CN103664893A - Novel DPP-4 inhibitor, and preparation method and medicinal application thereof - Google Patents

Abstract

The invention relates to the field of medicaments, and particularly relates to a compound shown in Formula I, a preparation method thereof and application thereof as a novel DPP-4 inhibitor medicament, especially application in preparation of antidiabetic medicaments. In vivo/in vitro pharmacodynamic experiment results indicate that the compound shown in Formula I has obvious inhibiting action on DPP-4, shows superior blood sugar reducing effect and has higher acting effect in comparison with a similar medicament linagliptin. Meanwhile, the compound provided by the invention is short in preparation route, accessible in raw materials, simple in process and suitable for industrial large-scale production, and has great developing prospects.

Description

A kind of novel DPP-4 inhibitor, its preparation method and medicinal use

Technical field

The present invention relates to pharmacy field, be specifically related to 1-(2-butyne-1-yl)-6-[3 (R)-amino piperidine-1-yl]-3-[(4-methyl quinazoline-2-yl) methyl]-2,4-(1H, 3H) pyrimidine dione, the invention also discloses preparation method and the medicinal use of this compound, particularly as DPP-4 inhibitor, for the preparation of the purposes of anti-diabetic and complication medicine aspect thereof.

Background technology

Diabetes are a kind of slow progressivity diseases, according to estimates, approximately there is diabetic subject 2.4 hundred million in the whole world at present, its sickness rate rapidly increases, expect 2025 and will reach 3.8 hundred million, in addition, approximately have every year 3800000 people to die from diabetes, AIDS patient's number is suitable with dying from, and has become the disease that is only second to tumour, cardiovascular diseases the third-largest harm humans health afterwards.In diabetic subject, more than 90% be diabetes B, therefore at present main antidiabetic medicine research all launches for diabetes B.Diabetes B is a kind ofly mainly due to insulin resistant, to follow relative insufficient insulin, defect of insulin secretion with or without insulin resistant, cause the metabolic trouble of chronic hyperglycemia.Traditional antidiabetic drug mainly contains euglycemic agent, Regular Insulin succagoga, the large class of alpha-glucosidase inhibitor 3.Will there is the untoward reactions such as body weight increase, hypoglycemia and drug effect reduce gradually in these drug mains, therefore in the urgent need to research and development novel blood sugar lowing medicine.

Dipeptidyl peptidase (DPP-4) inhibitor belongs to the antidiabetic medicine of latest generation, is the medicine based on glucagon-like-peptide-1 (GLP-1), has to control blood sugar and do not put on weight, and can not cause the advantages such as hypoglycemia.At present, the research of DPP-4 inhibitor has caused the concern of world Ge great drugmaker, and dissimilar small molecules DPP-4 inhibitor continues to bring out.The most representative is sitagliptin (sitagliptin), BMS-477118 (saxagliptin), BI 1356 (linagliptin), the Egelieting (alogliptin) of having gone on the market.

A synthetic 8-(3-the amino piperidine)-xanthine derivative (WO 2004018468/CN 1675212) of BI 1356 Shi You Boehringer Ingelheim company design is the very strong DPP-4 inhibitor (IC of activity ₅₀=1nmol/L), there is highly selective, long-acting and orally active feature, there is good security and tolerance simultaneously, in May, 2011, obtain FDA approval listing, be used for the treatment of diabetes B.

The pyrimidine dione class DPP-4 inhibitor (WO 2005095381/CN 1926128) of Egelieting Ze Shiyouwu field company exploitation, its DPP-4 suppresses active IC ₅₀for 7nmol/L.In April, 2010, in Japan, go on the market, for dietary control add physical activity or dietary control add a-Glycosylase inhibitory drugs for treatment glycemic control still not good adult diabetes B patient single therapy or with thiazolidinediones combination therapy.Because its DPP-4 suppresses, active BI 1356 is poor relatively, quantity also more greatly every day 12.5-25mg.In addition, the security of this medicine need to confirm, in June, 2009, FDA once with the insufficient refusal Wu Tian company that serves as reasons of cardiovascular safety data about the application for quotation of Egelieting, the said firm in July, 2011 again submitted Egelieting with and with the application for quotation of pioglitazone combined preparation.Recently, FDA issues statement, the Egelieting of temporarily refusing the research and development of Japanese Wu Tian company with and with the application for quotation of pioglitazone combined preparation, require to provide more validity and security evidence.Thereby the application prospect of Egelieting need to observe.

Sitagliptin (sitagliptin) Saxagliptin (BMS-477118)

Alogliptin (Egelieting) Linagliptin (BI 1356)

The object of the invention is exploitation active higher, selectivity is stronger and the lower novel antidiabetic drug of cardiovascular security risk.

Summary of the invention

The inventor is surprised to find, designed, synthetic suc as formula the 1-shown in I (2-butyne-1-yl)-6-[3 (R)-amino piperidine-1-yl]-3-[(4-methyl quinazoline-2-yl) methyl]-2,4-(1H, 3H) pyrimidine dione, its drug effect and security are all better than Egelieting and BI 1356, and being expected to exploitation becomes a new drug that can be applied to treat diabetes.

The invention discloses formula I compound, its tautomer, enantiomer, its mixture, prodrug and salt thereof, acceptable salt on or organic sour physiology inorganic with it particularly, solvate, polymorphic form, it has significant restraining effect for dipeptidyl peptidase-4 (DPP-4) activity, it maybe can prevent or palliate a disease or the purposes, particularly I type or type ii diabetes of symptom by reducing DPP-4 activity in prevention or the treatment disease relevant with increasing DPP-4 activity form or symptom.

The form of the salt of the formula I compound that the present invention is included is pharmacy acceptable salt, includes but not limited to the acid salt that the compounds of this invention and following acid form: hydrochloric acid, Hydrogen bromide, trifluoroacetic acid, sulfuric acid, citric acid, tartrate, phosphoric acid, lactic acid, acetic acid, toxilic acid, fumaric acid, oxysuccinic acid, tussol, camphorsulfonic acid, methylsulfonic acid, Phenylsulfonic acid, oxalic acid, phenylformic acid, succsinic acid.

Another object of the present invention has been to provide the preparation method of formula I compound, following reaction formula:

Specifically comprise the following steps:

(1) formula II compound reacts under alkali (consumption is 1～5 equivalent of formula II compound) exists with the bromo-2-butyne of 1-(their mol ratio is 1: 1.2), obtain formula III compound, the alkali adopting is selected from triethylamine, pyridine, diisopropylethylamine, potassium hydride KH, sodium hydride, salt of wormwood, sodium carbonate, sodium bicarbonate, saleratus; The reaction solvent adopting is selected from one or more mixed solvents in ethyl acetate, acetone, ethanol, methyl alcohol, methylene dichloride, tetrahydrofuran (THF), acetonitrile, DMF, methyl-sulphoxide, preferably DMF and methyl-sulphoxide; Temperature of reaction is 0 ℃ to 120 ℃, and preferably temperature is 0 ℃ to 50 ℃;

(2) formula III compound and 2-chloromethyl-4-methyl quinazoline (their mol ratio is 1: 1.1) carry out alkylated reaction under protection of inert gas, obtain formula IV compound, the alkali adopting is selected from potassium hydride KH, sodium hydride, salt of wormwood, sodium carbonate, sodium methylate, potassium methylate, sodium ethylate, potassium ethylate, sodium tert-butoxide, potassium tert.-butoxide, potassium hydroxide, sodium hydroxide; The catalyzer adopting is selected from lithium chloride, lithiumbromide, sodium iodide, potassiumiodide; The solvent adopting is selected from one or more mixed solvents in acetone, ethanol, methyl alcohol, the trimethyl carbinol, Virahol, tetrahydrofuran (THF), acetonitrile, water, DMF, methyl-sulphoxide; Temperature of reaction is 0 ℃ to 160 ℃, preferably 50 ℃ to 100 ℃;

(3) formula IV compound carries out substitution reaction with (R)-3-amino piperidine dihydrochloride (their mol ratio is 1: 1.1), obtain Compound I, the alkali adopting is selected from salt of wormwood, sodium carbonate, sodium bicarbonate, saleratus, sodium hydroxide, potassium hydroxide; The catalyzer adopting is selected from molecular sieve, diatomite, anhydrous magnesium sulfate, preferred molecular sieve; The solvent adopting is selected from one or more mixed solvents in acetone, propyl carbinol, n-propyl alcohol, ethanol, methyl alcohol, the trimethyl carbinol, Virahol, dioxane, tetrahydrofuran (THF), acetonitrile, water, DMF, methyl-sulphoxide; Temperature of reaction is 50 ℃ to 160 ℃, preferably 80 ℃ to 120 ℃;

Aforesaid method provided by the invention carrys out preparation I compound, and syntheti c route is brief, and raw material is cheap and easy to get, and technique is simple, is applicable to very much large-scale industrialization and produces.

Another object of the present invention has been to provide 1-(2-butyne-1-yl)-6-[3 (R)-amino piperidine-1-yl]-3-[(4-methyl quinazoline-2-yl) methyl]-2,4-(1H, 3H) pyrimidine dione is the application in preparation treatment diabetes medicament as DPP-4 inhibitor of formula I compound.

Pharmacodynamics and toxicologic study result show, it is active that formula I compound has superpower inhibition to DPP-4, higher than Egelieting and BI 1356, and shows significant hypoglycemic and increase the effect of insulin sensitivity.Toxicologic study result shows, formula I compound has good security, and toxic side effect is less than positive control drug.Can predict after the compounds of this invention exploitation patent medicine, its drug effect and security all will be better than Egelieting and BI 1356.

Accompanying drawing explanation

Fig. 1 is the proton nmr spectra of formula III compound;

Fig. 2 is the proton nmr spectra of formula IV compound;

Fig. 3 is the proton nmr spectra of formula I compound;

Fig. 4 is the L MALIC ACID salt proton nmr spectra of formula I compound.

Embodiment

Below by embodiment, illustrate content of the present invention.In the present invention, the embodiment of the following stated is in order better to set forth the present invention, is not for limiting the scope of the invention.Any simple modification, equivalent variations and modification that every foundation technical spirit of the present invention is done above embodiment, all fall into protection scope of the present invention.

Embodiment 1:1-(2-butyne-1-yl)-6-is chloro-2,4-(1H, 3H) pyrimidine dione (III) synthetic

By 6-chlorouracil (71g, 480mmol), add 200ml DMF, DIPEA (110ml, 624mmol), is cooled to 0 ℃, drip the bromo-2-butyne of 1-(48ml, 528mmol), stirred overnight at room temperature, add water 200ml to separate out solid, filter, washing, ethyl acetate is washed, be dried to obtain pale solid 81g, yield 85%.mp：216-217℃； ¹H?NMR(300MHz，CDCl ₃)δ9.36(s，1H)，5.90(s，1H)，4.75(s，2H)，1.81(s，3H).HRMS-ESI(m/z)calcd?for?C ₈H ₈N ₂O ₂Cl[M+H] ⁺：199.0274，found?199.0276

The chloro-3-[(4-methyl quinazoline-2-of embodiment 2:1-(2-butyne-1-yl)-6-yl) methyl]-2,4-(1H, 3H) pyrimidine dione (IV)

Get 1-(2-butyne-1-yl)-6-chloro-2,4-(1H, 3H) pyrimidine dione (III) (14.6g, 73.5mmol), 2-chloromethyl-4-methyl quinazoline (15.6g, 80.8mmol), lithiumbromide (5.69g, 58.8mmol), 60% sodium hydride (3.82g, 95.6mmol), adds after DMF 150ml, the lower 80 ℃ of reactions of nitrogen environment are spent the night, DMF is removed in underpressure distillation, adds ethyl acetate 100ml, 200ml isopropyl ether, be cooled to-20 ℃ and separate out solid, filter, ethyl acetate is washed, and isopropyl ether is washed, obtain faint yellow solid 21g, yield 80%.mp：153℃； ¹H?NMR(300MHz，CDCl ₃)δ8.08(d，J＝8.3Hz，1H)，7.94(d，J＝8.2Hz，1H)，7.85(ddd，J＝8.4，6.9，1.3Hz，1H)，7.60(ddd，J＝8.1，6.9，1.2Hz，1H)，6.12(s，1H)，5.54(s，2H)，4.89(dd，J＝4.5，2.2Hz，2H)，2.95(s，3H)，1.89(t，J＝2.3Hz，3H).HRMS-ESI(m/z)calcd?for?C ₁₈H ₁₆N ₄O ₂Cl[M+H] ⁺：355.0962，found?355.0965

Embodiment 3:1-(2-butyne-1-yl)-6-[3 (R)-amino piperidine-1-yl]-3-[(4-methyl quinazoline-2-yl) methyl]-2,4-(1H, 3H) pyrimidine dione (I)

Get the chloro-3-[(4-methyl quinazoline-2-of 1-(2-butyne-1-yl)-6-yl) methyl]-2,4-(1H, 3H) pyrimidine dione (IV) (8g, 22.5mmol), NaHCO ₃(9.45g, 112.5mmol), 4A molecular sieve (2.6g), (R)-3-amino piperidine dihydrochloride (4.67g, 27mmol), add 150ml Virahol, react 2 hours at 100 ℃, be cooled to room temperature, solids removed by filtration, concentrating under reduced pressure, column chromatography (methylene chloride/methanol/triethylamine 100: 0.5: 0.5) obtains faint yellow solid 8g, yield 85%. ¹H?NMR(300MHz，CDCl ₃)δ8.00(d，J＝8.2Hz，1H)，7.87(d，J＝8.3Hz，1H)，7.76(t，J＝7.5Hz，1H)，7.51(t，J＝7.5Hz，1H)，5.47(s，2H)，5.32(s，1H)，4.58(d，J＝1.9Hz，2H)，3.39(d，J＝10.9Hz，1H)，3.27(d，J＝11.7Hz，1H)，3.13-2.98(m，1H)，2.98-2.82(m，3H)，2.76(dd，J＝15.4，6.2Hz，1H)，2.65-2.47(m，1H)，2.05-1.86(m，5H)，1.80(s，3H)，1.76-1.62(m，1H)，1.41-1.20(m，1H).HRMS-ESI(m/z)calcd?for?C ₂₃H ₂₇N ₆O ₂[M+H] ⁺：419.2195，found?419.2199

Embodiment 4:1-(2-butyne-1-yl)-6-[3 (R)-amino piperidine-1-yl]-3-[(4-methyl quinazoline-2-yl) methyl]-2, the L MALIC ACID salt of 4-(1H, 3H) pyrimidine dione

Get 1-(2-butyne-1-yl)-6-[3 (R)-amino piperidine-1-yl]-3-[(4-methyl quinazoline-2-yl) methyl]-2,4-(1H, 3H) pyrimidine dione (I) (1g, 2.39mmol), L MALIC ACID (336mg, 2.5mmol), adds 20ml dissolve with ethanol, stirring at room, after 1 hour, separate out gradually solid, after stirring is spent the night, filter, ethanol is washed to obtain 1.08g yellow solid, yield 82%. ¹H?NMR(500MHz，DMSO)δ8.31-8.18(m，1H)，7.93(m，1H)，7.80(d，J＝8.3Hz，1H)，7.76-7.60(m，1H)，5.34(s，1H)，5.24(s，2H)，4.70-4.43(m，2H)，3.87(dd，J＝10.2，3.8Hz，1H)，3.32(d，J＝11.3Hz，2H)，3.15(s，1H)，3.01(s，1H)，2.91-2.78(m，4H)，2.51-2.48(dd，J＝15.5，J＝10.2，2H)2.32(dd，J＝15.5，3.8Hz，1H)，1.92(d，J＝22.7Hz，2H)，1.83-1.73(m，3H)，1.68(dd，J＝9.1，3.8Hz，1H)，1.56(s，1H).

Embodiment 5: external activity test experiments

For measuring DPP-4 inhibiting external test method active and testing compound, be known, by those of ordinary skills, can measure at an easy rate the protease inhibiting activity of DPP-4 inhibitor.

1 experiment purpose

Mensuration formula I compound is active to the inhibition of DPP-4 enzyme.

2 experiment materials

DPP-4 enzyme (Cat:SE434-9090, Biomol), GP-AMC (Cat:P189-9090, Biomol), black 96 orifice plates, super microplate reader, the analysis buffer of DPP-4 (50mM Tris, pH=7.5), AMC standard substance (7-amino-4-methylcoumarin) (Cat:KI-107, Biomol)

3 experimental techniques

(1) DPP-4 enzyme assay

96 hole blackboards are equilibrated to room temperature, use 1: 50 times of dilution 500 μ M AMC substrates of reaction buffer, the reaction final concentration that makes AMC substrate is 5 μ M, the substrate solution that each hole needs 50 μ l to dilute.To all contrast I hole, in DPP-4 sample well and contrast II hole, add respectively 50 μ l, the reaction buffer of 35 μ l and 85 μ l.Use 1: 50 times of dilution DPP-4 enzyme of reaction buffer, in all reacting holes except contrast I hole, the DPP-4 that adds 15 μ l to dilute, the enzyme amount that makes DPP-4 in reacting hole is 0.26MU/ hole.The AMC substrate solution that adds 50 μ l to dilute in all reacting holes except contrast II hole carrys out initial action.Continuous-reading Ex:380nm/Em:460nm in super microplate reader, the whole dynamic read time is 20 minutes, at interval of 1 minute reading 1 time.

(2) mensuration of AMC typical curve

96 hole blackboards are equilibrated to room temperature, use reaction buffer by 2 times of dilutions of AMC standard substance, the concentration obtaining is followed successively by 15 μ M, 7.5 μ M, 3.75 μ M, 1.875 μ M, 0.938 μ M, 0.469 μ M, 0.235 μ M, 0.117 μ M, 0 μ M.Then to the AMC standard substance that add successively different weaker concns in all reacting holes.To all reacting holes, add 35 μ l reaction buffers, use 1: 50 times of dilution DPP-4 enzyme of reaction buffer, to the DPP-4 that adds 15 μ l to dilute in all reacting holes, making DPP-4 enzyme amount in reacting hole is 0.26MU/ hole.Normal temperature was hatched after 5 minutes, reading Ex:380nm/Em:460nm in super microplate reader.

(3) DPP-4 inhibitor enzymic activity test

96 hole blackboards are equilibrated to room temperature, use 1: 50 times of dilution 500 μ M AMC substrates of reaction buffer, the reaction final concentration that makes AMC substrate is 5 μ M, the substrate solution that each hole needs 50 μ l to dilute.With reaction buffer dilution 3 testing compounds (the compounds of this invention, BI 1356, Egelieting), making its final screening concentration is that 10 μ M start, 4 times of gradient dilutions, and totally 10 concentration point, each extent of dilution is done a multiple hole.To the testing compound of all different concns on 96 hole Sptting plates, measure in hole and add 25 μ l reaction buffers, and add successively the compound of the different concns that 10 μ l have diluted in corresponding reacting hole.For negative control hole, add 25 μ l reaction buffers and 10 μ l P32/98, all reference compound holes and control wells are done a multiple hole.Use 1: 50 times of dilution DPP-4 enzyme of reaction buffer, in all reacting holes except blank hole, the DPP-4 that adds 15 μ l to dilute, making the DPP-4 enzyme amount in reacting hole is 0.26MU/ hole.To the AMC substrate solution that adds 50 μ l to dilute in all reacting holes, carry out initial action.Normal temperature was hatched after 10 minutes, reading in super microplate reader, Ex:380nm/Em:460nm.

(4) data analysis: use GraphPad-Prism software analysis

The calculating of formula I compound inhibiting rate: calculate the average signal value of each sample, the signal value of each concentration of specimens deducts average background signal value.Calculate the inhibiting rate of each sample, 100% active hole reading is deducted respectively after each testing compound different concns corresponding aperture reading, divided by 100% active hole reading, then be multiplied by 100 inhibiting rates that obtain respectively each testing compound different concns.Finally, the inhibiting rate calculating is mapped for different testing compound concentration.Inhibiting rate=[100% active hole-sample aperture]/100% active hole * 100.

4 experimental results and discussion

Experimental result is in Table 1, BI 1356 IC ₅₀=0.43nM, Egelieting IC ₅₀=2.6nM, formula I Compound I C ₅₀=0.27nM, its inhibition activity to DPP-4 is better than Egelieting and BI 1356 as seen.

Table 1 test compounds is active to the inhibition of DPP-4

Embodiment 6: activity in vivo test experiments

1 experiment purpose

Utilize mouse glucose tolerance model, relatively the blood sugar reducing function of the compounds of this invention and positive control drug BI 1356.

2 experiment materials

Kunming mice, male, 18-22g, is provided random packet by Yangzhou University's comparative medicine animal center.Credit number: SCXK (Soviet Union) 2007-0001; Reagent kit of glucose is purchased from graceful bio tech ltd, Witter, Nanjing.

3 experimental techniques

3.1 compounds affect normal glucose tolerance in mice

According to body weight, be divided at random 6 groups, 10 every group.Animal overnight fasting before administration, positive controls is 1mg/kg, the high, medium and low dosage group of test-compound is respectively 1mg/kg, 0.3mg/kg and 0.1mg/kg, and blank group and model group mouse are isopyknic solvent (0.5%CMCNa).After administration, blood sampling is designated as 0min at once.After administration, 45min is to each group gavage 2g/kg (5ml/kg) glucose solution.Then respectively in 30min, 60min, 120min blood sampling.By adopt the blood centrifuging and taking supernatant of each time point for Reagent kit of glucose measure blood-sugar content wherein.

3.2 compounds affect diabetic mice sugar tolerance

Set up diabetes model: mouse fasting be can't help after water spends the night, according to 75mg/kg dosage tail vein injection 2% tetraoxypyrimidine solution, after 7 days, gather mouse fasting blood and measure its serum blood-sugar content, with blood sugar >=11.0mmol/L and≤33.0mmol/L is considered as modeling success.After modeling success, sugar tolerance experiment is carried out in grouping, and method is tested with normal mouse sugar tolerance.

4 experimental results

4.1 compounds affect normal glucose tolerance in mice

Result shows, Normal group mouse gives after glucose, and 30min blood glucose value significantly raises, and 60min blood glucose value starts to decline.With Normal group comparison, positive drug group, the high, medium and low dosage group of test-compound are when 30min, and blood sugar significantly reduces (P < 0.01).Test-compound on the impact of normal mouse oral glucose tolerance in Table 2.

Table 2: the impact of test-compound on normal mouse oral glucose tolerance

4.2 impacts of test-compound successive administration on diabetic mice sugar tolerance

As can be seen from Table 3, test-compound successive administration 7 days, to diabetic mice sugar tolerance due to tetraoxypyrimidine at 30min, the tool effect of being significantly improved during 120min.

The table 3 test-compound successive administration 7 days impact on diabetic mice sugar tolerance

Embodiment 7: toxicological experiment

1 experiment purpose

According to the requirement of < < chemicals acute toxicity test technical director principle > >, observe the compounds of this invention I and once give acute toxic reaction and the death condition after ICR mouse.

2 experiment materials

Given the test agent: given the test agent compound is faint yellow powdery solid, normal temperature is preserved, and by China Medicine University's new drug research center, is provided, and uses 0.5%CMC-Na to dissolve, now with the current.Animal subject: ICR mouse, purchased from Yangzhou University's comparative medicine center Experimental Animal Center, laboratory animal production licence: SCXK (Soviet Union) 2007-0001; Age in days: 6-8w; Body weight: 18-22g; Sex: male and female half and half; Number of animals: 10 every group, totally 80.

3 experimental techniques

Preliminary experiment and result

With given the test agent 4g/kg, 2g/kg, 1g/kg, give 4 male ICR mouse gavages respectively.Experimental result shows: dead 3 of 4g/kg, and dead 1 of 2g/kg, 1g/kg does not have death.

Anxious poison experiment group and dosage

According to preliminary experiment result, divide into groups as follows: with agent distance 0.7,5 dosage are set, are respectively 4g/kg, 3g/kg, 2.25g/kg, 1.69g/kg, 1.27g/kg.

Experiment grouping and observation

During test, animal is divided into 10 groups at random, 10 every group, animal fasting 12h before administration, by body weight, give gavage, after administration, observe mouse skin, eye, mucous membrane, activity, ingest, drinking-water, defecation, respiratory variations and poisoning manifestations and death condition totally 7 days.

4 results

Formula I compound gives mouse a gavage, and 4g/kg organizes all mouse and occurs the tic of faintness property, and miosis is prostrate motionless, after about 5min, twitches and disappears, but still have, pant, and prostrate motionless phenomenon, interior dead 8 of 5-30min, 10 all death of 24h; 3g/kg group, all mouse occur, and the phenomenon of panting is prostrate motionless, wherein has 4 with faintness, to twitch, and 5 death in 30min, during 48h dead 7 altogether, are survived 3 after 7 days by during 24h dead 7; 2.25g/kg group, there is the phenomenon of panting in all mouse, prostrate motionless, wherein has 2 with faintness, to twitch, and 4 death in 30min, during 48h dead 6 altogether, during 72h dead 6 altogether, are survived 4 after 7 days by during 24h dead 6; After 1.69g/kg administration, interior dead 4 24h of 30min interior dead 7, after 7 days, survive 3,1.27g/kg, and toxicity reaction is not obvious, dead 3 of 24h.According to above-mentioned result of study the BI 1356 toxicological experiment data that provide in conjunction with FDA, show, the acute toxicity of formula I compound is less than BI 1356.

Embodiment 7: the coated tablet that contains 5mg formula I compound

1 tablet core core contains

Formula I compound 5mg HPMC 15mg

Calcium phosphate 90mg Magnesium Stearate 1.5mg

W-Gum 35mg Polyvinylpyrolidone (PVP) 10mg

Total amount 166.5mg

Preparation:

Half magnesium stearate of formula I compound and calcium phosphate, W-Gum, Polyvinylpyrolidone (PVP), HPMC and specified amount is mixed.In pelleter, manufacture the sheet of 13 mm dias, then use suitable machine, make its friction through thering is the screen cloth of 1.5 millimeters of size of mesh, and mix with all the other Magnesium Stearates.In pelleter, suppress this particle, to form, wanted the tablet of shape.

Core core weight: 166.5mg drift: 9 millimeters, convex

The tablet core core of so making is used to the film coating substantially being formed by HPMC.By the thin membrane coated tablet finally completing with beeswax polishing.

The weight of coated tablet: 175mg.

Claims (6)

1. the 1-shown in formula I (2-butyne-1-yl)-6-[3 (R)-amino piperidine-1-yl]-3-[(4-methyl quinazoline-2-yl) methyl]-2,4-(1H, 3H) pyrimidine dione and pharmacy acceptable salt thereof:

2. a preparation method for formula I compound described in claim 1, following reaction formula:

Comprise the following steps::

(1) formula II compound reacts under alkali exists with the bromo-2-butyne of 1-, obtains formula III compound, and the alkali adopting is selected from triethylamine, pyridine, diisopropylethylamine, potassium hydride KH, sodium hydride, salt of wormwood, sodium carbonate, sodium bicarbonate, saleratus; The reaction solvent adopting is selected from one or more mixed solvents in ethyl acetate, acetone, ethanol, methyl alcohol, methylene dichloride, tetrahydrofuran (THF), acetonitrile, DMF, methyl-sulphoxide, preferably DMF and methyl-sulphoxide; Temperature of reaction is 0 ℃ to 120 ℃, and preferably temperature is 0 ℃ to 50 ℃;

(2) formula III compound and 2-chloromethyl-4-methyl quinazoline carry out alkylated reaction under protection of inert gas, obtain formula IV compound, the alkali adopting is selected from potassium hydride KH, sodium hydride, salt of wormwood, sodium carbonate, sodium methylate, potassium methylate, sodium ethylate, potassium ethylate, sodium tert-butoxide, potassium tert.-butoxide, potassium hydroxide, sodium hydroxide; The catalyzer adopting is selected from lithium chloride, lithiumbromide, sodium iodide, potassiumiodide; The solvent adopting is selected from one or more mixed solvents in acetone, ethanol, methyl alcohol, the trimethyl carbinol, Virahol, tetrahydrofuran (THF), acetonitrile, water, DMF, methyl-sulphoxide; Temperature of reaction is 0 ℃ to 160 ℃, preferably 50 ℃ to 100 ℃;

(3) formula IV compound carries out substitution reaction with (R)-3-amino piperidine dihydrochloride, obtains formula I compound, and the alkali adopting is selected from salt of wormwood, sodium carbonate, sodium bicarbonate, saleratus, sodium hydroxide, potassium hydroxide; The catalyzer adopting is selected from molecular sieve, diatomite, anhydrous magnesium sulfate, preferred molecular sieve; The solvent adopting is selected from one or more mixed solvents in acetone, propyl carbinol, n-propyl alcohol, ethanol, methyl alcohol, the trimethyl carbinol, Virahol, dioxane, tetrahydrofuran (THF), acetonitrile, water, DMF, methyl-sulphoxide; Temperature of reaction is 50 ℃ to 160 ℃, preferably 80 ℃ to 120 ℃.

3. formula I compound or its salt benefits from the purposes in the medicine of the disease that DPP-4 suppresses in preparation treatment or prevention as claimed in claim 1, described benefits from that the disease that DPP-4 suppresses is selected from diabetes, diabetic ester blood is abnormal, glucose tolerance lowers (IGT) disease, fasting plasma glucose lowers (IFG) disease, metabolic acidosis, ketosis, appetite stimulator, obesity, various cancer, nervous system disorders, disease of immune system.

4. as claimed in claim 3, wherein diabetes are diabetes Bs.

5. a pharmaceutical composition, it comprises formula I compound or its pharmacy acceptable salt described in claim 1, and one or more pharmaceutically acceptable auxiliary materials.

6. a pharmaceutical solid preparation, it comprises formula I compound or its pharmacy acceptable salt described in claim 1, and the normally used additive of one or more field of pharmaceutical preparations.

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