CN105503873B - Preparation method of chemical compound serving as DPP-IV (dipeptidyl peptidase-4) inhibitor - Google Patents

Abstract

The invention relates to a preparation method of a biguanide derivative serving as a DPP-IV (dipeptidyl peptidase-4) inhibitor. The biguanide derivative can be used for treating all symptoms or diseases which can be relieved if the activity of DPP-IV is inhibited, such as I-type diabetes, II-type diabetes, diabetic complications and other related diseases.

Description

A kind of preparation method of the compound as dipeptidyl peptidase-4 inhibitors

Divisional application of the application for the application for a patent for invention of Application No. 201410829322.7, the applying date of original application For on December 26th, 2014, Publication No. CN104592234 A, publication date were on 05 06th, 2015, entitled " a kind of As the preparation method of the compound of dipeptidyl peptidase-4 inhibitors ".

Technical field

The present invention relates to a kind of preparation method of the compound as dipeptidyl peptidase-4 inhibitors, the compound can use Can be such as I types and type ii diabetes, diabetes complicated because suppressing DPP-IV activity and all symptoms that be benefited or disease in treatment Disease etc..

Background technology

Dipeptidyl peptidase-4 (DPP-4) is a kind of high specific serine protease, and its natural substrate is pancreas hyperglycemia Plain sample peptide 1 (GLP-1) and glucose pancreotropic hormone polypeptide (GIP).GLP-1 has different physiological roles, can increase Portugal in pancreas The grape sugar insulin secretion for relying on, the secretion for suppressing glucagon, make d cell hypertrophy；Stomach after the meal can be delayed in gastrointestinal tract Emptying, so that delay intestinal glucose absorption.GIP has insulin secretion accelerating function.GLP- in DPP-4 energy fast degradation bodies L and GIP, is allowed to inactivate.GLP-1 in healthy human body is mainly secreted by the L cells of ileum and colon, basic dense in the human body Degree about 5～10pmol/L, postprandial concentration can raise 2～3 times.Only about 2 minutes its biological activity half-life.DPP-4 Inhibitor reduces the catalysis activity of enzyme, so as to suppress the degraded of GLP-1 and GIP by competitive binding DPP-4 active sites.

DPP-4 is widely present in the in-vivo tissues such as blood plasma, gastrointestinal tract, kidney, lymph node and connective tissue, wherein kidney At most.Its family member includes DPP-1, DPP-2, DPP-3, DPP-4, DPP-8, DPP-9 and fibroblast activation protein-α (FAP).The molecular weight of DPP-4 is 220kD, and active body is dimeric forms, and each subunit includes two domains, controls bottom The gateway of thing positioned at the large-scale cave that a size between two domains is 30～45A, the bag shaped structure in which is The active site of DPP-4, is that the polypeptide of proline (Pro) or alanine (Ala) is all DPP- on the N-terminal second of every structure 4 main substrates for playing activity.

Due to the rapid Degradation of internal DPP-4, GLP-1 in vivo the half-life it is very short (<2min), research finds to work as After DPP-4 activity inhibiteds, the action time of GLP-1 just effectively extends, and reduces blood sugar level effect so as to play.DPP-4 suppresses The agent mechanism of action is similar to natural substrate based on its structure, containing Xaa-Pro similar structures, being capable of competitive binding DPP- 4 active sites, and affinity is much larger than natural substrate, thus the conformation of DPP-4 is changed, reduce catalysis activity.Experiment card Bright DPP-4 inhibitor can reversibly suppress about 90% DPP-4 enzymatic activitys in 24h.Thus DPP-4 inhibitor can lead to Cross and improve internal GLP-1 concentration, extend its blood sugar reducing function time, and glucagon suppression secretion, extend GLP-1 to islets of langerhans The stimulus duration of element secretion.Because GLP-1 is presented strict blood sugar concentration dependency to the adjustment effect of insulin secretion, Only in blood glucose rise, GLP-1 can just increase insulin secretion, so DPP-4 inhibitor will not cause hypoglycemia that wind occurs Danger.DPP-4 inhibitor its particular mechanism and good security features, become the hot fields of Glucovance research. Research finds that some DPP-4 inhibitor and metformin therapeutic alliance diabetes can significantly improve blood sugar level, and low blood Sugar, increased weight and other adverse events risks are relatively low.But still lack hypoglycemia, increased weight and other adverse events risks Low single compound.

The content of the invention

The present invention provides a kind of preparation method of the Biguanide derivative as DPP-IV inhibitor, described Biguanide derivative Can be used to prevent or treat with DPP-IV inhibitory action be benefited relevant disease, with hypoglycemia, increased weight and its His low incidence rate of adverse events risk.Specifically, the present invention provides compound of the preparation as shown in following formula (I), its stereoisomerism Body, or the method for its pharmaceutically acceptable salt, methods described include making formula (II) compound contact to obtain with dicyandiamide Formula (I)：

Wherein,

R¹Represent C_1-8Alkyl, the C_1-8Alkyl can be by R¹¹Or R¹²- CO- replaces, and R¹¹Represent C_6-10Aryl, quinoline Base, isoquinolyl, quinazolyl, cinnolines base or indyl, R¹²Represent two (C_1-6Alkyl) amino, pyrrolidin-1-yl, piperidines -1- Base or C_6-10Aryl；

R²Represent C_1-8Alkyl or C_6-10Aryl；

R³Represent benzyl, C_2-8Thiazolinyl, C_2-8Alkynyl or C_3-8Phenyl ring in cycloalkenylmethyl, wherein benzyl can be by 1 or many Individual halogen or cyano group replace；

R⁴Represent C_1-8Alkyl, C_3-8Cycloalkyl, C_3-8Circle heterocycles alkyl, C_6-10Aryl or C_5-10Unit's heteroaryl；

R⁵Represent hydrogen or amino protecting group.

Wherein, described amino protecting group is it is known in the art that including but not limited to methoxycarbonyl group, carbethoxyl group, uncle Butoxy carbonyl, benzyloxycarbonyl group, 2- xenyls -2- propylene carbonyl oxygens (BPoc), p-toluenesulfonyl (Tosyl), trifluoroacetyl group, Tablet held before the breast by officials methoxycarbonyl group (Fmoc), allyloxycarbonyl (Alloc), trimethylsilyl ethoxycarbonyl (Teoc), phthalyl (Pht), P-toluenesulfonyl (Tos), trifluoroacetyl group (Tfa), adjacent or p-nitrophenyl sulfonyl (Ns), pivaloyl group, benzoyl, three Benzyl (Trt), 2,4- dimethoxy-benzyls (Dmb), to methoxy-benzyl (PMB), benzyl etc..

As R in formula (II)⁵When representing amino protecting group, need described amido protecting before contacting with dicyandiamide in advance Base is removed.

In a preferred embodiment, R¹Represent dimethyl-aminocarbonyl methyl, pyrrolidin-1-yl carbonvlmethyl, phenyl Carbonvlmethyl, benzyl, quinolyl methyl, tetrahydroisoquinolinylmethyl, quinazolyl methyl or cinnolines ylmethyl.

In a further preferred embodiment, R²Represent methyl, isopropyl or phenyl.

In a further preferred embodiment, R³Represent 2- methyl -2- propylene -1- bases, 2-butylene -1- bases, 2,3- diformazans Base -2-butylene -1- bases, 2-butyne -1- bases, 1- cyclopentenes -1- ylmethyls, 1- cyclohexene -1- ylmethyls, benzyl, 2- cyano group benzyls Base, 2,6- dicyano benzyls.

In a further preferred embodiment, R⁴Represent normal-butyl, cyclohexyl, phenyl, piperidyl or pyridine radicals.

In one embodiment, including making formula (III) compound contact with formula (IV) compound the step of, to obtain R⁵ Represent formula (II) compound of amino protecting group：

Wherein when needed in order to obtain R⁵Formula (II) compound of hydrogen is represented, alternatively by amino protecting group Pr¹Removing.

Wherein formula (IV) compound can be by 3- amino piperidines (formula (V) compound) amino protecting group Pr Jing different successively¹、 Pr²Protection, then introduces R⁴Group, reselection removing Pr²Protection group and be obtained：

Wherein Pr¹And Pr²Represent amino protecting group.Amino protecting group as herein described be it is well known in the art those, including But it is not limited to methoxycarbonyl group, carbethoxyl group, tert-butoxycarbonyl, benzyloxycarbonyl group, 2- xenyls -2- propylene carbonyl oxygens (BPoc), right Tosyl (Tosyl), trifluoroacetyl group, tablet held before the breast by officials methoxycarbonyl group (Fmoc), allyloxycarbonyl (Alloc), trimethyl silicane ethoxy Carbonyl (Teoc), phthalyl (Pht), p-toluenesulfonyl (Tos), trifluoroacetyl group (Tfa), adjacent or p-nitrophenyl sulphur Acyl group (Ns), pivaloyl group, benzoyl, trityl (Trt), 2,4- dimethoxy-benzyls (Dmb), to methoxy-benzyl (PMB), benzyl etc..

Formula (III) compound can be by 8- halos xanthine derivative successively by containing R¹The halides of group and R³The halogen of group Replace for thing and be obtained：

In formula, each X is identical or different, and represents halogen atom.

The salt of the compounds of this invention includes all of acid-addition salts and all salt formed with alkali.Especially, the salt includes Water-insoluble salt and water soluble salt.The mineral acid for being suitable to the acid-addition salts of the present invention includes but is not limited to hydrochloric acid, hydrobromic acid, phosphorus Acid, sulphuric acid etc..The organic acid of acid-addition salts for being suitable to the present invention includes, for example and is not limited to, citric acid, maleic acid, fumaric acid, Succinic acid, lactic acid, tartaric acid, methanesulfonic acid etc..Preferably, the salt of compound of the invention could be for formula (II) compound or The salt of the isolated or purified of the free cpds of its stereoisomer or their salt.Therefore, formed with mineral acid or organic acid Salt may include but be not limited to hydrochlorate, hydrobromate, phosphate, sulfate, citrate, maleate, fumarate, amber Amber hydrochlorate, lactate, tartrate, metilsulfate, mandelate etc..

Specific isomeric forms unless expressly indicated, all isomeric forms of formula (II) compound it is contemplated that this In invention, including all regional isomer forms and stereoisomer form, such as all Chiral forms, the enantiomerism bodily form Formula, diastereomeric form, racemic form, tautomeric forms and all geometric isomer forms.It will be apparent that it is preferred that Isomer that is pharmaceutically most effective and most having no side effect.

It should be understood that the compound of the present invention contains at least one, carbon atom of two or more Jing Asymmetrical substitutes, and Can be separated with optical active forms or racemic form as pure diastereomer or non-enantiomer mixture.

The present invention covers all possible stereoisomer, the diastereomer being especially mentioned above and enantiomerism Body, such as with substantially pure form, with enriched form and/or in the form of any mixed proportion, including racemic form, and Its salt.

In general, substantially pure stereoisomer can be obtained according to composition principle well known to those skilled in the art, For example by the separation of respective mixtures, close by using starting material pure in spatial chemistry and/or by stereo selectivity Into acquisition.It is known in the art how to prepare optical active forms, such as by the fractionation of racemic form or for example, by from optics Active starting materials synthesize and/or by using chiral reagent preparing.

The compound of the enantiomeric pure of the present invention can be appropriate for example by preparing and separating by asymmetric synthesis Compound/the intermediate of diastereomer, and/or by using Chirality Reaction component (such as chiral reagent, chiral catalysis Agent, chiral ligand, chiral synthon, chiral building block etc.) prepare, the compound/intermediate of wherein described diastereomer can By known such as chiral chromatogram separate or from suitable solvent fractional crystallization and it is isolated.

Additionally, those skilled in the art knows how from corresponding racemic mixture to prepare the change of enantiomeric pure Compound, such as by the corresponding racemic compound of chromatographic isolation on chiral separation post；Or tried by using appropriate fractionation Agent resolution of racemic compound；Diastereomer is formed with optically active acid or alkali by racemic compound for example Salt, the compound needed for subsequently splitting the salt and discharging from the salt；Or this is derived by using chiral auxiliary corresponding outer disappear Rotation compound, is subsequently isolated diastereomer and removes the chiral auxiliary group；By the kinetic resolution (example of racemate Such as pass through Enzymatic Resolution)；By under optimum conditions from the aggregation enantioselective crystallization of mirror image crystal；Or by chirality In the presence of auxiliary agent, from suitable solvent prepared by fractional crystallization.

The compound of resulting formula (II) is separable into its enantiomer and/or diastereomer.For example, it is suitable Formula/trans mixture is separable into their cis or trans isomer, and the change with least one optical activity carbon atom Compound may be split into their enantiomer.

Enantiomer is preferably separated by the following method：Post separation in chiral phase, or the weight from optical activity solvent Crystallization, or the derivant of especially sour and its activation or alcohol reaction forming salt or derivant such as ester or acyl with optically active substance Amine, and separate the non-enantiomer mixture of thus obtained salt or derivant, such as based on their deliquescent differences；Together When can be discharged from pure diastereomeric salt or derivant by the effect of Suitable agents, free enantiomer.Conventional optics Active acid is such as D and L-type tartaric acid or dibenzoyl tartaric acid, two-o-tolyl tartaric acid, malic acid, mandelic acid, Camphora Sulfonic acid, glutamic acid, aspartic acid or quinic acid.Optical activity alcohol can be, for example, the optics in (+) or (-) menthol and amide Active acyl, for example, can be (+) or (-) menthyl Epoxide carbonyl.

Those skilled in the art should be understood that organic compound or their salt can be together separated with solvent molecule or can The solvent that contacts with them, they react be located solvent, they are therefrom detached solvent (for example, precipitation, crystallization, lyophilizing etc.) Etc. forming complex.According to the understanding of those skilled in the art, such as when obtaining in solid form or separating, the one of the present invention A little compounds can the solvent containing variable or fixed amount (including aqueouss and/or non-aqueous solvent).Therefore, chemical combination of the invention The solvate (hydrate including hydrate, organic solvent compound and mixing/organic solvent compound) of thing is included in the present invention In the range of.The solvate of the compounds of this invention may include that stoichiometric solvate or non-stoichiometric solvent are closed The solvate of thing, the solvate combined closely or weak binding, and homogeneous solvent compound or xenogenesis solvate.It is preferred that Ground, solvent used is pharmaceutically acceptable solvent, such as water and/or low-molecular-weight fatty alcohol such as ethanol etc..Implement at one In scheme, the solvate of the compounds of this invention may include, for example, hydrate or alcohol adduct, and or the hydrate of mixing/alcohol closes Thing.The present invention includes unsolvated form and all of solvation form.Similarly, the present invention includes any solvate, non- Solvate, hydrate, anhydride, hygroscopicity and/or non-hygroscopic form.

Suppress DPP-IV activity and regulation blood as formula (I) compound of the invention, its stereoisomer and its salt have The ability of sugar level, which is suitable to treat all symptoms or the disease that can be benefited because suppressing DPP-IV activity.Therefore, it is contemplated that according to The compound of the present invention will be suitable for disease or the symptom prevented or treat, such as I types and type ii diabetes, diabetic complication (example Such as retinopathy, nephropathy or neuropathy), metabolism acidosis or ketosis, reactional hypoglycemia, insulin resistant, metabolism Syndrome, the lipodystrophy of separate sources, arthritis, atherosclerosiss and relevant disease, obesity, alloplast move Osteoporosis caused by plant and calcitonin.Additionally, these materials are also suitable for preventing beta cell degeneration, for example pancreas beta cell Apoptosises or necrosis.The material is also suitable for improving or recover the function of pancreatic cell, and increase the number of limb beta cell with Size.Additionally, and the link that suppresses based on the effect because of glucagon-like peptide such as GLP-1 and GLP-2 and its with DPP-IV, equally Ground, compound of the invention are applied to and reach other calm or anxiety and alleviate effect, and also advantageously affect to perform the operation or Katabolism state after the response of hormone stress, or reduce mortality rate or sickness rate after myocardial infarction.It is also suitable for treatment It is relevant with effect referred to above, and by GLP-1 or GLP-2 introduced symptomatology.Compound of the invention Can act also as diuretic or hypotensive agent is used, and be applied to prevention with treatment acute renal failure.Additionally, of the invention Compound can be used to treat respiratory tract inflammatory diseasess.Equally it is also suitable for prevention with treatment chronic inflammatory bowel disease, such as zest Bowel syndrome (IBS), clone disease or ulcerative colitiss and pancreatitiss.Similarly, which can be used for all wounds of gastrointestinal Evil species or infringement, such as colitis and enteritis.Also be expected DPP-IV inhibitor and therefore compound of the invention, also can be In the mankind or mammal be used for treat infertility or improve fertility, spy be when infertility be with insulin resistant or When polycystic ovary syndrome is relevant.On the other hand, these materials apply to affect the motility of sperm, and therefore can be used as man Property contraceptive use.The material also is adapted for treating and short and small relevant growth hormone deficiency in addition, and is also advantageously used for which In can using growth hormone any symptom.Compound of the invention, based on its inhibitory action to DPP-IV, is also suitable for Various autoimmune diseases are treated, for example rheumatic arthritis, multiple sclerosiss, thyroid disease and Ba Saidushi diseases etc..Which is also Can be used to treat virus disease, and for example in HIV, stimulate blood manufacture, in benign prostatic hyperplasia, gingivitiss In, and treatment neuron defects, and neurodegenerative disease, for example alzheimer's disease.The compound also can be used for Fat tumor is treated, is particularly changed tumor and is invaded and shift；Here example is which in treatment T- cell lymphomas, acute lymphoblastic Cell lymphoma leukemia, with the purposes in the thyroid carcinoma of cell base, basal cell carcinoma or breast carcinoma.During other indications are Wind, the myocardial ischemia of various origins, parkinson and migraine.Additionally, other indications include folliculitises and keratinization of epidermis Excessively, increase keratinocyte hyperplasia, psoriasiss, brain ridge inflammation, glomerulonephritiies, lipodystrophy and it is all difference into The somatics of cause, depressed and neuropsychopathy disease.

Formula (I) compound or its pharmaceutically acceptable salt of the present invention can be used as medicine, such as enteral, gastrointestinal The form of the pharmaceutical composition of outer or local administration.They can be given with the available any generally accepted administering mode in this area Medicine, such as oral administration, such as with tablet, coated tablet, sugar coated tablet, hard gelatin capsule and Perle, solution, Emulsion Or the form of suspension, per rectum administration, such as in the form of suppository, parenterally it is administered (including Jing veins), such as with note The form of solution or infusion solution is penetrated, or local is administered, such as in the form of ointment, ointment or oil.In possible administration In mode, preferably oral and IV delivery.

The pharmaceutical composition of the present invention can be generally from about 0.05 to 80 weight % or from about 0.1 to 50 weight containing total amount Formula (I) compound of at least one present invention of amount %, optionally with pharmaceutically acceptable carrier and/or excipient.

Formula (I) of the present invention in addition to one or more excipient, in being contained in dosage form of the invention or pharmaceutical composition The amount of compound or its tautomer or salt, can be at least 0.1% to 0.5%, or at least 0.5% to 1.5%, or at least 1% to 3%.

Those skilled in the art is familiar with pharmaceutically acceptable excipient, such as diluent, load based on his/her Professional knowledge Body, binding agent, disintegrating agent, surfactant, lubricant, carrier, auxiliary agent, adjuvant and/or, it is other known be suitable to prepare medicine group The additive of compound.

Used as pharmaceutically acceptable excipient, the commonly known any excipient for being suitable to pharmaceutical composition is among consideration. The example is included but is not limited to, diluent, filler, binding agent, disintegrating agent, lubricant, fluidizer, solvent, dispersant, emulsifying It is agent, solubilizing agent, gel former, ointment base, antioxidant, preservative, stabilizer, carrier, thickening agent, chelating agent, slow Electuary, pH adjusting agent (for example, with obtain neutrality, alkalescence or acidic formulation), penetration enhancer, polymer, coating materials, propulsion Agent, tension regulator, surfactant, coloring agent, flavoring agent, sweeting agent and dyestuff.

Generally, suitable carrier material is not only inorganic carrier material, is also organic support material.Thus, for example Lactose, Starch (such as corn starch) or derivatives thereof, Talcum, silicon dioxide, polyvinylpyrrolidone, stearic acid or its salt can be used as piece The carrier material of agent, coated tablet, sugar coated tablet and hard gelatin capsule.Suitable carrier material for Perle is, Such as vegetable oil, wax, fat and semi-solid and liquid polyol.For the suitable carrier material that solution and syrup are produced For such as water, polyhydric alcohol, sucrose, Nulomoline etc..Suitable carrier material for injecting or being transfused solution is, such as water, Alcohol, polyhydric alcohol, glycerol and vegetable oil.Suitable carrier material for suppository is, such as neutral oil or fixed oil, wax, fat And semiliquid or liquid polyol or Polyethylene Glycol.Suitable carrier material for topical formulations is glyceride, semi-synthetic Glyceride, hydrogenated oil and fat, liquid wax, liquid paraffin, liquid aliphatic alcohol, sterol, Polyethylene Glycol and cellulose derivative with synthesis.

Using the excipient of the type of pharmaceutical composition, formula or preparation and required administering mode needed for being adapted to, carrier And/or diluent.

The pharmaceutical composition of the present invention can by by the compound of one or more formula (I) or its pharmaceutically acceptable salt with For example known inert diluent of suitable excipient, carrier, disintegrating agent, adjuvant, surfactant, binding agent and/or lubrication Agent is obtained by mixing.The tablet can be also constituted by several layers.The compositionss of the present invention can also contain other active substances.

Therefore, compositionss of the invention can be prepared by known per se and method familiar to the person skilled in the art, example Such as by by the solid or liquid of the compound of described formula (I) or its optionally with one or more routine of pharmaceutically acceptable salt Body carrier and/or diluent, mix to conventional formulation such as conventional tablet or coated tablet, capsule, powder agent, suspensoid or bolt Prepare in agent.The example of the carrier include but is not limited to corn starch, Lactose, glucose, Microcrystalline Cellulose, magnesium stearate, Polyvinylpyrrolidone, citric acid, tartaric acid, water, water/ethanol, water/glycerol, water/Sorbitol, water/Polyethylene Glycol, poly- second two Alcohol, cetearyl alcohol, carboxymethyl cellulose or fatty material such as Hard Fat or its suitable mixture.

The example of the suitable diluent of the compounds of this invention may include cellulose powder, calcium hydrogen phosphate, erythritol, low take The hydroxy propyl cellulose in generation, mannitol, pregelatinized Starch or xylitol.

The property of disease treated or prevent depending on the compound, institute that are given and seriousness, age of patient and individual Body situation and administering mode and frequency, the dosage of the compounds of this invention can change in a wide range, and meet certainly each concrete Individual demand in case.Generally, the amount of the compounds of this invention considers the conventional order of magnitude of DPP-IV inhibitor.When with vein way When footpath is administered, the commonly required dosage of the compounds of this invention can be 0.001mg to 10mg, or 0.01mg to 10mg, or 0.1mg is extremely 10mg, such as 0.25mg to 5mg, and when being orally administered, can be 0.005mg to 100mg, or 0.05mg to 100mg, or 0.5mg to 100mg, such as 2.5mg to 50mg or 0.5mg to 10mg, preferably 2.5mg to 10mg or 1mg to 5mg, in various situations One day 1 to 4 time administration.Depending on the dosage, it is probably easily to give daily dose with some dosage units.

Specific embodiment

The preparation of formula (III) compound

1. the bromo- 7- of 8- (2-butyne -1- bases) -3- methyl isophthalic acids-((4- methylquinazolin -2- bases) methyl) -3,7- xanthine The preparation of (compound III-1)

(1)3- methyl -7- (2-butyne -1- bases) the xanthic preparations of the bromo- 3,7- of -8-

N,N-dimethylformamide solution of the 3- methyl -8- bromo- xanthine with N- ethyl diisopropyl amines (DIPEA) is mixed Close, add the bromo- 2-butyne of 1-, and be stirred overnight at room temperature.For being processed, reactant mixture is poured into water.To be analysed The precipitate sucking filtration for going out, with water washing, and is dried, obtains 3- methyl -7- (2-butyne -1- bases) bromo- xanthine (molecular formula of -8-： C₁₀H₉BrN₄O₂)。

Mass spectrum (ESI⁺)：M/z=298 [M+H]⁺

Elementary analysiss：C,40.43；H,3.05；Br,26.89；N,18.86；O,10.77

(2)The bromo- 7- of 8- (2-butyne -1- bases) -3- methyl isophthalic acids-((4- methylquinazolin -2- bases) methyl) -3,7- xanthine Preparation

By 2- (2- bromoethyls) -4- methylquinazolins add to 3- methyl -7- (2-butyne -1- bases) the chloro- xanthine of -8- with In the N,N-dimethylformamide mixed solution of potassium carbonate.Reactant mixture is stirred at room temperature 8 hours.Aqueous solution process Afterwards, crude product is made through silicon stock column chromatography purification, with methylene chloride/methanol eluting, obtain the bromo- 7- of 8- (2-butyne -1- bases) -3- first Base -1- ((4- methylquinazolin -2- bases) methyl) -3,7- xanthine (molecular formula：C₂₀H₁₇BrN₆O₂)。

Mass spectrum (ESI⁺)：M/z=454 [M+H]⁺

Elementary analysiss：C,52.99；H,3.78；Br,17.63；N,18.54；O,7.06

¹H-NMR(d6-DMSO)：8.12(1H),7.84(1H),7.82(1H),7.58(1H),4.42(4H),3.41 (3H),2.94(3H),1.80(3H)。

2. the bromo- 7- of 8- (2- cyanobenzyls) -3- methyl isophthalic acids-(quinoline -2- ylmethyls) -3,7- xanthine (compound III- 2) preparation

(1)The bromo- 7- of 8- (2- cyanobenzyls) the xanthic preparations of -1,3- dimethyl -3,7-

N,N-dimethylformamide solution of the 3- methyl -8- bromo- xanthine with N- ethyl diisopropyl amines (DIPEA) is mixed Close, add 2- cyano-benzyl bromides, and be stirred overnight at room temperature.For being processed, reactant mixture is poured into water.To be analysed The precipitate sucking filtration for going out, with water washing, and is dried, obtains the bromo- xanthine (molecular formula of 3- methyl -7- (2- cyanobenzyls) -8-： C₁₄H₁₀BrN₅O₂)。

Mass spectrum (ESI⁺)：M/z=36 [1M+H]⁺；

Elementary analysiss：C,46.69；H,2.80；Br,22.19；N,19.44；O,8.88.

(2)The bromo- 7- of 8- (2- cyanobenzyls) -3- methyl isophthalic acids-(quinoline -2- ylmethyls) xanthic preparations of -3,7-

2- (2- bromoethyls)-quinoline is added to the 3- methyl -7- chloro- xanthine of (2- cyanobenzyls) -8- and potassium carbonate In N,N-dimethylformamide mixed solution.Reactant mixture is stirred at room temperature 8 hours.After aqueous solution process, thick product is made Thing obtains 1- (quinoline -2- ylmethyls) -3- methyl -7- (2- cyano group through silicon stock column chromatography purification with methylene chloride/methanol eluting Benzyl) the bromo- 3,7- xanthine (molecular formula of -8-：C₂₄H₁₇BrN₆O₂)。

Mass spectrum (ESI⁺)：M/z=502 [M+H]⁺；

Elementary analysiss：C,57.50；H,3.42；Br,15.94；N,16.76；O,6.38.

¹H-NMR(d6-DMSO)：8.07(1H),8.01(1H),7.79(1H),7.64(1H),7.48-7.55(4H), 7.32(2H),5.47(2H),4.76(2H),3.40(3H)

3. the bromo- 7- of 8- (1- cyclohexenyl methyls) -3- methyl isophthalic acids-(dimethyl-aminocarbonyl methyl) -3,7- xanthine (is changed Compound III-3) preparation

(1)The bromo- xanthic preparations of 3- methyl -7- (1- cyclohexenyl methyls) -8-

N,N-dimethylformamide solution of the 3- methyl -8- bromo- xanthine with N- ethyl diisopropyl amines (DIPEA) is mixed Close, add 1- cyclohexenyl methyl bromines, and be stirred overnight at room temperature.For being processed, reactant mixture is poured into water.Will The precipitate sucking filtration for being separated out, with water washing, and is dried, and obtains 3- methyl -7- (1- cyclohexenyl methyls) the bromo- xanthine of -8- and (divides Minor：C₁₃H₁₅BrN₄O₂)。

Mass spectrum (ESI⁺)：M/z=340 [M+H]⁺；

Elementary analysiss：C,46.03；H,4.46；Br,23.56；N,16.52；O,9.43.

(2)The bromo- 7- of 8- (1- cyclohexenyl methyls) -3- methyl isophthalic acids-(dimethyl-aminocarbonyl methyl) xanthic systems of -3,7- It is standby

N, N- dimethyl -2- bromo acetamides are added to 3- methyl -7- (1- cyclohexenyl methyls) the chloro- xanthine of -8- In the N,N-dimethylformamide mixed solution of potassium carbonate.Reactant mixture is stirred at room temperature 8 hours.Aqueous solution process Afterwards, crude product is made through silicon stock column chromatography purification, with methylene chloride/methanol eluting, obtain 1- (dimethyl-aminocarbonyl methyl) -3- first The bromo- 3,7- xanthine (molecular formula of base -7- (1- cyclohexenyl methyls) -8-：C₁₇H₂₂BrN₅O₃)。

Mass spectrum (ESI⁺)：M/z=425 [M+H]⁺；

Elementary analysiss：C,48.12；H,5.23；Br,18.83；N,16.51；O,11.31.

¹H-NMR(d6-DMSO)：5.40(1H),4.41(2H),4.29(2H),3.42(3H),2.99(6H),1.95- 1.99(4H),1.75(2H),1.61(2H)。

4. 1- (pyrrolidin-1-yl carbonvlmethyl) -3- methyl -7- (2-butyne -1- bases) the bromo- 3,7- xanthine of -8- (is changed Compound III-4) preparation

1- (2- Bromoacetyls) pyrrolidine is added to 3- methyl -7- (2-butyne -1- bases) the chloro- xanthine of -8- and carbon In the N,N-dimethylformamide mixed solution of sour potassium.Reactant mixture is stirred at room temperature 6 hours.After aqueous solution process, Crude product is made through silicon stock column chromatography purification, with methylene chloride/methanol eluting, 1- (pyrrolidin-1-yl carbonvlmethyl) -3- first is obtained Base -7- (2-butyne -1- bases) the bromo- 3,7- xanthine (molecular formula of -8-：C₁₆H₁₈BrN₅O₃)。

Mass spectrum (ESI⁺)：M/z=409 [M+H]⁺；

Elementary analysiss：C,47.07；H,4.44；Br,19.57；N,17.15；O,11.76.

¹H-NMR(d6-DMSO)：4.42(2H),4.29(2H),3.42(3H),3.10(4H),1.84(4H),1.80 (3H)。

5. the bromo- 7- of 8- (2-butyne -1- bases) -3- methyl isophthalic acids-((4- methylquinazolin -2- bases) methyl) -3,7- xanthine The preparation of (compound III-5)

(1)3- methyl -7- (2,3- methyl-2-butene -1- bases) the xanthic preparations of the bromo- 3,7- of -8-

N,N-dimethylformamide solution of the 3- methyl -8- bromo- xanthine with N- ethyl diisopropyl amines (DIPEA) is mixed Close, add bromo- 2, the 3- methyl-2-butenes of 1-, and be stirred overnight at room temperature.Reactant mixture is poured into water, is filtered and is separated out Precipitate, with water washing, and be dried, obtain 3- methyl -7- (2, the 3- methyl-2-butene -1- bases) bromo- xanthine (molecules of -8- Formula：C₁₂H₁₅BrN₄O₂)。

Mass spectrum (ESI⁺)：M/z=328 [M+H]⁺；

Elementary analysiss：C,44.05；H,4.62；Br,24.42；N,17.12；O,9.78.

(2)The bromo- 7- of 8- (2,3- methyl-2-butene -1- bases) -3- methyl isophthalic acids-((4- methylquinazolin -2- bases) methyl) - The xanthic preparations of 3,7-

2- (2- bromoethyls) -4- methylquinazolins are added to 3- methyl -7- (2,3- methyl-2-butene -1- bases) -8- In the N,N-dimethylformamide mixed solution of chloro- xanthine and potassium carbonate.Reactant mixture is stirred at room temperature 8 hours. After aqueous solution process, crude product is made through silicon stock column chromatography purification, with methylene chloride/methanol eluting, obtain 8- bromo- 7- (2,3- first Base -2-butylene -1- bases) -3- methyl isophthalic acids-((4- methylquinazolin -2- bases) methyl) -3,7- xanthine (molecular formula： C₂₂H₂₃BrN₆O₂)。

Mass spectrum (ESI⁺)：M/z=484 [M+H]⁺；

Elementary analysiss：C,54.67；H,4.80；Br,16.53；N,17.39；O,6.62.

¹H-NMR(d6-DMSO)：8.14(1H),7.85(1H),7.82(1H),7.57(1H),4.43(2H),4.39 (2H),3.42(3H),2.95(3H),1.78(9H)。

The preparation of formula (IV) compound

1. the preparation of N- normal-butyls-N- ((R)-piperidines -3- bases) t-butyl carbamate (compound IV-1)

(1) (R) preparation of -3- (t-butoxycarbonyl amino) piperidines -1- benzyl carboxylates

At 0-5 DEG C, successively use triethylamine and benzyl chloroformate to process the THF of (R) -3- (t-butoxycarbonyl amino) piperidines Solution, and stir 24 hours at such a temperature.Then reactant mixture is evaporated in vacuo to about 1/4 volume, and in acetic acid second Distribute between ester and 1M hydrochloric acid solutions.Organic faciess are separated, in order with the 1M hydrochloric acid solutions of other part, saturation NaHCO₃It is water-soluble Liquid and aqueous salt solu-tion.Then organic faciess part is dried with anhydrous magnesium sulfate, is filtered, and vacuum evaporation obtains (R) -3- (t-butoxycarbonyl amino) piperidines -1- benzyl carboxylate (molecular formula：C₁₈H₂₆N₂O₄)。

Mass spectrum (ESI⁺)：M/z=335 [M+H]⁺；

Elementary analysiss：C,64.65；H,7.84；N,8.38；O,19.14.

(2) (R) preparation of -3- (tertbutyloxycarbonyl (normal-butyl) amino) piperidines -1- benzyl carboxylates

(R) -3- (t-butoxycarbonyl amino) piperidines -1- benzyl carboxylates are dissolved in into DMF at 0-5 DEG C, and with sodium hydride The process of 60% suspension.Reactant mixture is warming up to into room temperature 10min, is then cooled down again, and add n-butyl bromide.After 2 hours, Room temperature is warming up to, the n-butyl bromide of additional quantity is added, and reactant mixture is stirred 16 hours.Then reactant mixture vacuum is steamed ～1/4 volume is sent to, and is distributed between ethyl acetate and water.Then by organic faciess part aqueous salt solu-tion, with anhydrous Magnesium sulfate is dried, and filters, and vacuum evaporation obtains residue, and which is carried out silica gel column chromatography to obtain N- with ethyl acetate/hexane N-butyl product (molecular formula：C₂₂H₃₄N₂O₄)。

Mass spectrum (ESI⁺)：M/z=391 [M+H]⁺；

Elementary analysiss：C,67.66；H,8.78；N,7.17；O,16.39.

(3) preparation of N- normal-butyls-N- ((R)-piperidines -3- bases) t-butyl carbamate

N- N-butyl products are dissolved in ethanol, are processed with 10% palladium carbon, and hydrogenation 6 is little under 60-70psi hydrogen When.Then crude product mixture is filtered with kieselguhr, is evaporated in vacuo, and is again dissolved in ethanol, passed through nylon syringe Formula filter filters to remove residual catalyst.Mixture is evaporated to obtain title compound (molecular formula：C₁₄H₂₈N₂O₂)。

Mass spectrum (ESI⁺)：M/z=257 [M+H]⁺；

Elementary analysiss：C,65.59；H,11.01；N,10.93；O,12.48.

¹H-NMR(d6-DMSO)：3.61(1H),3.09(1H),2.95(2H),2.85(1H),2.75(2H),2.0(1H), 1.86(1H),1.61(1H),1.51(3H),1.45(1H),1.42(9H),1.30(2H),0.89(3H)。

2. the preparation of (R)-N- cyclohexyl-N- (piperidines -3- bases) t-butyl carbamate (compound IV-2)

(R) -3- (t-butoxycarbonyl amino) piperidines -1- benzyl carboxylates are dissolved in DMF, and are suspended with the 60% of sodium hydride Liquid process.Reactant mixture is warming up to into room temperature 15 minutes, is then cooled down again, and add bromocyclohexane.After 2 hours, rise Warm to room temperature, add the bromocyclohexane of additional quantity, and reactant mixture is stirred 16 hours.Then by reactant mixture vacuum ～1/4 volume is evaporated to, and is distributed between ethyl acetate and water.Then by organic faciess part aqueous salt solu-tion, with nothing Water magnesium sulfate is dried, and filters, and vacuum evaporation obtains residue, and which is carried out silica gel column chromatography to obtain with ethyl acetate/hexane The intermediate of N- cyclohexyls.The intermediate of N- cyclohexyls is dissolved in ethanol, is processed with 10% palladium carbon, and in 60- Hydrogenate 6 hours under 70psi hydrogen.Then crude product mixture is filtered with kieselguhr, is evaporated in vacuo, and is again dissolved in ethanol, Filter to remove residual catalyst.Mixture is evaporated to obtain title compound (molecular formula：C₁₆H₃₀N₂O₂)。

Mass spectrum (ESI⁺)：M/z=283 [M+H]⁺；

Elementary analysiss：C,68.04；H,10.71；N,9.92；O,11.33.

¹H-NMR(d6-DMSO)：3.61(1H),3.55(1H),3.09(1H),2.85(1H),2.74(2H),2.0(1H), 1.86(1H),1.72(2H),1.61(1H),1.51(2H),1.48(2H),1.45(2H),1.43(9H),1.16(4H)。

3. the preparation of (R)-N- phenyl-N- (piperidines -3- bases) t-butyl carbamate (compound IV-3)

(R) -3- (t-butoxycarbonyl amino) piperidines -1- benzyl carboxylates are dissolved in into DMF at 0-5 DEG C, with sodium hydride The process of 60% suspension.Reactant mixture is warming up to into room temperature 30 minutes, is then cooled down again, and add trifluoromethanesulfonic acid benzene Ester.After 3 hours, room temperature is warming up to, adds the trifluoromethanesulfonic acid phenyl ester of additional quantity, and reactant mixture is stirred 18 hours.So Afterwards reactant mixture is evaporated in vacuo to～1/4 volume, and is distributed between ethyl acetate and water.Then organic faciess part is used Aqueous salt solu-tion, is dried with anhydrous magnesium sulfate, is filtered, and vacuum evaporation obtains residue, and which is entered with ethyl acetate/hexane Row silica gel column chromatography is obtaining the intermediate of N- phenylatings.The intermediate of N- phenylatings is dissolved in ethanol, is processed with 10% palladium carbon, And hydrogenate 6 hours under 60-70psi hydrogen.Then crude product mixture is filtered with kieselguhr, is evaporated in vacuo, and it is molten again In ethanol, filter to remove residual catalyst.Mixture is evaporated to obtain title compound (molecular formula：C₁₆H₂₄N₂O₂)。

Mass spectrum (ESI⁺)：M/z=277 [M+H]⁺；

Elementary analysiss：C,69.53；H,8.75；N,10.14；O,11.58.

¹H-NMR(d6-DMSO)：7.72(2H),7.32(2H),6.99(1H),3.60(1H),3.28(1H),3.03 (1H),2.75(2H),2.03(1H),2.01(1H),1.77(1H),1.46-1.50(2H),1.42(9H)。

4. 3- (tertbutyloxycarbonyl ((3R)-piperidines -3- bases) amino) piperidines -1- carboxylic acid tert-butyl esters (compound IV-4) Prepare

(R) -3- (t-butoxycarbonyl amino) piperidines -1- benzyl carboxylates are dissolved in into DMF at 0-5 DEG C, with sodium hydride The process of 60% suspension.Reactant mixture is warming up to into room temperature 10min, is then cooled down again, and add 3- bromine piperidines -1- carboxylic acids The tert-butyl ester.After 2 hours, room temperature is warming up to, adds the 3- bromine piperidines -1- carboxylic acid tert-butyl esters of additional quantity, and reactant mixture is stirred Mix 16 hours.Then reactant mixture is evaporated in vacuo to～1/4 volume, and is distributed between ethyl acetate and water.Then will Organic faciess part aqueous salt solu-tion, is dried with anhydrous magnesium sulfate, is filtered, and vacuum evaporation obtains residue.

Residue is dissolved in ethanol, is processed with 10% palladium carbon, and hydrogenated 6.5 hours under 60-70psi hydrogen.Then Crude product mixture is filtered with kieselguhr, is evaporated in vacuo, and is again dissolved in ethanol, filter to remove residual catalyst.Will be mixed Compound evaporates, and carries out silica gel column chromatography to obtain title compound (molecular formula with ethyl acetate/hexane：C₂₀H₃₇N₃O₄)。

Mass spectrum (ESI⁺)：M/z=384 [M+H]⁺；

Elementary analysiss：C,62.63；H,9.72；N,10.96；O,16.69.

¹H-NMR(d6-DMSO)：3.75(2H),3.61(1H),3.54(2H),3.49(1H),3.08(1H),2.86 (1H),2.74(2H),2.12(1H),1.87(2H),1.70(2H),1.61(2H),1.49(2H),1.42(18H)。

The preparation of formula (II) compound

1. l- ((4- methyl-quinazoline -2- bases) methyl) -3- methyl -7- (2-butyne -1- bases) -8- (positive fourths of (R) -3- Amino-piperadine -1- bases)-xanthine (compound II-1)

(R) -3- tertbutyloxycarbonyls (normal-butyl) amino piperidine (compound IV-1) is added to 1- ((4- methyl-quinoline azoles Quinoline -2- bases) methyl) -3- methyl -7- (2-butyne -1- bases) the chloro- xanthine of -8- (compound III-1) and sodium carbonate diformazan it is sub- In sulfone mixed solution.Reactant mixture is stirred 18 hours at 60 DEG C.For being processed, which is mixed with water, and sucking filtration institute The precipitate of formation.Solid sediment is dissolved in dichloromethane, mix with trifluoracetic acid, and stir half an hour at room temperature. For being processed, by reactant mixture with dchloromethane, and washed with unsaturated carbonate potassium solution, organic faciess anhydrous slufuric acid Sodium is dried, and is evaporated, and through silica gel column chromatography, with methylene chloride/methanol (1:0 to 4:1) eluting, obtains (R) -8- (3- normal-butyls Amino piperidine -1- bases) -7- (2-butyne -1- bases) -3- methyl isophthalic acids-((4- methylquinazolin -2- bases) methyl) -3,7- xanthine (molecular formula：C₂₉H₃₆N₈O₂)。

Mass spectrum (ESI⁺)：M/z=529 [M+H]⁺

Elementary analysiss：C,65.89；H,6.86；N,21.20；O,6.05；

¹H NMR(d6-DMSO)：8.12(1H),7.84(2H),7.59(1H),4.43(4H),3.49(1H),3.38 (3H),3.33(1H),3.22-3.28(3H),2.95(3H),2.65(1H),2.53(2H),1.80(3H),1.70(1H), 1.30-1.58(7H),0.89(3H)。

2. l- (quinoline -2- ylmethyls) -3- methyl -7- (2- cyanobenzyls) -8- ((R) -3- n-butylamino-piperidines - 1- yls)-xanthine (compound II-2) preparation

(R) -3- tertbutyloxycarbonyls (normal-butyl) amino piperidine (compound IV-1) is added to 1- ((quinoline -2- bases) first Base) -3- methyl -7- (2- cyanobenzyls) the chloro- xanthine of -8- (compound III-2) and sodium carbonate dimethyl sulfoxide mixed solution In.Reactant mixture is stirred 18 hours at 60 DEG C.For being processed, which is mixed with water, and the precipitation formed by sucking filtration Thing.Solid sediment is dissolved in dichloromethane, mix with trifluoracetic acid, and stir half an hour at room temperature.For being processed, By reactant mixture with dchloromethane, and washed with unsaturated carbonate potassium solution, organic faciess anhydrous sodium sulfate drying, steamed It is dry, and through silica gel column chromatography, with methylene chloride/methanol (1:0 to 4:1) eluting, obtain (R) -8- (3- n-butylamino piperidines - 1- yls) -7- (2- cyanobenzyls) -3- methyl isophthalic acids-((quinoline -2- bases) methyl) -3,7- xanthine (molecular formula：C₃₃H₃₆N₈O₂；).

Mass spectrum (ESI⁺)：M/z=577 [M+H]⁺；

Elementary analysiss：C,68.73；H,6.29；N,19.43；O,5.55；

¹H-NMR(d6-DMSO)：8.07(1H),8.02(1H),7.76(1H),7.61(1H),7.48-7.59(4H), 7.32(2H),5.46(2H),4.75(2H),3.38(1H),3.35(3H),3.32(1H),3.27(3H),2.63(1H),2.53 (2H),1.70(1H),1.50(1H),1.42(2H),1.35(4H),0.89(3H)。

3. l- (dimethylaminocarbonylmethyl) -3- methyl -7- (cyclohexene -1- ylmethyls) -8- ((R) -3- cyclohexyl Amino-piperadine -1- bases)-xanthine (compound II-3) preparation

(R) -3- tertbutyloxycarbonyls (cyclohexyl) amino piperidine (compound IV-2) is added to 1- (Dimethylaminocarbonyls Methyl) dimethyl sulfoxide of -3- methyl -7- (cyclohexene -1- ylmethyls) the chloro- xanthine of -8- (compound III-3) and sodium carbonate mixes Close in solution.Reactant mixture is stirred 18 hours at 55 DEG C.For being processed, which is mixed with water, and sucking filtration is formed Precipitate.Solid sediment is dissolved in dichloromethane, mix with trifluoracetic acid, and stir half an hour at room temperature.For entering Row is processed, and by reactant mixture with dchloromethane, and is washed with unsaturated carbonate potassium solution, and organic faciess are dry with anhydrous sodium sulfate It is dry, it is evaporated, and through silica gel column chromatography, with methylene chloride/methanol (1:0 to 4:1) eluting, obtains (R) -1- (dimethylamino carbonyls Ylmethyl) -3- methyl -7- (cyclohexene -1- ylmethyls) -8- (3- n-butylamino piperidin-1-yls) -3,7- xanthine (molecules Formula：C₂₈H₄₃N₇O₃；).

Mass spectrum (ESI⁺)：M/z=526 [M+H]⁺；

Elementary analysiss：C,63.97；H,8.25；N,18.65；O,9.13；

¹H-NMR(d6-DMSO)：5.41(1H),4.40(2H),4.28(2H),3.48(1H),3.41(3H),3.34 (1H),3.27(2H),3.21(1H),2.98(6H),2.63(1H),2.55(1H),1.99(2H),1.92(2H),1.74(2H), 1.71(1H),1.61(4H),1.57(1H),1.47(4H),1.35(2H),1.21(2H),1.10(2H)。

4. l- (pyrrolidin-1-yl carbonvlmethyl) -3- methyl -7- (2-butyne -1- bases) -8- ((R) -3- phenyl aminos - Piperidin-1-yl)-xanthine (compound II-4) preparation

(R) -3- tertbutyloxycarbonyls (phenyl) amino piperidine (compound IV-3) is added to 1- (pyrrolidin-1-yl carbonyls Methyl) -3- methyl -7- (2-butyne -1- bases) the chloro- xanthine of -8- (compound III-1) mix molten with the dimethyl sulfoxide of sodium carbonate In liquid.Reactant mixture is stirred 20 hours at 60 DEG C.For being processed, which is mixed with water, and sucking filtration formed it is heavy Starch.Solid sediment is dissolved in dichloromethane, mix with trifluoracetic acid, and stir half an hour at room temperature.For being located Reason, by reactant mixture with dchloromethane, and is washed with unsaturated carbonate potassium solution, organic faciess anhydrous sodium sulfate drying, It is evaporated, and through silica gel column chromatography, with methylene chloride/methanol (1:0 to 4:1) eluting, obtains l- (pyrrolidin-1-yl carbonyl first Base) -3- methyl -7- (2-butyne -1- bases) -8- ((R) -3- phenyl aminos-piperidin-1-yl)-xanthine (molecular formula： C₂₇H₃₃N₇O₃；).

Mass spectrum (ESI⁺)：M/z=504 [M+H]⁺；

Elementary analysiss：C,64.39；H,6.61；N,19.47；O,9.53.

¹H-NMR(d6-DMSO)：7.08(2H),6.83(2H),6.67(1H),5.15(1H),4.42(2H),4.28 (2H),3.58(1H),3.42(3H),3.34(1H),3.28(2H),3.09(4H),2.63(1H),1.83(4H),1.81(1H), 1.78(3H),1.55(2H),1.46(1H)。

5. l- (4- quinazoline -2- ylmethyls) -3- methyl -7- (2,3- dimethyl but-2-ene -1- bases) -8- ((R) -3- (piperidines -3- base amino) piperidin-1-yl)-xanthine (compound II-5) preparation

(R) -3- tertbutyloxycarbonyls (normal-butyl) amino piperidine (compound IV-1) is added to 1- ((4- methyl-quinoline azoles Quinoline -2- bases) methyl) -3- methyl -7- (2,3- dimethyl but-2-ene -1- bases) the chloro- xanthine of -8- (compound III-1) and carbon In the dimethyl sulfoxide mixed solution of sour sodium.Reactant mixture is stirred 20 hours at 55 DEG C.For being processed, which is mixed with water Close, and the precipitate formed by sucking filtration.Solid sediment is dissolved in dichloromethane, mix with trifluoracetic acid, and at room temperature Stirring half an hour.For being processed, by reactant mixture with dchloromethane, and washed with unsaturated carbonate potassium solution, it is organic Anhydrous sodium sulfate drying is mutually used, is evaporated, and through silica gel column chromatography, with methylene chloride/methanol (1:0 to 4:1) eluting, obtains l- (4- quinazoline -2- ylmethyls) -3- methyl -7- (2,3- dimethyl but-2-ene -1- bases) -8- ((R) -3- (piperidines -3- base ammonia Base) piperidin-1-yl)-xanthine (molecular formula：C₃₂H₄₃N₉O₂；).

Mass spectrum (ESI⁺)：M/z=586 [M+H]⁺；

Elementary analysiss：C,65.62；H,7.40；N,21.52；O,5.46；

¹H-NMR(d6-DMSO)：8.11(1H),7.85(2H),7.59(1H),4.43(2H),4.39(2H),3.49 (1H),3.39(3H),3.31(1H),3.27(2H),3.22(1H),2.94(3H),2.87(1H),2.76(2H),2.71(1H), 2.63(2H),2.02(1H),1.79(9H),1.71(2H),1.58(4H),1.48(2H)。

Following compound can be also prepared in a similar fashion：

Table 1

The preparation of formula (I) compound

1. l- ((4- methyl-quinazoline -2- bases) methyl) -3- methyl -7- (2-butyne -1- bases) -8- ((R) -3- are prepared (N1- normal-butyl biguanide bases)-piperidin-1-yl)-xanthine (compound I-1)

Dicyandiamide is dissolved in isopropanol, (R) -8- (3- n-butylamino piperidin-1-yls) -7- (2-butyne -1- are added Base) -3- methyl isophthalic acids-((4- methylquinazolin -2- bases) methyl) -3,7- xanthine (compound II-1), pH is adjusted with 36%HCl 5～6, control temperature is reacted 6 hours at 80～100 DEG C, stands cooling, separate out a large amount of white crystals after the completion of reaction, and sucking filtration is washed Wash, dehydrated alcohol recrystallization is used after being dried, obtain title compound (molecular formula：C₃₁H₄₀N₁₂O₂；).

Mass spectrum (ESI⁺)：M/z=613 [M+H]⁺；

Elementary analysiss：C,60.77；H,6.58；N,27.43；O,5.22；

¹H-NMR(d6-DMSO)：8.12(1H),7.85(2H,br),7.80(2H),7.59(1H),6.63(2H,br), 4.43(4H),3.59(1H),3.38(3H),3.34(1H),3.30(2H),3.28(2H),2.95(3H),2.65(1H),2.0 (1H,br),1.80(3H),1.70(1H),1.30-1.58(7H),0.89(3H)。

2. ((N1- is just for (R) -3- for -3- methyl -7- (2- cyanobenzyls) -8- for l- ((4- methyl-quinazoline -2- bases) methyl) Butyl biguanide base)-piperidin-1-yl)-xanthine (compound I-2) preparation

Dicyandiamide is dissolved in isopropanol, (R) -8- (3- n-butylamino piperidin-1-yls) -7- (2-butyne -1- are added Base) -3- methyl isophthalic acids-((4- methylquinazolin -2- bases) methyl) -3,7- xanthine (compound II-1), pH is adjusted with 36%HCl 5～6, control temperature is reacted 6 hours at 80～100 DEG C, stands cooling, separate out a large amount of white crystals after the completion of reaction, and sucking filtration is washed Wash, dehydrated alcohol recrystallization is used after being dried, obtain title compound (molecular formula：C₃₅H₄₀N₁₂O₂)。

Mass spectrum (ESI⁺)：M/z=661 [M+H]⁺

Elementary analysiss：C,63.62；H,6.10；N,25.44；O,4.84.

¹H-NMR(d6-DMSO)：8.06(1H),8.01(1H),7.83(2H),7.75(1H),7.63(1H),7.52 (3H),7.46(1H),7.30(2H),6.62(2H),5.47(2H),4.76(2H),3.61(1H),3.39(3H),3.34(1H), 3.30(2H),3.25(2H),2.63(1H),2.15(1H),1.83(1H),1.48-1.58(5H),1.30(2H),0.89(3H)。

Following compound is prepared in a similar fashion

Table 2

Biological examples

1st, DPP-IV detections

Originated as DPP-IV using the extract of human colon cancer cells system Caco-2.For inducing DPP-IV expression, carefully Document is pressed in the differentiation of born of the same parents, and (Reiher etc., " the increase expression of enterocyte system Caco-2 ", in Proc.Natl.Acad.Sci. the 90th Volume, the 5757-5761 page (1993)) described in carry out.Cell extraction thing is at 4 DEG C, by being centrifuged 30 points under 35,000g Clock (to remove cell debris), be dissolved in buffer agent (10 μm of Tns HC1,0.15M NaCl, 0.04t.i.u. AKOLINEs, 0.5%Nonidet-P40, pH8.0) in and obtain.

Acylamino- -4- trifluoromethyl coumarin (AFC) matrix solutions that 50 microlitres of final concentrations are 100 μm are placed on black In color titer plate.Detect that (final concentration is 50 μm of Tns-HCl, 7.8,50 μm of NaCl of pH, 1% to buffer by 20 microlitres DMSO) drawn with pipette.Reaction is that (every hole final concentration is 0.14 by adding 30 microlitres of Caco-2 protein for having dissolved Micrograms of protein) start.Test substance that generally will be studied, beforehand dilution detect buffer and add again at 20 microlitres, Wherein detect that the volume of buffer is accordingly reduced.Reaction is to carry out at room temperature, is cultivated 60 minutes.Then, exist Fluorescent is measured in Victor14Multilabel enumerators, and excitation wavelength is 405 millimicrons, and launch wavelength is 535 millimicrons. Blank assay reading (active equivalent to 0%) is in the mixed of no any Caco-2 protein (volume by detect buffer put) Obtain in compound, control value (active equivalent to 100%) is obtained in the mixture for be not added with material.It is to be measured in discussion The drug effect of material, with IC₅₀Value is represented, and is calculated by dosage/activity curve, and which includes 11 measurement points in each situation.

The IC of embodiments of the invention compound I-1～13 (Biguanide derivative)₅₀Value≤10nM, and its IC₅₀Value is notable Less than corresponding aminoderivative, such as less than about 1/5～1/10.Detailed results see the table below 2.

Table 2：Inhibitory action of the compounds of this invention to DPP-IV

Compound number	IC50(nM)
		Compound I-1	0.5
Compound I-2	0.3
		Compound I-3	0.06
Compound I-4	0.2
		Compound I-5	0.05
Compound I-6	0.01
		Compound I-7	0.08
Compound I-8	0.5
		Compound I-9	0.09
Compound I-10	0.3
		Compound I-11	0.2
Compound I-12	0.15
		Compound I-13	0.10

2. impact of the compounds of this invention to alloxan diabetess mouse blood sugar

Healthy mice 60 is selected, 10 is separated at random and is only used as Normal group (normal saline 0.5ml/ only), remaining 50 Mice fasting can't help water 24 hours, the equal lumbar injection alloxan 180mg/kg of each Mus, and eye socket blood sampling in 72 hours thereafter determines blood Sugar, blood sugar concentration are diabetic mice higher than 11.1mmol/L person.50 diabetic mices are randomly divided into into 5 groups： Artificial diabeteses blank group (normal saline 0.5ml/ is only), Li Gelieting groups (5mg/kg), the high dose of the compounds of this invention Group (5mg/kg), middle dose group (1mg/kg), low dose group (0.2mg/kg).Every morning 8:00～9:00 gastric infusion, even Continuous 30d.Fasting 12h takes blood from rear mouse orbit rear vein beard and surveys blood glucose value.

As a result show, embodiments of the invention compound I-1～13 (Biguanide derivative) are shown in various degree than phase The stronger hypoglycemic activity of the aminoderivative answered and the action time for significantly extending, and it is less prone to hypoglycemia.For example, The low dose group of the compound of embodiment I-1～13 is shown with the higher and more longlasting blood sugar lowering energy of Billy's Ge Lieting groups Power, with significant difference (p ＜ 0.05).

Claims (6)

1. a kind of compound prepared shown in formula (I), its stereoisomer, or the method for its pharmaceutically acceptable salt, bag Include the step of making formula (II) compound contact with dicyandiamide：

Wherein,

R¹Represent C_1-8Alkyl, the C_1-8Alkyl can R¹²- CO- replaces, and R¹²Represent two (C_1-6Alkyl) amino, pyrrolidine -1- Base, piperidin-1-yl or C_6-10Aryl；

R²Represent C_1-8Alkyl or C_6-10Aryl；

R³Represent benzyl, C_2-8Thiazolinyl, C_2-8Alkynyl or C_3-8Phenyl ring in cycloalkenylmethyl, wherein benzyl can be by one or more halogen Element or cyano group replace；

R⁴Represent C_1-8Alkyl, C_3-8Cycloalkyl, C_3-8Circle heterocycles alkyl, C_6-10Aryl or, C_5-10Unit's heteroaryl；

R⁵Represent hydrogen or amino protecting group；And

Work as R⁵When representing amino protecting group, also include the step of described amino protecting group is removed before contacting with dicyandiamide.

2. method according to claim 1, it is characterised in that described amino protecting group is selected from methoxycarbonyl group, ethoxy carbonyl Base, tert-butoxycarbonyl, benzyloxycarbonyl group, 2- xenyl -2- propylene carbonyl oxygens, allyloxycarbonyl, trimethylsilyl ethoxycarbonyl, adjacent benzene Diformyl, p-toluenesulfonyl, trifluoroacetyl group, neighbour or p-nitrophenyl sulfonyl, pivaloyl group, benzoyl, triphen first Base, 2,4- dimethoxy-benzyls, to methoxy-benzyl or benzyl.

3. method according to claim 1 and 2, it is characterised in that also including making formula (III) compound and formula (IV) chemical combination The step of thing is contacted：

Wherein Pr¹Represent amino protecting group；

When needed in order to obtain R⁵Formula (II) compound of hydrogen is represented, alternatively by amino protecting group Pr¹Removing,

Wherein R¹-R⁵As defined in claim 1.

4. method according to claim 3, it is characterised in that also including formula (III) compound is prepared the step of：Including making 8- halos xanthine derivative with contain R³The halides contact of group, then makes products therefrom and contains R¹The halides contact of group：

Wherein R¹-R³As defined in claim 1, and X represents halogen atom.

5. method according to claim 3, it is characterised in that also including formula (IV) compound is prepared the step of, including The amino protecting group Pr for making 3- amino piperidines (formula (V) compound) Jing different successively¹、Pr²Protection, then with containing R⁴The halogen of group Contact for thing, reselection removing Pr²Amino protecting group：

Wherein R¹-R⁵As defined in claim 1, Pr¹And Pr²It is independent of each other to represent identical or different amino protecting group.

6. the method according to claim 4 or 5, it is characterised in that Pr¹And Pr²It is identical or different, it is independent of each other to represent first Oxygen carbonyl, carbethoxyl group, tert-butoxycarbonyl, benzyloxycarbonyl group, 2- xenyl -2- propylene carbonyl oxygens, allyloxycarbonyl, trimethyl silicane Carbethoxyl group, phthalyl, p-toluenesulfonyl, trifluoroacetyl group, neighbour or p-nitrophenyl sulfonyl, pivaloyl group, benzene Formoxyl, trityl, 2,4- dimethoxy-benzyls, to methoxy-benzyl or benzyl.