.For this reason, the expressed oil of any gentleness can be used, comprise monoglyceride and the triglyceride of synthesis.In addition, can in injectable formulation, use fatty acids as oleic acid.

Compound of the present invention can also be used for rectal administration with the form of suppository.These compositions can be prepared by being mixed with suitable nonirritant excipient by medicine, and these compositions are solid at normal temperatures, but it is liquid under anus temperature, therefore melt in the rectum to discharge medicine.These materials are theobroma oil and polyoxyethylene glycol.

For topical, can use containing the emulsion of the compounds of this invention, ointment, gelifying agent, solution or suspension etc.(concerning this application, topical should comprise mouth wash shua and mouthwass).Pharmaceutical composition of the present invention and method are containing comprising other therapeutical active compound described herein, and it is generally used in the treatment of above-mentioned pathological state.

Need to suppress in the illness of dipeptidyl peptidase-IV enzymic activity in treatment or prevention, suitable dosage level is about 0.01-500mg every kg weight in patients every day usually, and it can with single dose or multiple dose administration.Preferably, described dosage level is about 0.1-and is about 250mg/kg every day; Be more preferably 0.5-and be about 100mg/kg every day.Suitable dosage level can be about 0.01-250mg/kg every day, about 0.05-100mg/kg every day, or about 0.1-50mg/kg every day.Within the scope of this, described dosage can be 0.05-0.5,0.5-5 or 5-50mg/kg every day.For oral administration, described composition preferably provides with the form of tablet, this tablet contains the activeconstituents of 1.0-1000 milligram, particularly the activeconstituents of 1.0,5.0,10.0,15.0,20.0,25.0,50.0,75.0,100.0,150.0,200.0,250.0,300.0,400.0,500.0,600.0,750.0,800.0,900.0 and 1000.0 milligrams; According to the symptom adjust dosages of treated patient.Described compound can with every day 1-4 time, the preferably Dosage Regimens Dosage of once a day or twice.

When treat prevent diabetes and/or hyperglycemia or hypertriglyceridemia or compound of the present invention treatable Other diseases time, when compound of the present invention is with the per daily dose administration of about 0.1 milligram of-Yue 100 milligrams per kilogram of the weight of animals, preferred single per daily dose or the divided dose administration of 2-6 time every day, or with sustained release forms administration.For the Mammals that great majority are larger, total per daily dose is about 1.0 milligrams of-Yue 1000 milligrams, preferably about 1 milligram of-Yue 50 milligrams.When the grownup of 70kg body weight, total per daily dose is about 7 milligrams of-Yue 350 milligrams usually.This dosage can be regulated to provide best therapeutic response.

But, be appreciated that, the concrete dosage level of any particular patient and administration number of times can change, and depend on that various factors comprises the activity of used particular compound, the metabolic stability of this compound and acting duration, age of host, body weight, general health situation, sex, diet, administering mode and the severity of number of times, drainage rate, drug combination and concrete illness and the treatment of host's experience.

Scheme and embodiment illustrate several preparation methods of the compounds of this invention below.Parent material can be prepared according to methods known in the art or method as herein described.

Compound of the present invention can as shown in scheme 1, by beta amino acids intermediate e such as formula those and the substituted heterocyclic radical intermediate e of IV such as formula those of V, VI and VII, use standard peptide coupling conditions, then deprotection preparation.

These intermediates can have been bought.

Scheme 1

Intermediate compound IV and V are as shown in scheme 1 under standard peptide coupling conditions, such as, use 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), I-hydroxybenzotriazole (HOBT) and alkali, usual diisopropylethylamine, at solvent such as N, coupling 3-48 hour at ambient temperature in dinethylformamide (DMF) or methylene dichloride, obtains intermediate 1.Then, removing protecting group, such as when Boc, trifluoroacetic acid or methanolic hydrochloric acid (methanolichydrogenchloride) remove protecting group, obtain required amine I.If necessary, by recrystallization, development, preparative thin layer chromatography, as W.C.Still etc., J.Org.Chem., 43, the flashchromatography on silica gel described in 2923 (1978) or HPLC, described product removes unwanted by product.The compound of being purified by HPLC can with the isolated in form of corresponding salt.The purification of intermediate is carried out equally in an identical manner.

In some cases, before removing protecting group, the intermediate 1 obtained by coupled reaction described in scheme 1 can be modified further, such as, by the substituting group on conversion X or Y or Z.These conversion can include, but are not limited to, the known reduction of those skilled in the art, oxidation, alkylation, acidylate and hydrolysis reaction.

In some cases, the order of carrying out above-mentioned reaction scheme can change promote reaction or avoid unwanted reaction product.There is provided the following examples, to understand the present invention more fully.These embodiments are only illustrative, should not be construed and are construed as limiting the present invention by any way.

Embodiment

Embodiment 1

(R)-3-amino-4-(2,5-difluorophenyl)-1-(5,6-glyoxalidine is [1,5-a] pyrazine-7 (8H)-Ji also) butane-1-ketone dihydrochloride

Steps A. (R)-1-(2,5-difluorophenyl)-4-(5,6-glyoxalidine is [1,5-a] pyrazine-7 (8H)-Ji also)-4-oxo-butanes-2-carbamate

At 0 DEG C, to 5,6,7,8-imidazolidine is [1,5-a] pyrazine (50.0mg also, 0.40mmol) with Boc-(R)-3-amino-4-(2,5-difluorophenyl) add HOBT (54.5mg, 0.42mmol) in the solution of butyric acid (128.0mg, 0.40mmol) in methylene dichloride (5mL).This reaction stirs 10min at 0 DEG C, then adds EDC (96.6mg, 0.50mmol).After removing ice bath, this reaction is stirred 14h at ambient temperature.Mixture is concentrated and to purify (Gilson with HPLC; YMC-PackProC18 post, 100x20mmI.D.; Solvent Gradient from 10% acetonitrile, 90% water and 0.1% trifluoroacetic acid to 90% acetonitrile, 10% water and 0.1% trifluoroacetic acid), obtain the title compound of 103mg foaming solid form.

1HNMR(400MHz，CDC13)1.38(s，9H)，2.61(m，2H)，2.85(m，2H)3.80(bs，1H)，3.86(m，1H)，4.05(m，4H).4.65(m，2H)5.30-5.38(m，1H)6.65-6.95(m，4H)，7.28(s，1H)。LC/MS421(M+1)。

Step B. (R)-3-amino-4-(2,5-difluorophenyl)-1-(5,6-glyoxalidine is [1,5-a] pyrazine-7 (8H)-Ji also) butane-1-ketone dihydrochloride

This title compound is by (the R)-1-(2 saturated with hydrogenchloride in 1.5mL methyl alcohol, 5-difluorophenyl)-4-(5,6-glyoxalidine also [1,5-a] pyrazine-7 (8H)-Ji)-4-oxo-butanes-2-carbamate (100.0mg, 0.24mmol, from steps A) preparation.Described reaction mixture is stirred 1h at ambient temperature.Steaming desolventizes, and obtains the title compound of 78mg foam solid form.1HNMR(400MHz，CD30D)3.10-3.17(m，2H)，2.89-2.99(m，2H)，3.94-4.22(m，4H)，4.33(m，1H)，4.91-5.48(m，2H)，6.89-7.20(m，3H)，7.48(s，1H)。ESI-MS321(M+1)。

Substantially according to step listed by embodiment 1, compound listed in obtained table 1.

Table 1

Although by some specific embodiments, invention has been described and explanation, but those skilled in the art are appreciated that, without departing from the spirit and scope of the present invention, various amendment, change, modification, replacement, deletion or increase can be carried out to method of the present invention and scheme.Such as, effective dose can be regarded as the result of variations of the response for the treatment of mammiferous any described symptom with above-mentioned compound of the present invention, instead of the foregoing concrete dosage of this paper.According to depend on selected concrete active compound or whether containing pharmaceutical carrier and preparation type used and administering mode, can be observed the response of concrete pharmacology should change, and in these results desired change or difference and object of the present invention with realize consistent.Therefore, the present invention is defined by claims below, and these claims are as far as possible with extensive interpretation.

Carry out detecting with the biological activity measuring compound as above as follows.

Pharmacodynamics test

Test the mensuration of DPP IV (DP-IV) inhibit activities in 1 rat plasma

Testing compound is dissolved in the DMSO of 100%, uses 10%DMSO/H ₂o is diluted to required concentration, as need testing solution.Using the Wistar rat freely raised as animal for research, per os gives carrier and enzyme inhibitors.At a series of time point, get blood from the eye socket rear vein beard of rat, front isofluranum of taking a blood sample carries out anaesthetic treatment to rat, and blood sampling time to continue up to after administration 24 hours.The blood plasma gathered adds after EDTA and carries out freezing treatment, is used for measuring the activity of DPP IV (DP-IV), during mensuration, first hatches 10 minutes at 37 degree, then carry out 5 times of dilutions to blood plasma, then measure.

According to the people such as DooseopKim (JournalofMedicinalChemistry, vol.48, pp.141-151,2005) method, the enzyme substrates of Gly-Pro-AMC as this research is chosen in this test, this substrate generates after the cutting of DP-IV enzyme has epipolic AMC group, uses continuous fluorescence assay method, the activity of monitoring DP-IV peptase.In typical enzyme reaction, reaction solution cumulative volume is 70 μ L, comprising rat plasma 35 μ L, and the Gly-Pro-AMC of damping fluid (HEPES of 100mM, the BSA of pH7.5,0.1mg/mL) and 50 μMs.After adding the Gly-Pro-AMC substrate of 50 μMs, enzyme reaction starts to carry out.The AMC group of reaction release adopts 96 orifice plate luminescent counter (BMG-Fluostar; BMG-Technologies) METHOD FOR CONTINUOUS DETERMINATION is carried out.Excitation wavelength is set as 360nm, and emission wavelength is set as 460nm.Absorbance increase (Δ ODc) by measuring absorbancy and increment (Δ ODs) thereof, when this reaction mixture does not conform to test compound, and this reaction mixture does not contain the absorbance increase (Δ ODb) of test compound and enzyme, and be calculated as follows the suppression percentage ratio (%) of dipeptidyl peptidase-iv:

{1-[(ΔODs-ΔODb)/(ΔODc-ΔODb)]}×100

The DPP IV inhibit activities of test compound group is expressed as IC ₅₀value (nM), lists in measurement result in table 2 below.

The IC of table 2 different tests compound ₅₀value

Test compound sequence number	IC ₅₀Value (nM)	Test compound sequence number	IC ₅₀Value (nM)
				1	455	21	465
2	126	22	162
				3	468	23	600
4	442	24	210
				5	132	25	531
6	87	26	179
				7	79	27	564
8	17	28	147
				9	144	29	456
10	76	30	137
				11	118	31	510
12	67	32	149
				13	410	33	135
14	156	34	85
				15	453	35	499
16	412	36	127
				17	141	37	431
18	63	38	221
				19	417	39	439
20	125	40	165

In addition, according to above-mentioned experimental technique, also measured were the IC of sitagliptin ₅₀value, test-results shows the IC of this compound ₅₀value is 18nM.

As implied above, compound of the present invention all has excellent DPP IV inhibit activities, and wherein the activity of compound 8 and sitagliptin is the strongest, is obviously better than other compound; Compound 8 is strong compared with the inhibit activities of Sitagliptin.Generally speaking, above-mentioned 40 kinds of compounds can be used as the medicament of prevention or treatment diabetes etc.

Test the oral glucose tolerance test of 2 mouse

From the test of above-mentioned DP-IV inhibition of enzyme activity, visual compounds 8 has excellent DPP IV inhibit activities, has therefore carried out the resistance to quantity research of oral glucose further to it.

Trial drug: compound 8.

Laboratory animal is male mice in kunming, 7-12 week age, body weight is 20-25g.

Experimental technique: mouse is divided into 6 groups at random, often organizes 10, is divided into 1 normal group (loading group), 1 control group (give loading and water, do not give glucose) and 4 trial-product groups.Before mouse experiment, fasting 24 hours, can't help water.Test first 1 hour, from the tail venous blood sampling of mouse, measure the baseline glucose concentration of mouse.Then, per os gives 2 groups of mouse carriers (methylcellulose gum of 0.25%, 5mL/kg), gives 4 groups of above-mentioned trial drug (3,1,0.3,0.1mg/kg of mouse; 5mL/kg).After administration 1 hour, measure blood sugar concentration.Then, per os gives mouse glucose (5g/kg, 10mL/kg), but gives water by the mouse of 1 group of oral carrier thing, as a control group.When giving after glucose 20,40,60 and 120 minutes, measure blood sugar concentration from tail vein blood.Draw blood sugar concentration from t=0min to t=120min through time curve, be used for measuring area under curve (AUC).

Test-results: as illustrated in figs. ia and ib.

Fig. 1 a shows compound 8 takes the glucose level after glucose impact on its mouse oral.

Fig. 1 b shows the blood glucose AUC value (from 0min to 120min) of different dosing group small mouse.

From upper Fig. 1 a, 4 dosage groups of this test setting all significantly can reduce the sugar tolerance of mouse, and Fig. 1 b demonstrates dosage and the inhibit activities of medicine to DP-IV enzyme presents positive correlation.Result shows, this compound significantly can suppress blood sugar concentration, can be used to treat diabetes.

In a word, the sugar tolerance to compound 8 of above-mentioned test 1 Chinese traditional medicine to the research of DP-IV inhibit activities and test 2 is studied, and all shows that above-mentioned 40 kinds of compounds have the biological activity suppressing blood sugar concentration, therefore can as the medicine for the treatment of diabetes.

Claims

1. compound, it is:

Or its pharmacy acceptable salt.

2. pharmaceutical composition, it comprises compound and the inert support of claim 1.

3. the purposes of compound in the medicine for the preparation of suppression dipeptidyl peptidase-IV enzymic activity of claim 1.

4. claim 1 compound for the preparation for the treatment of, control or prevent diabetes medicine in purposes.

5. claim 1 compound for the preparation of need this treatment Mammals in treatment, control or prevention non-insulin-dependent (II type) diabetes medicine in purposes.

6. claim 1 compound for the preparation of need this treatment Mammals in treatment, control or prevention hyperglycemia medicine in purposes.

7. claim 1 compound for the preparation of need this treatment Mammals in treatment, control or prevention insulin resistance medicine in purposes.