CN104016993A

CN104016993A - Compounds for inhibiting DPP-IV and intermediate of compounds

Info

Publication number: CN104016993A
Application number: CN201410062710.7A
Authority: CN
Inventors: 李进; 斯托克斯迈克尔; 窦登峰; 万金桥; 潘飞; 宋宏梅; 胡晓; 易磊
Original assignee: Chengdu Lead Drug Development Corp Ltd
Current assignee: Chengdu Pioneer Drug Development Co Ltd
Priority date: 2013-02-22
Filing date: 2014-02-24
Publication date: 2014-09-03
Anticipated expiration: 2034-02-24
Also published as: CN104016993B; WO2014127747A1

Abstract

The invention provides compounds shown in formulas IA and IB or pharmaceutically acceptable salts of the compounds, as well as a preparation method and application thereof. The invention further provides an intermediate of the compounds and a preparation of the intermediate. The compounds can effectively inhibit the activity of DPP-IV. Compared with the commercially available medicine Januvia, the compound 1 has the similar inhibiting effect on DPP4, and but has lower inhibiting effect on other DPP family members (such as DPP2, DPP8 and DPP9); therefore, the compound can effectively inhibit the pharmacodynamics activity of DPP4, and can further reduce the inhibiting activity to other members of DPP family; the toxic and side effects are reduced, and the medicinal safety is better.

Description

A kind of compound and intermediate thereof that suppresses DPP-IV

Technical field

The present invention relates to compound and the intermediate thereof of a kind of DPP-IV of inhibition.

Background technology

Diabetes are a kind of metabolic disturbance diseases that caused by inherited genetic factors and environmental factors acting in conjunction, serious threat human health and life security.In China, along with the raising of quality of life and the change of mode of life, the morbidity of diabetes sharply increases.The diabetes epidemic data demonstration that IDF (IDF) announces for 2013, Chinese Adult diabetic subject number has reached 9,840 ten thousand people, occupies first place, the whole world, and the situation is tense in prevention and control.In the face of the huge market requirement, the research and development of Novel diabetes medicine enjoy the concern of domestic and international pharmaceutical manufacturer always, and dipeptidyl peptidase-IV (DPP-IV) inhibitor is an important research object in this field.

DPP-IV, the t cell surface antigen CD26 that is otherwise known as, is a kind of II type transmembrane glycoprotein, is made up of 766 amino-acid residues.DPP-IV is distributed widely in Various Tissues and organ in vivo; be included in (the Abbott CA such as kidney, liver, lung, small intestine, lymphocyte and vascular endothelial cell; Baker E; Sutherland GR; McCaughan GW.Genomic organization; exact localization, and tissue expression of the human CD26 (dipeptidyl peptidase IV) gene.Immunogenetics.1994; 40 (5): 331-8), part is present in (Mentlein R.Dipeptidyl-peptidase IV (CD26)--role in the inactivation of regulatory peptides.Regul Pept.1999 in blood plasma with soluble form; 85 (1): 9-24.).DPP-IV is a species specific serine protease, and its substrate is the polypeptide that the 2nd of N end inverse exists proline(Pro) (Pro) or L-Ala (Ala), can take off dipeptides from the N end check of this class polypeptide.The pharmacological action of DPP-IV inhibitor is mainly that the concentration by improving activity in vivo GLP-1 (GLP-1) realizes.GLP-1 is synthetic and secretion by small intestine L cell, is the strongest intestines peptide hormone of promoting insulin secretion of having found.Food digestion can promote GLP-1 to secrete and be released into blood, and brings into play physiological function after special GLP-1 receptor acting.Research shows; GLP-1 can bring into play blood sugar reducing function (Gautier JF from many aspects; Fetita S; Sobngwi E, Sala ü n-Martin C.Biological actions of the incretins GIP and GLP-1and therapeutic perspectives in patients with type2diabetes.Diabetes Metab.2005; 31 (3Pt1): 233-42.): 1) promote the insulin secretion of dependence on the glucose, increase the picked-up of tissue to glucose; 2) by promoting transcribing and strengthen mRNA stability its biosynthesizing being increased of proinsulin gene; 3) secretion of glucagon suppression, reduces glycogen and discharges; 4) promote beta Cell of islet proliferation and differentiation, suppress β apoptosis; 5) by suppressing stomach emptying, appetite control, reduces blood sugar, reduces the risk that body weight increases simultaneously.The treatment that the above-mentioned physiological function of GLP-1 is diabetes B provides an important target spot.But, GLP-1 two amino-acid residues hydrolysis and then the inactivation from N end by DPP-IV very easily in vivo, its, less than 2 minutes, seriously limited its clinical application plasma half-life.DPP-IV is one of key enzyme impelling in body GLP-1 degraded, inactivation, and selectivity inhibition DPP-IV can improve the blood plasma level of active GLP-1.Therefore, the research and development of DPP-IV inhibitor class medicine provide a new way for treating glycosuria.

2006, the Xi Gelieting (Sitagliptin) of Merck (Merck) company exploitation obtained FDA (Food and Drug Adminstration) (FDA) approval, became the DPP-IV inhibitor class medicine of first listing.In addition the DPP-IV inhibitor having gone on the market at present, also comprises Vildagliptin (vildagliptin), four kinds of BMS-477118 (saxagliptin), Egelieting (alogliptin) and BI 1356s (linagliptin) etc.Carrying out at present the clinical study of domestic III phase by China medicine independent research novel DPP-IV inhibitor Rui Gelieting of enterprise (retagliptin).Clinical study shows; DPP-IV inhibitor has good hypoglycemic activity (Argyrakopoulou G, Doupis J. DPP4 inhibitors:from sitagliptin monotherapy to the new alogliptin-pioglitazone combination therapy. Adv Ther. 2009 in diabetes B patient body; 26 (3): 272-80.); compared with traditional diabetes medicament, [Scheen AJ. DPP-4 inhibitors in the management of type 2 diabetes:a critical review of head-to-head trials. Diabetes Metab.2012 have the following advantages; 38 (2): 89-101.; Gallwitz B. Emerging DPP-4 inhibitors:focus on linagliptin for type 2diabetes. Diabetes Metab Syndr Obes. 2013; 6:1-9.]: 1) oral administration; 2) have the dependent incretin secretion of blood sugar, hypoglycemia risk is little; 2) can protect, improve islet beta cell function, stop β cell degradation, contribute to fundamentally to contain the process of diabetes B; 3) body weight of not putting on weight.Therefore, DPP-IV inhibitor is being brought into play more and more important effect in the treatment of diabetes B, becomes the study hotspot of current antidiabetic thing.

Summary of the invention

The object of the present invention is to provide compound of a kind of DPP-IV of inhibition and its production and use.Another object of the present invention is to provide the intermediate of this compound.

The invention provides the compound shown in formula I or its pharmacy acceptable salt,

R ¹be selected from and replace or unsubstituted phenyl; R ²be selected from the alkyl of C1-5 or substituted alkyl, 1-5 containing assorted alkyl or replace containing assorted alkyl or 1-5 heteroatoms or replacement heteroatoms; R ³be selected from H, CN or C1-10 alkyl or substituted alkyl; X is selected from N or CH; Y is selected from N or CR ⁶, wherein, R ⁶be selected from H, CN, carboxyl or ester group.

Wherein, R ¹in, the substituting group of described substituted-phenyl is 1-5 R ⁴, wherein R ⁴be selected from the alkoxyl group of the alkoxy or halogen replacement of the alkyl of CN, halogen, C1-6 or the alkyl of halogen replacement or C1-6;

R ²in, the substituting group of described substituted alkyl is selected from halogen, CN, OH, R ⁵, OR ⁵, NHSO ₂r ⁵, SO ₂r ⁵, COOH or CO ₂r ⁷, replacing heteroatoms or replacing containing the substituting group of assorted alkyl is halogen, CN, OH, R ⁵, OR ⁵, NHSO ₂r ⁵, SO ₂r ⁵, COOH or CO ₂r ⁷;

R ³in, the substituting group of described substituted alkyl is selected from 1-5 halogen;

R ⁶in, described carboxyl is COOH, described ester group is CO ₂r ⁷;

Wherein, heteroatoms is N, O or S; R ⁵for alkyl or the substituted alkyl of C1-6, its substituting group is 1-5 halogen, COOH or CO ₂r ⁷; R ⁷for C1-6 alkyl.

Further, described is the alkyl that contains 1 heteroatomic C1-4 containing assorted alkyl.

Further, R ¹be selected from the phenyl that halogen replaces; R ²be selected from containing assorted alkyl, a 1-2 heteroatoms or C1-2 alkyl; R ³be selected from the alkyl that halogen replaces; R ⁶be selected from CO ₂r ⁷.

Further, described halogen is F or Cl.

Further, described compound is

Wherein, R ²be selected from the alkyl of C1-5 or substituted alkyl or 1-5 containing assorted alkyl or replace containing assorted alkyl or 1-5 heteroatoms or replacement heteroatoms; R ³be selected from H, CN or C1-10 alkyl or substituted alkyl; X is selected from N or CH; Y is selected from N or CR ⁶, wherein, R ⁶be selected from H, CN, carboxyl or ester group.

Further, R ²in, the substituting group of described substituted alkyl is selected from halogen, CN, OH, R ⁵, OR ⁵, NHSO ₂r ⁵, SO ₂r ⁵, COOH or CO ₂r ⁷, described heteroatoms is selected from N, S or O, and replacing heteroatoms or replacing containing the substituting group of assorted alkyl is halogen, CN, OH, R ⁵, OR ⁵, NHSO ₂r ⁵, SO ₂r ⁵, COOH or CO ₂r ⁷;

Wherein, R ⁵for alkyl or the substituted alkyl of C1-6, its substituting group is 1-5 halogen, COOH or CO ₂r ⁷; R ⁷for C1-6 alkyl.

Preferably, described compound or its pharmacy acceptable salt are:

Activity to DPP-IV inhibitor of substituted five-membered fragrant heterocycle tetrahydrochysene pyrazine and other members' of DPP (DPP2, DPP8, DPP9) selectivity play main contributions, and tetrahydrochysene pyrazine is wherein replaced to the activity and selectivity that can regulate compound.But, increase substituting group and will additionally produce multiple chiral isomers, add to the difficulties to compou nd synthesis and discriminating.The present invention finds in research process, introduces substituted radical by bridge joint mode, not only can reduce the complicacy of chipal compounds and increase chirality controllability, has increased compound rigidity simultaneously, can change the activity and selectivity of compound.But, find in building-up process in early stage, five yuan of fragrant heterocycles that replace tetrahydrochysene pyrazine are carried out to bridging, form bridge compound in parallel and there is very strong challenge on synthetic, need to solve a lot of technical difficult problems, for example the bridged ring of chirality control is difficult to effective formation, bridge ring cannot directly form (especially shorter to bridged ring situation) by corresponding and ring, five yuan of fragrant heterocycle building processs can be subject to the impact of bridged ring tension force and cannot complete, amino acid is subject to steric influence to be connected difficulty with the secondary amine of bridge ring, the problems such as bridge the ring intermediate poor stability in the final compound process of formation.Contriver is through a large amount of experimental exploring and analysis, finally work up comparatively desirable synthesis path, effectively overcome the difficult problem existing in building-up process, bridge tetrahydrochysene pyrazine substituted five-membered fragrant heterocycle are successfully synthesized, and, through experimental verification, these compounds have good restraining effect and selectivity to DPP4.

The present invention also provides above-claimed cpd or its pharmacy acceptable salt, in the purposes for the preparation of in dipeptidyl peptidase-iv inhibitor compounds.

Further, described dipeptidyl peptidase-iv inhibitor is that treatment is or/and the medicine of prevent diabetes, hyperglycemia, insulin resistance.

The present invention also provides the preparation method of above-mentioned formula IIA or IIB compound, and reactions steps is as follows:

Comprising following steps:

Step1:f1 and diphenyl phosphate azide, diisopropyl azodiformate, triphenylphosphine reaction, the product obtaining is as next step reactant;

Step2: previous step product is sloughed ester group and obtained next step reaction product under alkaline condition;

Step3: previous step product deaminize blocking group under acidic conditions obtains next step product;

Step4: previous step product reacts and obtains amido protecting product with fluorenes methoxy dicarbonyl chloride under neutrallty condition;

Step5: previous step product and R ³the formyl hydrazine and the condensing agent that replace react in solvent, and the product obtaining is as next step reactant;

Step6: previous step product, trichlorine phosphine oxide react in solvent, and the product obtaining is as next step reactant;

Step7: the reaction of previous step product hydrogenation, the product obtaining is as next step reactant;

Step8: the intramolecular reaction in solvent of previous step product, the product obtaining is as next step reactant;

Step9: react in previous step product and basic solution, the product obtaining is as next step reactant;

Step10: previous step product and R ¹the protection β aminobutyric acid reaction replacing, the product obtaining is as next step reactant;

Step11: previous step product adds acid-respons, obtains product IIA;

Or according to above-mentioned reactions steps, the starting raw material that uses two chiral centres to reverse prepares Compound I IB simultaneously; Or, use raceme starting raw material, after above-mentioned steps reaction, split optical isomer and can obtain Compound I IA or IIB.

Further, in Step1, solvent for use is non-protonic solvent, and described aprotic solvent is preferably tetrahydrofuran (THF), and temperature of reaction is 0 DEG C to 30 DEG C, and the reaction times is preferably 6 to 18 hours;

In Step2, solvent for use is water and tetrahydrofuran (THF) mixed solvent, and alkali used is inorganic strong alkali, and described inorganic strong alkali is preferably lithium hydroxide or sodium hydroxide, and temperature of reaction is 20 DEG C to 30 DEG C, and the reaction times is 2 to 12 hours;

In Step3, solvent for use is polar solvent, and described polar solvent is preferably methylene dichloride, ethyl acetate or methyl alcohol, and acid used is organic acid, is preferably trifluoroacetic acid or hydrochloric acid, and temperature of reaction is 20 DEG C to 30 DEG C, and the reaction times is 0.5-2 hour;

Solvent described in Step4 is preferably tetrahydrofuran (THF) and water mixed solvent, and alkali used is weak base, and described weak base is preferably triethylamine, and temperature of reaction is 0 DEG C to 30 DEG C, and the reaction times is 4 to 12 hours;

Solvent described in Step5 is non-protonic solvent, described non-protonic solvent is preferably methylene dichloride, and alkali used is organic bases, and described organic bases is preferably N, N-diisopropylethylamine, described condensing agent is preferably 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride and I-hydroxybenzotriazole, or 2-(7-azo benzotriazole)-N, N, N', N'-tetramethyl-urea phosphofluoric acid ester, temperature is 20 DEG C to 30 DEG C, the reaction times is 2 to 12 hours;

In Step6, solvent for use is polar solvent, and described polar solvent is preferably acetonitrile, and temperature of reaction is 70 DEG C to 90 DEG C, and the reaction times is 12 to 24 hours;

Solvent described in Step7 is polar solvent, and described polar solvent is preferably methyl alcohol, and hydrogenation catalyst used is 10% palladium carbon, and hydrogenation time is 2 to 4 hours;

Described in Step8, solvent is preferably acetic acid, and temperature of reaction is 110 DEG C to 120 DEG C, and the reaction times is 18 to 24 hours;

In Step9, solvent for use is tetrahydrofuran (THF) and water mixed solvent, described alkali is mineral alkali or organic bases, described mineral alkali is preferably sodium hydroxide or lithium hydroxide, described organic bases is preferably piperidines, and temperature of reaction is 20 DEG C to 30 DEG C, and the reaction times is 0.5 to 2 hour;

In Step10, solvent for use is non-protonic solvent, described non-protonic solvent is preferably methylene dichloride, N, dinethylformamide or tetrahydrofuran (THF), alkali used is organic bases, described organic bases is preferably N, N-diisopropylethylamine, condensing agent is 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride and I-hydroxybenzotriazole, or 2-(7-azo benzotriazole)-N, N, N', N'-tetramethyl-urea phosphofluoric acid ester, temperature of reaction used is 0 DEG C to 30 DEG C, and the reaction times is 0.5 to 2 hour;

Described in Step11, acid is proton strong acid, described proton strong acid is preferably hydrochloric acid or trifluoroacetic acid, and solvent for use is non-protonic solvent, and described non-protonic solvent is preferably methylene dichloride or ethyl acetate, temperature of reaction is 0 DEG C to 30 DEG C, and the reaction times is 0.5 to 4 hour.

The present invention also provides the preparation method of above-mentioned formula III A or IIIB compound, and reactions steps is as follows:

Wherein: Step1:g1 and diphenyl phosphate azide, diisopropyl azodiformate, triphenylphosphine reaction, the product obtaining is as next step reactant;

Step2: previous step product and HR ⁶under highly basic, react, the product obtaining is as next step reactant;

Step3: previous step product and acid-respons, the product obtaining is as next step reactant;

Step4: previous step product reacts with fluorenes methoxy dicarbonyl chloride under weak basic condition, and the product obtaining is as next step reactant;

Step5: previous step product carries out reacting with tert-Butyl dicarbonate after hydrogenation, and the product obtaining is as next step reactant;

Step6: previous step product and sodium nitrite solution react under acidic conditions, the product obtaining is as next step reactant;

Step7: previous step product and zinc powder, anhydride reaction, the product obtaining is as next step reactant;

Step8: previous step product reacts under acidic conditions, the product obtaining is as next step reactant;

Step9: previous step product and hmds, ammonium sulfate reacts under acidic conditions, and the product obtaining is as next step reactant;

Step10: previous step product and R ¹the protection β aminobutyric acid replacing carries out condensation reaction under alkaline condition, and the product obtaining is as next step reactant;

Step11: previous step product adds under strong acidic condition and reacts, and obtains product;

Or according to above-mentioned reactions steps, the starting raw material that uses two chiral centres to reverse prepares compound III B simultaneously; Or, use raceme starting raw material, after above-mentioned steps reaction, split optical isomer and can obtain compound III A or IIIB.

Further, solvent described in Step1 is non-protonic solvent, is preferably tetrahydrofuran (THF), and temperature of reaction is 0 DEG C to 30 DEG C, and the reaction times is 8 to 12 hours;

In Step2, solvent for use is non-protonic solvent, described non-protonic solvent is preferably tetrahydrofuran (THF), and described alkali is non-nucleophilicity organic alkali, and described non-nucleophilicity organic alkali is preferably lithium diisopropyl amido, temperature of reaction is subzero 70 to spend to minus 20 degrees, and the reaction times is 2 hours;

Step3 acid used is proton strong acid, and described proton strong acid is preferably trifluoroacetic acid or hydrochloric acid, and solvent for use is preferably methylene dichloride or ethyl acetate, and temperature of reaction is 0 DEG C to 30 DEG C, and the reaction times is 0.5 to 4 hour;

Step4 solvent for use is polar solvent, is preferably methylene dichloride or water, and alkali used is weak base, is preferably triethylamine, and temperature of reaction is 0 DEG C to 30 DEG C, and the reaction times is 2 to 8 hours;

In Step5, used catalyst is metal catalyst, is optimized for 10% palladium carbon, and solvent is protic solvent, is preferably methyl alcohol, and temperature of reaction is 20 DEG C to 30 DEG C, and the reaction times is 1 to 2 hour;

In Step6, solvent for use is water and acetic acid, and temperature of reaction is 0 degree to 30 DEG C, and the reaction times is 12 hours;

Solvent described in Step7 is acetic acid, and temperature of reaction is 30 DEG C, and the reaction times is 2 hours;

Step8 acid used is proton strong acid, and described proton strong acid is preferably trifluoroacetic acid or hydrochloric acid, and solvent for use is preferably methylene dichloride or ethyl acetate, and temperature of reaction is 0 DEG C to 30 DEG C, and the reaction times is 0.5 to 4 hour;

In Step9, solvent for use is high boiling point non-protonic solvent, and described high boiling point non-protonic solvent is preferably toluene, and used catalyst is ammonium sulfate, and temperature of reaction is 110 DEG C to 130 DEG C, and the reaction times is 4 to 12 hours;

In Step10, solvent for use is non-protonic solvent, described non-protonic solvent is preferably as methylene dichloride, tetrahydrofuran (THF) or N, dinethylformamide, alkali used is organic bases, described organic bases is preferably diisopropylethylamine, condensing agent used is 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride and I-hydroxybenzotriazole or 2-(7-azo benzotriazole)-N, N, N', N'-tetramethyl-urea phosphofluoric acid ester, range of reaction temperature used is 0 DEG C to 30 DEG C, and the reaction times is 0.5 to 2 hour;

Step11 acid used is proton strong acid, and described proton strong acid is preferably trifluoroacetic acid or hydrochloric acid, and solvent for use is preferably methylene dichloride or ethyl acetate, and temperature of reaction is 0 DEG C to 30 DEG C, and the reaction times is 0.5 to 4 hour.

Wherein, Step12:h1 carries out sulfonylation under alkaline condition, and the product obtaining is as next step reactant;

Step13: previous step reaction product is hydrolyzed under alkaline condition, the product obtaining is as next step reactant;

Step14: previous step reaction product is reacted with oxalyl chloride, gained compound and trimethyl silicone hydride diazomethane reaction, and then add hydrobromic acid solution reaction, the product obtaining is as next step reactant;

Step15: previous step reaction product and R ³substituted formamidine reaction, the product obtaining is as next step reactant;

Step16: previous step reaction product self is closed ring under strong alkaline condition, and the product obtaining is as next step reactant;

Step17: previous step reaction product is carried out catalytic hydrogenation, the product obtaining is as next step reactant;

Step18: previous step product and R ¹the protection β aminobutyric acid replacing carries out condensation reaction under alkaline condition, and the product obtaining is as next step reactant;

Step19: previous step reaction product is deprotection under strong acidic condition, obtains product IIIA after purifying;

Further, in Step12, solvent for use is non-protonic solvent, and described non-protonic solvent is preferably tetrahydrofuran (THF) or pyridine, alkali used is organic bases, described organic bases is preferably N-Methylimidazole, and temperature of reaction is 20 DEG C to 30 DEG C, and the reaction times is 4 to 24 hours;

In Step13, solvent for use is water and tetrahydrofuran (THF) mixed solvent, described water and tetrahydrofuran (THF) mixed solvent ratio are 1:1(V/V), alkali used is mineral alkali, described mineral alkali is preferably lithium hydroxide or sodium hydroxide, temperature of reaction is 20 DEG C to 30 DEG C, and the reaction times is 2 to 4 hours;

In step14, solvent for use is preferably acetonitrile, tetrahydrofuran (THF), normal hexane or three's mixed system, and temperature of reaction is 0 DEG C to 30 DEG C, and the reaction times is 8 to 12 hours;

In Step15, solvent for use is protic solvent, and described non-protonic solvent is preferably ethanol, and temperature of reaction is 70 to 90 DEG C, and the reaction times is 3 to 6 hours;

In Step16, solvent for use is non-protonic solvent, and described non-protonic solvent is preferably DMF, and described alkali is inorganic strong alkali, and described inorganic strong alkali is preferably sodium hydride or potassium hydride KH, and temperature is 70 DEG C to 80 DEG C, and the reaction times is 3 to 6 hours;

In Step17, used catalyst is metal catalyst, is optimized for palladium carbon, and solvent is protic solvent, is preferably methyl alcohol, and temperature of reaction is 20 DEG C to 30 DEG C, and the reaction times is 1 to 2 hour;

In Step18, solvent for use is aprotic solvent, described aprotic solvent is preferably methylene dichloride, tetrahydrofuran (THF) or N, dinethylformamide, alkali used is diisopropylethylamine or other organic bases, condensing agent is 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride and I-hydroxybenzotriazole or 2-(7-azo benzotriazole)-N, N, N', N'-tetramethyl-urea phosphofluoric acid ester, temperature of reaction used is 0 DEG C to 30 DEG C, and the reaction times is 0.5 to 2 hour;

Step19 acid used is proton strong acid, and described proton strong acid is preferably trifluoroacetic acid or hydrochloric acid, and solvent for use is preferably methylene dichloride or ethyl acetate, and temperature of reaction is 0 DEG C to 30 DEG C, and the reaction times is 0.5 to 4 hour.

The present invention also provides the intermediate of above-mentioned formula IA and IB compound, and its structural formula is as follows:

Wherein, R ²be selected from the alkyl of C2-5 or substituted alkyl or 2-5 heteroatoms or replace heteroatoms or 2-5 containing assorted alkyl or replace containing assorted alkyl; R ³be selected from H, CN or C1-10 alkyl or substituted alkyl; X is selected from N or CH; Y is selected from N or CR ⁶, wherein, R ⁶be selected from H, CN, carboxyl or ester group.

The present invention also provides midbody compound, and described intermediate is:

The present invention also provides the preparation method of above-mentioned formula IIAA and IIBA compound, and reactions steps is as follows:

Compound I IBA uses the starting raw material that two chiral centres reverse simultaneously to obtain; Compound I IAA also can prepare and separate acquisition with IIBA with identical synthetic method with corresponding raceme starting raw material.

The present invention also provides the preparation method of above-mentioned formula III AA and IIIBA compound, and reactions steps is as follows:

Compound III BA uses the starting raw material that two chiral centres reverse simultaneously to obtain; Compound III AA also can prepare and separate acquisition with IIIBA with identical synthetic method with corresponding raceme starting raw material.

The present invention also provides the preparation method of above-mentioned formula IA and IB compound salt, and step is as follows:

In aqueous methanol (the approximately 20% water) solution of about 30mM free amino group IA or IB, accurately add the acid (48%HBr, phosphoric acid, succinic acid, acetic acid, oxysuccinic acid, toxilic acid, fumaric acid, tartrate etc.) of equimolar amount, and at room temperature stir 1 hour.Then methyl alcohol is reduced pressure and steamed at 25 DEG C, after residual aqueous solution freeze-drying, obtain corresponding IA or IB compound salt.

The compound providing herein and derivative can be according to IUPAC (International Union of Pure and Applied Chemistry(IUPAC)) or the names of CAS (chemical abstracts service, Columbus, OH) naming system.

About the definition of use term of the present invention.

Except as otherwise noted, the original definition that group or term provide is herein applicable to this group or the term of entire description.For the term being not specifically defined herein, should be according to disclosure and context, provide those skilled in the art and can give their implication.

" replacement " refers to that the hydrogen atom in molecule replaced by other different atom or molecule.

In hydrocarbon group, the minimum value of carbon content and maximum value are by prefix designates, and for example, prefix (Ca-b) alkyl shows any " a " alkyl to " b " individual carbon atom that contains.Therefore, for example, (C1 – 4) alkyl refers to the alkyl that comprises 1-4 carbon atom.

Term " pharmaceutically acceptable " refers to certain carrier, load, thinner, auxiliary material, and/or the salt forming conventionally chemically or physically with form certain pharmaceutical dosage form other becomes phase-splitting compatibility, and on physiology with acceptor compatibility mutually.

Term " salt " and " pharmaceutically useful salt " are by compound or its steric isomer, and acid and/or subsalt with inorganic and/or organic bronsted lowry acids and bases bronsted lowry forms, also comprise zwitter-ion salt (inner salt), also comprises quaternary ammonium salt, for example alkylammonium salt.These salt can be directly to obtain in the last separation of compound and purifying.Also can be by by compound, or its steric isomer, with the acid of some amount or alkali suitably (for example equivalent) be obtained by mixing.These salt may form precipitation and collect with filter method in solution, or after solvent evaporation, reclaim and obtain, or in water medium, react postlyophilization and make.Salt described in the present invention can be hydrochloride, vitriol, citrate, benzene sulfonate, hydrobromate, hydrofluoride, phosphoric acid salt, acetate, propionic salt, succinate, oxalate, malate, succinate, fumarate, maleate, tartrate or the trifluoroacetate of compound.

In one embodiment of the present invention, present invention includes isotope-labeled formula (IA) or (IB) compound, described compound isotopically labelled refers to listed Compound Phase is same herein, but one or more atom is replaced by another atom, the atomic mass of this atom or total mass number are different from the common atomic mass of occurring in nature or total mass number.Can drawing-in system (IA) or (IB) isotropic substance in compound comprise hydrogen, carbon, nitrogen, oxygen, sulphur, i.e. 2H, 3H, 13C, 14C, 15N, 17O, 18O, 35S.Compound and the steric isomer thereof of the formula (I) that contains above-mentioned isotropic substance and/or other atom isotope, and within the pharmaceutically useful salt of this compound, steric isomer all should be included in the scope of the invention.

Key intermediate in the present invention separates and purifying with compound, and the mode using is that Isolation and purification method conventional in organic chemistry and the example of described method comprise filtration, extract, are dried, are spin-dried for and various types of chromatograms.Selectively, can make that intermediate is not purified carries out next step reaction.

In some embodiments, one or more compounds of the present invention use that can combine with one another.Also can select compound of the present invention to be combined with any other active agent, for the preparation of medicine or the pharmaceutical composition of regulating cell function or treatment disease.If what use is one group of compound, can be by these compounds simultaneously, respectively or in an orderly manner study subject be entered

The compounds of this invention can effectively suppress DPP-IV activity, compared with marketed drugs Jie Nuowei, compound 1 is suitable to the restraining effect of DPP4, but prompt Novi is lower to other DPP family members' (DPP2, DPP8, DPP9) inhibition specific activity, show that the compounds of this invention not only can effectively suppress DPP4 performance drug activity, can also reduce the inhibition activity to other families of DPP, reduce toxic side effect, medicinal security is better.

The compounds of this invention can be used for the multiple treatment with DPP-4 relative disease such as diabetes, and better drug safety is provided.

Embodiment

Embodiment 1(R)-3-amino-1-((5S, 8S)-3-(trifluoromethyl)-5,6-dihydro-5,8-bridge methyl [1,2,4] triazole also [4,3-a] pyrazine-7(8H)-alkyl)-4-(2,4,5-trifluorophenyl) preparation of-1-butanone hydrochloride (compound 1)

Step 1:N-tertiary butyl oxycarbonyl-cis-4-nitrine-L-PROLINE methyl esters (compound 1b)

Under room temperature, 1a (34.7g, 164mmol) and triphenyl phosphorus (54g, 204mmol) are dissolved in tetrahydrofuran (THF) (500mL), stir and nitrogen replacement.Under ice bath, diisopropyl azodiformate (44g, 212mmol) is slowly added dropwise in tetrahydrofuran solution through syringe.Then, then diphenyl phosphate azide (56g, 204mmol) slowly splash into.After adding, reaction system slowly rises to 25 degrees Celsius and stirs 12 hours.Then solvent underpressure distillation is removed, then obtain product 1b, yellow oil 35g, yield 72% through column chromatography (silica gel, sherwood oil: ethyl acetate=10:1).

MS(ESI)m/z:215(M-56+1); ¹HNMR(400MHz,CDCl ₃):δ4.44-4.31(m,1H),4.16-4.13(m,1H),3.74(s,3H),3.52-3.445(m,1H),2.49-2.44(m,1H),2.19-2.15(m,1H),1.66(s,1H),1.47-1.25(m,9H)。

Step 2:N-tertiary butyl oxycarbonyl-cis-4-nitrine-L-PROLINE (compound 1c)

At room temperature, hydronium(ion) is oxidized to lithium and adds 1b(1.6g, 5.9mmol) tetrahydrofuran (THF) and the aqueous solution in and stir 4h.Then under ice bath, use the hydrochloric acid acidizing reaction liquid of 1N to pH2, then with ethyl acetate extraction (25mL × 3).Merge organic phase, dry, filter, the concentrated 1c, colorless oil compounds 1.2g, yield 83% of obtaining.Products obtained therefrom does not need to be further purified, and is directly used in next step reaction.MS(ESI)m/z:257(M+1)。

Step 3: cis 4-nitrine-L-PROLINE (compound 1d)

Under ice bath, to 1c(1.0g, 3.9mmol) dichloromethane solution in drip trifluoroacetic acid, stir.Slowly rise to room temperature and continue to stir 2h.Then obtain 1d by concentrated reaction solution, yellow oil 1.5g, yield is greater than 100%(and remains trifluoroacetic acid).Without being further purified, be directly used in next step reaction.MS(ESI)m/z:157(M+1)

Step 4:N-9-fluorenylmethyloxycarbonyl--cis 4-nitrine-L-PROLINE (compound 1e)

Under ice bath, to 1d (1.5g, in tetrahydrofuran (THF) (20mL) 3.9mmol) and water (10mL) mixing solutions, slowly add fluorenes methoxy dicarbonyl chloride (1.5g, tetrahydrofuran (THF) (10mL) solution 5.8mmol), and allowing reaction solution to rise to room temperature, stirring is spent the night.After having reacted, first tetrahydrofuran (THF) underpressure distillation is removed, then extract excessive fluorenes methoxy dicarbonyl chloride with methyl tertiary butyl ether.Under ice bath with the hcl acidifying water of 1N to pH2, be then extracted with ethyl acetate (15mL × 3).Merge organic phase, with saturated common salt washing (10mL × 3).Organic phase drying, filtration, concentratedly obtain compound 1e, white solid 1.0g, yield 68%.

MS(ESI)m/z:379(M+1); ¹HNMR(400MHz,DMSO-d ₆):δ12.8(br,1H),7.92-7.90(m,2H),7.69-7.64(m,2H),7.45-7.30(m,4H),4.45-4.18(m,5H),3.69-3.65(m,1H),2.59-2.54(m,1H),2.13-2.00(m,1H)。

Step 5:(2S, 4S)-4-nitrine-2-(2-trifluoroacetyl) hydrazine carbonyl pyrrolidine alkane-1-formic acid 9-fluorenes methyl esters (compound 1f)

Under stirring at room temperature, to 1e(0.6g, in dichloromethane solution 1.6mmol), add successively N, N-diisopropylethylamine (0.8g, 6.4mmol), 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride (0.46g, 2.4mmol) and I-hydroxybenzotriazole (0.32g, 2,4mmol).After 10 minutes, trifluoroacetyl hydrazine (0.25g, 1.9mmol) is added in above-mentioned reaction solution, under room temperature, stir and spend the night.After having reacted, solvent underpressure distillation is removed.Crude product obtains 1f, white solid 0.6g, yield 76% through column chromatography (silica gel, sherwood oil: ethyl acetate=8:1).

MS(ESI)m/z:489(M+1); ¹HNMR(400MHz,DMSO-d ₆):δ11.57(s,1H),10.30(s,1H),7.92-7.88(m,2H),7.70-7.61(m,2H),7.45-7.29(m,4H),4.56-4.28(m,5H),3.79-3.72(m,1H),2.67-2.55(m,1H),2.06-1.96(m,1H)。

Step 6:(2S, 4S)-4-nitrine-2-(5-Trifluoromethyl-1,3,4-oxazole-2-alkyl) tetramethyleneimine-1-formic acid 9-fluorenes methyl esters (compound 1g)

To compound 1f(0.6g, 1.2mmol) 20mL acetonitrile solution in add phosphorus oxychloride (1.8g, 12mmol) stirring and refluxing 24h.Reaction is cooled to after room temperature, adds appropriate ice cancellation in ice bath downhill reaction liquid, stirs 0.5h complete to ensure cancellation.Then acetonitrile underpressure distillation is removed, then use methylene dichloride (15mL × 3) extraction.After dry organic phase, concentrated, crude product obtains compound 1g, white solid 0.4g, yield 69% through column chromatography (silica gel, sherwood oil: ethyl acetate=6:1).

MS(ESI)m/z:471(M+1); ¹HNMR(400MHz,DMSO-d ₆):δ7.91-7.80(m,2H),7.68-7.66(m,1H),7.51-7.23(m,5H),5.36-5.34(d,0.5H),5.18-5.20(d,0.5H),4.57-4.51(m,1H),4.40-4.38(m,1H),4.33-4.31(m,1.5H),4.14-4.12(m,0.5H),3.76-3.72(m,1H),3.46-3.39(m,1H),2.77-2.70(m,1H),2.36-2.29(m,1H)。

Step 7:(2S, 4S)-4-amino-2-(5-Trifluoromethyl-1,3,4-oxazole-2-alkyl) tetramethyleneimine-1-formic acid 9-fluorenes methyl esters (compound 1h)

To 1g(0.4g, 0.85mmol) 15mL methanol solution in add 10% palladium carbon (50mg) and pass into hydrogen (2atm).Under room temperature, stir after 2h the filtering of palladium carbon, then add 10mL glacial acetic acid in filtrate.Being evaporated to resid vol is the acetic acid solution that 1mL obtains 1h.This solution is directly used in next step reaction.MS(ESI)m/z:445(M+1)。

Step 8:(5S, 8S)-3-(trifluoromethyl)-5,6-dihydro-5, also [4,3-a] pyrazine-7(8H of 8-methyl [1,2,4] triazole)-formic acid 9-fluorenes methyl esters (compound 1i)

In residue after concentrated to the 8th step, add 15mL glacial acetic acid, stirring and refluxing 18h.React complete, except desolventizing, residue is purified and is obtained compound 1i, white solid 40mg, yield 17% with preparing silica-gel plate.

MS(ESI)m/z:427(M+1); ¹HNMR(400MHz,DMSO-d ₆):δ7.90-7.86(m,3H),7.50-7.25(m,5H),5.59-5.45(m,2H),4.37-4.21(m,3H),3.70(s,1H),3.17-3.12(m,0.5H),2.92-2.90(m,0.5H),2.71-2.59(m,1H),2.45-2.43(m,1H)。

Step 9:(5S, 8S)-3-(trifluoromethyl)-5,6,7,8-tetrahydrochysene-5, also [4,3-a] pyrazine (compound 1j) of 8-methyl [1,2,4] triazole

At room temperature, to 1i(40mg, 0.09mmol) 3mL tetrahydrofuran solution in add the aqueous sodium hydroxide solution (0.2mL) of 1N, and stir 0.5h.After having reacted, with the hcl acidifying of 1N, to pH2, reconcentration is to dry.In resistates, add 3mL water, then with methyl tertiary butyl ether extracting impurities, the aqueous solution obtains compound 1j, faint yellow solid 25mg, yield 100% after being concentrated into and doing.Products obtained therefrom does not need to be further purified, and is directly used in next step reaction.

MS(ESI)m/z:205(M+1)。

Step 10:((R)-4-oxygen-4-((5S, 8S)-3-(trifluoromethyl)-5,6-dihydro-5,8-methyl [1,2,4] triazole also [4,3-a] pyrazine-7(8H) alkyl)-1-(2,4,5-trifluorophenyl) fourth-2-alkyl) t-butyl carbamate (compound 1k)

To (3R)-N-tertbutyloxycarbonyl-3-amino-4-(2,4,5-trifluorophenyl) butyric acid (33.3mg, in 5mL dichloromethane solution 0.1mmol), add successively N, N-diisopropylethylamine (48mg, 0.37mmol), 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride (27mg, 0.14mmol) and I-hydroxybenzotriazole (19mg, 0.14mmol). stir after 10min, add 1j(25mg, 0.1mmol), stirred overnight at room temperature.Underpressure distillation is except after desolventizing, and crude product obtains compound 1k by preparative HPLC purifying, white solid 22mg, yield 45%.MS(ESI)m/z:520(M+1)。

Step 11:(R)-3-amino-1-((5S, 8S)-3-(trifluoromethyl)-5,6-dihydro-5, also [4,3-a] pyrazine-7(8H of 8-bridge methyl [1,2,4] triazole)-alkyl)-4-(2,4,5-trifluorophenyl)-1-butanone hydrochloride (compound 1)

1k is dissolved in 2mL2N hydrochloric ethyl acetate solution to stirring at room temperature 0.5h.Then concentrating under reduced pressure, crude product obtains target compound 1, white solid 22mg, yield 81% after preparative HPLC purifying.

MS(ESI)m/z:420(M+1); ¹HNMR(400MHz,DMSO-d ₆):δ8.00-7.94(br,2H),7.59-7.44(m,2H),5.80-5.60(m,2H),3.83-3.80(m,1H),3.71-3.66(m,1H),3.30-3.28(m,1H),3.09-3.01(m,1H),3.00-2.93(m,1.5H),2.90-2.88(m,0.5H),2.85-0.75(m,0.5H),2.66-2.64(m,0.5H),2.60-2.58(m,0.5H),2.46-2.36(m,0.5H)。

(R)-3-amino-1-((5R, 8R)-3-(trifluoromethyl)-5,6-dihydro-5, also [4,3-a] pyrazine-7(8H of 8-bridge methyl [1,2,4] triazole)-alkyl)-4-(2,4,5-trifluorophenyl) preparation of-1-butanone hydrochloride (compound 1 ')

The 1a that two chiral centres of preparation use of compound 1 ' reverse simultaneously, as starting raw material, uses identical experimental procedure to obtain.

Embodiment 2:(5S; 8S)-7-((R)-3-amino-4-(2; 4; 5-trifluorophenyl) butyryl radicals)-3-(trifluoromethyl)-5; 6,7,8-tetrahydrochysene-5; 8-bridge Methylimidazole is the preparation of [1,5-a] pyrazine-1-methyl-formiate trifluoroacetate (compound 2) also

Step 1:(2S, 4S)-4-nitrine-2-(3-methoxyl group-3-carbonyl propionyl) tetramethyleneimine-1-t-butyl formate (compound 2b)

Under room temperature, methyl acetate (26.1g, 296.0mmol) is dissolved in to anhydrous tetrahydro furan (300mL), after nitrogen protection, is cooled to-70 DEG C, then at this temperature, drip LDA (148mL, 2M/L) and stir 1 hour.Keep at this temperature, the tetrahydrofuran (THF) of compound 2a (20g, 74mmol) (20mL) solution is added drop-wise in reaction solution slowly, then reaction solution is warming up to-20 ° and at this temperature, stir 1 hour slowly.Then for reaction solution, after saturated ammonium chloride solution (100mL) cancellation, concentrating under reduced pressure is removed after most tetrahydrofuran (THF), then uses ethyl acetate (100mL × 3) extraction.Merge organic phase, with saturated common salt washing (300mL).Organic phase drying, filtration, concentrated, then obtain compound 2b, yellow oil 7.4g, yield 27% through column chromatography (silica gel, sherwood oil: ethyl acetate=8:1).

MS(ESI)m/z:335(M+23); ¹HNMR(400MHz,CDCl ₃):δ4.42-4.29(m,1H),4.19(s,1H),3.74(s,3H),3.81-3.69(m,4H),3.68-3.60(m,1H),3.59-3.38(m,1H),2.45-2.27(m,2H),1.66(s,1H),1.48-1.37(m,9H)。

Step 2:3-((2S, 4S)-4-nitrine pyrroles-2-alkyl)-3-carbonyl propionic acid methyl esters (compound 2c)

Under ice bath, in methylene dichloride (50mL) solution of compound 2b (6g, 22.2mmol), drip trifluoroacetic acid (24mL), stir.After dripping off, slowly rise to room temperature and continue to stir 3h.Then reaction solution is concentrated to the trifluoroacetate 6.4g that obtains compound 2c, brown oil, yield 100%, without being further purified, is directly used in next step reaction.

MS(ESI)m/z:213(M+1)。

Step 3:(2S, 4S)-4-nitrine-(3-methoxyl group-3-carbonyl propionyl) tetramethyleneimine-1-formic acid (9H-fluorenes-9) methyl esters (compound 2d)

Under ice bath, to the trifluoroacetate (5.4g of compound 2c, in methylene dichloride (20mL) solution 17mmol), add triethylamine (7.9g, 27.6mmol), and then slowly add fluorenes methoxy dicarbonyl chloride (6g, methylene dichloride (10mL) solution 18.6mmol), and react 1h under ice bath.React rear water (30mL) and washed separatory, methylene dichloride for water (30mL × 2) back extraction.Merge organic phase, with saturated common salt washing (90mL).Organic phase drying, filtration, concentratedly obtain compound 2d, yellow oil 3g, yield 40% through column chromatography (silica gel, sherwood oil: ethyl acetate=5:1) again.

MS(ESI)m/z:435(M+1); ¹HNMR(400MHz,CDCl ₃):δ7.81-7.71(m,2H),7.62-7.45(m,2H),7.42-7.38(m,2H),7.36-7.31(m,2H),4.67-4.57(m,1H),4.45-4.40(m,1H),4.26-4.11(m,2H),3.73-3.52(m,4H),3.51-3.44(m,1H),3.44-3.32(m,1H),3.29-2.98(m,1H),2.34-2.04(m,1H),1.33-1.24(m,2H)。

Step 4:(2S, 4S)-4-(tertiary butyloxycarbonyl amino)-2-(3-methoxyl group-3-carbonyl propionyl)-tetramethyleneimine-1-formic acid (9H-fluorenes-9) methyl esters (compound 2e)

To compound 2d(2.4g, 5.5mmol) 20mL methanol solution in add 10% palladium carbon (0.24g) and Boc ₂o (1.4g, 6mmol) also passes into hydrogen.Under room temperature, stir after 1h the filtering of palladium carbon, filtrate decompression is concentrated obtains target compound 2e, colorless oil 2.4g, yield 86%.

MS(ESI)m/z:509(M+1); ¹HNMR(400MHz,CDCl ₃):δ7.81-7.71(m,2H),7.62-7.45(m,2H),7.42-7.38(m,2H),7.36-7.31(m,2H),5.32-5.30(m,1H),5.06-5.00(m,1H),4.48-4.46(m,1H),4.44-4.13(m,4H),3.88-3.80(m,5H),3.47-3.45(m,1H),2.60-2.53(m,1H),2.00-1.92(m,1H),1.44-1.25(m,12H)。

Step 5:(2S, 4S)-4-tertiary butyloxycarbonyl amido-2-((Z)-2-oximido-3-methoxyl group-3-carbonyl propionyl-tetramethyleneimine-1-formic acid (9H-fluorenes-9) methyl esters (compound 2f)

Under ice bath to water (20mL) solution that slowly drips Sodium Nitrite (0.64g, 9.20mmol) in acetic acid (40mL) solution of compound 2e (2.4g, 4.7mmol).After dripping, reaction solution risen to slowly to room temperature and stir and spend the night.Then regulate pH to 8 left and right with saturated sodium bicarbonate solution (30mL) concentrated reaction solution removing after acetic acid, then use ethyl acetate (30mL × 3) extraction.Merge organic phase, with saturated common salt washing (90mL).Organic phase drying, filtration, concentrated, then obtain compound 2f, yellow oil 1.6g, yield 64% through column chromatography (silica gel, sherwood oil: ethyl acetate=5:1).

MS(ESI)m/z:538(M+1); ¹HNMR(400MHz,CDCl ₃)δ7.81-7.71(m,2H),7.62-7.45(m,2H),7.42-7.38(m,2H),7.36-7.31(m,2H),5.00-4.70(m,1H),4.38-4.31(m,2H),3.85(s,3H),3.83-3.74(m,2H),3.55-3.39(m,1H),2.43-2.26(m,3H),1.47-1.43(m,9H)。

Step 6:(2S, 4S)-4-tertiary butyloxycarbonyl amido-2-(3-methoxyl group-3-carbonyl-2-(2,2,2-trifluoroacetamido) propionyl) tetramethyleneimine-1-formic acid (9H-fluorenes-9) methyl esters (compound 2g)

Under room temperature, in acetic acid (20mL) solution of compound 2f (0.8g, 1.5mmol), add zinc powder (0.2g, 3mmol) and trifluoroacetic anhydride (0.4g, 1.9mmol).Reaction solution at room temperature stirs after 2 hours and filters, and after filtrate is concentrated, obtains compound 2g, yellow oil 0.4g, yield 43% again through column chromatography (silica gel, sherwood oil: ethyl acetate=5:1).

MS(ESI)m/z:620(M+1); ¹HNMR(400MHz,CDCl ₃):δ7.81-7.71(m,2H),7.62-7.45(m,2H),7.42-7.38(m,2H),7.36-7.31(m,2H),5.31-4.80(m,1H),4.65-4.58(m,2H),4.35-4.56(m,2H),3.88-3.68(m,3H),3.48(s,3H),1.86-1.70(m,2H),1.47-1.43(m,9H)。

Step 7:(2S, 4S)-4-amino-2-(3-methoxyl group-3-carbonyl-2-(2,2,2-trifluoroacetamido)-propionyl) tetramethyleneimine-1-formic acid (9H-fluorenes-9) methyl esters (compound 2h)

Under ice bath to compound 2g (150mg, in methylene dichloride (3mL) solution 0.24mmol), drip trifluoroacetic acid (2mL), stir after 1 hour the concentrated reaction solution compound 2h that obtains, yellow oily compounds 152mg, yield 100%, without being further purified, be directly used in next step reaction.

MS（ESI)m/z:520(M+1)。

Step 8:(5S, 8S)-3-trifluoromethyl-5,6,7,8-tetrahydrochysene-5,8-endo-methylene group-imidazo [1,5-a] pyrazine-tetramethyleneimine-1-methyl-formiate (compound 2i)

Compound 2h (152mg, 0.24mmol) is dissolved in after dimethylbenzene (5mL) stirring and refluxing 2 hours, is cooled to the concentrated brown oil that obtains after room temperature.This oily matter is dissolved in to the hmds of 20mL, then adds acetic acid (1.5mg, 0.02mmol) and ammonium sulfate (2.3mg, 0.02mmol), stir and flow through next time night.Then reaction solution concentrated and add 5mL water, then using ethyl acetate (5mL × 3) extraction.Merge organic phase, with saturated common salt washing (15mL).Organic phase drying, filtration, concentratedly obtain crude compound 2i, yellow oil 10mg, yield 16%.Without being further purified, be directly used in next step reaction.MS(ESI)m/z:262(M+1)。

Step 9:(5S, 8S)-7-((R)-3-tertiary butyloxycarbonyl amido-4-(2,4; 5-trifluorophenyl) butyryl radicals)-3-trifluoromethyl-5,6,7; 8-tetrahydrochysene-5,8-endo-methylene group-imidazo [1,5-a] pyrazine-1-methyl-formiate (compound 2j)

To (3R)-N-tertbutyloxycarbonyl-3-amino-4-(2,4,5-trifluorophenyl) add successively DIPEA (17.5mg in the 3mL dichloromethane solution of butyric acid (18.8mg, 0.56mmol), 0.13mmol), EDCI (13mg, 0.067mmol) and HOBT (9.2mg, 0.067mmol). stir after 10min, add compound 2i (10mg, 0.045mmol), stirred overnight at room temperature.Underpressure distillation is except after desolventizing, and crude product obtains compound 2j through preparative HPLC purifying, colorless oil 5mg, yield 19%.MS(ESI)m/z:577(M+1)。

Step 10:(5S; 8S)-7-((R)-3-amino-4-(2; 4; 5-trifluorophenyl) butyryl radicals)-3-(trifluoromethyl)-5; 6,7,8-tetrahydrochysene-5; 8-endo-methylene group-imidazo [1,5-a] pyrazine-1-methyl-formiate trifluoroacetate (compound 2)

Under ice bath, to compound 2j (5mg, in methylene dichloride (2mL) solution 0.009mmol), drip trifluoroacetic acid (1mL), stir after 1 hour reaction solution is concentrated, crude product obtains the trifluoroacetate of target compound 2 after preparative HPLC purifying, colorless oil 3mg, yield 64%.

MS(ESI)m/z:477(M+1); ¹HNMR(400MHz,DMSO-d ₆):δ7.81-7.71(m,2H),7.62-7.45(m,2H),7.42-7.38(m,2H),7.36-7.31(m,2H),5.31-4.80(m,1H),4.65-4.58(m,2H),4.35-4.56(m,2H),3.88-3.68(m,3H),3.48(s,3H),1.86-1.70(m,2H),1.47-1.43(m,9H)。

Embodiment 3:

((5S, 8S)-7-((R)-3-amino-4-(2,4; 5-trifluorophenyl) butyryl radicals)-3-methyl-5,6,7; 8-tetrahydrochysene-5, the preparation of 8-endo-methylene group-imidazo [1,5-a] pyrazine-1-methyl-formiate (compound 3)

Step 1:(2S, 4S)-2-(2-acetylaminohydroxyphenylarsonic acid 3-methoxyl group-3-carbonyl propionyl)-4-(tertiary butyloxycarbonyl amido) tetramethyleneimine-1-formic acid (9H-fluorenes-9) methyl esters (compound 3b)

Under room temperature, in acetic acid (20mL) solution of compound 3a (2f) (0.8g, 1.48mmol), add zinc powder (0.2g, 3mmol) and aceticanhydride (0.2g, 1.9mmol).Reaction solution at room temperature stirs after 2 hours and filters, and after filtrate is concentrated, obtains compound 3b, yellow oil 0.5g, yield 59% again through column chromatography (silica gel, sherwood oil: ethyl acetate=5:1).

MS(ESI)m/z:566(M+1); ¹HNMR(400MHz,CDCl ₃):δ7.81-7.71(m,2H),7.62-7.45(m,2H),7.42-7.38(m,2H),7.36-7.31(m,2H),5.48-4.32(m,1H),5.05-4.77(m,2H),4.40-4.34(m,2H),4.33-4.21(m,2H),3.85-3.82(m,3H),3.79-3.74(m,1H),3.74-3.39(m,1H),2.61-2.19(m,1H),2.09(s,3H),2.08-2.04(m,1H),1.47-1.39(m,9H)。

Step 2:(2S, 4S)-2-(2-acetylaminohydroxyphenylarsonic acid 3-methoxyl group-3-carbonyl propionyl)-4-amino-pyrrolidine-1-formic acid (9H-fluorenes-9) methyl esters (compound 3c)

Under ice bath to compound 3b (150mg, in methylene dichloride (3mL) solution 0.27mmol), drip trifluoroacetic acid (2mL), stir after 1 hour the concentrated reaction solution compound 3c that obtains, yellow oil 156mg, yield 100%, without being further purified, be directly used in next step reaction.

MS(ESI)m/z:466(M+1)。

Step 3:(5S, 8S)-3-methyl-5,6,7,8-tetrahydrochysene-5,8-endo-methylene group-imidazo [1,5-a] pyrazine-1-formic acid (9H-fluorenes-9) methyl esters (compound 3d)

The trifluoroacetate (156mg, 0.27mmol) of compound 3c is dissolved in after dimethylbenzene (5mL) stirring and refluxing 2 hours, is cooled to the concentrated brown oil that obtains after room temperature.This oily matter is dissolved in to the hmds of 20mL, then adds acetic acid (1.5mg, 0.02mmol) and ammonium sulfate (2.3mg, 0.02mmol), stir and flow through next time night.Then reaction solution concentrated and add 5mL water, then using ethyl acetate (5mL × 3) extraction.Merge organic phase, with saturated common salt washing (15mL).Organic phase drying, filtration, concentratedly obtain compound 3d, yellow oil 20mg, yield 36%, without being further purified, is directly used in next step reaction.MS(ESI)m/z:208(M+1)。

Step 4:(5S, 8S)-7-((R)-3-tertiary butyloxycarbonyl amido-4-(2,4; 5-trifluorophenyl) butyryl radicals)-3-methyl-5,6,7; 8-tetrahydrochysene-5,8-endo-methylene group-imidazo [1,5-a] pyrazine-1-methyl-formiate (compound 3e)

To (3R)-N-tertbutyloxycarbonyl-3-amino-4-(2,4,5-trifluorophenyl) butyric acid (18.8mg, in 3mL dichloromethane solution 0.55mmol), add successively N, N-diisopropylethylamine (17.5mg, 0.13mmol), EDCI (13mg, 0.067mmol) and HOBT (9.2mg, 0.067mmol).Stir after 10min, add compound 5 (10mg, 0.048mmol), stirred overnight at room temperature.Underpressure distillation is except after desolventizing, and crude product obtains compound 3e through preparative HPLC purifying, and colorless oil 15mg receives 60%.MS(ESI)m/z:523(M+1)。

Step 5:((5S; 8S)-7-((R)-3-amino-4-(2; 4; 5-trifluorophenyl) butyryl radicals)-3-methyl-5; 6,7,8-tetrahydrochysene-5; 8-endo-methylene group-imidazo [1,5-a] pyrazine-1-methyl-formiate trifluoroacetate (compound 3)

Under ice bath, in methylene dichloride (2mL) solution of compound 3e (15mg, 0.028mmol), drip trifluoroacetic acid (1mL), stir after 1 hour reaction solution is concentrated, crude product obtains target compound 3, colorless oil 8mg, yield 48.5% after preparative HPLC purifying.

MS(ESI)m/z:423(M+1); ¹HNMR(400MHz,CD ₃OD):δ7.31-7.15(m,2H),5.94-5.55(m,2H),3.95-3.88(m,3H),3.87-3.80(m,3H),3.44-3.41(m,1H),3.18-3.17(m,1H),3.14-3.06(m,2H),3.05-2.90(m,1H),2.70-2.62(m,1H),2.61-2.52(m,3H),2.50-2.44(m,3H)。

Embodiment 4:(R)-3-amino-1-((5S, 8S)-3-trifluoromethyl-5,6-dihydro-5,8-endo-methylene group-imidazo [1,5-a] pyrazine-7 (8H)-alkyl)-4-(2,4,5-trifluorophenyl) preparation of-1-butanone trifluoroacetate (compound 4)

Step 1:(2S, 4R)-4-tolysulfonyl oxygen base-tetramethyleneimine-1,2-dioctyl phthalate-1-benzyl-2-methyl esters (compound 4b)

Under ice bath, in tetrahydrofuran (THF) (5mL) solution of compound p-toluenesulfonyl imidazoles (10g, 45.2mmol), slowly drip trifluoromethanesulfonic acid methyl esters (7.9g, 48.2mmol).Ice bath reaction, after 1 hour, is added dropwise to tetrahydrofuran (THF) (20mL) solution of compound 4a (8.4g, 30.1mmol) and N-Methylimidazole (3.7g, 45.2mmol) in reaction mixture.Reaction solution stirs 24 hours after slowly rising to room temperature, and then solvent is removed in decompression, is adding 40mL water, with ethyl acetate (30mL × 3) extraction.Merge organic phase, with saturated common salt washing (90mL).Organic phase drying, filtration, concentrated again through column chromatography (silica gel, sherwood oil: acetoacetic ester=3:1) compound 4b, white solid 7.3g, yield 56%.

MS(ESI)m/z:434(M+1); ¹HNMR(400MHz,DMSO-d ₆):δ7.78-7.75(m,2H),7.38-7.29(m,7H),5.20-4.98(m,3H),4.49-4.42(m,1H),3.74(s,1.5H),3.72-3.62(m,2H),3.52(s,1.5H),2.59-2.49(m,1H),2.46-2.43(m,3H),2.22-2.15(m,1H)。

Step 2:(2S, 4R)-1-benzyloxy carbonyl acyl group-4-tolysulfonyl oxygen base tetramethyleneimine-2-formic acid (compound 4c)

Under ice bath by lithium hydroxide aqueous solution (20mL, 0.8g, 20mmol) add in the methyl alcohol (100mL) of compound 4b (7.3g, 16.9mmol) and tetrahydrofuran (THF) (20mL) solution and stir 1h, then rising to room temperature reaction and spend the night.Reaction solution is concentrated adds 20mL water except after desolventizing, uses the hydrochloric acid acidizing reaction liquid of 2N to pH3 under ice bath, then with ethyl acetate extraction (30mL × 3).Merge organic phase, dry, filter, the concentrated compound 4c, colorless oil compounds 6.2g, yield 88% of obtaining.

MS(ESI)m/z:420(M+1); ¹HNMR(400MHz,CDCl ₃):δ7.93(br,1H),7.77-7.75(m,2H),7.38-7.26(m,7H),5.12-5.05(m,3H),4.50-4.44(m,1H),3.79-3.72(t,1H),3.66-3.59(m,1H),3.59-3.43(m,4H),2.28-2.21(m,1H)。

Step 3:(2S, 4R)-1-benzyloxy carbonyl acyl group-2-(2-acetyl bromide)-4-tolysulfonyl oxygen base tetramethyleneimine (compound 4d)

Under ice bath to compound 4c (2.0g, in methylene dichloride (30mL) solution 4.8mmol), drip oxalyl chloride (1.2g, 9.5mmol) He 2 DMF. at room temperature react after 1h concentrated dry after, residuum is dissolved in to 20mL tetrahydrofuran (THF) and 20mL acetonitrile, drips TMSN after being cooled to 0 DEG C ₃the hexane solution of (4.77mL, 2N, 9.54mmol), and then under room temperature, reaction is spent the night.Then under ice bath, the HBr aqueous solution (48%, 1mL) is added drop-wise in reaction solution, and at room temperature anti-1h. will add saturated NaHCO after concentrated solvent doing ₃(30mL), with ethyl acetate (30mL × 3) extraction.Merge organic phase, with saturated common salt washing (90mL).Organic phase drying, filtration, concentratedly obtain compound 4d, yellow oil 1.8g, yield 76% through column chromatography (silica gel, sherwood oil: acetoacetic ester=3:1) again.

MS(ESI)m/z:496(M+1),498(M+23); ¹HNMR(400MHz,CDCl ₃):δ7.78-7.76(m,2H),7.37-7.26(m,7H),5.11-4.97(m,3H),4.77-7.69(m,1H),4.33-4.11(m,1H),3.97-3.89(m,1H),3.79-3.76(m,1H),3.68-3.60(m,1H),2.61-2.44(m,4H),2.16-2.07(m,1H)。

Step 4:(2S, 4R)-1-benzyloxy carbonyl acyl group-4-tolysulfonyl oxygen base-2-(2-Trifluoromethyl-1 H-imidazoles-5-alkyl) tetramethyleneimine (compound 4e)

Compound 4d (0.47g, 0.947mmol) and trifluoro ethanamidine (0.22g, 1.89mmol) are dissolved in 10mL ethanol.After reflux 2h, add anhydrous MgSO ₄(0.2g), continue backflow 3h.Reaction solution after filtration, concentrated obtain compound 4e, yellow oil 0.11g, yield 23% through column chromatography (silica gel, sherwood oil: acetoacetic ester=2:1) again.

MS(ESI)m/z:510(M+1); ¹HNMR(400MHz,CDCl ₃):δ7.73-7.71(d,2H),7.28-7.19(m,6H),7.06(s,2H),5.07-4.97(m,4H),3.81-3.78(m,1H),3.57-3.58(m,1H),2.44(br,2H),2.37(s,3H)

Step 5:(5S, 8S)-7 (8H)-benzyloxy carbonyl acyl group-3-trifluoromethyl-5,6-dihydro-5,8-endo-methylene group-imidazo [1,5-a] pyrazine (compound 4f)

Under ice bath, in DMF (10mL) solution of compound 4e (0.11g, 0.216mmol), add NaH (17.3mg, 0.43mmol, 60%).Reaction solution stirs after 3h at 70 DEG C, concentrates and removes DMF, obtains target compound 4f, colorless oil 40mg, yield 55% after preparative HPLC purifying.

MS(ESI)m/z:338(M+1); ¹HNMR(400MHz,CD ₃OD):δ7.27-7.15(m,5H),6.82-6.65(d,1H),5.34-5.27(m,2H),5.08-5.90(m,2H),3.69-3.61(m,1H),2.96-2.92(m,1H),2.38(s,2H)。

Step 6:(5S, 8S)-3-trifluoromethyl-5,6,7,8-tetrahydrochysene-5,8-endo-methylene group-imidazo [1,5-a] pyrazine (compound 4g)

Pd/C (10mg, 10%) joins ethyl acetate (6mL) solution of compound 4f (30mg, 0.089mmol), stirs 2h in hydrogen.Reaction solution after filtration, concentrated obtain compound 4g, colorless oil 20mg, yield 100%.

MS(ESI)m/z:204(M+1)

Step 7:(R)-3-tertiary butyloxycarbonyl amido-1-((5S, 8S)-3-trifluoromethyl-5,6-dihydro-5,8-endo-methylene group-imidazo [1,5-a] pyrazine-7 (8H)-alkyl)-4-(2,4,5-trifluorophenyl)-1-butanone (compound 4h)

To (3R)-N-tertbutyloxycarbonyl-3-amino-4-(2,4,5-trifluorophenyl) add successively EDCI (25.9mg, 0.135mmol), N in the 5mL dichloromethane solution of butyric acid (45mg, 0.135mmol), N-diisopropylethylamine (34.90mg, 0.27mmol) and HOBT (18.24mg, 0.135mmol). stir after 30min, add compound 4g (18.29mg, 0.09mmol), stirred overnight at room temperature.Underpressure distillation is except after desolventizing, and crude product obtains compound 4h through preparative HPLC purifying, colorless oil 21mg, yield 44%.MS(ESI)m/z:519(M+1)

Step 8:(R)-3-amino-1-((5S, 8S)-3-trifluoromethyl-5,6-dihydro-5,8-endo-methylene group-imidazo [1,5-a] pyrazine-7 (8H)-alkyl)-4-(2,4,5-trifluorophenyl)-1-butanone trifluoroacetate (compound 4)

Under ice bath, in methylene dichloride (3mL) solution of compound 4h (21mg), drip trifluoroacetic acid (1mL), stir after 1 hour reaction solution is concentrated, crude product obtains the trifluoroacetate of target compound 4 after preparative HPLC purifying, colorless oil 18mg, yield 45%.

MS(ESI)m/z:419(M+1); ¹HNMR(400MHz,CD3OD):δ7.22-7.20(m,2H),6.93-6.92(d,1H),5.69-5.52(m,2H),3.91-3.88(m,2H),3.16-3.06(m,3H),2.95-2.90(m,1H),2.69-2.50(m,3H)。

Embodiment 5:(R)-3-amino-1-((5S, 8S)-3-methyl-5,6-dihydro-5,8-endo-methylene group-imidazo [1,2-a] pyrazine-7 (8H)-alkyl)-4-(2,4,5-trifluorophenyl)-1-butanone trifluoroacetate (compound 5)

Step 1 (2S, 4R)-1-benzyloxy carbonyl acyl group-2-chlorine acyl group-4-tolysulfonyl oxygen base-tetramethyleneimine (compound 5a)

Under ice bath, to compound 4c(11.0g, 28.37mmol) add the DMF of oxalyl chloride (4.87mL, 7.20g, 56.74mmol) and catalytic amount, then rise to stirring at room temperature 2 hours.After reaction solution underpressure distillation, obtain crude product compound 5a, brown oil 14g.This crude product is unprocessed, enters next step reaction.

Step 2 (2S, 4R)-1-benzyloxy carbonyl acyl group-2-carbamyl-4-tolysulfonyl oxygen base-tetramethyleneimine (compound 5b)

Crude product 5a (14g) in upper step is dissolved in to dry tetrahydrofuran (THF) (30mL), under ice bath, drips ammoniacal liquor (100mL).After 1 hour, with ethyl acetate (80mL × 3) extractive reaction liquid, the organic phase of merging, with dry after saturated common salt water washing, then boils off solvent, obtains compound 5b, faint yellow solid 10.5g.Yield 78%.MS(ESI)m/z:419(M+H)。

Step 3 (2S, 4R)-1-benzyloxy carbonyl acyl group-2-(imido grpup (methoxyl group) methyl)-4-tolysulfonyl oxygen base-tetramethyleneimine (compound 5c)

To in the methylene dichloride of compound 5b (3.00g, 7.17mmol) (50mL) solution, add trimethylammonium oxygen Tetrafluoroboric acid (2.12g, 14.34mmol), stirred overnight at room temperature.After underpressure distillation, obtain crude product compound 5c, faint yellow solid 5.2g.This crude product is unprocessed enters next step reaction.MS(ESI)m/z:433(M+H)。

Step 4 (2S, 4R)-1-benzyloxy carbonyl acyl group-2-amidino-4-tolysulfonyl oxygen base-tetramethyleneimine (compound 5d)

Compound 5c (5.20g) is dissolved in 20mL methyl alcohol, and adds methanolic ammonia solution (20mL, 40mmol).Under room temperature, stir after 4 hours, by whole excess of ammonia methanol solution decompression go out, obtain crude product compound 5d faint yellow solid 5.50g.This crude product is unprocessed enters next step reaction.

MS(ESI)m/z:418(M+H) ¹H-NMR(400Hz,CD ₃OD):δ7.82-7.79(d,2H),7.49-7.47(d,2H),7.39-7.35(m,5H),5.25-5.09(m,3H),4.72-4.70(m,1H),3.76-3.52(m,2H),2.73-2.67(m,1H),2.48(s,3H),2.27-2.23(m,1H)。

Step 5 (2S, 4R)-1-benzyloxy carbonyl acyl group-2-(5-methyl isophthalic acid H-imidazoles-2-alkyl)-4-tolysulfonyl oxygen base-tetramethyleneimine (compound 5e)

Compound 5d (1.10g, 2.63mmol) is dissolved in 60mL ethanol, adds successively salt of wormwood (1.09g, 7.89mmol) and martonite (720mg, 5.26mmol), and under agitation reflux 4 hours.After reacting liquid filtering, by filtrate evaporate to dryness.Distillation residue are dissolved in to ethyl acetate (20mL), use saturated common salt water washing.Then after organic phase is dry, be concentrated into dry, with silicagel column purify (methylene dichloride: methyl alcohol=100:1) obtain compound 5e, brown oil 130mg, yield 11%.

MS(ESI)m/z:456(M+H); ¹HNMR(400Hz,CD ₃OD):δ7.85-7.81(m,2H),7.50-7.44(m,2H),7.37-7.33(m,3H),7.09-7.05(m,1H),5.22-5.06(m,4H),3.86-3.76(m,2H),2.75-2.70(m,1H),2.49-2.45(d,3H),2.26-2.32(m,2.5H),2.18(s,1.5H)。

Step 6 (5S, 8S)-3-methyl-5,6-dihydro-7 (8H)-benzyloxy carbonyl acyl group-5,8-endo-methylene group-imidazo [1,2-a] pyrazine (compound 5f)

Compound 5e (130mg, 285mmol) is dissolved in to DMF (20mL), adds sodium hydride (25mg, 1.04mmol), be heated to 70 DEG C of reactions 1 hour.After the near room temperature of reaction solution, add 5mL methyl alcohol cancellation reaction.Reaction solution is concentrated into dry, with silicagel column purify (methylene dichloride: methyl alcohol=100:1) obtain brown oily compound 5f, yellow oil 35mg, yield 43%.MS(ESI)m/z:284(M+H)。

Step 7 (5S, 8S)-3-methyl-5,6,7,8-tetrahydrochysene-5,8-endo-methylene group-imidazo [1,2-a] pyrazine (compound 5g)

Compound 5f (35mg, 124 μ mol) is dissolved in 5mL methyl alcohol, adds 10% palladium carbon (15mg), room temperature hydrogenolysis 2 hours.By after the filtering of palladium carbon, reaction is concentrated into dry, obtains compound 5g, white solid 18mg, yield 97%.This crude product is unprocessed enters next step reaction.MS(ESI)m/z:150(M+H)。

Step 8 (R)-3-benzyloxy carbonyl amido-1-((5S, 8S)-3-methyl-5,6-dihydro-5,8-endo-methylene group-imidazo [1,2-a] pyrazine-7 (8H)-alkyl)-4-(2,4,5-trifluorophenyl)-1-butanone (compound 5h)

By (3R)-N-tertbutyloxycarbonyl-3-amino-4-(2,4,5-trifluorophenyl) butyric acid (40.2mg, 121 μ mol) be dissolved in 5mL methylene dichloride, add successively 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride (34.7mg, 181 μ mol), I-hydroxybenzotriazole (24.5mg, 181 μ mol) and N, N-diisopropylethylamine (62.3mg, 483 μ mol), and at room temperature stir 1 hour.Then add compound 5g (18mg, 120 μ mol), under room temperature, reaction is spent the night.Reaction solution is concentrated into dry, with silicagel column purify (methylene dichloride: methyl alcohol=100:1) obtain compound 5h, white solid 35mg, yield 62%.

MS(ESI)m/z:465(M+H),409（M+H- ^tBu） ⁺。

Step 9 (R)-3-amino-1-((5S, 8S)-3-methyl-5,6-dihydro-5,8-endo-methylene group-imidazo [1,2-a] pyrazine-7 (8H)-alkyl)-4-(2,4,5-trifluorophenyl)-1-butanone trifluoroacetate (compound 5)

Under ice bath, compound 5h (12mg) is added to the trifluoroacetic acid of 2mL, then allow reaction solution slowly rise to room temperature reaction half an hour.After trifluoroacetic acid underpressure distillation is removed, add the freeze-drying of 1mL water, obtain compound 5, white solid 15mg.

MS(ESI)m/z:365(M+H); ¹HNMR(400Hz,CD ₃OD):δ7.24-7.05(m,3H),5.78(s,0.5H),5.50(s,0.5H),5.43(s,0.5H),3.79-3.70(m,2H),3.10-3.08(m,0.5H),2.99-2.84(m,3H),2.51-2.46(m,2.5H),2.36-2.29(m,1H),2.27-1.95(m,3H)。

The preparation of embodiment 6 compound 6a-h

Compound 6a-h universal synthesis method

The beta amino acid (0.15mmol) of corresponding N-tertbutyloxycarbonyl protection is dissolved in to 4mL methylene dichloride; add successively DIPEA (63.2mg; 0.49mmol), EDCI (28.2mg, 0.15mmol) and HOBT (19.9mg, 0.15mmol).Stir after 30 minutes, by (5S, 8S)-3-(trifluoromethyl)-5,6,7,8-tetrahydrochysene-5,8-methyl [1,2,4] triazole also [4,3-a] pyrazine (compound 1j) (45.0mg, 0.098mmol) adds reaction solution, and stirs and spend the night.After solvent underpressure distillation, by resistates with silicagel column purify (methylene dichloride: methyl alcohol=100:1) obtain the compound 6a-g of corresponding N-tertbutyloxycarbonyl protection.Then it is placed in respectively under ice bath, adds trifluoroacetic acid (2mL), and stir 30 minutes.Then trifluoroacetic acid is removed in underpressure distillation, repeatedly dissolves after evaporate to dryness to obtain corresponding compound 6a-h with solvent.

(R)-3-amino-1-((5S, 8S)-3-(trifluoromethyl)-5,6-dihydro-5, also [4,3-a] pyrazine-7(8H of 8-bridge methyl [1,2,4] triazole)-alkyl)-4-(4-bromophenyl)-1-butanone trifluoroacetate (compound 6a)

MS(ESI)m/z:444(M+H)； ¹HNMR(400Hz,CD ₃OD)δ7.57-7.50(m,2H),7.27-7.17(m,2H),5.88(s,0.5H),5.72(s,0.5H),5.64-5.61(d,1H),3.89-3.77(m,2H),3.31-3.23(m,1H),3.11-3.05(d,0.5H),3.01-2.91(m,2H),2.82-2.78(d,0.5H),2.74-2.57(m,2H),2.56-2.48(m,1H)。

(R)-3-amino-1-((5S, 8S)-3-(trifluoromethyl)-5,6-dihydro-5, also [4,3-a] pyrazine-7(8H of 8-bridge methyl [1,2,4] triazole)-alkyl)-4-(3-chloro-phenyl-)-1-butanone trifluoroacetate (compound 6b)

MS(ESI)m/z:400(M+H)； ¹HNMR(400Hz,CD ₃OD)δ7.42-7.31(m,3H),7.27-7.25(d,0.5H),7.20-7.18(d,0.5H),5.90(s,0.5H),5.72(s,0.5H),5.64-5.61(d,1H),3.94-3.78(m,2H),3.33-3.24(m,1H),3.13-3.08(d,0.5H),3.01-2.91(m,2H),2.82-2.78(d,0.5H),2.74-2.57(m,2H),2.56-2.49(m,1H)。

(R)-3-amino-1-((5S, 8S)-3-(trifluoromethyl)-5,6-dihydro-5, also [4,3-a] pyrazine-7(8H of 8-bridge methyl [1,2,4] triazole)-alkyl)-4-(2-aminomethyl phenyl)-1-butanone trifluoroacetate (compound 6c)

MS(ESI)m/z:380(M+H)； ¹H?NMR(400Hz,CD ₃OD)δ7.25-7.14(m,4H),5.88(s,0.5H),5.68(s,0.5H),5.6(s,1H),3.90-3.79(m,2H),3.26-3.23(m,1H),3.10-3.03(m,2H),2.98-2.92(m,0.5H),2.81-2.79(m,0.5H),2.74-2.38(m,3H),2.39(m,1.5H),2.34(m,1.5H)。

(R)-3-amino-1-((5S, 8S)-3-(trifluoromethyl)-5,6-dihydro-5, also [4,3-a] pyrazine-7(8H of 8-bridge methyl [1,2,4] triazole)-alkyl)-4-(3-trifluoromethyl)-1-butanone trifluoroacetate (compound 6d)

MS(ESI)m/z:434(M+H)； ¹H?NMR(400Hz,CD ₃OD)δ7.69-7.54(m,4H),5.90(s,0.5H),5.72(s,0.5H),5.64-5.61(d,1H),3.95-3.71m,2H),3.32-3.24(m,1H),3.14-3.05(m,2.5H),2.82-2.78(d,0.5H),2.75-2.56(m,2H),2.54-2.48(m,1H)。

(R)-3-amino-1-((5S, 8S)-3-(trifluoromethyl)-5,6-dihydro-5, also [4,3-a] pyrazine-7(8H of 8-bridge methyl [1,2,4] triazole)-alkyl)-4-(3-cyano-phenyl)-1-butanone trifluoroacetate (compound 6e)

MS(ESI)m/z:391(M+H)； ¹H?NMR(400Hz,CD ₃OD)δ7.67-7.52(m,4H),5.91(s,0.5H),5.73(s,0.5H),.65-5.61(d,1H),3.95-3.71(m,2H),3.32-3.24(m,1H),3.14-3.05(m,2.5H),2.82-2.78(d,0.5H),2.75-2.56(m,2H),2.54-2.49(m,1H)。

(R)-3-amino-1-((5S, 8S)-3-(trifluoromethyl)-5,6-dihydro-5, also [4,3-a] pyrazine-7(8H of 8-bridge methyl [1,2,4] triazole)-alkyl)-4-(2-fluorophenyl)-1-butanone trifluoroacetate (compound 6f)

MS(ESI)m/z:384(M+H)； ¹H?NMR(400Hz,CD ₃OD)δ7.40-7.10(m,4H),5.89(s,0.5H),5.70(s,0.5H),5.6(s,1H),3.90-3.77(m,2H),3.32-3.23(m,1H),3.13-3.06(m,2H),3.03-3.01(m,0.5H),2.81-2.78(m,0.5H),2.74-2.48(m,3H)。

(R)-3-amino-1-((5S, 8S)-3-(trifluoromethyl)-5,6-dihydro-5, also [4,3-a] pyrazine-7(8H of 8-bridge methyl [1,2,4] triazole)-alkyl)-4-cyano-phenyl-1-butanone trifluoroacetate (compound 6g)

MS(ESI)m/z:366(M+H) ¹HNMR(400Hz,CD ₃OD)δ7.41-7.29(m,4H),7.25-7.23(m,1H),5.89(s,0.5H),5.70(s,0.5H),5.61(s,1H),3.83-3.78(m,2H),3.26-3.23(m,1H),3.13-3.06(m,2H),3.03-3.01(m,0.5H),2.81-2.78(m,0.5H),2.74-2.50(m,3H)。

(R)-3-amino-1-((5S, 8S)-3-(trifluoromethyl)-5,6-dihydro-5, also [4,3-a] pyrazine-7(8H of 8-bridge methyl [1,2,4] triazole)-alkyl)-4-(2,5-difluorophenyl)-1-butanone trifluoroacetate (compound 6h)

MS(ESI)m/z:402(M+H) ¹HNMR(400Hz,CD ₃OD)δ7.22-7.05(m,3H),5.89(s,0.5H),5.72-5.70(m,0.5H),5.65-5.61(m,1H),3.95-3.78(m,2H),3.30-3.20(m,1H),3.15-2.97(m,2.5H),2.85-2.80(m,0.5H),2.74-2.70(m,1H),2.67-2.60(m,1H),2.56-2.42(m,1H)。

Embodiment 7(R)-3-amino-1-((5S, 8S)-3-(trifluoromethyl)-5,6-dihydro-5, also [4,3-a] pyrazine-7(8H of 8-bridge methyl [1,2,4] triazole)-alkyl)-4-(2,4,5-trifluorophenyl) the preparation of-various salt of 1-butanone

Compound 7a

By being dissolved in 2mL methyl alcohol with the compound 1 (26mg) of sodium hydroxide neutralization, carefully add 48% Hydrogen bromide of 0.5mL, under room temperature, stir 1 hour.Add 3mL water, remove methyl alcohol under reduced pressure rear freeze-drying, obtain faint yellow compound 7a (32mg).MS(ESI)m/z:420(M+H); ¹HNMR(400Hz,CD ₃OD)δ7.28-7.21(m,2H),5.90(s,0.5H),5.75(s,0.5H),5.66-5.62(d,1H),3.99-3.79(m,2H),3.35-3.24(m,1H),3.14-3.13(m,0.5H),3.06-2.99(m,2H),2.83-2.80(m,0.5H),2.75-2.55(m,2.5H),2.53-2.52(m,0.5H)

Compound 7b

By being dissolved in 2mL Virahol with the compound 1 (45mg) of sodium hydroxide neutralization, carefully add the 0.5mL aqueous isopropanol (having solid to separate out) of phosphoric acid (0.11mL, 10.52mg), under room temperature, stir 2 hours.After evaporated under reduced pressure, obtain faint yellow compound 7b (56mg).

MS(ESI)m/z:420(M+H); ¹H-NMR(400Hz,CD ₃OD)δ7.27-7.22(m,1H),7.13-7.05(m,1H),5.77(s,0.5H),5.64(s,0.5H),5.52-5.49(d,1H),3.91-3.90(d,0.5H),3.88-3.67(m,1.5H),3.20-3.18(m,1H),3.14-2.92(m,2.5H),2.71-2.67(m,0.5H),2.64-2.42(m,3H)。

Compound 7c

The preparation of 1M succinic acid solution: succinic acid (590mg, 5mmol) is dissolved in to methyl alcohol: the solution of water (v/v=4/1), for subsequent use.By being dissolved in 2mL methyl alcohol with the compound 1 (28mg, 0.066mmol) of sodium hydroxide neutralization, add the 1M succinic acid solution of 0.066mL, stir 30 minutes.After evaporated under reduced pressure, obtain white solid look compound 7c (36mg).

MS(ESI)m/z:420(M+H); ¹HNMR(400Hz,CD ₃OD)δ7.26-7.05(m,2H),5.77(s,0.5H),5.62(s,0.5H),5.52-5.49(d,1H),3.85-3.82(m,0.5H),3.67-3.60(m,1.5H),3.15-3.12(m,0.5H),2.99-2.82(m,2.5H),2.70-2.44(m,2H),2.43-2.39(m,6H)。

Compound 7d

With succinate with legal system for white solid compound 7d(33mg).

MS(ESI)m/z:420(M+H); ¹HNMR(400Hz,CD ₃OD)δ7.19-7.04(m,2H),5.77(s,0.5H),5.62(s,0.5H),5.52-5.49(d,1H),3.84-3.81(d,0.5H),3.70-3.66(m,1H),3.70-3.51(m,0.5H),3.15-3.12(m,0.5H),2.92-2.84(m,1.5H),2.79-2.75(m,1H),2.70-2.67(m,0.5H),2.61-2.59(m,0.5H),2.53-2.47(m,1H),2.48-2.33(m,2H),1.82(s,3H)。

Compound 7e

With succinate with legal system for white solid compound 7e (36mg).

MS(ESI)m/z:420(M+H); ¹HNMR(400Hz,CD ₃OD)δ7.35-7.03(m,2H),5.77(s,0.5H),5.62(s,0.5H),5.52-5.49(d,1H),4.21-4.17(m,1H),3.85-3.82(m,0.5H),3.67-3.60(m,1.5H),3.25(s,1H),3.15-3.12(m,0.5H),3.03-2.94(m,1.5H),2.91-2.84(m,1H),2.71-2.53(m,3H),2.50-2.39(m,3H)。

Compound 7f

With succinate with legal system for white solid compound 7f (35mg).

MS(ESI)m/z:420(M+H); ¹HNMR(400Hz,CD ₃OD)δ7.16-7.08(m,2H),6.15(s,2H),5.78(s,0.5H),5.61(s,0.5H),5.53-5.50(d,1H),3.82-3.67(m,2H),3.15-3.12(m,0.5H),3.04-2.86(m,2.5H),2.71-2.68(m,0.5H),2.63-2.37(m,3H)。

Compound 7g

With succinate with legal system for white solid fumarate compound 7g (22mg).

MS(ESI)m/z:420(M+H); ¹HNMR(400Hz,CD ₃OD)δ7.26-7.07(m,2H),6.59(s,2H),5.77(s,0.5H),5.61(s,0.5H),5.52-5.50(d,1H),3.83-3.80(m,1H),3.70-3.67(m,1H),3.15-3.12(m,0.5H),3.04-2.86(m,2.5H),2.71-2.68(m,0.5H),2.63-2.37(m,3H)。

Compound 7h

With succinate with legal system (40mg) for white solid tartrate compound 7h.

MS(ESI)m/z:420(M+H); ¹HNMR(400Hz,CD ₃OD):δ7.39-7.17(m,2H),5.89(s,0.5H),5.75(s,0.5H),5.63-5.61(d,1H),4.45(s,2H),4.00-4.01(m,0.5H),3.89-3.79(m,1.5H),3.29-3.23(m,1H),3.16-2.98(m,2.5H),2.82-2.79(m,0.5H),2.74-2.52(m,3H)。

Illustrate beneficial effect of the present invention by test example below.

Test example 1

The member of DPP-4, DPP-2, DPP-8, DPP-9 Jun Shi DPP family, research is found, if suppress other enzyme except DPP-4, can have side effects to health, as when suppressing DPP-8, DPP-9 enzyme and living, to cause dormancy T necrocytosis, a series of toxic side effect such as stomach toxicity and body's immunity, therefore, be DPP-IV to DPP-4(at present) in the exploitation of enzyme inhibitors, require to improve the selectivity of inhibitor to DPP-4, reduce the selectivity to other families of DPP simultaneously, thereby reduce the toxic side effect of inhibitor.

The present invention measures the inhibition of the compounds of this invention to DPP family by following test.

DPP-4, DPP-2, DPP-8, DPP-9 external activity detect

Testing compound is dissolved in to methyl-sulphoxide, then uses buffered soln (DPP4:100mM HEPES, pH7.5,0.1mg/mL BSA; DPP2:100mM HEPES, pH5.5,0.1mg/mL BSA; DPP8:50mM Tris-HCl, pH7.5,0.1mg/mL BSA; DPP9:25mM Tris-HCl, pH7.5,0.1mg/mL BSA) be diluted to a series of working solution.Recombinant human DPP-4(ultimate density is about to 180ng/mL) or DPP-2(ultimate density be about 100ng/mL) or DPP-8(ultimate density be about 200ng/mL) or DPP-9(ultimate density be about 50ng/mL) mix with above-mentioned a series of compound working solution, then (ultimate density DPP4 is 50 μ M to add Gly-Pro-AMC, DPP2/8/9 is 20 μ M) (total reaction volume is 100 μ L), the AMC(excitation wavelength 360nm that continuous detecting discharges at once, emission wavelength 460nm) 15 minutes.With SigmaPlot computed in software half-inhibition concentration IC50, result sees table.

Table 1 compound is to DPP-IV inhibition

The inhibition activity of table 2 compound to DPP family

The above results shows, the compounds of this invention can effectively suppress DPP-IV activity, compared with marketed drugs Jie Nuowei, compound 1 is suitable to the restraining effect of DPP4, but prompt Novi is lower to other DPP family members' (DPP2, DPP8, DPP9) inhibition specific activity, show that the compounds of this invention not only can effectively suppress DPP4 performance drug activity, can also reduce the inhibition activity to other families of DPP, reduce toxic side effect, medicinal security is better.

Above-claimed cpd provided by the invention can be used for the multiple treatment with DPP-4 relative disease such as diabetes, and better drug safety is provided.

Claims

1. the compound shown in formula IA and IB or its pharmacy acceptable salt,

2. compound according to claim 1 or its pharmacy acceptable salt, is characterized in that:

R ¹in, the substituting group of described substituted-phenyl is 1-5 R ⁴, wherein R ⁴be selected from the alkoxyl group of the alkoxy or halogen replacement of the alkyl of CN, halogen, C1-6 or the alkyl of halogen replacement or C1-6;

R ⁶in, described carboxyl is COOH, described ester group is CO ₂r ⁷;

3. compound according to claim 1 and 2 or its pharmacy acceptable salt, is characterized in that: R ¹be selected from the phenyl that halogen replaces; R ²be selected from containing assorted alkyl, a 1-2 heteroatoms or C1-2 alkyl; R ³be selected from the alkyl that halogen replaces; R ⁶be selected from CO ₂r ⁷.

4. compound according to claim 1 and 2 or its pharmacy acceptable salt, is characterized in that: described is the alkyl that contains 1 heteroatomic C1-4 containing assorted alkyl.

5. compound according to claim 3 or its pharmacy acceptable salt, is characterized in that: described halogen is F or Cl.

6. according to compound or its pharmacy acceptable salt described in claim 1-5 any one, it is characterized in that: described compound is

Wherein, R ²be selected from the alkyl of C1-5 or substituted alkyl or 1-5 containing assorted alkyl or replace containing assorted alkyl or 1-5 heteroatoms or replacement heteroatoms; R ³be selected from H, CN or C1-10 alkyl or substituted alkyl; X is selected from N or CH; R ⁶be selected from H, CN, carboxyl or ester group.

7. compound according to claim 6 or its pharmacy acceptable salt, is characterized in that:

R ²in, the substituting group of described substituted alkyl is selected from halogen, CN, OH, R ⁵, OR ⁵, NHSO ₂r ⁵, SO ₂r ⁵, COOH or CO ₂r ⁷, described heteroatoms is selected from N, S or O, and replacing heteroatoms or replacing containing the substituting group of assorted alkyl is halogen, CN, OH, R ⁵, OR ⁵, NHSO ₂r ⁵, SO ₂r ⁵, COOH or CO ₂r ⁷;

Wherein, R ⁵for alkyl or the substituted alkyl of C1-6, its substituting group is 1-5 halogen, COOH or CO ₂r ⁷; R ⁶in, described carboxyl is COOH, described ester group is CO ₂r ⁷; R ⁷for C1-6 alkyl.

8. according to compound or its pharmacy acceptable salt described in claim 1-7 any one, it is characterized in that: described compound or its pharmacy acceptable salt are:

9. according to compound or its pharmacy acceptable salt described in claim 1-8 any one, it is characterized in that: hydrochloride, vitriol, citrate, benzene sulfonate, hydrobromate, hydrofluoride, phosphoric acid salt, acetate, propionic salt, succinate, oxalate, malate, succinate, fumarate, maleate, tartrate or trifluoroacetate that described pharmacy acceptable salt is compound.

10. compound or its pharmacy acceptable salt described in claim 1-9 any one, in the purposes for the preparation of in dipeptidyl peptidase-iv inhibitor class medicine.

11. purposes according to claim 10, is characterized in that: described dipeptidyl peptidase-iv inhibitor is that treatment is or/and the medicine of prevent diabetes, hyperglycemia, insulin resistance.

The preparation method of formula IIA or IIB compound described in 12. claims 6, is characterized in that: reactions steps is as follows:

Comprising following steps:

Step11: previous step product adds acid-respons, obtains product IIA;

13. according to the preparation method who prepares Compound I IA or formula IIB described in claim 12, it is characterized in that:

In Step1, solvent for use is non-protonic solvent, and described aprotic solvent is preferably tetrahydrofuran (THF), and temperature of reaction is 0 DEG C to 30 DEG C, and the reaction times is preferably 6 to 18 hours;

In Step3, solvent for use is polar solvent, and described polar solvent is preferably methylene dichloride, ethyl acetate or methyl alcohol, and acid used is strong acid, is preferably trifluoroacetic acid or hydrochloric acid, and temperature of reaction is 20 DEG C to 30 DEG C, and the reaction times is 0.5 to 2 hour;

Solvent described in Step5 is non-protonic solvent, described non-protonic solvent is preferably methylene dichloride, and alkali used is organic bases, and described organic bases is preferably N, N-diisopropylethylamine, described condensing agent is preferably 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride and I-hydroxybenzotriazole, or 2-(7-azo benzotriazole)-N, N, N', N'-tetramethyl-urea phosphofluoric acid ester, temperature of reaction is 20 DEG C to 30 DEG C, the reaction times is 2 to 12 hours;

In Step9, solvent for use is tetrahydrofuran (THF) and water mixed solvent, described alkali is mineral alkali or organic bases, and described mineral alkali is preferably sodium hydroxide or lithium hydroxide, and described organic bases is preferably piperidines, temperature of reaction is 20 DEG C to 30 DEG C, and the reaction times is 0.5 to 2 hour;

The preparation method of formula III A or IIIB compound described in 14. claims 6, is characterized in that: reactions steps is as follows:

Wherein:

Step1:g1 and diphenyl phosphate azide, diisopropyl azodiformate, triphenylphosphine reaction, the product obtaining is as next step reactant;

Step9: previous step product reacts under acidic conditions with hmds, ammonium sulfate, and the product obtaining is as next step reactant;

15. according to the preparation method who prepares compound formula III A or formula III B described in claim 14, it is characterized in that:

Solvent described in Step1 is non-protonic solvent, is preferably tetrahydrofuran (THF), and temperature of reaction is 0 DEG C to 30 DEG C, and the reaction times is 8 to 12 hours;

The preparation method of formula III A or IIIB compound described in 16. claims 6, is characterized in that: reactions steps is as follows:

17. according to method described in claim 16, it is characterized in that:

In Step12, solvent for use is non-protonic solvent, and described non-protonic solvent is preferably tetrahydrofuran (THF) or pyridine, and alkali used is organic bases, and described organic bases is preferably N-Methylimidazole, and temperature of reaction is 0 DEG C to 30 DEG C, and the reaction times is 4 to 24 hours;

In Step13, solvent for use is water and tetrahydrofuran (THF) mixed solvent, described water and tetrahydrofuran (THF) mixed solvent ratio are 1:1V/V, and alkali used is mineral alkali, and described mineral alkali is preferably lithium hydroxide or sodium hydroxide, temperature of reaction is 20 DEG C to 30 DEG C, and the reaction times is 2 to 4 hours;

In Step15, solvent for use is protic solvent, and described protic solvent is preferably ethanol, and temperature of reaction is 70 to 90 DEG C, and the reaction times is 3 to 6 hours;

In Step16, solvent for use is non-protonic solvent, and described non-protonic solvent is preferably DMF, and described highly basic is inorganic strong alkali, and described inorganic strong alkali is preferably sodium hydride or potassium hydride KH, and temperature is 70 DEG C to 80 DEG C, and the reaction times is 3 to 6 hours;

In Step17, used catalyst is metal catalyst, is optimized for palladium carbon, and solvent is protic solvent, is preferably ethyl acetate, and temperature of reaction is 20 DEG C to 30 DEG C, and the reaction times is 1 to 2 hour;

In Step18, solvent for use is aprotic solvent, described aprotic solvent is preferably methylene dichloride, tetrahydrofuran (THF) or N, dinethylformamide, alkali used is organic bases, described organic bases is preferably diisopropylethylamine, condensing agent is 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride and I-hydroxybenzotriazole or 2-(7-azo benzotriazole)-N, N, N', N'-tetramethyl-urea phosphofluoric acid ester, temperature of reaction used is 0 DEG C to 30 DEG C, and the reaction times is 0.5 to 2 hour;

The intermediate of formula IA and IB compound described in 18. claims 1, its structural formula is as follows:

19. intermediates according to claim 18, is characterized in that: R ²in, the substituting group of described substituted alkyl is selected from halogen, CN, OH, R ⁵, OR ⁵, NHSO ₂r ⁵, SO ₂r ⁵, COOH or CO ₂r ⁷, described heteroatoms is selected from N, S or O, and replacing heteroatoms or replacing containing the substituting group of assorted alkyl is halogen, CN, OH, R ⁵, OR ⁵, NHSO ₂r ⁵, SO ₂r ⁵, COOH or CO ₂r ⁷;

20. according to the intermediate described in claim 18 or 19, it is characterized in that: described intermediate is:

The preparation method of formula IIAA and IIBA compound described in 21. claims 20, is characterized in that: reactions steps is as follows:

The preparation method of formula III AA and IIIBA compound described in 22. claims 21, is characterized in that: reactions steps is as follows: