CN103044392A - Preparation method of high-efficient DPP (dipeptidyl peptidase)-IV inhibitor - Google Patents

Abstract

The invention belongs to the technical field of medicines, relates to a preparation method of a high-efficient DPP (dipeptidyl peptidase)-IV inhibitor, and specifically relates to a compound taking uracil as a mother nucleus, or a pharmaceutically acceptable salt thereof, a preparation method, a composition and an application of the compound as the DPP-IV inhibitor in prevention or treatment of diseases which benefit from DPP-IV inhibition. The compound disclosed by the invention has a very good selective inhibition effect against DPP-IV and has very low cardiotoxicity, thereby being a DPP-IV inhibitor type medicament with great prospects.

Description

A kind of preparation method of efficient DPP-IV inhibitor

Technical field

The invention belongs to medical technical field, relate to particularly a kind of compound or pharmaceutically acceptable salt thereof, its preparation method, composition and this compounds take uracil as parent nucleus and benefit from purposes in the disease that DPP-IV suppresses as dipeptidyl peptidase (DPP-IV) inhibitor in prevention or treatment.

Background technology

Diabetes are because the absolute or relative deficiency of Regular Insulin causes blood sugar increasing, thereby cause serious complication, finally cause disabling or causing death of patient.Clinically diabetes are divided into I type and II type.Type i diabetes is because pancreaticβ-cell is destroyed, lacks insulin secretion, thereby causes blood sugar increasing, and this class patient can only depend on exogenous insulin; Type ii diabetes is that its sickness rate accounts for more than 90% of all diabetic subject's numbers owing to insulin secretion relative deficiency or the unsound initiation hyperglycemia of insulin action link.Present drug research mainly is aimed at the type ii diabetes expansion.

Traditional ofhypoglycemic medicine is of a great variety, mainly is divided into euglycemic agent (such as biguanides, thiazolidinediones etc.) and Regular Insulin succagoga (such as sulfonylurea and non-sulphonyl class medicine), etc.The problem of the toxic side effect such as but these medicines can not stop the deterioration of diabetes, and exist the body weight increase, hypoglycemia and the final forfeiture of drug effect.Thereby the antidiabetic medicine of development of new overcomes above many deficiencies, and it is extremely urgent task that prevention even reverse disease worsen.

Dipeptidyl peptidase (DPP-IV) is a kind of glycoprotein that is distributed widely in the human body, and its function class is similar to serine protease, makes its inactivation by the shearing to polypeptide, thereby reaches the effect of regulation of physiological functions.DPP-IV is constant to the shearing position of substrate, is proline(Pro) or the L-Ala of its N end penultimate.Glucagon-like-peptide-1 (GLP-1) is a kind of endogenic hormone, along with the rising of postprandial blood sugar, is produced by the L emiocytosis in the small intestine, and then stimulates the secretion of Regular Insulin.Therefore, the intake of the secretion of GLP-1 and blood sugar is closely related.Can effectively control blood sugar and do not put on weight based on the treatment plan of GLP-1, can not produce the untoward reactions such as hypoglycemia.But GLP-1 is as the substrate of DPP-IV, and the transformation period is very short, and secretion will be sheared rapidly by DPP-IV within rear 1-2 minute, inactivation.Therefore can adopt the strategy of two kinds of new drug developments based on the mechanism of action of GLP-1: the GLP-1 analogue of exploitation DPP-IV tolerance and exploitation DPP-IV inhibitor.The inventor finds that according to this thinking the uracil compound is a kind of effective DPP-IV inhibitor, and effectively lowering blood glucose does not cause that body weight increases and the hypoglycemia equivalent risk simultaneously, and finishes the present invention based on this.

Summary of the invention

One aspect of the present invention provides the compound or pharmaceutically acceptable salt thereof of a kind of formula I

Wherein, R ₁Choose generation or unsubstituted alkyl, cycloalkyl, aryl, heteroaryl or Heterocyclylalkyl; Substituting group is selected from alkyl, alkoxyl group, halogen, cyano group, amino, hydroxyl, nitro, carbonyl, alkylsulfonyl alkyl, amido, carbonylic alkyl, aryl, aryloxy, Heterocyclylalkyl, heteroaryl, heteroaryloxy, cycloalkyl, cycloalkylalkyl, alkylsulfonyl or sulfinyl; R ₁The preferred replacement or unsubstituted aryl or heteroaryl;

R ₂Be selected from:

(1) hydrogen;

(2) cyano group;

(3) alkyl that does not replace or replace;

(4) do not replace or by 1-5 the phenyl that replace of substituting group independently separately, substituting group is selected from halogen, cyano group, OH, alkyl, alkoxyl group, NHSO ₂R ₃, N (alkyl) SO ₂R ₃, SO ₂R ₃, SO ₂NR ₄R ₅, NR ₄R ₅, CONR ₄R ₅, COOH and carboxyalkyl;

(5)OH；

(6) alkoxyl group;

(7)NR ₄R ₅；

R ₂Preferred hydrogen, alkyl or alkoxyl group;

R ₃Be selected from and replace or unsubstituted alkyl, substituting group is selected from 1-5 separately independently halogen or COOH;

R ₄And R ₅Individually independently be selected from separately:

(1) hydrogen,

(2) phenyl that does not replace or replace, substituting group is selected from halogen, OH, alkyl or alkoxyl group,

(3) do not replace or by the C that replaces of substituting group independently separately _3-6Cycloalkyl, substituting group are selected from halogen, OH, alkyl or alkoxyl group,

(4) alkyl that does not replace or replace, substituting group is selected from:

(a) halogen, or

(b) do not replace or by 1-5 the phenyl that replace of substituting group independently separately, substituting group is selected from halogen, OH, alkyl or alkoxyl group.

Formula I compound of the present invention, R ₁The preferred replacement or unsubstituted aryl or heteroaryl,, substituting group is selected from alkyl, alkoxyl group, halogen, cyano group, amino, hydroxyl, nitro or carbonyl;

R ₂Preferred hydrogen, alkyl or alkoxyl group, more preferably hydrogen.

For example formula I compound provided by the invention is preferred, R ₁Be selected from replacement or unsubstituted aryl or heteroaryl, substituting group is selected from alkyl, alkoxy or halogen, R ₂Be selected from hydrogen.

R ₁Further preferably replace or unsubstituted phenyl, quinazolyl, benzimidazolyl-, quinolyl, pyrimidyl, indyl, quinoxalinyl, isoquinoline 99.9 or aza-phenanthrenes.

R for example ₁Be selected from phenyl, 4-methyl quinazoline-2 base, benzimidazolyl-2 radicals-Ji, quinoline-2-base, 6-bromoquinoline-2-base, 6-chloroquinoline-2-base, 6-fluorine quinoline-2-base, 6-toluquinoline-2-base, 7-chloroquinoline-2-base, 7-fluorine quinoline-2-base, 6-methoxy quinoline-2-base, 4-chloroquinoline-2-base, 3-methyl-quinoxaline-2-base, quinolyl-4, quinoline-3-base, pyrimidine-2-base, indol-3-yl, quinoxaline-2-base, isoquinolyl-1 or 1-aza-phenanthrenes-2-base, R ₂Be selected from hydrogen.

Term " alkyl " refers to the saturated aliphatic hydrocarbon group of the straight or branched that is comprised of carbon atom and hydrogen atom, and it is connected with the rest part of molecule by singly-bound, and it has 1-6 carbon atom usually also can be expressed as C _1-6Alkyl preferably has the C of 1-4 carbon atom _1-4Alkyl.Described alkyl can be non-substituted or be replaced by one or more substituting groups that are selected from alkyl, alkoxyl group, aryl, halogen, amino, hydroxyl, nitro or carboxyl etc.The limiting examples of non-substituted alkyl includes but not limited to such as methyl, ethyl, propyl group, sec.-propyl, normal-butyl, isobutyl-, tert-butyl, just-amyl group, 2-methyl butyl, neo-pentyl, n-hexyl or 2-methyl hexyl etc.

Term " alkoxyl group " refers to have that alkyl replaces contains the oxygen part, namely-the O-alkyl group, usually formed by oxygen and the alkyl that contains 1-6 carbon atom, namely-O-C _1-6Alkyl, preferred-O-C _1-4Alkyl, concrete example include but not limited to methoxyl group, oxyethyl group, propoxy-, isopropoxy, butoxy, isobutoxy, tert.-butoxy, n-pentyloxy, 2-methyl butoxy, neopentyl oxygen, positive hexyloxy or 2-methyl hexyloxy etc.

Term " Heterocyclylalkyl " refers to heteroatomic five yuan or hexa-atomic replacement or unsubstituted monocycle non-aromatic cyclic groups such as one or two nitrogen, oxygen, sulphur, concrete example includes but not limited to piperazine, piperidines, Pyrrolidine or morpholine etc., and substituting group can be selected from cyano group, halogen, hydroxyl, amino or amido etc.

Term " aryl " refers to contain full carbon monocycle or the bicyclic carbocyclic aromatic nucleus system of 6-10 carbon atom, has the π-electron system of total conjugated, the limiting examples of aryl such as phenyl, xenyl or naphthyl etc.Aryl can be that replace or unsubstituted, and substituting group is selected from alkyl, alkoxyl group, aryl, halogen, amino, hydroxyl, nitro or carboxyl etc.

Term " heteroaryl " refers to monocycle, dicyclo or the three cyclophane bases of 5-14 atom, and it contains 1 heteroatoms that is selected from N, O or S at least, and remaining atom is C.Heteroaryl can be that replace or unsubstituted, and substituting group is selected from alkyl, alkoxyl group, aryl, hydroxyl or amino etc.The limiting examples of unsubstituted heteroaryl such as pyrroles, furans, thiophene, imidazoles, oxazole, pyrazoles, pyridine, pyrimidine, indoles, quinoline, isoquinoline 99.9, oxazoline, benzoglyoxaline, quinoxaline, aza-phenanthrenes or quinazoline etc.

" halogen " refers to fluorine, chlorine, bromine or iodine to term.

Term " amino " refers to-the NH2 group ,-NH (alkyl) group or-N (alkyl) ₂Amino object lesson includes but not limited to-NH ₂,-NHCH ₃,-N (CH ₃) ₂,-NHC ₂H ₅Or-N (C ₂H ₅) ₂Deng.

Term " alkylsulfonyl alkyl " refers to the alkyl of the straight or branched that contains 1-6 carbon atom that replaced by alkylsulfonyl; such as the alkylsulfonyl methyl; alkylsulfonyl ethyl or 1-alkylsulfonyl-2-methylethyl etc.; alkyl can be that replace or unsubstituted, and substituting group is selected from alkyl, alkoxyl group, aryl, halogen, amino, hydroxyl, nitro or carboxyl etc.

Term " cycloalkyl " refers to contain the saturated cyclic hydrocarbons of 3-6 carbon atom, include but not limited to cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, cycloalkyl can be that replace or unsubstituted, and substituting group is selected from alkyl, alkoxyl group, aryl, halogen, amino, hydroxyl, carboxyl, amido or cyano group etc.

Term " cycloalkylalkyl " refers to the straight or branched alkyl group that contains 1-6 carbon atom that is substituted by cycloalkyl, as encircles the third methyl, ethyl cyclopentane or 1-cyclohexyl-3-ethyl-butyl etc.

The unsaturated alkyl that term " thiazolinyl " refers to contain 2-6 carbon atom and contains at least the straight or branched of 1 two key, thiazolinyl can be non-substituted or selectedly to replace from following substituting group: alkyl, alkoxyl group, hydroxyl, amino or halogen etc., the object lesson of non-substituted thiazolinyl include but not limited to vinyl, propenyl, 2-propenyl, 1-butylene base, crotyl, 1-pentenyl or 1-hexenyl etc.

Term " alkynyl " refers to contain the straight or branched unsaturated alkyl of 2-6 carbon atom and at least 1 triple bond, alkynyl can be non-substituted or selectedly to replace from following substituting group: alkyl, alkoxyl group, hydroxyl, amino or halogen etc., the object lesson of non-substituted alkynyl is such as including but not limited to ethynyl, 1-proyl, 2-propynyl, ethyl acetylene base or 2-butyne base etc.

Term " carboxyl " refers to contain-the straight or branched group of the 1-7 carbon atom of COO-group, include but not limited to HOOC-, CH ₃OOC-or CH ₃CH ₂OOC-etc.

Particular compound provided by the present invention is exemplified below, but is not limited to following compounds or its pharmacologically acceptable salt:

Further aspect of the present invention provides the formula I preparation method of compound, comprises the steps:

(1) formula A compound and formula B compound react production C compound in organic solvent in the presence of alkali:

(2) formula C compound and R-3-amino piperidine dihydrochloride reaction production I compound:

Wherein, R ₁, R ₂Identical with the definition in the above-mentioned formula I compound, Hal is chlorine or bromine.

In the step (1), alkali is selected from sodium hydride, salt of wormwood or yellow soda ash; Preferred sodium hydride; Organic solvent is selected from the mixed solvent of glycol dimethyl ether and DMF.

In a specific embodiment of the present invention, reaction method is exemplified as:

(1) mixed solvent of alkali with glycol dimethyl ether and DMF mixed 0 ℃ of lower stirring 10 minutes, adding formula A compound, 0 ℃ of lower stirring 20 minutes adds anhydrous lithium bromide, stirring at room 30 minutes adds formula B compound, stirred overnight at room temperature.React complete, add trash ice, dichloromethane extraction, dry, concentrated after the water washing of gained organic phase usefulness saturated common salt, namely obtain formula C compound behind the column chromatography purification;

(2) formula C compound is mixed the rear ethanol that adds with R-3-amino piperidine dihydrochloride, mix rear adding sodium bicarbonate, oil bath reflux 6 hours.React complete, reaction solution is concentrated, and resistates namely obtains formula I compound through column chromatography purification.

Wherein, the alkali described in the step (1) is selected from sodium hydride, salt of wormwood or yellow soda ash; Preferred sodium hydride; Glycol dimethyl ether and DMF are anhydrous solvent, and the two volume ratio is 2: 1.

Wherein, formula A compound can adopt method commonly used in this area to prepare.Such as, work as R ₂During for H, formula A can prepare from following method:

The concrete preparation method of formula A compound is exemplified as:

Add DMF (DMF) in the 6-chlorourea pyrimidine, add DIPEA (DIEA) after the dissolving, the rear dropping 1-bromo-2-butyne that stirs, 25 ℃ of stirrings of reaction solution are spent the night.Add entry after reacting completely, separate out solid, suction filtration, filter cake drying after water, ether are washed had both got target compound.

Wherein, 6-chlorourea pyrimidine and 1-bromo-2-butyne mol ratio are 1: 1.05-1: 2, and preferred 1: 1.1; Reaction times is 3-36 hour, preferred 24 hours; Temperature of reaction 10-40 ℃, preferred 25 ℃.

Formula B compound can have been bought from the market or adopt method commonly used in this area to synthesize and obtain, and is exemplified below without limitation:

Concrete reaction method is exemplified as: add Diisopropyl azodicarboxylate (AIBN) after methyl compound unsubstituted or that replace mixes with N-bromo-succinimide (NBS), add tetracol phenixin, the oil bath reacting by heating.React complete, the filtering insolubles, filtrate is spin-dried for by column chromatography purification and namely obtains target compound.

Wherein, methyl compound and N-bromo-succinimide mol ratio unsubstituted or that replace are 1: 1-1: 2, and preferred 1: 1; Reaction times is 1-12 hour, preferred 6 hours; Temperature of reaction 50-80 ℃, preferred 80 ℃.Methyl compound and Diisopropyl azodicarboxylate mol ratio unsubstituted or that replace are 1: 0.01-1: 0.1, and preferred 1: 0.02.

In the specific embodiment of the present invention, the preparation method is exemplified below:

(1) formula A ' compound and formula B compound react production C ' compound in the mixed solvent of glycol dimethyl ether and DMF in the presence of sodium hydride:

(2) formula C ' compound and R-3-amino piperidine dihydrochloride reaction production I ' compound:

Wherein, R ₁Identical with the definition in the above-mentioned formula I compound, Hal is chlorine or bromine.

In a specific embodiment of the present invention, reaction method is exemplified as:

(1) sodium hydride is mixed 0 ℃ of lower stirring 10 minutes with the mixed solvent of glycol dimethyl ether/DMF, adding formula A ' compound, 0 ℃ of lower stirring 20 minutes adds anhydrous lithium bromide, stirring at room 30 minutes adds formula B compound, stirred overnight at room temperature.React complete, add trash ice, dichloromethane extraction, dry, concentrated after the water washing of gained organic phase usefulness saturated common salt, namely obtain formula C ' compound behind the column chromatography purification.

(2) formula C ' compound is mixed the rear ethanol that adds with R-3-amino piperidine dihydrochloride, mix rear adding sodium bicarbonate, oil bath reflux 6 hours.React complete, reaction solution is concentrated, and resistates namely obtains formula I ' compound through column chromatography purification.

Wherein, glycol dimethyl ether and DMF are anhydrous solvent, and the two volume ratio is 2: 1.

Uracil compound provided by the invention can form free with it or salt exist, when the compounds of this invention possesses the form of free alkali, make free alkali form and the pharmaceutically acceptable inorganic or organic acid reaction of compound, the acid salt that can prepare the compounds of this invention, these salt include but not limited to: hydrochloride, hydrobromate, hydriodate, phosphoric acid salt, vitriol, nitrate, esilate, tosylate and benzene sulfonate, acetate, maleate, tartrate, succinate, Citrate trianion, benzoate, ascorbate salt and salicylate, malonate, adipate, hexanoate, arginic acid salt, fumarate, nicotinate, phthalate, oxalate etc.

Further aspect of the present invention provides the purposes of formula I compound in the medicine for preparing the disease for the treatment of or prevent to benefit from the DPP-IV inhibition.The disease that the described DPP-IV of benefiting from suppresses is selected from type ii diabetes, diabetic dyslipidaemia, glucose tolerance attenuating (IGT) disease, fasting plasma glucose attenuating (IFG) disease, metabolic acidosis, ketosis, appetite stimulator, obesity, various cancer, neurological conditions, disorder of immune system etc., preferably include type ii diabetes and obesity, more preferably comprise type ii diabetes.

Further aspect of the present invention provides a kind of pharmaceutical composition, comprises formula I compound or pharmaceutically acceptable salt thereof of the present invention and one or more pharmaceutically acceptable auxiliary materials.Composition of the present invention can be liquid, semiliquid or solid form, prepares according to the mode that is suitable for used route of administration.Composition of the present invention can be according to following administering mode administration: in oral, parenteral, intraperitoneal, intravenously, transdermal, hypogloeeis, intramuscular, rectum, oral cavity, the nose, the mode such as liposome.

Oral compositions can be solid, gel or liquid.The example of solid preparation includes but not limited to tablet, capsule, granule and pulvis in bulk.These preparations can selectively contain tackiness agent, thinner, disintegrating agent, lubricant, glidant, sweeting agent and correctives etc.The example of tackiness agent includes but not limited to Microcrystalline Cellulose, glucose solution, mucialga of arabic gummy, gelatin solution, sucrose and starch paste; The example of lubricant includes but not limited to talcum, starch, Magnesium Stearate, calcium stearate, stearic acid; The example of thinner includes but not limited to lactose, sucrose, starch, mannitol, Si Liaodengji dicalcium phosphate feed grade; The example of glidant includes but not limited to silicon-dioxide; The example of disintegrating agent includes but not limited to croscarmellose sodium, primojel, alginic acid, W-Gum, yam starch, methylcellulose gum, agar and carboxymethyl cellulose.

Give the present composition with parenteral, generally take injection as main, comprise subcutaneous, intramuscular or intravenous injection.Injection can be made into any conventionally form, such as liquor or suspension, be suitable for being dissolved or suspended in solid form or emulsion in the liquid before injection.The example that can be used for the pharmaceutically receivable carrier of injection of the present invention includes but not limited to aqueous carrier, non-aqueous carrier, biocide, isotonic agent, buffer reagent, oxidation inhibitor, suspension and dispersion agent, emulsifying agent, sequestrant and other pharmaceutically acceptable material.The example of aqueous carrier comprise sodium chloride injection, Lin Geshi injection liquid, etc. ooze glucose injection, sterilized water injection liquid, glucose and lactic acid ringer's inj; The example of non-aqueous carrier comprises fixedly oil, Oleum Gossypii semen, Semen Maydis oil, sesame oil and the peanut oil of plant origin; The example of biocide comprises meta-cresol, benzylalcohol, butylene-chlorohydrin, benzalkonium chloride etc.; The example of isotonic agent comprises sodium-chlor and glucose; Buffer reagent comprises phosphoric acid salt and Citrate trianion.

The present composition can also be prepared into aseptic lyophilized injectable powder, compound is dissolved in buffer solution of sodium phosphate, wherein contain glucose or other vehicle that is fit to, subsequently under standard conditions well known by persons skilled in the art with solution sterile filtration, succeeded by lyophilize, obtain required preparation.

Above-mentioned heterocycle hepyramine compounds preparation technology provided by the invention is simple, raw material is easy to get, be fit to large-scale industrialization production, and experimental verification in external and body, the compounds of this invention has extraordinary selective inhibitory to DPP-IV, in establishment DPP-IV activity, on the almost not impact of activity of DPP-VIII and DPP-IX, toxicity will be lower after can predicting the compounds of this invention exploitation patent medicine, have extraordinary application prospect.

Embodiment

Compound provided by the invention can synthesize by multiple preparation method, and the exemplary process of synthetic these compounds only is provided among the embodiment.Here be noted that free acid and/or alkali form regardless of the compounds of this invention of developing in which way, or the form of salt, scope of the present invention all belonged to.The purpose of specific embodiment is to further specify content of the present invention but do not mean that to limit the invention.

The initial feed of using in the specific embodiment of the invention, reaction reagent etc. are the commercially available prod.

The reagent abbreviation letter that uses among the present invention is this area phraseology commonly used, and implication is as follows:

DME: glycol dimethyl ether

DMF:N, dinethylformamide

DIEA:N, the N-diisopropylethylamine

Synthesizing of embodiment 1. compounds 4

Compound 4-0's is synthetic:

In the 500mL eggplant-shape bottle, add 6-chlorourea pyrimidine (15g, 102.4mmol) and 250mL DMF, add 15mL DIEA after the dissolving, the rear dropping 1-bromo-2-butyne (9.85mL, 112.64mmol) that stirs, 25 ℃ of stirrings of reaction solution are spent the night.React completely, add frozen water, suction filtration, filter cake dry after water, ether are washed and are target compound.

1H NMR(400MHz，CDCl ₃)δ8.85(s，1H)，5.91(s，1H)，4.75(d，J＝2.0Hz，2H)，1.82(t，J＝2.4Hz，3H).MS 199.82[M+H]+.

Compound 4-1's is synthetic:

Under the nitrogen protection; in the 100mL eggplant-shape bottle, add 60%NaH (54mg; 3.78mmol); 0 ℃ of lower anhydrous DME/DMF (2: 1) solution that adds; 0 ℃ was stirred after 10 minutes; the anhydrous DME/DMF of 10ml (2: the 1) solution that adds 1-(2-butyne)-6-chlorourea pyrimidine (500mg, 2.52mmol).Finish, 0 ℃ of stirring added anhydrous lithium bromide (263mg, 3.02mmol) after 20 minutes, and stirring at room adds 2-chloromethyl-4-methyl quinazoline (534mg, 2.77mmol), reaction solution stirred overnight at room temperature after 30 minutes.React completely, add trash ice, dichloromethane extraction, use the saturated common salt water washing after merging organic phase, organic phase anhydrous sodium sulfate drying after the washing filters, and filtrate decompression is concentrated, resistates namely obtains compound 4-1 with silica gel column chromatography (petrol ether/ethyl acetate=1/1 wash-out, UV colour developing) purifying.

¹H NMR(400MHz，CDCl ₃)δ8.03(d，J＝8.0Hz，1H)，7.89(d，J＝8.0Hz，1H)，7.80(t，J＝7.2Hz，1H)，7.55(t，J＝8.0Hz，1H)，6.06(s，1H)，5.47(s，2H)，4.83(d，J＝2.0Hz，2H)，2.90(s，3H)，1.83(t，J＝2.4Hz，3H).MS 355.80[M+H] ⁺.

Synthesizing of compound 4:

In the 50mL eggplant-shape bottle, add compound 4-1 (306mg, 0.86mmol), sodium bicarbonate (361mg, 4.30mmol), R-3-amino piperidine dihydrochloride (224mg, 1.29mmol) and 25mL ethanol, oil bath was heated to return stirring 6 hours.React completely, the reaction solution concentrating under reduced pressure, resistates namely obtains compound 4 with silica gel column chromatography (methylene chloride/methanol=15/1+1% ammoniacal liquor wash-out, UV colour developing) purifying.

¹H NMR(400MHz，CDCl ₃)δ7.94(d，J＝8.0Hz，1H)，7.81(d，J＝8.4Hz，1H)，7.70(t，J＝7.2Hz，1H)，7.45(t，J＝7.2Hz，1H)，5.41(s，2H)，5.26(s，1H)，4.52(d，J＝1.6Hz，2H)，3.33(m，1H)，3.22(m，1H)，3.00(m，1H)，2.81(s，3H)，2.72(m，1H)，2.50(m，1H)，1.92(m，1H)，1.82(m，1H)，1.74(s，3H)，1.64(m，1H)，1.26(m，1H).MS 419.50[M+H] ⁺.

Synthesizing of embodiment 2. compounds 3

Compound 3-1's is synthetic:

Take cylite as raw material, the synthetic method of compound 4-1 in the reference example 1 prepares compound 3-1.

¹H NMR(400MHz，CDCl ₃)δ7.43-7.19(m，5H)，5.97(s，1H)，5.42(s，2H)，4.60(s，2H)，1.77(s，3H).MS 289.07[M+H] ⁺.

Synthesizing of compound 3:

The compound 3-1 that obtains in the previous step is as raw material, and the synthetic method of compound 4 in the reference example 1 prepares compound 3.

¹H NMR(400MHz，CDCl ₃)δ7.46-7.22(m，5H)，5.33(s，2H)，5.20(s，1H)，4.40(s，2H)，3.22(m，3H)，2.96(m，1H)，2.62(m，1H)，2.42(m，1H)，1.90(m，1H)，1.77(s，3H)，1.60(m，1H)，1.22(m，1H).MS 353.2[M+H] ⁺.

Synthesizing of embodiment 3. compounds 5

Compound 5-1's is synthetic:

Take the 2-chloromethyl benzimidazole as raw material, the synthetic method of compound 4-1 in the reference example 1 prepares compound 5-1.

¹H NMR(400MHz，CDCl ₃)δ7.58(m，2H)，7.23(m，2H)，5.99(s，1H)，5.40(s，2H)，4.75(d，J＝2.4Hz，2H)，1.79(t，J＝2.4Hz，3H).MS 329.10[M+H] ⁺.

Synthesizing of compound 5:

The compound 5-1 that obtains in the previous step is as raw material, and the synthetic method of compound 4 in the reference example 1 prepares compound 5.

¹H NMR(400MHz，CDCl ₃)δ7.47(m，2H)，7.14(m，2H)，5.31(s，2H)，5.18(s，1H)，4.40(s，2H)，3.21(m，3H)，2.95(m，1H)，2.61(m，1H)，2.43(m，1H)，1.92(m，1H)，1.75(s，3H)，1.61(m，1H)，1.21(m，1H).MS 393.45[M+H] ⁺.

Synthesizing of embodiment 4. compounds 6

Compound 6-1's is synthetic:

Take the 2-chloromethyl quinoline as raw material, the synthetic method of compound 4-1 in the reference example 1 prepares compound 6-1.

¹H NMR(400MHz，CDCl ₃)δ8.10(d，J＝8.4Hz，1H)，8.00(d，J＝8.4Hz，1H)，7.77(d，J＝8.4Hz，1H)，7.65(m，1H)，7.48(m，1H)，7.32(d，J＝8.4Hz，1H)，6.05(s，1H)，5.43(s，2H)，4.81(m，2H)，1.82(t，J＝2.4Hz，3H).MS 340.90[M+H] ⁺.

Synthesizing of compound 6:

The compound 6-1 that obtains in the previous step is as raw material, and the synthetic method of compound 4 in the reference example 1 prepares compound 6.

¹H NMR(400MHz，CDCl ₃)δ8.05(d，J＝8.8Hz，1H)，8.00(d，J＝8.8Hz，1H)，7.73(d，J＝8.0Hz，1H)，7.62(t，J＝7.2Hz，1H)，7.44(t，J＝7.2Hz，1H)，7.29(t，J＝8.4Hz，1H)，5.40(s，2H)，5.28(s，1H)，4.53(d，J＝2.4Hz，2H)，3.34(m，1H)，3.23(m，1H)，3.00(m，1H)，2.72(m，1H)，2.50(m，1H)，1.95(m，1H)，1.77(m，1H)，1.72(s，3H)，1.66(m，1H)，1.27(m，1H).MS 404.49[M+H] ⁺.

Synthesizing of embodiment 5. compounds 7

Compound 7-1's is synthetic:

Take 2-chloromethyl pyrimidine as raw material, the synthetic method of compound 4-1 in the reference example 1 prepares compound 7-1.

¹H NMR(400MHz，CDCl ₃)δ8.56(d，J＝4.8Hz，2H)，7.09(t，J＝4.8Hz，1H)，5.94(s，1H)，5.28(s，2H)，4.72(m，2H)，1.73(t，J＝2.4Hz，3H).MS 291.78[M+H] ⁺.

Synthesizing of compound 7:

The compound 7-1 that obtains in the previous step is as raw material, and the synthetic method of compound 4 in the reference example 1 prepares compound 7.

¹H NMR(400MHz，CDCl ₃)δ8.59(d，J＝4.8Hz，2H)，7.09(t，J＝4.8Hz，1H)，5.32(s，2H)，5.25(s，1H)，4.51(d，J＝2.0Hz，2H)，3.35(m，1H)，3.24(m，1H)，2.99(m，1H)，2.72(m，1H)，2.50(m，1H)，1.95(m，1H)，1.84(m，1H)，1.76(s，3H)，1.64(m，1H)，1.28(m，1H).MS 355.45[M+H] ⁺.

Synthesizing of embodiment 6. compounds 8

Compound 8-1's is synthetic:

Take 3-brooethyl indoles as raw material, the synthetic method of compound 4-1 in the reference example 1 prepares compound 8-1. ¹H NMR (400MHz, CDCl ₃) 7.55 (m, 2H), 7.28 (m, 2H), 7.22 (s, 1H), 5.97 (s, 1H), 5.41 (s, 2H), 4.74 (d, J=2.4Hz, 2H), 1.80 (t, J=2.4Hz, 3H) .MS 328.08[M+H] ⁺.

Synthesizing of compound 8:

The compound 8-1 that obtains in the previous step is as raw material, and the synthetic method of compound 4 in the reference example 1 prepares compound 8. ¹H NMR (400MHz, CDCl ₃) 7.46 (m, 2H), 7.22 (m, 2H), 7.20 (s, 1H), 5.34 (s, 2H), 5.19 (s, 1H), (4.41 s, 2H), 3.25 (m, 3H), 2.97 (m, 1H), 2.61 (m, 1H), 2.42 (m, 1H), (1.93 m, 1H), 1.73 (s, 3H), (1.62 m, 1H), 1.25 (m, 1H) .MS 392.2[M+H] ⁺.

Synthesizing of embodiment 7. compounds 9

Compound 9-1's is synthetic:

Take 2-brooethyl quinoxaline as raw material, the synthetic method of compound 4-1 in the reference example 1 prepares compound 9-1.

¹H NMR(400MHz，CDCl ₃)δ8.34(s，1H)，8.07(m，1H)，8.00(m，1H)，7.71(m，2H)，6.04(s，1H)，5.47(s，2H)，4.79(m，2H)，1.81(t，J＝2.4Hz，3H).MS 341.78[M+H] ⁺.

Synthesizing of compound 9:

The compound 9-1 that obtains in the previous step is as raw material, and the synthetic method of compound 4 in the reference example 1 prepares compound 9.

¹H NMR(400MHz，CDCl ₃)δ8.77(s，1H)，7.98(m，2H)，7.64(m，2H)，5.41(s，2H)，5.25(s，1H)，4.49(d，J＝1.2Hz，2H)，3.31(m，1H)，3.20(m，1H)，2.96(m，1H)，2.68(m，1H)，2.47(m，1H)，1.91(m，1H)，1.80(m，1H)，1.74(s，3H)，1.63(m，1H)，1.23(m，1H).MS 405.50[M+H] ⁺.

Synthesizing of embodiment 8. compounds 10

Compound 10-1's is synthetic:

Take the 4-bromomethyl quinoline as raw material, the synthetic method of compound 4-1 in the reference example 1 prepares compound 10-1.

¹H NMR(400MHz，CDCl ₃)δ8.82(d，J＝4.8Hz，1H)，8.18(d，J＝8.4Hz，1H)，8.13(d，J＝8.4Hz，1H)，7.73(m，1H)，7.61(m，1H)，7.16(d，J＝4.4Hz，1H)，6.06(s，1H)，5.61(s，2H)，4.79(m，2H)，1.82(t，J＝2.4Hz，3H).MS 340.90[M+H] ⁺.

Synthesizing of compound 10:

The compound 10-1 that obtains in the previous step is as raw material, and the synthetic method of compound 4 in the reference example 1 prepares compound 10.

¹H NMR(400MHz，CDCl ₃)δ8.78(d，J＝4.4Hz，1H)，8.19(d，J＝8.4Hz，1H)，8.09(d，J＝8.0Hz，1H)，7.69(t，J＝7.2Hz，1H)，7.57(t，J＝7.2Hz，1H)，7.16(d，J＝4.4Hz，1H)，5.58(s，2H)，5.28(s，1H)，4.52(d，J＝2.0Hz，2H)，3.33(m，1H)，3.23(m，1H)，2.99(m，1H)，2.71(m，1H)，2.51(m，1H)，1.95(m，1H)，1.86(m，1H)，1.78(s，3H)，1.67(m，1H)，1.88(m，1H).MS 404.50[M+H] ⁺.

Synthesizing of embodiment 9. compounds 11

Compound 11-1's is synthetic:

Take the 3-bromomethyl quinoline as raw material, the synthetic method of compound 4-1 in the reference example 1 prepares compound 11-1.

¹H NMR(400MHz，CDCl ₃)δ9.04(d，J＝2.0Hz，1H)，8.25(d，J＝1.2Hz，1H)，8.06(d，J＝8.4Hz，1H)，7.76(t，J＝8.0Hz，1H)，7.66(m，1H)，7.49(m，1H)，6.00(s，1H)，5.24(s，2H)，4.72(d，J＝2.4Hz，2H)，1.77(t，J＝2.4Hz，3H).MS 340.78[M+H] ⁺.

Synthesizing of compound 11:

The compound 11-1 that obtains in the previous step is as raw material, and the synthetic method of compound 4 in the reference example 1 prepares compound 11.

¹H NMR(400MHz，CDCl ₃)δ9.05(d，J＝2.0Hz，1H)，8.28(d，J＝1.6Hz，1H)，8.03(d，J＝8.4Hz，1H)，7.76(t，J＝8.0Hz，1H)，7.64(m，1H)，7.47(m，1H)，5.24(s，2H)，5.22(s，1H)，4.48(d，J＝1.6Hz，2H)，3.29(m，1H)，3.18(m，1H)，2.97(m，1H)，2.66(m，1H)，2.47(m，1H)，1.94(m，1H)，1.84(m，1H)，1.77(s，3H)，1.64(m，1H)，1.27(m，1H).MS 404.51[M+H] ⁺.

Synthesizing of embodiment 10. compounds 12

Compound 12-1's is synthetic:

Take 1-brooethyl isoquinoline 99.9 as raw material, the synthetic method of compound 4-1 in the reference example 1 prepares compound 12-1.

¹H NMR(400MHz，CDCl ₃)δ8.34(d，J＝5.6Hz，1H)，8.15(d，J＝8.4Hz，1H)，7.83(d，J＝8.0Hz，1H)，7.69(m，1H)，7.63(m，1H)，7.52(d，J＝6.0Hz，1H)，6.06(s，1H)，5.78(s，2H)，4.82(m，2H)，1.84(t，J＝2.4Hz，3H).MS 340.90[M+H] ⁺.

Synthesizing of compound 12:

The compound 12-1 that obtains in the previous step is as raw material, and the synthetic method of compound 4 in the reference example 1 prepares compound 12.

¹H NMR(400MHz，CDCl ₃)δ8.32(d，J＝5.6Hz，1H)，8.14(d，J＝8.4Hz，1H)，7.77(d，J＝7.6Hz，1H)，7.63(m，1H)，7.58(m，1H)，7.47(d，J＝5.6Hz，1H)，5.75(s，2H)，5.30(s，1H)，4.54(d，J＝2.4Hz，2H)，3.35(m，1H)，3.25(m，1H)，2.98(m，1H)，2.72(m，1H)，2.49(m，1H)，1.93(m，1H)，1.83(m，1H)，1.77(t，J＝2.0Hz，3H)，1.65(m，1H)，1.26(m，1H).MS 404.49[M+H] ⁺.

Synthesizing of embodiment 11. compounds 13

Compound 13-1's is synthetic:

Take 2-brooethyl-6-bromoquinoline as raw material, the synthetic method of compound 4-1 in the reference example 1 prepares compound 13-1.

¹H NMR(400MHz，CDCl ₃)δ8.01(d，J＝8.8Hz，1H)，7.93(d，J＝2.0Hz，1H)，7.86(d，J＝8.8Hz，1H)，7.71(m，1H)，7.34(d，J＝8.4Hz，1H)，6.05(s，1H)，5.40(s，2H)，4.81(m，2H)，1.82(t，J＝2.4Hz，3H).MS 419.77[M+H] ⁺.

Synthesizing of compound 13:

The compound 13-1 that obtains in the previous step is as raw material, and the synthetic method of compound 4 in the reference example 1 prepares compound 13.

¹H NMR(400MHz，CDCl ₃)δ7.95(d，J＝8.8Hz，1H)，7.87(m，2H)，7.67(m，1H)，7.31(d，J＝8.4Hz，1H)，5.37(s，2H)，5.28(s，1H)，4.54(d，J＝2.4Hz，2H)，3.35(m，1H)，3.24(m，1H)，3.01(m，1H)，2.72(m，1H)，2.53(m，1H)，1.96(m，1H)，1.86(m，1H)，1.77(t，J＝2.4Hz，3H)，1.69(m，1H)，1.30(m，1H).MS 483.48[M+H] ⁺.

Synthesizing of embodiment 12. compounds 14

Compound 14-1's is synthetic:

Take 2-brooethyl-6-chloroquinoline as raw material, the synthetic method of compound 4-1 in the reference example 1 prepares compound 14-1.

¹H NMR(400MHz，CDCl ₃)δ8.00(d，J＝8.4Hz，1H)，7.92(d，J＝8.8Hz，1H)，7.74(d，J＝2.4Hz，1H)，7.57(m，1H)，7.34(d，J＝8.4Hz，1H)，6.04(s，1H)，5.40(s，2H)，4.80(m，2H)，1.82(t，J＝2.4Hz，3H).MS 357.21[M+H] ⁺.

Synthesizing of compound 14:

The compound 14-1 that obtains in the previous step is as raw material, and the synthetic method of compound 4 in the reference example 1 prepares compound 14.

¹H NMR(400MHz，CDCl ₃)δ7.95(d，J＝8.8Hz，1H)，7.91(d，J＝8.8Hz，1H)，7.70(d，J＝2.4Hz，1H)，7.53(m，1H)，7.30(d，J＝8.4Hz，1H)，5.37(s，2H)，5.28(s，1H)，4.53(d，J＝2.4Hz，2H)，3.34(m，1H)，3.23(m，1H)，3.00(m，1H)，2.71(m，1H)，2.52(m，1H)，1.95(m，1H)，1.85(m，1H)，1.76(t，J＝2.4Hz，3H)，1.67(m，1H)，1.29(m，1H).MS438.80[M+H] ⁺.

Synthesizing of embodiment 13. compounds 15

Compound 15-1's is synthetic:

Take 2-brooethyl-6-fluorine quinoline as raw material, the synthetic method of compound 4-1 in the reference example 1 prepares compound 15-1.

¹H NMR(400MHz，CDCl ₃)δ8.04(d，J＝8.4Hz，1H)，7.99(dd，J＝5.2Hz，1H)，7.39(m，3H)，6.05(s，1H)，5.41(s，2H)，4.81(m，2H)，1.82(t，J＝2.0Hz，3H).MS 358.78[M+H] ⁺.

Synthesizing of compound 15:

The compound 15-1 that obtains in the previous step is as raw material, and the synthetic method of compound 4 in the reference example 1 prepares compound 15.

¹H NMR(400MHz，CDCl ₃)δ7.98(m，2H)，7.35(m，3H)，5.37(s，2H)，5.28(s，1H)，4.53(d，J＝2.4Hz，2H)，3.34(m，1H)，3.23(m，1H)，3.01(m，1H)，2.71(m，1H)，2.51(m，1H)，1.95(m，1H)，1.85(m，1H)，1.77(t，J＝2.0Hz，3H)，1.66(m，1H)，1.28(m，1H).MS 422.50[M+H] ⁺.

Synthesizing of embodiment 14. compounds 16

Compound 16-1's is synthetic:

Take 2-brooethyl-6-toluquinoline as raw material, the synthetic method of compound 4-1 in the reference example 1 prepares compound 16-1.

¹H NMR(400MHz，CDCl ₃)δ8.01(d，J＝8.4Hz，1H)，7.95(d，J＝8.8Hz，1H)，7.87(d，J＝1.6Hz，1H)，7.79(m，1H)，7.26(d，J＝8.4Hz，1H)，5.99(s，1H)，5.25(s，2H)，4.72(m，2H)，2.72(s，3H)，1.80(t，J＝2.4Hz，3H).MS 354.89[M+H] ⁺.

Synthesizing of compound 16:

The compound 16-1 that obtains in the previous step is as raw material, and the synthetic method of compound 4 in the reference example 1 prepares compound 16.

¹H NMR(400MHz，CDCl ₃)δ7.98(d，J＝8.4Hz，1H)，7.89(m，2H)，7.81(m，1H)，7.21(d，J＝8.4Hz，1H)，5.23(m，3H)，4.49(d，J＝1.2Hz，2H)，3.30(m，1H)，3.19(m，1H)，2.98(m，1H)，2.68(s，3H)，2.47(m，1H)，1.94(m，1H)，1.83(m，1H)，1.78(t，J＝2.4Hz，3H)，1.64(m，2H)，1.28(m，1H).MS 418.66[M+H] ⁺.

Synthesizing of embodiment 15. compounds 17

Compound 17-1's is synthetic:

Take 2-brooethyl-6-methoxy quinoline as raw material, the synthetic method of compound 4-1 in the reference example 1 prepares compound 17-1.

¹H NMR(400MHz，CDCl ₃)δ7.98(d，J＝8.4Hz，1H)，7.90(d，J＝9.2Hz，1H)，7.29(m，2H)，7.03(d，J＝2.8Hz，1H)，6.04(s，1H)，5.39(s，2H)，4.80(m，2H)，3.90(s，3H)，1.82(t，J＝2.4Hz，3H).MS 370.89[M+H] ⁺.

Synthesizing of compound 17:

The compound 17-1 that obtains in the previous step is as raw material, and the synthetic method of compound 4 in the reference example 1 prepares compound 17.

¹H NMR(400MHz，CDCl ₃)δ7.94(d，J＝8.4Hz，1H)，7.89(d，J＝9.2Hz，1H)，7.26(m，2H)，7.00(d，J＝2.4Hz，1H)，5.37(s，2H)，5.28(s，1H)，4.53(d，J＝1.6Hz，2H)，3.86(s，3H)，3.33(m，1H)，3.23(m，1H)，3.01(m，1H)，2.71(m，1H)，2.52(m，1H)，1.96(m，1H)，1.85(m，1H)，1.78(s，3H)，1.67(m，1H)，1.28(m，1H).MS 434.49[M+H] ⁺.

Synthesizing of embodiment 16. compounds 18

Compound 18-1's is synthetic:

Take 2-brooethyl-7-fluorine quinoline as raw material, the synthetic method of compound 4-1 in the reference example 1 prepares compound 18-1.

¹H NMR(400MHz，CDCl ₃)δ8.08(d，J＝8.8Hz，1H)，7.75(m，1H)，7.61(m，1H)，7.27(m，2H)，6.05(s，1H)，5.41(s，2H)，4.82(m，2H)，1.83(t，J＝2.4Hz，3H).MS 358.78[M+H] ⁺.

Synthesizing of compound 18:

The compound 18-1 that obtains in the previous step is as raw material, and the synthetic method of compound 4 in the reference example 1 prepares compound 18.

¹H NMR(400MHz，CDCl ₃)δ8.05(d，J＝8.8Hz，1H)，7.73(m，1H)，7.63(m，1H)，7.25(m，2H)，5.40(s，2H)，5.31(s，1H)，4.56(d，J＝1.6Hz，2H)，3.37(m，1H)，3.26(m，1H)，3.04(m，1H)，2.75(m，1H)，2.55(m，1H)，1.99(m，1H)，1.88(m，1H)，1.80(s，3H)，1.74(m，1H)，1.32(m，1H).MS 422.50[M+H] ⁺.

Synthesizing of embodiment 17. compounds 19

Compound 19-1's is synthetic:

Take 2-brooethyl-7-chloroquinoline as raw material, the synthetic method of compound 4-1 in the reference example 1 prepares compound 19-1.

¹H NMR(400MHz，CDCl ₃)δ8.06(d，J＝8.4Hz，1H)，8.00(s，1H)，7.69(d，J＝8.8Hz，1H)，7.42(m，1H)，7.32(d，J＝8.4Hz，1H)，6.04(s，1H)，5.40(s，2H)，4.81(m，2H)，1.83(t，J＝2.4Hz，3H).MS 375.21[M+H] ⁺.

Synthesizing of compound 19:

The compound 19-1 that obtains in the previous step is as raw material, and the synthetic method of compound 4 in the reference example 1 prepares compound 19.

¹H NMR(400MHz，CDCl ₃)δ8.03(d，J＝8.4Hz，1H)，7.99(s，1H)，7.67(d，J＝8.4Hz，1H)，7.40(m，1H)，7.30(d，J＝8.4Hz，1H)，5.39(s，2H)，5.30(s，1H)，4.56(d，J＝2.0Hz，2H)，3.37(m，1H)，3.26(m，1H)，3.04(m，1H)，2.75(s，3H)，2.55(m，1H)，1.98(m，1H)，1.88(m，1H)，1.79(s，3H)，1.73(m，1H)，1.32(m，1H).MS 438.98[M+H] ⁺.

Synthesizing of embodiment 18. compounds 20

Compound 20-1's is synthetic:

Take 2-chloromethyl-4-chloroquinoline as raw material, the synthetic method of compound 4-1 in the reference example 1 prepares compound 20-1.

¹H NMR(400MHz，CDCl ₃)δ8.17(d，J＝8.0Hz，1H)，8.02(d，J＝8.4Hz，1H)，7.70(m，1H)，7.58(m，1H)，7.41(s，1H)，6.05(s，1H)，5.39(s，2H)，4.81(m，2H)，1.82(t，J＝2.4Hz，3H).MS 375.21[M+H] ⁺.

Synthesizing of compound 20:

The compound 20-1 that obtains in the previous step is as raw material, and the synthetic method of compound 4 in the reference example 1 prepares compound 20.

¹H NMR(400MHz，CDCl ₃)δ8.13(d，J＝8.4Hz，1H)，8.02(d，J＝8.4Hz，1H)，7.68(t，J＝7.6Hz，1H)，7.55(t，J＝7.6Hz，1H)，7.38(s，1H)，5.37(s，2H)，5.30(s，1H)，4.55(s，2H)，3.37(m，1H)，3.25(m，1H)，3.03(m，1H)，2.74(m，1H)，2.53(m，1H)，1.97(m，1H)，1.88(m，1H)，1.79(s，3H)，1.70(m，1H)，1.30(m，1H).MS 438.90[M+H] ⁺.

Synthesizing of embodiment 19. compounds 21

Compound 21-1's is synthetic:

Take 2-chloromethyl-3-methyl-quinoxaline as raw material, the synthetic method of compound 4-1 in the reference example 1 prepares compound 21-1.

¹H NMR(400MHz，CDCl ₃)δ8.18(d，J＝8.0Hz，1H)，8.06(d，J＝8.4Hz，1H)，7.69(m，1H)，7.57(m，1H)，6.00(s，1H)，5.41(s，2H)，4.82(m，2H)，1.80(t，J＝2.4Hz，3H).MS 354.09[M+H] ⁺.

Synthesizing of compound 21:

The compound 21-1 that obtains in the previous step is as raw material, and the synthetic method of compound 4 in the reference example 1 prepares compound 21.

¹H NMR(400MHz，CDCl ₃)δ8.16(d，J＝8.4Hz，1H)，8.07(d，J＝8.4Hz，1H)，7.66(m，1H)，7.55(m，1H)，5.37(s，2H)，5.32(s，1H)，4.58(s，2H)，3.37(m，1H)，3.25(m，1H)，3.03(m，1H)，2.74(m，1H)，2.53(m，1H)，2.43(s，3H)，1.95(m，1H)，1.86(m，1H)，1.78(s，3H)，1.73(m，1H)，1.32(m，1H).MS 419.2[M+H] ⁺.

Synthesizing of embodiment 20. compounds 22

Compound 22-1's is synthetic:

Take 1-azepine-2-chloromethyl phenanthrene as raw material, the synthetic method of compound 4-1 in the reference example 1 prepares compound 22-1.

¹H NMR(400MHz，CDCl ₃)δ8.34(m，1H)，8.21(m，1H)，8.06(m，1H)，7.88(m，1H)，7.69(m，2H)，7.57(m，2H)，5.99(s，1H)，5.38(s，2H)，4.83(m，2H)，1.79(t，J＝2.4Hz，3H).MS 390.10[M+H] ⁺.

Synthesizing of compound 22:

The compound 22-1 that obtains in the previous step is as raw material, and the synthetic method of compound 4 in the reference example 1 prepares compound 22.

¹H NMR(400MHz，CDCl ₃)8.35(m，1H)，8.23(m，1H)，8.08(m，1H)，7.89(m，1H)，7.70(m，2H)，7.58(m，2H)，6，00(s，1H)，5.36(s，2H)，4.81(m，2H)，3.37(m，1H)，3.25(m，1H)，3.01(m，1H)，2.76(m，1H)，2.53(m，1H)，2.41(s，3H)，1.94(m，1H)，1.86(m，1H)，1.79(s，3H)，1.72(m，1H)，1.31(m，1H).MS 454.2[M+H] ⁺.

The experiment of embodiment 21. external activities

Compound provided by the invention can be measured with the homogeneous luminescent detection system (DPP-IV-Glo Protease Assay, Promega cat#G8350) of DPP-IV-Glo proteolytic ferment the inhibiting rate of DPP-IV.This system contains the Laemmli buffer system Laemmli of the amino luciferin of DPP-IV substrate Gly-Pro-and luciferase activity detection, after being cut by DPP-IV, DPPIV-Glo can activate the luciferase reaction, " glow-type " type of generation luminous signal detects the activity that luminous signal can characterize DPP-IV with Turner Ver ita s microwell plate luminometer again.

1, experiment purpose

Measure the compounds of this invention to DPP-IV enzymeinhibition activity and selective inhibitory.

2, experiment material

Embodiment of the invention compound;

DPP-IV enzyme, DPP-VIII enzyme, DPP-IX enzyme, GP-AMC (BioMol), black 96 orifice plates, super microplate reader;

The analysis buffer of DPP-IV and DPP-VIII: 100mmol/l Tris/HCl buffer, pH 8.0,0.1mg/ml BSA;

The analysis buffer of DPP-IX: 100mmol/l Tris/HCl buffer, pH 7.4,0.1mg/mlBSA.

3, experimental technique

Determining of a, enzymic activity:

GP-AMC is diluted in separately the damping fluid, and concentration is 100umol/L, every hole 25ul; Enzyme gradient dilution, initial concentration are respectively DPP-VIII, DPP-IX:0.01ug/ul, DPP-IV:0.01mU/ul, and by 5 times of dilutions, every hole 25ul, mixing; 37 ℃, 360/460nm measures the dynamic change of fluorescent value, measures 30 minutes; Take absorbancy linearly rise, the enzyme concn of S/B 〉=5 is as working concentration.

B, inhibitor activity are measured:

All enzymes, inhibitor, GP-AMC arrange without the compound contrast, contrast without enzyme liquid all with the analysis buffer preparation.

Press the working concentration preparation enzyme liquid of enzyme, every hole 25ul; Gradient dilution inhibitor (10 times or 5 times of dilutions), every hole 25ul, mixing; Add the good GP-AMC solution 50ul of dilution, mixing; 37 ℃ were reacted 20 minutes, and 360/460nm measures fluorescent value.

C, data analysis: use the GraphPad-Prism software analysis.

4, experimental result

Embodiment of the invention compound 3-22 to three kinds of enzymeinhibition activity datas shown in following table one.

The external activity of table one and selective data

Common DPP-IV inhibitor is when suppressing the DPP-IV activity, with some DPP-IV relevant enzyme, activity such as DPP-II, DPP-VIII, DPP-IX has also suppressed, wherein DPP-VIII and DPP-IX are considered to the cytosol enzyme, the suppressed toxic action that may cause some DPP-IV inhibitor of their effect, as bald, thrombopenia (disease), anaemia, splenomegaly and many lesion tissues etc.More satisfactory state is to develop DPP-IV is had the high selectivity restraining effect, simultaneously the activity of DPP-VIII and DPP-IX is not had influential compound.

The experimental result explanation: embodiment of the invention compound has extraordinary selective inhibitory to DPP-IV, in establishment DPP-IV activity, almost not impact of activity on DPP-VIII and DPP-IX, side effect is lower after can predicting the compounds of this invention exploitation patent medicine, has extraordinary using value.

Embodiment 22. external restraining effect to hERG

Non-heart class medicine might cause by suppressing hERG (IKr) passage the prolongation of myocardial action potential time-histories, increases the possibility that life-threatening torsades de pointes (TdP) ventricular arrhythmia occurs.It is host cell that the HEK293 cell without endogenous IKr electric current is adopted in this experiment, and this clone is widely used in the detection of hERG.

HERG clone cellar culture goes down to posterity in the DMEM substratum that contains 10% foetal calf serum and 250g/ml G418.A culture dish is taken out in each experiment, cleans twice with extracellular fluid, is positioned on the inverted microscope Stage microscope.The experiment of full cell patch pincers is at room temperature carried out, and used borosilicate glass microelectrode tip resistance is 3-5M Ω.

After forming full cell record pattern, membrane potential is clamped down at-80mV, giving cell+50mV depolarize voltage every 30s stimulates, and continues behind the 2s repolarization to-50mV, continues 3s, can draw the hERG tail current.Depolarize voltage gives first cell 50ms before stimulating ,-50mV repolarization voltage, and the electric current that records under this voltage is as the baseline that calculates the hERG tail current.Before adding compound, hERG tail current stable recording 3 minutes at least in extracellular fluid.Change less than 5% the time when hERG tail current amplitude after the perfusion administration, be considered to drug effect and reach stable state.If electric current did not reach stable state in 6 minutes, then also finish this concentration compound test.

Testing compound calculates according to following equation the inhibiting rate of hERG electric current: % inhibiting rate={ 1-(electric current residue amplitude)/(contrast current amplitude) } * 100

Obtain a plurality of concentration of embodiment of the invention compound to the inhibiting rate of hERG electric current according to above-mentioned method of calculation, use the logistic equation that data are carried out match, obtain IC ₅₀Value.Embodiment of the invention compound is to inhibiting rate and the IC of hERG ₅₀Data as shown in Table 2.

Table two. external restraining effect to hERG

The compound title	Highest detection concentration	The maximum concentration inhibiting rate	IC 50Value	Standard deviation
					Compound 4	300μM	79.0±1.2％	114.2μM	7.3μM
Compound 6	300μM	91.7±1.6％	47.9μM	4.5μM
					Compound 9	300μM	84.9±8.3％	62.7μM	9.2μM

This research at the HEK293 cell observation of exogenous expression hERG potassium-channel the effect of compound to the hERG electric current.Experimental result shows, a little less than the inhibition of compound 4, compound 6 and 9 couples of hERG of compound, point out each compound to cause the prolongation of myocardial action potential time-histories by suppressing hERG (IKr) passage, the possibility that increases the generation of life-threatening torsades de pointes (TdP) ventricular arrhythmia is very low, be that cardiac toxic will be lower after the present invention is developed to the medicine for the treatment of diabetes.

Claims (10)

1. the preparation method of a formula I compound comprises the steps:

(1) formula A compound and formula B compound react production C compound in organic solvent in the presence of alkali:

(2) formula C compound and R-3-amino piperidine dihydrochloride reaction production I compound:

Wherein, Hal is chlorine or bromine,

R ₁Choose generation or unsubstituted alkyl, cycloalkyl, aryl, heteroaryl or Heterocyclylalkyl; Substituting group is selected from alkyl, alkoxyl group, halogen, cyano group, amino, hydroxyl, nitro, carbonyl, alkylsulfonyl alkyl, amido, carbonylic alkyl, aryl, aryloxy, Heterocyclylalkyl, heteroaryl, heteroaryloxy, cycloalkyl, cycloalkylalkyl, alkylsulfonyl or sulfinyl;

R ₂Be selected from:

(1) hydrogen;

(2) cyano group;

(3) alkyl that does not replace or replace;

(4) do not replace or by 1-5 the phenyl that replace of substituting group independently separately, substituting group is selected from halogen, cyano group, OH, alkyl, alkoxyl group, NHSO ₂R ₃, N (alkyl) SO ₂R ₃, SO ₂R ₃, SO ₂NR ₄R ₅,

NR ₄R ₅, CONR ₄R ₅, COOH and carboxyalkyl;

(5)OH；

(6) alkoxyl group;

(7)NR ₄R ₅；

R ₃Be selected from and replace or unsubstituted alkyl, substituting group is selected from 1-5 separately independently halogen or COOH; R ₄And R ₅Individually independently be selected from separately:

(1) hydrogen,

(2) phenyl that does not replace or replace, substituting group is selected from halogen, OH, alkyl or alkoxyl group,

(3) do not replace or by the C that replaces of substituting group independently separately _3-6Cycloalkyl, substituting group are selected from halogen, OH, alkyl or alkoxyl group,

(4) alkyl that does not replace or replace, substituting group is selected from:

(a) halogen, or

(b) do not replace or by 1-5 the phenyl that replace of substituting group independently separately, substituting group is selected from halogen, OH, alkyl or alkoxyl group.

2. the preparation method of formula I compound claimed in claim 1, wherein, R ₁Be selected from replacement or unsubstituted aryl or heteroaryl, substituting group is selected from alkyl, alkoxyl group, halogen, cyano group, amino, hydroxyl, nitro or carbonyl, R ₂Be selected from hydrogen, alkyl or alkoxyl group.

3. the preparation method of formula I compound claimed in claim 2, wherein, R ₁Be selected from replacement or unsubstituted aryl or heteroaryl, substituting group is selected from alkyl, alkoxyl group, halogen, R ₂Be selected from hydrogen.

4. the preparation method of formula I compound claimed in claim 3, wherein, R ₁Be selected from and replace or unsubstituted phenyl, quinazolyl, benzimidazolyl-, quinolyl, pyrimidyl, indyl, quinoxalinyl, isoquinoline 99.9 or aza-phenanthrenes.

5. the preparation method of formula I compound claimed in claim 4, wherein, R ₁Be selected from phenyl, 4-methyl quinazoline-2-base, benzimidazolyl-2 radicals-Ji, quinoline-2-base, 6-bromoquinoline-2-base, 6-chloroquinoline-2-base, 6-fluorine quinoline-2-base, 6-toluquinoline-2-base, 7-chloroquinoline-2-base, 7-fluorine quinoline-2-base, 6-methoxy quinoline-2-base, 4-chloroquinoline-2-base, 3-methyl-quinoxaline-2-base, quinolyl-4, quinoline-3-base, pyrimidine-2-base, indol-3-yl, quinoxaline-2-base, isoquinolyl-1 or 1-aza-phenanthrenes-2-base.

6. the preparation method of each described formula I compound among the claim 1-5, wherein, in the step (1), alkali is selected from sodium hydride, salt of wormwood, yellow soda ash.

7. the preparation method of each described formula I compound among the claim 1-5, wherein, in the step (1), organic solvent is selected from the mixed solvent of glycol dimethyl ether and DMF.

8. the preparation method of each described formula I compound among the claim 1-7 comprises the steps:

(1) alkali is mixed 0 ℃ of lower stirring 10 minutes with the mixed solvent of glycol dimethyl ether/DMF, adding formula A compound, 0 ℃ of lower stirring 20 minutes adds anhydrous lithium bromide, stirring at room 30 minutes adds formula B compound, stirred overnight at room temperature.React complete, add trash ice, dichloromethane extraction, dry, concentrated after the water washing of gained organic phase usefulness saturated common salt, namely obtain formula C compound behind the column chromatography purification.

(2) formula C compound is mixed the rear ethanol that adds with R-3-amino piperidine dihydrochloride, mix rear adding sodium bicarbonate, oil bath reflux 6 hours.React complete, reaction solution is concentrated, and resistates namely obtains formula I compound through column chromatography purification.

9. the preparation method of formula I compound claimed in claim 8, wherein, alkali is selected from sodium hydride, salt of wormwood, yellow soda ash in the step (1); Glycol dimethyl ether and DMF are anhydrous solvent, and the two volume ratio is 2: 1.

10. the preparation method of a formula I ' compound comprises the steps:

(1) formula A ' compound and formula B compound react production C ' compound in the mixed solvent of glycol dimethyl ether and DMF in the presence of sodium hydride:

(2) formula C ' compound and R-3-amino piperidine dihydrochloride reaction production I ' compound:

Wherein, R ₁Identical with the definition in the claim 1, Hal is chlorine or bromine.