CN106565684A

CN106565684A - 6-pyrromonazole substituted quinazoline compound, and derivative, synthetic method and application thereof

Info

Publication number: CN106565684A
Application number: CN201610935466.XA
Authority: CN
Inventors: 万山河; 王广发; 伍小云; 田元新; 叶玲; 李中皇; 张婷婷; 朱正光; 金宏; 张嘉杰
Original assignee: Southern Medical University
Current assignee: Southern Medical University
Priority date: 2016-03-18
Filing date: 2016-10-25
Publication date: 2017-04-19
Anticipated expiration: 2036-10-25
Also published as: CN106565684B

Abstract

The invention discloses a compound as shown in the formula (I), (II) or (III), or salt of the compound acceptable in pharmacy, aquo-complex, a solvate, a polymorphic substance, a tautomer or a prodrug; a synthetic method of the formula (I) comprises the following steps: 5-bromo-2-aminobenzoic acid is subjected to cyclization and chlorine substituting sequentially, a product reacts with benzyl alcohol or benzyl amine, and a product is coupled with boric acid ester; a synthetic method of the formula (II) comprises the following steps: 2-aminonicotinic acid is subjected to bromine substituting, cyclization and chlorine substituting sequentially, a product reacts with benzyl alcohol or benzyl amine, and a product is coupled with boric acid ester; a synthetic method of the formula (III) comprises the following steps: 5-bromo-2-aminobenzoic acid is subjected to cyclization with dicyandiamide, deamidination is carried out, a product is condensed with benzyl alcohol or benzyl amine, and then a product is coupled with boric acid ester. A medicine composition comprises at least one of the following active components: an active compound, salt of the active compound acceptable in pharmacy, the aquo-complex, the solvate, the polymorphic substance, the tautomer or the prodrug. The invention also discloses an application of the active component in preparing a tyrosine kinase inhibitor, or an application of the active component in preparing an anti-tumor drug. Synthetic series compounds and derivatives thereof have higher tyrosine kinase inhibitory activity.

Description

6- pyrazoles substituted quinazoline class compound and its derivative, synthetic method and its application

Technical field

The present invention relates to 6- pyrazoles substituted quinazoline class compound and its derivative, synthetic method and its application.

Background technology

Malignant tumour seriously threatens human health, and in present illness, the death rate that malignant tumour causes is only second to painstaking effort Guard system disease.The major fields that the efficient, antineoplastic of low toxicity, few side effects is current medical research and development are found, in recent years Research finds that the generation of tumour is tight with the abnormal activation of protein tyrosine kinase (Protein tyrosine kinase, PTK) It is related.Protein tyrosine kinase (PTK) is the superfamily protein for being currently known maximum, is that γ positions phosphoric acid turns on class catalysis ATP The kinases on protein-tyrosine residue is moved on to, the tyrosine residue phosphorylation of various substrate proteins can be catalyzed, maintaining cell Play an important role in growth, differentiation.Protein tyrosine kinase includes receptor type tyrosine kinase and non-receptor tyrosine kinase Two big class.

Since listing from first protein tyrosine kinase inhibitor Imatinib (Imatinib), existing tens effects List in succession in the medicine of LCK, and be increasingly becoming the first-line drug for the treatment of tumour.These pharmaceutically-active sides Formula is various, mainly includes：The modes such as closing part, blocking acceptor and suppression kinase activation.Closing part is by antibody or melts Hop protein is combined with part, and part is closed with receptor binding site, hinders the combination of part and acceptor.Blocking acceptor is By the antibody that can be combined with receptor-specific, the combination of acceptor and part is blocked.The medicine of closing part and blocking acceptor Belong to biotech drug, the medicine for having listed has kind more than 10.Suppress kinase activation to be tied by micromolecular compound and kinases Close, the kinase activation for suppressing ATP to be combined with kinases and produced.

Therefore, this area is in the urgent need to developing structure novelty, the strong Novel tyrosine kinase inhibitors of activity.

The content of the invention

It is an object of the invention to provide 6- pyrazoles substituted quinazoline class compound and its derivative, synthetic method and its should With.

The technical solution used in the present invention is：

The invention has the beneficial effects as follows：

Compound or its pharmaceutically acceptable salt, hydrate, solvent conjunction shown in formula (I) or (II) or (III) Thing, polymorph, dynamic isomer or prodrug,

In formula：

X is O or NH；

R₁Having structure unit can be selected from：

Wherein R₃, R₄It is each independently selected from H, F, Cl, Br, I, CH₃、OCH₃、NO₂、NH₂、SO₂NH₂、CF₃、OCF₃In It is a kind of；

R₂Selected from one of following construction unit：

Described compound is chosen in particular from any one in following compounds：

4- [(1- (the chloro- 3-fluorophenyls of 2,6- bis-) ethyoxyl]-6- (1- methyl isophthalic acid H- pyrazoles-4- bases) quinazoline,

N- [1- (the chloro- 3-fluorophenyls of 2,6- bis-) ethyl]-6- (1- piperidin-4-yl-1H- pyrazoles-4- bases) quinazoline-4- Amine,

4- [1- (the chloro- 3-fluorophenyls of 2,6- bis-) ethyoxyl]-6- (1- piperidin-4-yl-1H- pyrazoles-4- bases) quinazoline,

N- (quinoline -6- bases) -6- (1- methyl isophthalic acid H- pyrazoles -4- bases) quinazoline -4- amine,

4- (hydroxyl of quinoline -6) -6- (1- methyl isophthalic acid H- pyrazoles -4- bases) quinazoline,

N- (quinoline -6- bases) -6- (1- piperidin-4-yl -1H- pyrazoles -4- bases) quinazoline -4- amine,

N- (the chloro- 4-fluorobenzene of 3-) base-6-- (the 1- methyl isophthalic acid H- pyrazoles-4- bases) amine of quinazoline-4,

N- (the chloro- 4- fluorophenyls of 3-) -6- (1- piperidin-4-yl -1H- pyrazoles -4- bases) quinazoline -4- amine,

N- (3- methoxybenzyls) -6- (the 1- methyl isophthalic acid H- pyrazoles -4- bases) amine of quinazoline -4,

N- (3- ethynyl phenyls) -6- (the 1- methyl isophthalic acid H- pyrazoles -4- bases) amine of quinazoline -4,

N- [(1- (the chloro- 3-fluorophenyls of 2,6- bis-) ethyl)]-6- (1- piperidin-4-yl-1H- pyrazoles-4- bases) pyrido [2, 3-d] amine of pyrimidine -4,

N- [(1- (the chloro- 3-fluorophenyls of 2,6- bis-) ethyl)]-6- (1- methyl isophthalic acid H- pyrazoles-4- bases) pyrido [2,3-d] The amine of pyrimidine -4,

N- (3- nitrobenzyls) -6- (1- methyl isophthalic acid H- pyrazoles -4- bases) pyrido [2,3-d] amine of pyrimidine -4,

N- (3- methoxybenzyls) -6- (1- methyl isophthalic acid H- pyrazoles -4- bases) pyridines [2,3-d] and the amine of pyrimidine -4,

N- [1- (pyridin-4-yl) ethyl] -6- (1- methyl isophthalic acid H- pyrazoles -4- bases) pyrido [2,3-d] pyrimidine.

The synthetic method of compound shown in formula (I), step is：By the bromo- 2- aminobenzoic acids of initiation material 5- and formyl Amine cyclization, generates bromo- 4 (the 3H)-quinazolinones of 6-, in the presence of thionyl chloride, adds the DMF of catalytic amount, is heated to reflux carrying out 4 The chlorine of position replaces, then generates 4 substituted compounds with corresponding benzylalcohol or benzylamine, is coupled with borate ester compound Jing C-C respectively anti- Deserved target product.

The synthetic method of compound shown in formula (II), step is：By initiation material 2- amino-nicotinic acids and bromine reaction life Into the acid of 5- bromo- 2- amino-nicotinic acids, then with formamide cyclization, bromo- pyrido [2, the 3-d] pyrimidine of 4- hydroxyls -6 is generated, in protochloride In the presence of sulfone, the DMF of catalytic amount is added, be heated to reflux carrying out the chlorine replacement of 4, then taken with corresponding benzylalcohol or benzylamine generation 4 For compound, target product is obtained with borate ester compound Jing C-C coupling reactions respectively.

The synthetic method of compound shown in formula (III), step is：By the bromo- 2- aminobenzoic acids of initiation material 5- and double cyanogen Amine cyclization, obtains the compound containing guanidine radicals class, and deamidination obtains the bromo- quinazolines of 2- amino-4-hydroxy -6-, directly employs card Special condensing agent is condensed with corresponding benzylalcohol or benzylamine, then obtains target compound with borate ester compound Jing C-C coupling reactions.

A kind of pharmaceutical composition, including at least one in following active ingredients：A) compound, b) compound is in pharmacy Upper acceptable salt, c) hydrate of the compound, d) solvate of the compound, e) polymorph of the compound, f) The prodrug of the dynamic isomer of the compound, g) compound；Wherein, described compound is formula (I) or (II) or (III) Shown compound.

Also include excipient substance.

Application of the described active component in tyrosine kinase inhibitor is prepared.

Described active component is preparing treatment and/or prevention and/or is delaying and/or auxiliary treatment and/or process and junket Application in the medicine of the related disease of histidine kinase hyperactivity.

Application of the described active component in antineoplastic is prepared.

The invention has the beneficial effects as follows：

Series compound synthesized by the present invention has higher tyrosine-kinase enzyme inhibition activity；To human breast cancer cell with And lung adenocarcinoma cell etc. is with stronger inhibitory activity.

Specific embodiment

In formula：

X is O or NH；

R₁Having structure unit can be selected from：

R₂Selected from one of following construction unit：

Preferably, described compound is chosen in particular from any one in following compounds：

The synthetic method of compound shown in formula (I), step is：By the bromo- 2- aminobenzoic acids of initiation material 5- and formyl Amine cyclization, generates bromo- 4 (the 3H)-quinazolinones of 6-, in the presence of thionyl chloride, adds the DMF of catalytic amount, is heated to reflux carrying out 4 The chlorine of position replaces, then generates 4 substituted compounds with corresponding benzylalcohol or benzylamine, is coupled with borate ester compound Jing C-C respectively anti- Deserved target product；

Synthetic line is schematically as follows：

The synthetic method of compound shown in formula (II), step is：By initiation material 2- amino-nicotinic acids and bromine reaction life Into the acid of 5- bromo- 2- amino-nicotinic acids, then with formamide cyclization, bromo- pyrido [2, the 3-d] pyrimidine of 4- hydroxyls -6 is generated, in protochloride In the presence of sulfone, the DMF of catalytic amount is added, be heated to reflux carrying out the chlorine replacement of 4, then taken with corresponding benzylalcohol or benzylamine generation 4 For compound, target product is obtained with borate ester compound Jing C-C coupling reactions respectively；

Synthetic line is schematically as follows：

The synthetic method of compound shown in formula (III), step is：By the bromo- 2- aminobenzoic acids of initiation material 5- and double cyanogen Amine cyclization, obtains the compound containing guanidine radicals class, and deamidination obtains the bromo- quinazolines of 2- amino-4-hydroxy -6-, directly employs card Special condensing agent is condensed with corresponding benzylalcohol or benzylamine, then obtains target compound with borate ester compound Jing C-C coupling reactions；

Synthetic line is schematically as follows：

Also include excipient substance；

It is further preferred that described auxiliary material includes at least one in following material：It is solvent, propellant, solubilizer, steady Determine agent, glidant, flavouring, preservative, suspending agent, coating material, aromatic, anti-binder, integrated agent, penetration enhancer, PH value regulator, buffer, plasticizer, cosolvent, emulsifying agent, colouring agent, binder, disintegrant, filler, lubricant, profit Humectant, osmotic pressure regulator, surfactant, foaming agent, defoamer, thickener, inclusion agents, NMF, absorbent, dilution Agent, flocculant and deflocculant, filter aid, release retarding agent.

The pharmaceutical composition of the present invention can be made into various formulations：Decentralized system according to formulation is classified：Specifically, Following formulation can be made：Solution-type, colloidal solution type, emulsion-type, suspension type, gas decentralized, microdispersed form, solid point Scattered type；According to typoiogical classification, specifically, following formulation can be made：Liquid dosage form is (such as aromatic waters, solution, injection Agent, mixture, lotion, liniment etc.), gas formulation (such as aerosol, spray), solid dosage forms is (such as powder, pill, tablet, film Agent etc.), semisolid dosage form (such as ointment, suppository, paste)；Classify according to method of administration：Specifically, can make following Formulation：The formulation of Jing gastrointestinal administrations, without the formulation of gastrointestinal administration.

Compound or its pharmaceutically acceptable salt, hydrate, solvent conjunction shown in formula (I) or (II) or (III) The application of thing, polymorph, dynamic isomer or prodrug in tyrosine kinase inhibitor is prepared.

Preferably, formula (I) or the compound shown in (II) or (III) or its pharmaceutically acceptable salt, hydrate, Solvate, polymorph, dynamic isomer or prodrug are preparing treatment and/or prevention and/or are delaying and/or auxiliary treatment And/or the application in the medicine of process related disease too high to tyrosine kinase activity；

It is further preferred that formula (I) or the compound shown in (II) or (III) or its pharmaceutically acceptable salt, water The application of compound, solvate, polymorph, dynamic isomer or prodrug in antineoplastic is prepared；

Still more preferably, at least one in following compounds or its pharmaceutically acceptable salt, hydrate, molten The application of agent compound, polymorph, dynamic isomer or prodrug in antineoplastic is prepared；For example, anti-breast cancer is being prepared Application in medicine, or prepare the application in anti-lung cancer (such as adenocarcinoma of lung) medicine；

Preferably, " pharmaceutically acceptable salt " refers to the basic group in parent compound is converted into salt in the present invention Form.Pharmaceutically acceptable salt include but not limited to, the inorganic or acylate of basic group such as amine (ammonia) base Class.Pharmaceutically acceptable salt of the present invention can be synthesized by parent compound, i.e., the basic group in parent compound is worked as with 1-4 The acid of amount is reacted in a solvent system.Suitable salt is set forth in Remington ' s Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985,1418 and Journal of Pharmaceutical Science, in 66,2,1977.

Pharmaceutically acceptable acid-addition salts can be prepared by inorganic and organic acid.By the inorganic acid bag of derivative acid-addition salts Include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid etc..By derivative acid-addition salts organic acid include acetic acid, propionic acid, glycolic, third Ketone acid, oxalic acid, malic acid, malonic acid, butanedioic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, almond Acid, methanesulfonic acid, ethyl sulfonic acid, p-methyl benzenesulfonic acid, salicylic acid, benzene sulfonic acid etc..The inorganic acid and organic acid of derivative acid-addition salts is especially Selected from hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, perchloric acid, hydrobromic acid, acetic acid, benzoic acid and p-methyl benzenesulfonic acid.

Embodiment 1：

The synthesis of the bromo- quinazolines of the chloro- 6- of 4-

In equipped with agitator, the 250mL there-necked flasks of reflux condensing tube, the bromo- 2- aminobenzoic acids 21.6g of 5- are added (0.1mol), 100mL formamides are heated to reflux, and control 130 DEG C of temperature, after reaction about 5h, stop reaction, and reactant liquor is poured into In 800mL water, stirring has solid to separate out, and filters washing, is dried, and obtains white gray solid 18g, and yield 80% is not purified direct For the next step.

In equipped with agitator, the 250mL there-necked flasks of reflux condensing tube, bromo- 4 (the 3H)-quinazolinone 15g of 6- are added (0.067mol), 150mL (2.1mol) thionyl chloride, 3-4 drops DMF is added to be heated to reflux, reactant liquor clarification after about 1h continues After reaction 5h, Distillation recovery thionyl chloride, the thionyl chloride of vacuum distillation residual continuously adds the distillation of 50mL dichloromethane and takes out of The thionyl chloride of residual, repeatedly for three times, is finally heated to reflux several minutes with 180mL ethyl acetate, places to room temperature, filters, very Sky is dried, and obtains pale yellow needles solid 12.2g, yield 75%.¹H NMR(400MHz,DMSO-d₆)δ8.36(s,1H),8.22 (d, J=2.4Hz, 1H), 8.01 (dd, J=2.4,8.8Hz, 1H), 7.68 (d, J=8.4Hz, 1H), ESI-MS m/z:244.9 [M+H]⁺.

Embodiment 2：

The synthesis of N- [1- (the chloro- 3- fluorophenyls of the 2,6- bis-) ethylamino-] bromo- quinazoline -4- amine of -6-

In the 100mL single-necked flasks equipped with agitator, 1- (2,6- bis- chloro- 3- fluorophenyls) ethamine 3.12g is added (0.015mol), the chloro- quinazoline 2.43g (0.01mol) of the bromo- 4- of 6-, add Isosorbide-5-Nitrae-dioxane 25mL, stirred under room temperature At night, pour reactant liquor into 150mLH₂O, has solid to separate out, and filters, and is washed with 50% ethanol, obtains white solid thing 3.0g, yield 71%.

¹H NMR(400MHz,DMSO-d₆) δ 8.91 (d, J=2.0Hz, 1H), 8.76 (d, J=5.6Hz, 1H), 8.33 (s, 1H), 7.90 (dd, J=2.0,8.8Hz, 1H), 7.61 (d, J=9.2Hz, 1H), 7.45-7.42 (m, 1H), 7.32 (t, J= 8.8Hz, 1H), 5.85 (q, J=7.2Hz, 1H), 1.70 (d, J=7.2Hz, 3H) .ESI-MS m/z:416.4[M+H]⁺.

Embodiment 3：

The synthesis of 4- (1- (the fluoro- 2,6- dichlorophenyls of 3-) ethyoxyl) the bromo- quinazolines of -6-

In 100mL single-necked flasks, 1- (fluoro- 2, the 6- dichlorophenyls of 3-) ethanol 2.71g (0.013mol), 15mL1 are added, 4- dioxane, cools down in ice bath, controls at 1-5 DEG C, to add 1.2g sodium hydrides (60%0.03mol), about 15min to add The bromo- quinazolines of the chloro- 6- of 2.43g (0.01mol) 4-, pour reactant liquor into 100mL H after about 1h in room temperature reaction overnight₂In O, Neutrality is adjusted to 10% hydrochloric acid, is filtered, the washing of 50% ethanol obtains white solid thing 3.3g, yield 80%.¹H NMR (400MHz,DMSO-d₆) δ 8.73 (s, 1H), 8.36 (d, J=2.0Hz, 1H), 8.10 (dd, J=2.0,8.8Hz, 1H), 7.86 (d, J=9.2Hz, 1H), 7.54 (dd, J=5.0,9.2Hz, 1H), 7.43 (t, J=8.8Hz, 1H), 6.90 (q, J=6.8Hz, 1H), 1.86 (d, J=6.8Hz, 3H) .ESI-MS m/z:417.6[M+H]⁺.

Embodiment 4：

The synthesis of N- (the chloro- 4-fluorophenyls of the 3-) bromo- quinazoline-4- ammonia of-6-

In 100mL there-necked flasks, the chloro- quinazoline 2.45g (0.01mol) of the bromo- 4- of 6-, the chloro- 4- fluoroanilines 1.45g of 3- are added (0.01mol), acetone 20mL, H₂O 5mL, concentrated hydrochloric acid 2-3 drops are heated to reflux 3h, place room temperature, and with concentrated ammonia liquor neutrality is adjusted to, Filter, water washing, be dried, obtain khaki solids 2.8g, yield 80%.¹H NMR(400MHz,DMSO-d₆)δ11.73(s, 1H), 9.28 (d, J=2.0Hz, 1H), 8.97 (s, 1H), 8.23 (dd, J=2.0,8.8Hz, 1H), 8.10 (dd, J=2.4, 6.8Hz, 1H), 7.94 (d, J=9.2Hz, 1H), 7.80 (ddd, J=2.8,4.4,7.2Hz, 1H), 7.55 (t, J=9.0Hz, 1H).ESI-MS m/z:354.3[M+H]⁺.

Embodiment 5：

The synthesis of the synthesis of the bromo- quinazoline -4- amine of N- (3- methoxy-benzyls) -6-

The preparation method of reference implementation example 2.Yield 68%.¹H NMR(400MHz,DMSO-d₆) δ 8.90 (t, J=5.4Hz, 1H), 8.63 (s, 1H), 8.50 (s, 1H), 7.90 (d, J=8.8Hz, 1H), 7.65 (d, J=9.2Hz, 1H), 7.24 (t, J= 8.0Hz, 1H), 6.94 (d, J=6.4Hz, 2H), 6.82 (d, J=7.6Hz, 1H), 4.75 (d, J=5.6Hz, 2H), 3.73 (s, 3H).

ESI-MS m/z:344.4[M+H]⁺.

Embodiment 6：

(14) synthesis of intermediate N (the 3- acetylene phenyl) bromo- quinazoline -4- amine of -6-

The preparation method of reference implementation example 4.Yield 86%.¹H NMR(400MHz,DMSO-d₆)δ11.83(s,1H),9.34 (d, J=1.6Hz, 1H), 8.99 (s, 1H), 8.25 (dd, J=2.0,8.8Hz, 1H), 7.97 (d, J=8.8Hz, 1H), 7.94 (t, J=1.6Hz, 1H), 7.84-7.81 (m, 1H), 7.51 (t, J=8.0Hz, 1H), 7.44-7.42 (m, 1H), 4.30 (s, 1H).ESI-MSm/z:326.2[M+H]⁺.

Embodiment 7：

The synthesis of N- (quinoline -6- bases) bromo- quinazoline -4- amine of -6-

The preparation method of reference implementation example 4.Yield 74%.¹H NMR(400MHz,DMSO-d₆)δ12.19(s,1H),9.45 (d, J=1.6Hz, 1H), 9.19 (dd, J=1.2,4.8Hz, 1H), 9.03 (s, 1H), 8.99 (d, J=8.0Hz, 1H), 8.71 (d, J=2.0Hz, 1H), 8.47 (dd, J=2.0,9.2Hz, 1H), 8.41 (d, J=9.2Hz, 1H), 8.27 (dd, J=2.0, 8.8Hz, 1H), 8.01 (d, J=8.8Hz, 1H), 7.96 (dd, J=5.2,8.4Hz, 1H) .ESI-MS m/z:353.7[M+H ]⁺.

Embodiment 8：

The synthesis of 4- (quinoline -6- epoxides) the bromo- quinazolines of -6- (II B-2)

In 100mL single port bottles, 6- oxyquinoline 1.6g (0.011mol), Se are added₂CO₃7.2g (0.022mol), two The ring 20mL of oxygen six, stirring adds the bromo- quinazoline 2.42g (0.01mol) of the chloro- 6- of 4- after 1 hour, be stirred overnight at room temperature, and will react Liquid pours 200mL H into₂In O, neutrality is adjusted to hydrochloric acid, is filtered, water washing, be dried, obtain white solid thing 2.1g, yield 60%.¹H NMR(400MHz,DMSO-d₆) δ 8.96 (d, J=2.8Hz, 1H), 8.79 (s, 1H), 8.62 (d, J=2.0Hz, 1H), 8.41 (d, J=8.0Hz, 1H), 8.22 (dd, J=2.0,8.8Hz, 1H), 8.16 (d, J=9.2Hz, 1H), 8.00 (d, J=9.2Hz, 2H), 7.82 (dd, J=2.8,9.2Hz, 1H), 7.61 (dd, J=4.0,8.4Hz, 1H) .ESI-MS m/z:354.8[M+H]⁺.

Embodiment 9：

The synthesis of 4- [(1- (3-fluoro- 2,6- Dichloro-phenyls) ethyoxyl]-6- (1- methyl isophthalic acid H- pyrazoles-4- bases) quinazoline

In 100mL there-necked flasks, 4- (1- (fluoro- 2, the 6- Dichloro-phenyls of 3-) ethyoxyl) the bromo- quinazoline 208mg of -6- are added (0.5mmol), 1- methyl -4- (4,4,5,5- tetramethyl -1,3,2- dioxy pentaborane -2- bases) -1H- pyrazoles 108mg (0.52mmol), K₂CO₃207mg (3equiv), Isosorbide-5-Nitrae-dioxane 12mL, H₂O 0.5mL, the lower logical N of stirring₂About 15min, plus Enter double (diphenyl phosphine) ferrocene palladium chloride (II) chloride dichloromethane complex pd (dppf) Cl of catalyst 1,1'-₂ 40.1mg (10%equiv) logical N, is continued₂After about 15min, heating stirring and N under 85 DEG C of oil baths are put into₂Protection, after reaction 4h.Add 30mL H₂O, is extracted three times, combined ethyl acetate with 90mL ethyl acetate, and revolving reclaims ethyl acetate, uses silica gel mixed sample rapid column chromatography (Yi Suan Yi Zhi ︰ Shi You Mi ︰ methyl alcohol (containing 1% the ammonia)=︰ 0.05 of 40 ︰ 60) is isolated and purified, product 98mg is obtained, 47% is received.¹H NMR (400MHz,DMSO-d₆) δ 8.62 (s, 1H), 8.36 (s, 1H), 8.30 (d, J=2.0Hz, 1H), 8.18 (dd, J=2.0, 8.8Hz, 1H), 8.02 (s, 1H), 7.88 (d, J=8.8Hz, 1H), 7.53 (dd, J=4.8,8.8Hz, 1H), 7.42 (t, J= 8.8Hz, 1H), 6.94 (q, J=6.8Hz, 1H), 3.93 (s, 3H), 1.88 (d, J=7.2Hz, 3H).¹³C NMR(101MHz, DMSO-d₆)δ164.9,153.4,149.6,137.6,136.9,132.6,129.1,128.5,121.2,117.7,117.5, 117.3,116.6,71.5,39.2,25.4,18.5.ESI-MS m/z:417.3[M+H]⁺.HRMS,ESI⁺,m/z:Calcd forC₂₀H₁₅Cl₂FN₄O(M+H)⁺,417.0680；found,417.0685.

Embodiment 10：

N- [1- (the fluoro- 2,6- Dichloro-phenyls of 3-) ethyl] -6- (1- piperidin-4-yl -1H- pyrazoles -4- bases) quinazoline -4- The synthesis of amine

In 100mL there-necked flasks, N is added⁴- (1- (the fluoro- 2,6- Dichloro-phenyls of the 3-) ethyl) bromo- quinazoline -4- amine of -6- 457mg (1.1m), adds 4- [4- (4,4,5,5- tetramethyl -1,3,2- dioxy ring pentaborane -2- bases) -1H- pyrazol-1-yls] piperazine Pyridine -1- t-butyl formate 400mg (1.06mmol), K₂CO₃420mg (3equiv), Isosorbide-5-Nitrae-dioxane 12mL, H₂O 0.5mL, The lower logical N of stirring₂About 15min, adds catalyst 1, double (diphenyl phosphine) ferrocene palladium chloride (II) chloride dichloromethane complex of 1'- pd(dppf)Cl₂80.2mg (10%equiv) continues logical N₂After about 15min, heating stirring under 85 DEG C of oil baths, N are put into₂Protection, Reaction 6h.Add 30mL H₂O, 150mL ethyl acetate is extracted in three times, combined ethyl acetate, and revolving reclaims ethyl acetate and obtains white Color solids.(Yi Suan Yi Zhi ︰ Shi You Mi ︰ methyl alcohol (containing 1% the ammonia)=︰ of 40 ︰ 60 are isolated and purified with silica gel mixed sample rapid column chromatography 0.05), white solid is obtained.White solid thing is dissolved in 15mL dichloromethane, is stirred overnight under addition 3.5mL trifluoroacetic acids de- BOC, revolving removes organic solvent, uses acetone:Petroleum ether=1:3 stirrings have solid to separate out, and filtration drying obtains white product 381mg, yield 79%.¹H NMR (400MHz, MeOD) δ 8.65 (d, J=1.6Hz, 1H), 8.29 (s, 2H), 8.15 (s, 1H), 8.03 (dd, J=2.0,8.4Hz, 1H), 7.70 (d, J=8.8Hz, 1H), 7.35 (dd, J=4.8,8.8Hz, 1H), 7.13 (t, J=8.6Hz, 1H), 6.10 (q, J=7.2Hz, 1H), 4.64 (ddd, J=4.4,12.6,17.0Hz, 1H), 3.61 (dt, J= 3.2,16.4Hz, 2H), 3.27 (td, J=3.6,12.8Hz, 2H), 2.45-2.30 (m, 4H), 1.84 (d, J=7.2Hz, 3H) .¹³C NMR(101MHz,MeOD)δ158.6,158.4,155.9,154.0,146.8,140.0,136.9,130.8,130.5, 129.4,129.3,126.7,126.0,122.1,117.6,115.0,114.8,55.7,49.1,42.8,28.9,16.4.ESI- MS m/z:485.5[M+H]⁺.HRMS,ESI⁺,m/z:Calcd forC₂₄H₂₃Cl₂FN₆(M+H)⁺,485.1418；found, 485.1420.

Embodiment 11：

(21) 4- [1- (the fluoro- 2,6- Dichloro-phenyls of 3-) ethyoxyl] -6- (1- piperidin-4-yl -1H- pyrazoles -4- bases) quinoline azoles The synthesis of quinoline (I -3)

The preparation method of reference implementation example 10.Yield 50%.¹H NMR(400MHz,MeOD)δ8.56(s,1H),8.39 (s, 1H), 8.26 (s, 1H), 8.14 (dd, J=1.2,8.4Hz 1H), 7.99 (s, 1H), 7.85 (d, J=8.8Hz, 1H), 7.41 (dd, J=4.8,8.8Hz, 1H), 7.21 (t, J=8.6Hz, 1H), 7.02 (q, J=6.8Hz, 1H), 4.49-4.42 (m, 1H), 3.36 (d, J=7.2Hz, 1H), 3.29 (s, 1H), 2.96-2.90 (m, 2H), 2.25-2.01 (m, 5H), 1.94 (d, J= 6.8Hz,3H).¹³CNMR(101MHz,MeOD)δ165.3,158.5,156.0,153.1,148.9,138.5,137.5,136.3, 132.1,129.8,128.7,127.0,125.7,121.3,118.0,116.5,104.9,72.0,58.4,44.1,31.7, 17.0.ESI-MS m/z:486.6[M+H]⁺.HRMS,ESI⁺,m/z:Calcd for C₂₄H₂₂Cl₂FN₅O(M+H)⁺, 486.1258；found,486.1254.

Embodiment 12：

The synthesis of N- (quinoline -6- bases) -6- (1- methyl isophthalic acid H- pyrazoles -4- bases) quinazoline -4- amine

The preparation method of reference implementation example 10.Yield 79%.¹H NMR(400MHz,DMSO-d₆)δ10.02(s,1H), 8.84 (dd, J=1.6,4.0Hz, 1H), 8.78 (d, J=1.6Hz, 1H), 8.65 (s, 1H), 8.57 (d, J=2.0Hz, 1H), 8.37 (d, J=7.6Hz, 1H), 8.31 (s, 1H), 8.24 (dd, J=2.0,8.8Hz, 1H), 8.17-8.07 (m, 3H), 7.82 (d, J=8.8Hz, 1H), 7.53 (dd, J=4.0,8.0Hz, 1H), 3.95 (s, 3H).¹³C NMR(101MHz,DMSO-d₆)δ 157.8,154.1,149.7,148.7,145.4,137.7,136.9,135.9,131.4,131.2,129.5,128.8, 128.6,126.6,122.2,121.8,118.6,117.9,116.0,99.9,39.2.ESI-MS m/z:353.6[M+H]⁺ .HRMS,ESI⁺,m/z:Calcd forC₂₁H₁₆N₆(M+H)⁺,353.1509；found,353.1512.

Embodiment 13：

The synthesis of 4- (hydroxyl of quinoline -6) -6- (1- methyl isophthalic acid H- pyrazoles -4- bases) quinazoline

With reference to the preparation method for changing embodiment 10.Yield 83%.¹H NMR(400MHz,DMSO-d₆) δ 8.96 (dd, J= 1.6,4.0Hz, 1H), 8.67 (s, 1H), 8.55 (d, J=2.0Hz, 1H), 8.49 (s, 1H), 8.42 (dd, J=1.2,6.8Hz, 1H), 8.31 (dd, J=2.0,8.8Hz, 1H), 8.16 (t, J=4.6Hz, 2H), 8.02 (dd, J=4.4,7.2Hz, 2H), 7.83 (dd, J=2.4,8.8Hz, 1H), 7.61 (dd, J=4.0,8.4Hz, 1H), 3.92 (s, 3H).¹³C NMR(101MHz, DMSO-d₆)δ166.6,153.3,150.9,150.4,150.2,146.2,137.1,136.1,133.1,132.9,131.0, 129.4,128.9,128.6,126.1,122.4,121.1,119.4,117.6,116.6,39.2.MS:m/z(ESI+)354.5 [M+H]⁺.HRMS,ESI⁺,m/z:Calcdfor C₂₁H₁₅N₅O(M+H)⁺,354.1349；found,354.1348.

Embodiment 14：

The synthesis of N- (quinoline -6- bases) -6- (1- piperidin-4-yl -1H- pyrazoles -4- bases) quinazoline -4- amine

The preparation method of reference implementation example 10.Yield 63%.¹H NMR(400MHz,DMSO-d₆)δ10.02(s,1H), 8.84 (dd, J=1.6,4.4Hz, 1H), 8.78 (d, J=1.2Hz, 1H), 8.64 (s, 1H), 8.57 (d, J=2.0Hz, 1H), 8.39-8.35 (m, 2H), 8.25 (dd, J=2.4,9.2Hz, 1H), 8.16-8.12 (m, 2H), 8.07 (d, J=9.2Hz, 1H), 7.81 (d, J=8.4Hz, 1H), 7.52 (dd, J=4.0,8.0Hz, 1H), 4.29-4.24 (m, 1H), 3.09 (d, J= 12.0Hz, 2H), 2.63 (t, J=11.8Hz, 2H), 2.06-2.03 (m, 2H), 1.90-1.81 (m, 2H).¹³C NMR (101MHz,DMSO-d₆)δ157.8,154.0,149.6,148.7,145.4,137.7,136.6,135.9,131.6,131.3, 129.4,128.8,128.6,126.6,125.8,122.2,121.3,118.6,117.9,116.0,59.8,45.4, 33.9.ESI-MS m/z:422.7[M+H]⁺.HRMS,ESI⁺,m/z:Calcd for C₂₅H₂₃N₇(M+H)⁺,422.2088； found,422.2093.

Embodiment 15：

The synthesis of N- (the chloro- 4-fluorobenzene of 3-) base-6- (the 1- methyl isophthalic acid H- pyrazoles-4- bases) amine of quinazoline-4

The preparation method of reference compound embodiment 10.Yield 69%.¹H NMR(400MHz,DMSO-d₆)δ9.78(s, 1H), 8.61 (d, J=21.6Hz, 2H), 8.22 (d, J=23.6Hz, 2H), 8.05 (s, 2H), 7.82 (d, J=27.6Hz, 2H),7.46(s,1H),3.93(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ157.6,155.0,153.9,152.6, 148.6,136.9,131.2,128.8,124.1,122.9,121.8,119.4,119.2,117.7,117.1,116.9, 115.8,39.9.ESI-MS m/z:354.6[M+H]⁺.HRMS,ESI⁺,m/z:Calcd for C₂₁H₁₆N₆(M+H)⁺, 354.0916；found,354.0919.

Embodiment 16：

The synthesis of N- (the chloro- 4- fluorophenyls of 3-) -6- (1- piperidin-4-yl -1H- pyrazoles -4- bases) quinazoline -4- amine

The preparation method of reference compound embodiment 10.Yield 63%.¹H NMR(400MHz,DMSO-d₆)δ9.82(s, 1H), 8.67 (d, J=1.4Hz, 1H), 8.59 (s, 1H), 8.36 (s, 1H), 8.20 (dd, J=2.4,6.8Hz, 1H), 8.12 (dd, J=2.0,8.8Hz, 1H), 8.08 (s, 1H), 7.87 (ddd, J=2.4,5.6,7.6Hz, 1H), 7.79 (d, J= 8.8Hz, 1H), 7.47 (t, J=9.2Hz, 1H), 4.29-4.22 (m, 1H), 3.10-2.99 (m, 3H), 2.66-2.55 (m, 2H), 2.03 (d, J=10.0Hz, 2H), 1.89-1.82 (m, 2H).¹³C NMR(101MHz,DMSO-d₆)δ157.6,155.0 153.9,152.6,148.6,136.5,131.6,128.7,125.8,124.1,123.0,121.2,119.4,117.7, 117.1,116.9,115.8,59.8,45.4,33.9,33.7,22.9.ESI-MS m/z:423.5[M+H]⁺.HRMS,ESI⁺,m/ z:Calcd for C₂₄H₂₂Cl₂FN₅O(M+H)⁺,423.1495；found,423.1499.

Embodiment 17：

The synthesis of N- (3- methoxybenzyls) -6- (the 1- methyl isophthalic acid H- pyrazoles -4- bases) amine of quinazoline -4

The preparation method of reference implementation example 10.Yield 47%.¹H NMR(400MHz,DMSO-d₆)δ8.71(s,1H),8.53 (s, 1H), 8.41 (s, 1H), 8.20 (s, 1H), 7.99 (d, J=8.8Hz, 2H), 7.68 (d, J=8.8Hz, 1H), 7.25 (t, J =7.6Hz, 1H), 6.97 (s, 2H), 6.83 (d, J=8.4Hz, 1H), 4.81 (d, J=3.6Hz, 2H), 3.91 (s, 3H), 3.73(s,3H).¹³C NMR(101MHz,DMSO-d₆)δ159.8,159.6,154.8,148.0,141.5,136.7,130.6, 130.5,129.9,128.5,128.4,121.9,119.8,117.9,115.6,113.6,112.5,55.4,43.9, 39.2.ESI-MS m/z:346.2[M+H]⁺.HRMS,ESI⁺,m/z:Calcd for C₂₀H₁₉N₅O(M+H)⁺,346.1662； found,346.1666.

Embodiment 18：

The synthesis of N- (3- ethynyl phenyls) -6- (the 1- methyl isophthalic acid H- pyrazoles -4- bases) amine of quinazoline -4

The preparation method of reference implementation example 10.Crude product C18 reversed phase chromatography separations are purified, Jia Chun ︰ H₂O (0.05mol acetic acid Ammonia, formic acid adjusts pH=4.0)=35 ︰ 65, yield 37%.¹H NMR(400MHz,DMSO-d₆)δ9.79(s,1H),8.70(s, 1H), 8.60 (s, 1H), 8.28 (s, 1H), 8.10-8.06 (m, 3H), 7.96 (d, J=8.0Hz, 1H), 7.79 (d, J= 8.4Hz, 1H), 7.45 (t, J=7.8Hz, 1H), 7.27 (d, J=7.6Hz, 1H), 4.23 (s, 1H), 3.93 (s, 3H).¹³C NMR(101MHz,DMSO-d₆)δ157.7,153.9,148.3,139.8,136.9,131.4,131.3,129.4,128.8, 128.6,127.3,125.5,123.3,122.3,121.8,117.8,115.9,83.8,81.1,39.2.ESI-MS m/z: 326.6[M+H]⁺.HRMS,ESI⁺,m/z:Calcd for C₂₁H₁₆N₆(M+H)⁺,326.1400；found,326.1406.

Embodiment 19：

The synthesis of chloro- 6 bromo- pyrido [2,3-d] pyrimidines of 4-

In equipped with agitator, the 500mL there-necked flasks of reflux condensing tube, 2- amino-nicotinic acid 20.7g (0.15mol) is added, 80mL acetic acid, stirs under room temperature, and the mixed solution of 40mL acetic acid and 4.1mL bromines (0.08mol) is added dropwise with constant pressure funnel, Drip off continuation and stir 4-5h.Filter, washed with acetic acid, be dried, solids methyl alcohol 100mL is heated to reflux 10min and places room temperature, Filtration drying.Obtain white solid 31.2g, yield 96%.¹H NMR(400MHz,DMSO-d6)δ12.72(s,1H),9.05(d,J =2.4Hz, 1H), 8.63 (d, J=2.4Hz, 1H), 8.36 (s, 1H) .MS:m/z(ESI+)228.3[M+H]⁺。

The preparation method of the A of synthesized reference I of chloro- 6 bromo- pyrido [2,3-d] pyrimidines of 4-.Yield 73%.¹H NMR (400MHz,CDCl₃) δ 9.33 (d, J=7.6Hz, 2H), 8.80 (s, 1H), MS:m/z(ESI+)246.9[M+H]⁺。

Embodiment 20：

The synthesis of intermediate N (3- methoxy-benzyls) -6- bromo- pyrido [2,3-d] pyrimidine -4- amine

The preparation method of reference implementation example 2.Yield 58%.¹H NMR(400MHz,DMSO-d₆) δ 9.16 (t, J=5.6Hz, 1H), 9.11 (d, J=2.4Hz, 1H), 9.07 (d, J=2.4Hz, 1H), 8.64 (s, 1H), 7.25 (t, J=8.2Hz, 1H), (s, 3H) the .ESI-MS m/z of 6.96-6.95 (m, 2H), 6.86-6.85 (m, 1H), 4.77 (d, J=5.6Hz, 2H), 3.74: 347.1[M+H]⁺.

Embodiment 21：

The synthesis of N- (3- nitrobenzyls) -6- bromo- pyrido [2,3-d] pyrimidine -4- amine

The preparation method of reference implementation example 2.Yield 72%.¹H NMR(400MHz,DMSO-d₆)δ9.66(s,1H),9.14 (d, J=8.0Hz, 2H), 8.72 (s, 1H), 8.28 (s, 1H), 8.15 (d, J=7.6Hz, 1H), 7.88 (d, J=7.6Hz, 1H), 7.65 (t, J=7.8Hz, 1H), 4.95 (d, J=5.2Hz, 2H) .ESI-MS m/z:362.3[M+H]⁺

Embodiment 22：

N- (pyridin-4-yl) ethyl) the bromo- pyridines of -6- [2,3-d] and pyrimidine -4- amine synthesis

With reference to the preparation method for changing embodiment 2.Yield 76%.¹H NMR(400MHz,DMSO-d₆) δ 9.27 (d, J= 2.4Hz, 1H), 9.08 (d, J=2.4Hz, 1H), 8.90 (d, J=7.4Hz, 1H), 8.57 (s, 1H), 8.50 (dd, J=1.6, 4.8Hz, 2H), 7.43 (dd, J=1.2,4.4Hz, 2H), 5.50 (q, J=7.0Hz, 1H), 1.60 (d, J=7.2Hz, 3H) .ESI-MS m/z:332.5[M+H]⁺.

Embodiment 23：

The conjunction of intermediate N (1- (the chloro- 3- fluorophenyls of 2,6- bis-)-ethyl) -6- bromo- pyrido [2,3-d] pyrimidine -4- amine Into

With reference to the preparation method for changing embodiment 2.Yield 86%.¹H NMR(400MHz,DMSO-d₆) δ 9.40 (d, J= 2.4Hz, 1H), 9.07 (d, J=2.0Hz, 1H), 9.03 (d, J=5.2Hz, 1H), 8.48 (s, 1H), 7.47-7.44 (m, 1H), (d, J=7.2Hz, 3H) the .ESI-MS m/z of 7.35 (t, J=8.8Hz, 1H), 5.85 (q, J=7.0Hz, 1H), 1.71:417.4 [M+H]⁺.

Embodiment 24：

N- [(1- (the chloro- 3-fluorophenyls of 2,6- bis-) ethyl)]-6- (1- piperidin-4-yl-1H- pyrazoles-4- bases) pyrido [2, 3-d] amine of pyrimidine -4 synthesis

The preparation method of reference implementation example 10.There is solid to separate out with the stirrings of Bing Tong ︰ petroleum ether=1 ︰ 1, filtration drying obtains white Color product, yield 48%.¹H NMR(400MHz,DMSO-d₆)δ11.22(s,1H),10.13(s,1H),9.42(s,1H),9.21 (s, 1H), 8.78 (s, 2H), 8.39 (s, 1H), 7.49 (s, 1H), 7.39 (t, J=7.2Hz, 1H) 5.96 (s, 1H), 4.60 (s, 1H), (d, J=6.0Hz, 3H) the .13C NMR (101MHz, MeOD) of 3.61 (s, 2H), 3.12 (s, 2H), 2.27 (s, 4H), 1.87 δ161.0,158.5,156.0,155.1,151.8,146.4,138.1,137.1,130.4,129.9,128.1,127.6, 117.6,116.0,115.8,109.2,56.1,51.3,43.0,28.9,16.1.ESI-MS m/z:486.5[M+H]+.HRMS, ESI+,m/z:Calcd for C23H22Cl2FN7(M+H)+,486.1371；found,486.1368.

Embodiment 25：

N- [(1- (3-fluoro- 2,6- Dichloro-phenyls) ethyl)]-6- (1- methyl isophthalic acid H- pyrazoles-4- bases) pyrido [2,3-d] The synthesis of the amine of pyrimidine -4

The preparation method of reference implementation example 10.Silica gel chromatograph column separating purification, second acid second ester ︰ stones oil ether ︰ methyl alcohol (contains 1% Ammonia) ︰ 0.05 of=40 ︰ 60, yield 62%.¹H NMR (400MHz, MeOD) δ 9.20 (d, J=2.0Hz, 1H), 9.06 (d, J= 2.4Hz, 1H), 8.43 (s, 1H), 8.21 (s, 1H), 8.07 (s, 1H), 7.35 (dd, J=8.8,5.2Hz, 1H), 7.14 (t, J =8.6Hz, 1H), 6.07 (q, J=7.4Hz, 1H), 4.01 (s, 3H), 1.83 (d, J=7.2Hz, 3H).¹³C NMR(101MHz, MeOD)δ159.6,157.2,155.9,152.9,139.6,136.5,129.5,129.4,128.4,127.1,126.9, 118.8,115.2,114.9,109.6,49.3,37.8,16.3.ESI-MS m/z:417.5[M+H]⁺.HRMS,ESI⁺,m/z: Calcd for C₁₉H₁₅Cl₂FN₆(M+H)⁺,417.0792；found,417.0794..

Embodiment 26：

The synthesis of N- [1- (pyridin-4-yl) ethyl] -6- (1- methyl isophthalic acid H- pyrazoles -4- bases) pyrido [2,3-d] pyrimidine

The preparation method of reference implementation example 10.Yield 55%.¹H NMR (400MHz, MeOD) δ 9.20 (d, J=9.6Hz, 1H), 8.99 (s, 1H), 8.49 (d, J=4.8Hz, 3H), 8.20 (d, J=7.6Hz, 1H), 8.05 (d, J=7.2Hz, 1H), 7.52 (d, J=5.6Hz, 2H), 5.61 (q, J=5.6Hz, 1H), 4.00 (s, 3H), 1.74 (d, J=7.2Hz, 3H).¹³C NMR (101MHz,MeOD)δ160.3,157.1,156.0,154.4,153.1,148.8,136.5,128.4,127.1,126.9, 121.7,118.7,109.8,49.9,37.8,20.3.ESI-MS m/z:332.6[M+H]⁺.HRMS,ESI⁺,m/z:Calcd for C₁₈H₁₇N₇(M+H)⁺,332.1618；found,332.1621.

Embodiment 27：

The synthesis of N- (3- methoxybenzyls) -6- (1- methyl isophthalic acid H- pyrazoles -4- bases) pyrido [2,3-d] amine of pyrimidine -4

The preparation method of reference implementation example 10.Yield 67%.¹H NMR(400MHz,DMSO-d₆)δ9.28(s,1H),9.11 (s, 1H), 8.99 (s, 1H), 8.58 (s, 1H), 8.32 (s, 1H), 8.03 (s, 1H), 7.26 (t, J=8.0Hz, 1H), 6.98 (s, 2H), 6.85 (d, J=7.6Hz, 1H), 4.82 (d, J=5.6Hz, 2H), 3.92 (s, 3H), 3.74 (s, 3H).¹³C NMR (101MHz,DMSO-d₆)δ159.8,159.6,154.8,148.1,141.5,136.7,130.6,130.6,129.9,128.5, 128.4,121.9,119.8,117.9,115.7,113.6,112.5,55.4,43.9,39.2.ESI-MS m/z:347.4[M+ H]⁺.HRMS,ESI⁺,m/z:Calcd for C₁₉H₁₈N₆O(M+H)⁺,347.1615；found,347.1619.

Embodiment 28：

The synthesis of N- (3- nitrobenzyls) -6- (1- methyl isophthalic acid H- pyrazoles -4- bases) pyrido [2,3-d] amine of pyrimidine -4

The preparation method of reference implementation example 10.Yield 74%.¹H NMR(400MHz,DMSO-d₆)δ9.41(s,1H),9.28 (d, J=1.6Hz, 1H), 9.05 (s, 1H), 8.54 (s, 1H), 8.35 (s, 1H), 8.27 (s, 1H), 8.13 (d, J=8.0Hz, 1H), 8.06 (s, 1H), 7.88 (d, J=7.2Hz, 1H), 7.64 (t, J=7.8Hz, 1H), 4.94 (s, 2H), 3.93 (s, 3H) .¹³C NMR(101MHz,DMSO-d₆)δ161.0,157.8,157.0,153.6,148.3,142.1,136.8,134.6, 130.3,128.8,127.0,126.7,122.4,122.3,118.7,110.2,66.8,43.8.ESI-MS m/z:362.2[M+ H]⁺.HRMS,ESI⁺,m/z:Calcd for C₁₈H₁₅N₇O₂(M+H)⁺,362.1360；found,362.1363.

Embodiment 29：

The synthesis of the bromo- quinazolines of 2- amino-4-hydroxy -6-

In equipped with agitator, the 500mL there-necked flasks of reflux condensing tube, the bromo- benzoic acid 15g of 2- amino -5- are added (0.07mol) solution that, 150ml water and the 8mL concentrated sulfuric acids are mixed, after being heated to 100 DEG C, after about 15min dicyandiamide is added 8.75g (0.11mol), reacts 2h, is cooled to room temperature, filters to obtain white solid, the white solid NaOH of 100mL (4mol/L) It is basified, 15min is heated, place to room temperature, filter, with substantial amounts of water washing to neutrality, dry white solid thing 11.8g, yield 60% is not purified to be directly used in the next step.

In equipped with agitator, the 100mL there-necked flasks of reflux condensing tube, the bromo- quinazolines of 2- amidino groups -4- hydroxyl -6- are added 8g (0.028mol), KOH7.8g (0.14mol), ethylene glycol 20mL, 4- mercaptopyridine 6.2g (0.056mol), oil bath heating is arrived 170 DEG C of stir about 4h stop reaction, place to room temperature, pour 500mL H into₂In O, neutrality is adjusted to hydrochloric acid, there are a large amount of solids to analyse Go out, filter, water washing for several times, is washed with 50% ethanol, be vacuum dried, obtain off-white powder 4.6g, yield 68%.¹H NMR (400MHz,DMSO-d₆) δ 11.11 (s, 1H), 7.97 (d, J=2.4Hz, 1H), 7.69 (dd, J=2.8,8.8Hz, 1H), 7.57 (s, 2H), 7.30 (d, J=8.8Hz, 1H) .ESI-MS m/z:242.0[M+H]⁺.

Embodiment 30：

N⁴The synthesis of-(1- (the fluoro- 2,6- Dichloro-phenyls of the 3-) ethyl) bromo- quinazoline -2,4- diamines of -6-

In 100mL single port bottles, the bromo- quinazoline 1.2g (0.005mol) of 2- amino-4-hydroxy -6-, the nitrogen of benzo three are added (dimethylamino) phosphorus hexafluorophosphate (BOP) 2.9g (0.0065mol, 1.3equiv) the acetonitrile 20mL of azoles -1- bases epoxide three, It is stirred at room temperature, adds carbon -7- alkene (DBU) 1.1mL (0.0075mol, the 1.5equiv) solution of 1,8- diaza-bicyclos 11 to become clarification, There is solid to separate out after reaction 10min, add 1- (fluoro- 2, the 6- dichlorophenyls of 3-) ethamine 1.56g (0.0075mol, 1.5equiv), It is stirred overnight at room temperature, adds 100mL H₂O, is extracted three times, combined ethyl acetate with 90mL ethyl acetate, and revolving reclaims acetic acid second Ester, obtains white solid thing.(︰ 2 of Shi You Mi ︰ bis- Lv Jia Wan ︰ ethyl acetate=2 ︰ 1) purifying is separated with silica gel mixed sample rapid column chromatography Obtain 0.97g, yield 45%.¹H NMR(400MHz,DMSO-d₆₎δ 8.60 (d, J=2.4Hz, 1H), 8.46 (d, J=5.6Hz, 1H), 8.17 (s, 1H), 7.62 (dd, J=2.4,9.2Hz, 1H), 7.42 (dd, J=4.8,8.8Hz, 1H), 7.32 (t, J= 8.8Hz, 1H), 7.15 (d, J=8.8Hz, 1H), 5.94 (s, 1H), 5.82 (q, J=6.4Hz, 1H), 1.67 (d, J=7.4Hz, 3H).ESI-MS m/z:431.4[M+H]⁺。

Embodiment 31：

4- (1- (the fluoro- 2,6- Dichloro-phenyls of the 3-) ethyoxyl) bromo- quinazoline -2- amino of -6- into

In 100mL single port bottles, the bromo- quinazoline 1.2g (0.005mol) of 2- amino-4-hydroxy -6-, the nitrogen of benzo three are added (dimethylamino) phosphorus hexafluorophosphate (BOP) 2.9g (0.0065mol, 1.3equiv) of azoles -1- bases epoxide three, acetonitrile 20mL, is stirred at room temperature, and adds carbon -7- alkene (DBU) 1.1mL (0.0075mol, the 1.5equiv) solution of 1,8- diaza-bicyclos 11 to become Clarification, reaction 10min after have solid to separate out, add 1- (fluoro- 2, the 6- Dichloro-phenyls of 3-) ethanol 1.56g (0.0075mol, 1.5equiv), the mixed liquor of 0.4gNaH (60%) and 10mL acetonitriles composition, is stirred overnight at room temperature, and adds 200mL H₂O, uses 90mL ethyl acetate is extracted three times, combined ethyl acetate, and revolving reclaims ethyl acetate, obtains white solid thing.It is fast with silica gel mixed sample Fast column chromatography for separation (︰ 2 of Shi You Mi ︰ bis- Lv Jia Wan ︰ ethyl acetate=2 ︰ 1) purifies to obtain 1.2g, yield 55%.¹H NMR (400MHz,CDCl₃) δ 8.23 (d, J=2.0Hz, 1H), 7.70 (dd, J=9.2,2.4Hz, 1H), 7.35 (d, J=8.8Hz, 1H), 7.27 (dd, J=5.2,8.8Hz, 1H), 7.05 (t, J=8.4Hz, 1H), 6.86 (q, J=6.8Hz, 1H), 4.99 (s, 2H), 1.91 (d, J=6.9Hz, 3H) .ESI-MS m/z:432.4[M+H]⁺。

Embodiment 32：

N⁴- (1- (3-fluoro- 2,6- Dichloro-phenyls) ethyl)-6- (1- piperidin-4-yl-1H- pyrazoles-4- bases) quinazoline-2, The synthesis of 4- diamines

The preparation method of reference implementation example 10.Yield 74%.¹H NMR(400MHz,MeOD)δ8.69(s,1H),8.24 (s, 1H), 8.07-8.03 (m, 2H), 7.47-7.39 (m, 2H), 7.18 (dd, J=8.4,17.2Hz, 1H), 6.09 (q, J= 7.2Hz,1H),4.65–4.63(m,1H),3.64–3.53(m,3H),3.37(s,1H),3.28–3.17(m,2H),2.66(d,J =9.6Hz, 1H), 2.40-2.25 (m, 5H), 1.84 (d, J=7.2Hz, 3H).¹³C NMR(101MHz,MeOD)δ138.8, 137.2,136.6,132.4,129.9,129.7,127.9,125.8,121.2,119.3,117.2,115.6,115.4, 114.3,110.0,55.7,49.8,42.7,42.6,28.8,28.6,16.0.ESI-MS m/z:500.5[M+H]⁺.HRMS, ESI⁺,m/z:Calcd forC₂₄H₂₄Cl₂FN₇(M+H)⁺,500.1527；found,500.1528.

Embodiment 33：

4- (1- (3-fluoro- 2,6- Dichloro-phenyls) ethyoxyl)-6- (1- piperidin-4-yl-1H- pyrazoles-4- bases) quinazoline- The synthesis of 2- amino

The preparation method of reference implementation example 10.Yield 65%.¹H NMR(400MHz,DMSO-d₆)δ8.23(s,1H),8.06 (d, J=2.0Hz, 1H), 7.89-7.87 (m, 2H), 7.55 (dd, J=5.2,8.8Hz, 1H), 7.44 (t, J=8.8Hz, 1H), 7.34 (d, J=8.8Hz, 1H), 6.94 (q, J=6.8Hz, 1H), 6.39 (s, 2H), 4.34-4.28 (m, 1H), 3.16 (d, J= 14.4Hz, 3H), 2.76-2.70 (m, 2H), 2.06-1.91 (m, 4H), 1.85 (d, J=7.2Hz, 3H).¹³C NMR(101MHz, MeOD)δ166.2,159.5,158.6,151.2,137.5,136.3,132.0,129.9,128.9,126.8,125.3, 123.7,122.2,119.0,116.2,116.0,111.5,71.4,56.1,43.0,29.4,16.9.ESI-MS m/z:501.4 [M+H]⁺.HRMS,ESI⁺,m/z:Calcd for C₂₄H₂₃Cl₂FN₆O(M+H)⁺,501.1367；found,501.1369.

Embodiment 34：

N⁴- (1- (3-fluoro- 2,6- Dichloro-phenyls) ethyl)-6- (1- methyl isophthalic acid H- pyrazoles-4- bases)-2,4- quinazolines two The synthesis of amine

The preparation method of reference implementation example 10.Yield (51%)¹H NMR (400MHz, MeOD) δ 8.43 (d, J=2.0Hz, 1H), 8.01 (s, 1H), 7.96 (s, 1H), 7.81 (dd, J=1.6,8.4Hz, 1H), 7.38-7.30 (m, 2H), 7.14 (t, J= 8.6Hz, 1H), 6.04 (q, J=7.3Hz, 1H), 3.97 (s, 3H), 1.80 (d, J=7.2Hz, 3H).¹³C NMR(101MHz, MeOD)δ159.6,158.6,155.9,146.8,139.9,136.0,130.7,129.3,127.6,127.0,122.5, 122.4,118.3,115.1,114.9,110.7,48.9,37.6,16.2.ESI-MS m/z:431.5[M+H]⁺.HRMS,ESI⁺, m/z:Calcd forC₂₀H₁₇Cl₂FN₆(M+H)⁺,431.0949；found,431.0950.

Embodiment 35：

Tumor cell line inhibitory activity (IC₅₀)

Compound strain MDA-MB-231, MCF-7, BT-474, SK-BR-3 thin to human breast carcinoma, lung cancer are determined using mtt assay Five kinds of tumor cell line experiments of cell line A549.After five kinds of tumor cell line cell dissociations of exponential phase, list is blown and beaten into Cell suspension, is inoculated in respectively 96 well culture plates；Per hole 5 × 10₃Individual cell, the μ L of culture medium 200,37 DEG C, 5%CO are added per hole₂ Overnight incubation in incubator.After cell attachment, the test-compound and Gefitinib of various dose are separately added into, configuration is different The sample of concentration, with blank group as negative control group, with Gefitinib as positive controls, is further cultured for 72h in incubator.So Afterwards, add 20 μ L mass concentrations for the MTT liquid of 5mg/mL per hole, continuously cultivate 4h.Supernatant is sucked, 150 μ L bis- are added per hole First sulfoxide, culture plate is placed in micropore plate oscillator and vibrates 10min, dissolves crystal.ELIASA is used at 490nm wavelength Absorbance A value is surveyed, inhibiting rate is calculated；IC is calculated by Bliss methods₅₀.Result of the test see the table below.

Section Example numbering compound strain MDA-MB-231, MCF-7, BT-474, SK-BR-3 thin to human breast carcinoma, lung The inhibitory activity of JEG-3 A549 is shown in Table 1

Embodiment 36：

The test of kinase activity：

Reagent：EGFR (ErbB1) T790M L858R kinases (Life Technologies companies, article No.：PV4803)； EGFR (ErbB1) L858R kinases (Life Technologies companies, article No.：PV4128)；EGFR (ErbB1) kinases (LifeTechnologies companies, article No.：PV3872), ERBB2 (HER2) kinases (Life Technologies companies, goods Number：PV3366)；ERBB4 (HER4) kinases (Life Technologies companies, article No.：PV3626)；Tyrosine kinase activity Detection kit (Kinase Assay Kit, Life Technologies company, article No.：PV3190)；384 is micro- Hole dark circles bottom reaction plate (Corning Incorporated, article No.：3676)

Instrument：TECANM1000Pro all-wave length multi-function microplate readers

1.EGFR kinases existsThe optimization of optimum concentration in kinase activity detection reaction system

1. 1 × kinase buffer solution is prepared

2. the kinase solution of 10 concentration of two-fold dilution is prepared

3. the mixed liquor of 9 peptide substrates and ATP and ATP is prepared

4. Phosphorylated Peptide solution is prepared

5. differential protein hydrolysis enzyme solutions are prepared

6. the kinase solution of 5 μ L is sequentially added in 384 microwell plates, each concentration repeats 5 holes

7. 0% phosphorylation control wells add 1 × kinase buffer solution of 5 μ L

8. 100% phosphorylation control wells add the Phosphorylated Peptide of 5 μ L

9. 9 peptide substrates of 5 μ L and the mixed liquor of ATP and ATP are added in experimental port and control wells, are then shaken 30 seconds, Incubation at room temperature 1 hour.

10. add 5 μ L to prepare differential protein hydrolase, then shake 30 seconds, be incubated at room temperature 1 hour, then use TECANM1000Pro all-wave lengths multi-function microplate reader under conditions of excitation wavelength=400nm, Detection wavelength=445nm With the emitted luminescence intensity of wavelength=520nm.Transmitting ratio=E (wavelength=445nm)/E (wavelength=520nm)；Substrate phosphorylation hundred Fraction=1- (transmitting ratio * F100%-C100%)/[(C0-C100%)+transmitting ratio * (F100%-F0%)]

Substrate phosphorylation percentage is selected to be that 30% ± 10% corresponding kinase concentration is tyrosine kinase inhibitor screening The optium concentration of reaction system.

The screening of 2.5 kinds of kinase inhibitors

1. 1.33 × kinase buffer solution is prepared

2. the EGFR kinases and 9 peptide substrates solution for determining concentration is prepared

3. the solution of 4 × ATP is prepared

4. Phosphorylated Peptide solution is prepared

5. differential protein hydrolysis enzyme solutions are prepared

6. the kinase inhibition agent solution to be screened of 10 concentration gradients is prepared respectively with 4% DMSO solution, starting is dense Spend for 1000nM, with 3 times of dilution process each concentration is obtained, the variable concentrations that 2.5 μ L are sequentially added in 384 microwell plates swash Enzyme inhibitor solution, each concentration repeats 5 holes

7. control wells are suppressed 0%, 100% suppresses control wells, 100% phosphorylation control wells to be separately added into the 4% of 2.5 μ L DMSO solution

8. kinases and 9 peptide substrates solution that 5 μ L determine concentration are sequentially added

9. 100% phosphorylation control wells add 5 μ L Phosphorylated Peptide solution, and then 0% suppresses control wells and experimental port to add 4 The μ L of solution 2.5 of × ATP；100% suppresses control wells and 100% phosphorylation to add the μ L of 1.33 × kinase buffer solution 2.5, then Concussion 30 seconds, is incubated at room temperature 1 hour.

10. add 5 μ L to prepare differential protein hydrolase, then shake 30 seconds, be incubated at room temperature 1 hour, then use TECANM1000Pro all-wave lengths multi-function microplate reader under conditions of excitation wavelength=400nm, Detection wavelength=445nm With the emitted luminescence intensity of wavelength=520nm.Transmitting ratio=E (wavelength=445nm)/E (wavelength=520nm)；Substrate phosphorylation hundred Fraction=1- (transmitting ratio * F100%-C100%)/[(C0-C100%)+transmitting ratio * (F100%-F0%)]；Substrate phosphorylation Inhibiting rate=(100% phosphorylation control wells-experiment detection hole)/100% phosphorylation control wells.Have chosen compound concentration is During 1 μ Μ, when it suppresses more than 95% to kinase activity, the compound concentration (IC under 50% inhibitory action is carried out₅₀) experiment. Compound concentration (the IC under 50% inhibitory action is calculated using the softwares of GraphPad Prism 5₅₀).Experimental result is shown in Table 2：

Table 1：Compound on tumor cell proliferation results and the inhibiting rate (1 μ Μ) to kinases

Table 2：The IC that compound kinases EGFR biochemical activities suppress₅₀Value

Claims

1. formula (I) or the compound shown in (II) or (III) or its pharmaceutically acceptable salt, hydrate, solvate, Polymorph, dynamic isomer or prodrug,

In formula：

X is O or NH；

R₁Having structure unit can be selected from：

Wherein R₃, R₄It is each independently selected from H, F, Cl, Br, I, CH₃、OCH₃、NO₂、NH₂、SO₂NH₂、CF₃、OCF₃In one kind；

R₂Selected from one of following construction unit：

2. formula (I) according to claim 1 or the compound shown in (II) or (III) or it is pharmaceutically acceptable Salt, hydrate, solvate, polymorph, dynamic isomer or prodrug, it is characterised in that：Described compound is chosen in particular from Any one in following compounds：

N- [(1- (the chloro- 3-fluorophenyls of 2,6- bis-) ethyl)]-6- (1- piperidin-4-yl-1H- pyrazoles-4- bases) pyrido [2,3-d] The amine of pyrimidine -4,

N- [(1- (the chloro- 3-fluorophenyls of 2,6- bis-) ethyl)]-6- (1- methyl isophthalic acid H- pyrazoles-4- bases) pyrido [2,3-d] pyrimidine- 4 amine,

3. the synthetic method of compound shown in claim 1 formula of (I), it is characterised in that：Step is：By initiation material 5- Bromo- 2- aminobenzoic acids and formamide cyclization, generate bromo- 4 (the 3H)-quinazolinones of 6-, in the presence of thionyl chloride, add catalysis The DMF of amount, is heated to reflux carrying out the chlorine replacement of 4, then generates 4 substituted compounds with corresponding benzylalcohol or benzylamine, respectively with boron Esters of gallic acid compound Jing C-C coupling reactions obtain target product.

4. the synthetic method of compound shown in claim 1 formula of (II), it is characterised in that：Step is：By initiation material 2- The reaction of amino-nicotinic acid and bromine generates the bromo- 2- amino-nicotinic acids acid of 5-, then with formamide cyclization, generate the bromo- pyrido of 4- hydroxyls -6 [2,3-d] pyrimidine, in the presence of thionyl chloride, adds the DMF of catalytic amount, and the chlorine for being heated to reflux carrying out 4 replaces, then with accordingly Benzylalcohol or benzylamine generate 4 substituted compounds, obtain target product with borate ester compound Jing C-C coupling reactions respectively.

5. the synthetic method of compound shown in claim 1 formula of (III), it is characterised in that：Step is：By initiation material 5- Bromo- 2- aminobenzoic acids and dicyandiamide cyclization, obtain the compound containing guanidine radicals class, and deamidination obtains 2- amino-4-hydroxy -6- Bromo- quinazoline, directly employ card spy condensing agent and corresponding benzylalcohol or benzylamine condensation, then with borate ester compound Jing C-C idols Connection reacts to obtain target compound.

6. a kind of pharmaceutical composition, it is characterised in that：Including at least one in following active ingredients：A) compound, b) change Compound pharmaceutically acceptable salt, c) hydrate of the compound, d) solvate of the compound, e) compound is more The prodrug of the dynamic isomer of crystal formation thing, f) compound, g) compound；Wherein, described compound is claim 1-2 Any one of formula (I) or the compound shown in (II) or (III).

7. pharmaceutical composition according to claim 6, it is characterised in that：Also include excipient substance.

8. application of the active component described in claim 6 in tyrosine kinase inhibitor is prepared.

9. application according to claim 8, it is characterised in that：Described active component prepare treatment and/or prevent and/ Or delay and/or auxiliary treatment and/or the application in processing the medicine of too high to tyrosine kinase activity related disease.

10. application of the active component described in claim 6 in antineoplastic is prepared.