CN106831812A

CN106831812A - Heterocycle containing biaryl amide structure and pyrimidine or pyrazine compounds and its application

Info

Publication number: CN106831812A
Application number: CN201710033644.4A
Authority: CN
Inventors: 朱五福; 郑鹏武; 孙成钰; 陈晨; 徐珊; 唐启东; 王文惠; 王勤勤; 王操林
Original assignee: Jiangxi Science and Technology Normal University
Current assignee: Jiangxi Science and Technology Normal University
Priority date: 2016-08-09
Filing date: 2017-01-17
Publication date: 2017-06-13
Anticipated expiration: 2037-01-17
Also published as: CN106831812B

Abstract

The invention discloses a kind of heterocycle containing biaryl amide structure and pyrimidine or pyrazine compounds, as shown in formula I or II：

Description

Heterocycle containing biaryl amide structure and pyrimidine or pyrazine compounds and its application

Technical field

The present invention relates to quinazoline compounds, in particular to a kind of heterocycle containing biaryl amide structure and pyrimidine or pyrrole Piperazine class compound and its application.

Background technology

Malignant tumour is a kind of disease of serious harm human life and health, and its morbidity and mortality has exceeded heart and brain Vascular diseases, occupy the first place of all diseases, and cancer is to threaten first big " killer " of human health.Expect the year two thousand thirty whole world There to be 21,400,000 new cases, death toll reaches 13,200,000 people, wherein 70% occurs the developing country in middle and low income.With Continuing to develop for social economy, municipal pollution is increasingly severe, aging population development, in China, cancer mortality is year by year Rise, it has also become serious harm China human mortality quality, the major disease of obstruction social development.It is dead in three the whole country of China Because survey data shows, nearly 30 years, composition of the Cancer in China in the cause of the death was risen to by the 10.13% of 20 century 70s 22.32%.

Cancer is the disease caused because control growth and proliferation of cell mechanism is not normal, and the essence of cell carcinogenesis is cell letter The imbalance of number conducting system, so as to result in the fast-growth and infinite multiplication of cancer cell.By phosphinositides -3- kinases (phosphoinositide 3-kinase, PI3K) and protein kinase B (PKB/Akt) downstream, rapamycin target body albumen (mTOR) the PI3K-Akt-mTOR paths of composition are referred to as PI3K paths, have important work in the occurrence and development of tumour With micromolecular inhibitor of the key molecule as target spot has turned into the research of current antineoplastic medicine with PI3K/Akt signal paths Focus.

The cyclopean family that PI3K is made up of lipid and serine/threonine kinases, including several phosphinositides kinases and Protein kinase such as ATM, ATR and DNA-PK that DNA is relied on etc., it can make the 3rd di of phosphatidylinositols, produce Inositol lipid material --- phosphatidylinositols -3- phosphate esters (PIP3) with second messenger's effect.Second messenger PIP3 can make PI3K is combined with effector (particularly Akt) pairing in downstream, so as to cause film to be raised and phosphorylation.Research shows:PI3K family Numerous processes such as race and cell propagation, anti-apoptotic, cell migration, film bubble transhipment, cell cancerous transformation are related, these biological effects " anchor " molecule 3- phosphoinositides fat (PIP, PIP2, PIP3) to be formed mediation is mainly catalyzed by PI3K.Research discovery, PI3K paths are generally lacked of proper care in extensive human tumor spectrum, and the dysfunction or missing in the path caused by some gene mutations can draw Play normal cell turnover, promote tumor cell proliferation and survival and the invasion and attack and migration of mediate tumor cell, therefore be small molecule The favourable effects target position of inhibitor, for the treatment of cancer provides chance.

It was related to the patent (WO2009055730/WO2009036082) of the inhibitory action of PI3K and relevant report in recent years (Journal of Medicinal Chemistry,2008,51(18):5522-5532,Drugs ofthe Future, 2007,32(6):537-547.) sharply increase, it has now been found that the little molecules in inhibiting of various kinases in the signal path Agent, such as natural products wortmannin (Wortmannin) and Mongolian oak flavin compound L Y294002 are two kinds wide variety of PI3K inhibitor, the former can be combined with various hypotypes of PI3K；The inhibitor 1L-6-hydroxymethyl-chiro- of Akt Inositol2- (R) -2-O-methyl-3-O-octadecylcarbonate, the IC of its Selective depression Akt₅₀Value is about 5 μM, hence it is evident that suppress the IC of PI3K less than it₅₀90 μM of value；P70S6K is another feasibility target spot of PI3K-Akt signal paths, is exempted from Epidemic disease inhibitor Rapamycin (RPM) is widely used in clinical organ transplant, and research finds, RPM can make p70S6K remove phosphoric acid Change and suppress the activity of the kinases, so that suppress the growth of tumour cell, at present in various PTEN mutation or PI3K-Akt The selective antitumor activity of RPM is observed in the human tumour cell line that pathway activity is raised.

In PI3K families, I types PI3K can be activated by extracellular signal, therefore be studied so far in numerous PI3K hypotypes A most commonly used class.At present, the compound of many targeting I types PI3K has been enter into clinical investigation phase, such as：Natural products is wet graceful Penicillin, PX-866, LY-294002, TGX-115, TGX-155, PI-103, GDC-0941 etc., wherein most are PI3K- MTOR double inhibitors.

The GDC-0941 (shown in following structural formula) of document report belongs to thienopyrimidines, be by The oral PI3K inhibitor of Genentech companies exploitation, has completed Phase I clinical trial at present.GDC-0941 is to p110 α and δ IC₅₀Value reaches 3nM, is 10 times and 25 times of p110 β, γ, with good selectivity.Preclinical study shows, GDC- 0941 pair of various human tumor cell line (including glioblastoma, breast cancer cell, prostate gland cancer cell etc.) shows significantly Inhibit proliferaton effect, its IC₅₀Value reaches 0.009ug/mL.In Anticancer Activities in nude mouse, when oral dose is 75mg/ Kg, the growth inhibition rate to tumour reaches more than 80%.Double aryl urea structures are also widely used in anticancer small numerator inhibitor In research, Wyeth Venkatesan et al. reports a series of 1,3,5-triazines class PI3K inhibitor, wherein, PKI-587 Can well suppress growth of cancer cells and propagation in active testing in vitro and in vivo, and its apoptosis can be promoted；Thing PKI-402 It is that Wyeth transforms the substituted bisarylurea for obtaining like thing on the basis of PKI-587 and PKI-179, the compound has Good physicochemical property and pharmacokinetic property, in vitro and in vivo biomarker research find that PKI-402 can block PI3K- Akt-mTOR paths and energy cancer cell specific induction of apoptosis.

The content of the invention

It is an object of the invention to provide a kind of heterocycle containing biaryl amide structure and pyrimidine or pyrazine compounds and Its preparation method and application.

The present invention provide as shown in formula I the heterocycle containing biaryl amide structure simultaneously pyrimidine or pyrazine compounds, its is several What isomers and its pharmaceutically acceptable salt, hydrate, solvate or prodrug, structure is as shown in following formula I or II：

Wherein：

X=S, Y=CH or X=CH, Y=S；

Z is phenyl ring, five yuan or hexa-atomic heteroaromatic；

A, B, D, E, G are respectively CH or N；

R¹Selected from H,

R²Selected from H or-CH₃；

Ar is the heterocyclic radical of phenyl, naphthyl, 5~10 unit's heteroaryls, 5~10 yuan of saturations or fractional saturation, the phenyl, naphthalene Base, heteroaryl and heterocyclic radical contain 1~3 hetero atom selected from O, N and S, and optional 1~4 identical or different R of Ar³Take Generation；

R³Selected from hydrogen, halogen, hydroxyl, trifluoromethyl, trifluoromethoxy, amino, azido, nitro, cyano group, sulfydryl, (C₁ ~C₄) alkyl, (C₃~C₆) cycloalkyl, (C₂~C₄) alkenyl, (C₂~C₄) alkynyl, (C₁~C₄) alkoxy, (C₁~C₄) alkylthio group, Pi-allyl, (2- methyl) pi-allyl, (C₁~C₄) alkoxy methyl, (C₁~C₄) alkyl acyl or (C₁~C₃) alkylenedioxy group Substitution base.

R⁴、R⁵It is identical or different, separately selected from (C₁~C₆) alkyl or (C₃~C₆) cycloalkyl；Or R⁴And R⁵With The nitrogen-atoms connected with them forms 5~10 yuan of saturated heterocyclyls together, the saturated heterocyclyl except with R⁴And R⁵Connection Outside nitrogen-atoms, optionally containing 1~3 hetero atom selected from O, N and S.

Heterocycle and pyrimidine or Pyrazine chemical combination containing biaryl amide structure present invention is preferably related to such as above-mentioned formula I or II Thing, its geometric isomer and its pharmaceutically acceptable salt, hydrate, solvate or prodrug, wherein：

X=S, Y=CH or X=CH, Y=S；

Z is phenyl ring or pyridine ring；

A, B, D, E, G are respectively C or N；

R¹Selected from H,

R²Selected from H or-CH₃；

Ar is phenyl, naphthyl, pyridine radicals, pyrimidine radicals or thienyl, the phenyl, naphthyl, pyridine radicals, pyrimidine radicals, thiophene Optional 1~4 identical or different R of base³Substitution；

R³Selected from hydrogen, halogen, hydroxyl, trifluoromethyl, trifluoromethoxy, amino, azido, nitro, cyano group, sulfydryl, (C₁ ~C₄) alkyl, (C₃~C₆) cycloalkyl, (C₁~C₄) alkenyl, (C₁~C₄) alkynyl, (C₁~C₄) alkoxy, (C₁~C₄) alkylthio group, Pi-allyl, (2- methyl) pi-allyl, (C₁~C₄) alkoxy methyl, (C₁~C₄) alkyl acyl or (C₁~C₃) alkylenedioxy group Substitution base.

It is selected from：

In the present invention, the heterocycle containing biaryl amide structure and pyrimidine or pyrazine compounds of the formula I or II are Selected from the one kind in following compounds, but these compounds are not meant to any limitation of the invention：

4- (4- ethylphenyls)-N- (4- (4- morpholinoes thieno [2,3-d] pyrimidine -2-base) phenyl) picolinamide,

4- (2,4 difluorobenzene base)-N- (4- (4- morpholinoes thieno [2,3-d] pyrimidine -2-base) phenyl) picolinamide,

4- (3- fluorophenyls)-N- (4- (4- morpholinoes thieno [2,3-d] pyrimidine -2-base) phenyl) picolinamide,

N- (4- (4- morpholinoes thieno [2,3-d] pyrimidine -2-base) phenyl) -4- phenylpyridines acid amides,

4- (2,4- 3,5-dimethylphenyls)-N- (4- (4- morpholinoes thieno [2,3-d] pyrimidine -2-base) phenyl) pyridine acyl Amine,

6- (4- methoxyphenyls)-N- (4- (4- morpholinoes thieno [2,3-d] pyrimidine -2-base) phenyl) pyrimidine -4- first Acid amides,

N- (4- (4- morpholinoes thieno [2,3-d] pyrimidine -2-base) phenyl) -6- (p-methylphenyl) pyrimidine -4- formamides,

N- (4- (4- morpholinoes thieno [2,3-d] pyrimidine -2-base) phenyl) -6- (4- (trifluoromethyl) pyrimidines -4- Formamide,

5- (3- fluorophenyls)-N- (4- (4- morpholinoes thieno [2,3-d] pyrimidine -2-base) phenyl) picolinamide,

5- (2,4 difluorobenzene base)-N- (4- (4- morpholinoes thieno [2,3-d] pyrimidine -2-base) phenyl) picolinamide,

N- (4- (- morpholino imidazo [1,2-a] pyrazine -6- bases) phenyl) -4- phenylpyridines acid amides,

4- (2,4 difluorobenzene base)-N- (4- (8- morpholinoes imidazo [1,2-a] pyrazine -6- bases) phenyl) picolinamide,

4- (4- ethylphenyls)-N- (4- (8- morpholinoes imidazo [1,2-a] pyrazine -6- bases) phenyl) picolinamide,

4- (2,4- 3,5-dimethylphenyls)-N- (4- (8- morpholinoes imidazo [1,2-a] pyrazine -6- bases) phenyl) pyridine acyl Amine,

N- (4- (8- morpholinoes imidazo [1,2-a] pyrazine -6- bases) phenyl) -4- (p-methylphenyl) picolinamide,

5- (3- fluorophenyls)-N- (4- (8- morpholinoes imidazo [1,2-a] pyrazine -6- bases) phenyl) picolinamide,

5- (4- methoxyphenyls)-N- (4- (8- morpholinoes imidazo [1,2-a] pyrazine -6- bases) phenyl) picolinamide,

6- (4- methoxyphenyls)-N- (4- (8- morpholines imidazo [1,2-a] pyrazine -6- bases) phenyl) pyrimidine -4- formyls Amine,

6- (4- bromophenyls)-N- (4- (8- morpholines imidazo [1,2-a] pyrazine -6- bases) phenyl) pyrimidine -4- formamides,

N- (4- (8- morpholinoes imidazo [1,2-a] pyrazine -6- bases) phenyl) -6- phenyl pyrimidine -4- formamides,

N- (5- (4- morpholinoes thieno [3,2-d] pyrimidine -2-base) pyridine -2- bases) -4- phenylpyridines acid amides,

N- (5- (4- (3- methyl morpholines) thieno [3,2-d] pyrimidine -2-base) pyridine -2- bases) -4- phenylpyridines acid amides,

N- (5- (6- ((dimethylamino) methyl) -4- (3- methyl morpholines) thieno [3,2-d] pyrimidine -2-base) pyridine - 2- yls) -4- (thiophene -2- bases) picolinamide,

N- (5- (8- (3- methyl morpholines base) imidazo [1,2-a] pyrazine -6- bases) pyridine -2- bases) -4- (thiophene -2- bases) Picolinamide,

N- (5- (8- (3- methyl morpholines base) imidazo [1,2-a] pyrazine -6- bases) pyridine -2- bases) -4- phenylpyridine acyls Amine,

N- (5- (6- (2- hydroxyl propyl- 2- yls) -4- morpholinoes thieno [3,2-d] pyrimidine -2-base) pyridine -2- bases) -4- Phenylpyridine acid amides,

N- (5- (4- morpholinoes -6- (the fluoro- 2- hydroxyls propyl- 2- yls of 1,1,1- tri-) thieno [2,3-d] pyrimidine -2-base) pyrroles Pyridine -2- bases) -4- phenylpyridines acid amides,

N- (5- (4,6- dimorpholinos thieno [2,3-d] pyrimidine -2-base) pyridine -2- bases) -4- phenylpyridine acid amides.

Following synthetic route describes heterocycle and pyrimidine or pyrazine of the formula I or II of the present invention containing biaryl amide structure The preparation of class compound, all of raw material is all by way of described in synthetic route, by the common skill of organic chemistry filed It is prepared by method known to art personnel or commercially available.The final derivative of whole of the invention is all by described in synthetic route Method or prepared by similar method, these methods are organic chemistry fileds well-known to the ordinarily skilled artisan.Close Definition in the definition of the whole variable factor following articles applied into route or such as claim.

The target compound I of route 1 or II synthetic route

The synthetic intermediate 12 or 13 first of pass course of the present invention 1, then with different substituents (e.g., pyridine acids or pyrimidine Acids) side chain reaction, obtain target compound.

According to some usual methods of the art, in the present invention, the above-mentioned heterocycle containing biaryl amide structure is simultaneously Pyrimidine or pyrazine compounds can be with acid generation pharmaceutically acceptable salts.Pharmaceutically acceptable addition salts include inorganic acid and organic acid Addition salts, the salt with following sour addition is particularly preferred：Hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethyl sulfonic acid, to first Benzene sulfonic acid, benzene sulfonic acid, naphthalenedisulfonic acid, acetic acid, propionic acid, lactic acid, trifluoroacetic acid, maleic acid, citric acid, fumaric acid, oxalic acid, winestone Acid, benzoic acid etc..

Additionally, present invention additionally comprises the prodrug of derivative of the present invention.The prodrug of derivative of the present invention is above-mentioned containing biaryl The heterocycle and pyrimidine or pyrazine compounds of amide structure, their own may have weaker activity even without activity, but It is upon administration, to be converted to corresponding biology (such as by metabolism, solvolysis or other mode) in physiological conditions Activity form.

" halogen " refers to fluorine, chlorine, bromine or iodine generation in the present invention；" alkyl " refers to the alkyl of straight or branched；" alkylidene " It refer to the alkylidene of straight or branched；" cycloalkyl " refers to substituted or unsubstituted cycloalkyl；" the heterocycle of saturation or fractional saturation Base " refers to the heteroatomic monocyclic or polycyclic ring-type system that N, O, S are selected from containing one or more, such as pyrrolidinyl, morpholine Base, piperazinyl, piperidyl, pyrazolidinyl, imidazolidinyl and thiazolinyl etc..

The present invention can contain the above-mentioned heterocycle containing biaryl amide structure and pyrimidine or pyrazine compounds, and its pharmacy Upper acceptable salt, hydrate or solvate mix system as active ingredient with pharmaceutically acceptable carrier or excipients Standby and to be prepared into clinically acceptable formulation into composition, above-mentioned pharmaceutically acceptable excipients refer to any can be used for The diluent of pharmaceutical field, adjuvant and/or carrier.Derivative of the invention can be applied in combination with other active ingredients, only Them are wanted not produce other unfavorable effects, such as allergic reaction.

The above-mentioned heterocycle containing biaryl amide structure of the present invention and pyrimidine or pyrazine compounds are used for the clinical agent of patient Amount can basis：Active component therapeutic efficiency in vivo and bioavilability, their metabolism and discharge rate and patient's Age, sex, disease phase are suitably adjusted, but the daily dosage of adult typically should be 10~500mg, preferably 50 ~300mg.Therefore, when pharmaceutical composition of the invention is made into unit dosage forms, it is contemplated that above-mentioned effective dose, per unit system Agent should the heterocycle containing biaryl amide structure containing 10~500mg above formulas I and pyrimidine or pyrazine compounds, preferably 50 ~300mg.According to the guidance of doctor or pharmacist, these preparations can at certain intervals divide administration (preferably to six several times It is secondary).

Present invention also offers a kind of Pharmaceutical composition, including therapeutically effective amount above-mentioned formula I or II acyl containing biaryl It is the heterocycle of amine structure and pyrimidine or pyrazine compounds, its geometric isomer and its pharmaceutically acceptable salt, hydrate, molten Agent compound or prodrug are used as active component and pharmaceutically acceptable excipients.If Pharmaceutical composition of the invention can be configured to Dry kind of formulation, wherein containing some conventional excipient in drug field.Several formulation as described above can be using injection The drug forms such as agent, tablet, capsule, aerosol, suppository, film, pill, externally-applied liniment, ointment.

Carrier for pharmaceutical composition of the present invention is available common type in drug field, including：Adhesive, profit Lubrication prescription, disintegrant, cosolvent, diluent, stabilizer, suspending agent, non-pigment, flavouring, preservative, solubilizer and matrix etc.. Pharmaceutical preparation can by oral administration or parenteral (such as intravenous, subcutaneous, intraperitoneal or local) administration, if some drugses It is unstable under the conditions of stomach, enteric coated tablets can be configured to.

It has also been found that the above-mentioned heterocycle containing biaryl amide structure and pyrimidine or pyrazine compounds are preparing treatment And/or the application in prevention proliferative disease medicine.Reactive compound of the invention or its officinal salt and its solvate can It is used alone as unique anti-proliferate medicine, or can be used for the anti-proliferate Drug combination for having listed Treatment and/or prevention proliferative disease, such as psoriasis, benign prostatauxe, atherosclerosis and ISR.

It has also been found that the above-mentioned quinazoline compounds containing acylhydrazone structure are preparing the medicine for the treatment of and/or pre- anti-cancer Application in thing.The compounds of this invention has suppression tumor cell growth activity in vitro, therefore, it can serve as preparing treatment And/or the medicine of pre- anti-cancer, such as mammary gland, lung, liver, kidney, colon, rectum, stomach, prostate, bladder, uterus, pancreas, bone Marrow, testis, ovary, lymph, soft tissue, neck, thyroid gland, the cancer of esophagus and leukaemia, neuroblastoma etc..

It has also been found that the above-mentioned heterocycle containing biaryl amide structure and pyrimidine or pyrazine compounds, treat preparing And/or the application in prevention lung cancer, liver cancer, stomach cancer, colon cancer, the medicine of breast cancer.

Suppress lung carcinoma cell H460, Human Prostate Cancer Cells PC-3 and hepatoma cell strain SGC-7901 activity examination by external Test, people malignant glioblastoma cell U87MG, the compounds of this invention is thin to lung carcinoma cell, prostate gland cancer cell and liver cancer Born of the same parents have and significantly inhibit effect, it is especially useful in prepare treatment and/or prevent the medicine of prostate cancer, lung cancer and liver cancer.

Found by testing mTOR enzymatic activitys, the compounds of this invention has significant suppression mTOR kinase activities, right Lung carcinoma cell, Human Prostate Cancer Cells, glioblast of mTOR expression high etc. have stronger inhibitory action, it is especially useful in prepare Treatment and/or the medicine of prevention lung cancer.

Reactive compound of the invention or its officinal salt and its solvate can be independent as unique antineoplastic Use, or can be with antineoplastic (such as platinum medicine cis-platinum, camptothecine Irinotecan, the Changchun for having listed Flower bases medicine NVB, deoxidation born of the same parents' former times class medicine gemcitabine, etoposide, taxol etc.) it is used in combination.Therapeutic alliance is led to Cross each therapeutic component simultaneously, order or separate administration and realize.

Present invention design has synthesized a series of new heterocycles and pyrimidine (pyrazine) analog derivative, through in vitro to mTOR inhibitors Overexpression cell line carries out antitumor activity screening, as a result shows there is stronger antitumor activity and selectivity.

Specific embodiment

In order to preferably explain the present invention, below in conjunction with specific embodiment, the present invention is described in further detail, but They do not constitute to the present invention and limit.

Embodiment is intended to illustrate rather than limitation the scope of the present invention.The proton nmr spectra Bruker of derivative ARX-400 is determined, and mass spectrum is determined with Agilent 1100LC/MSD；It is pure or chemical pure that agents useful for same is analysis.

Heterocycle and pyrimidine or pyrazine compounds of the formula I or II containing biaryl amide structure：

The structural formula of the embodiment of the present invention 1~28 is as shown in table 1 below.

The structural formula of the embodiment 1~28 of table 1

Step A thieno [2,3-d] pyrimidine -2, the preparation of 4 (1H, 3H)-diketone (1)

Under nitrogen protection, 2- aminothiophene -3- methyl formates (10g) is well mixed with urea (30g), mechanical agitation Under be warming up to 180 DEG C, react 2h.Reaction is finished, and is cooled to less than 100 DEG C, and reactant is poured into the 1mol/L's equipped with 500mL In sodium hydroxide solution beaker, it is sufficiently stirred for, filters；Filtrate pH to 6 is adjusted with concentrated hydrochloric acid, pureed solid is separated out；Suction filtration, filtrate is cold Solid is separated out after hiding 12h, suction filtration obtains faint yellow product, i.e. thieno [2,3-d] pyrimidine -2,4 (1H, 3H)-diketone, yield： 56.1%.

1H NMR (400MHz, DMSO) δ 11.94 (s, 1H, CONH), 11.18 (s, 1H, CONH), 7.11 (d, J=16.0, 5.6Hz,2H,2Ar-H)；ESI-MS[M-H]+m/z:167.2.

The preparation of step B 2,4- dichloro-thiophenes simultaneously [2,3-d] pyrimidine (2)

To 1g dry thieno [2,3-d] pyrimidine -2,4 (1H, 3H)-diketone add 10g POCl3s, add 1 to drip DMF Catalysis, is warming up to 115 DEG C, reacts 5h, and reaction finishes, reactant is slowly added in trash ice, while being stirred vigorously, obtains brown color Solid, suction filtration, filter residue is dissolved in 50mL dichloromethane, to 1g activated carbons and 2g silica gel is added in solution, is warming up to 45 DEG C of backflows de- Color 1h, suction filtration, is evaporated off solvent while hot, obtains faint yellow solid, i.e. 2,4- dichloro-thiophenes simultaneously [2,3-d] pyrimidine, yield：56.0%.

¹H NMR (400MHz, DMSO) δ 8.16 (d, J=3.4Hz, 1H, Ar-H), 7.62 (d, J=3.4Hz, 1H, Ar- H).ESI-MS[M+H]+m/z:205.1。

The preparation of step C 4- (2- chlorothiophenes simultaneously [2,3-d] pyrimidine-4-yl) morpholine (3)

By 2, the 4- dichloro-thiophenes of 1g, simultaneously [2,3-d] pyrimidine is dissolved in 20mL methyl alcohol, 1mL morpholines is added dropwise under ice bath, gradually Be warmed to room temperature, react 0.5h, separate out solid, suction filtration obtains white solid, i.e. 4- (2- chlorothiophenes simultaneously [2,3-d] pyrimidine-4-yl) Quinoline, yield：86.1%.

¹H NMR(400MHz,DMSO)δ7.70(s,1H,Ar-H),7.69–7.65(m,1H,Ar-H),3.96–3.86(m, 4H,OCH₂), 3.75 (d, J=4.4Hz, 4H, NCH₂).ESI-MS[M+H]⁺m/z:256.1。

The preparation of step D 4- (4- morpholinoes thieno [2,3-d] pyrimidine -2-base) aniline (12)

By para-bromoaniline (2g, 0.01mol), double (pinacol conjunction) two boron (3.8g, 0.015mol), potassium acetate (2.9g, 0.03mol), catalyst bi triphenyl phosphorus palladium chloride (0.18g, 0.00025mol) is dissolved in three necks equipped with dioxane 50mL In flask, nitrogen 10min is continually fed into, is warming up to 80 DEG C, react 3h, solution blackening, to 20mL water is added in reaction system, instead Answer 2min, sequentially add sodium carbonate (2.1g, 0.02mol), catalyst bi triphenyl phosphorus palladium chloride (0.18g, 0.00025mol) and 4- (2- chlorothiophenes simultaneously [2,3-d] pyrimidine-4-yl) morpholine (1.2g, 0.006mol).100 DEG C are warming up to, instead Answer 8h.Reaction is finished, reaction solution vacuum distillation, removes partial solvent, to 100mL water is added in residual reaction liquid, separates out black Solid, suction filtration, filter residue is dissolved in dichloromethane：Methyl alcohol=5：In 1 solution 100mL, to addition 2g activated carbons and 5g silicon in solution Glue, is heated to reflux decolouring 1h, while hot suction filtration, and filtrate is evaporated to obtain faint yellow solid, i.e. (4- morpholinoes thieno [2,3-d] are phonetic for 4- Pyridine -2- bases) aniline.

Step E 4- (4- ethylphenyls)-N- (4- (4- morpholinoes thieno [2,3-d] pyrimidine -2-base) phenyl) pyridine acyl Amine

Pyridine acids side chain compound is dissolved in 8mL dichloromethane, the 1 dry DMF of drop is added, oxalyl chloride is dropwise added dropwise (1.5 times of equivalents), the DIPEA of the dichloromethane of 4- (4- morpholinoes thieno [2,3-d] pyrimidine -2-base) aniline and 1mL is mixed Conjunction solution is dropped in the dichloromethane solution of side chain, is finished reaction 10min, TLC and is detected that reacting completely rear stops reacting.

Reaction solution is washed 2 times with saturated sodium carbonate solution, takes organic layer, and solvent is evaporated off, and adds isopropanol stirring 30min, Suction filtration, filter residue is dissolved in 15mL dichloromethane, adds a small amount of silica gel, stirs 10min, and suction filtration takes filtrate, solvent is evaporated off, and is added 10mL isopropanols are washed, and suction filtration obtains neat solid product.

Embodiment 1

4- (4- ethylphenyls)-N- (4- (4- morpholinoes thieno [2,3-d] pyrimidine -2-base) phenyl) picolinamide

First according to step A~D synthetic intermediates 12, last intermediate 12 is by step E and pyridine acids side chain compound Reaction, obtains 4- (4- ethylphenyls)-N- (4- (4- morpholinoes thieno [2,3-d] pyrimidine -2-base) phenyl) picolinamide.

m.p.282-284℃；ESI-MS[M+H]⁺m/z:522.1；¹H NMR(400MHz,DMSO)δ10.88(s,1H), 8.78 (d, J=4.7Hz, 1H), 8.40 (d, J=7.2Hz, 3H), 8.08 (d, J=8.4Hz, 2H), 8.00 (s, 1H), 7.83 (d, J=7.6Hz, 2H), 7.64 (dd, J=18.7,6.0Hz, 2H), 7.41 (d, J=7.5Hz, 2H), 3.96 (s, 4H), 3.79 (s, 4H), 2.68 (d, J=7.4Hz, 2H), 1.23 (d, J=7.7Hz, 3H)

According to the method for embodiment 1, first according to step A~D synthetic intermediates 12, last intermediate 12 by step E with The side chain reaction of different substituents (e.g., pyridine acids or pyrimidine acids), is obtained the compound of embodiment 2~10 respectively.

Embodiment 2

4- (2,4 difluorobenzene base)-N- (4- (4- morpholinoes thieno [2,3-d] pyrimidine -2-base) phenyl) picolinamide

m.p.255-256℃；ESI-MS[M+H]⁺m/z:530.1；¹H NMR(400MHz,DMSO)δ10.90(s,1H), 8.86 (s, 1H), 8.40 (d, J=7.7Hz, 2H), 8.31 (s, 1H), 8.07 (d, J=7.1Hz, 2H), 7.89 (s, 2H), 7.64 (d, J=13.0Hz, 2H), 7.51 (s, 1H), 7.31 (s, 1H), 3.97 (s, 4H), 3.79 (s, 4H)

Embodiment 3

4- (3- fluorophenyls)-N- (4- (4- morpholinoes thieno [2,3-d] pyrimidine -2-base) phenyl) picolinamide

m.p.281-283℃；ESI-MS[M+H]⁺m/z:512.1；¹H NMR(400MHz,DMSO)δ10.90(s,1H), 8.83 (d, J=4.5Hz, 1H), 8.47-8.36 (m, 2H), 8.08 (d, J=8.6Hz, 3H), 7.85-7.71 (m, 2H), 7.71- 7.54 (m, 3H), 7.48 (d, J=6.0Hz, 1H), 7.37 (s, 1H), 6.60 (d, J=8.3Hz, 1H), 3.97 (s, 4H), 3.78 (s,4H).

Embodiment 4

N- (4- (4- morpholinoes thieno [2,3-d] pyrimidine -2-base) phenyl) -4- phenylpyridine acid amides

m.p.223-225℃；ESI-MS[M+H]⁺m/z:494.1；¹H NMR(400MHz,DMSO)δ10.87(s,1H), 8.79 (d, J=12.7Hz, 2H), 8.41 (d, J=9.2Hz, 2H), 8.08 (d, J=7.7Hz, 2H), 7.77 (d, J=7.0Hz, 2H), 7.64 (d, J=13.0Hz, 2H), 7.35 (dd, J=20.2,7.3Hz, 3H), 6.60 (d, J=8.2Hz, 1H), 3.97 (s,4H),3.79(s,4H).

Embodiment 5

4- (2,4- 3,5-dimethylphenyls)-N- (4- (4- morpholinoes thieno [2,3-d] pyrimidine -2-base) phenyl) picolinamide

m.p.277-279℃；ESI-MS[M+H]⁺m/z:522.2；¹H NMR(400MHz,DMSO)δ10.87(s,1H), 8.78 (d, J=4.7Hz, 1H), 8.66 (s, 1H), 8.06 (d, J=8.3Hz, 3H), 7.75-7.63 (m, 2H), 7.20 (dd, J =20.4,9.6Hz, 5H), 3.97 (s, 4H), 3.79 (s, 4H), 2.25 (d, J=7.9Hz, 6H)

Embodiment 6

6- (4- methoxyphenyls)-N- (4- (4- morpholinoes thieno [2,3-d] pyrimidine -2-base) phenyl) pyrimidine -4- first Acid amides

m.p.251-252℃；ESI-MS[M+H]⁺m/z:525.2；¹H NMR(400MHz,CDCl₃)δ10.50(s,1H), 10.06 (s, 1H), 9.26 (s, 1H), 8.48 (s, 2H), 8.22 (d, J=8.7Hz, 3H), 7.90 (d, J=8.5Hz, 1H), (s, the 8H) of 7.69 (d, J=8.2Hz, 1H), 7.07 (s, 3H), 3.91

Embodiment 7

N- (4- (4- morpholinoes thieno [2,3-d] pyrimidine -2-base) phenyl) -6- (p-methylphenyl) pyrimidine -4- formamides

m.p.244-245℃；ESI-MS[M+H]⁺m/z:509.2；¹H NMR(400MHz,DMSO)δ9.38(s,1H), 9.33 (s, 1H), 8.53 (s, 1H), 8.37 (d, J=8.4Hz, 2H), 8.19 (d, J=8.1Hz, 3H), 8.03 (d, J= 7.9Hz, 2H), 7.37 (d, J=7.0Hz, 3H), 3.93 (s, 4H), 3.75 (s, 4H), 2.36 (s, 3H)

Embodiment 8

N- (4- (4- morpholinoes thieno [2,3-d] pyrimidine -2-base) phenyl) -6- (4- (trifluoromethyl) pyrimidines -4- Formamide

m.p.266-267℃；ESI-MS[M+H]⁺m/z:563.1；¹H NMR(400MHz,DMSO)δ9.50(s,1H), 9.13 (s, 1H), 8.68 (s, 1H), 8.50 (d, J=8.1Hz, 2H), 8.37 (t, J=7.9Hz, 2H), 8.04 (d, J= 8.5Hz, 1H), 7.92 (d, J=7.5Hz, 1H), 7.85 (d, J=8.3Hz, 1H), 7.72 (s, 1H), 7.62 (s, 1H), 7.59 (s,1H),3.93(s,4H),3.75(s,4H).

Embodiment 9

5- (3- fluorophenyls)-N- (4- (4- morpholinoes thieno [2,3-d] pyrimidine -2-base) phenyl) picolinamide

m.p.262-263℃；ESI-MS[M+H]⁺m/z:512；¹H NMR(400MHz,DMSO)δ10.75(s,1H),8.79 (d, J=5.0Hz, 1H), 8.55 (s, 1H), 8.40 (s, 1H), 8.00 (s, 2H), 7.98 (s, 2H), 7.92 (s, 1H), 7.88 (d, J=7.3Hz, 2H), 7.56 (d, J=3.1Hz, 2H), 7.53 (s, 1H), 4.27 (s, 4H), 3.80-3.74 (m, 4H)

Embodiment 10

5- (2,4 difluorobenzene base)-N- (4- (4- morpholinoes thieno [2,3-d] pyrimidine -2-base) phenyl) picolinamide

m.p.287-289℃；ESI-MS[M+H]⁺m/z:530.1；¹H NMR(400MHz,DMSO)δ10.77(s,1H), 8.80 (d, J=4.6Hz, 1H), 8.55 (s, 1H), 8.40 (s, 1H), 8.00 (s, 2H), 7.98 (s, 2H), 7.92 (s, 2H), 7.62(s,2H),7.56(s,1H),4.27(s,4H),3.77(s,4H).

The preparation of the bromo- 2- Aminopyrazines (9) of step F 3,5- bis-

To addition dichloromethane (200mL) and pyridine in the three-necked bottle equipped with 2- Aminopyrazines (14.27g, 0.15mol) The mixed solution of (25.3mL, 0.315mol)；Under the conditions of lucifuge, 40 DEG C flow back, be slowly added dropwise bromine (16.2mL, Dichloromethane (100mL) solution 0.315mol), solution is changed into orange and is eventually become Chinese red from orange in course of reaction, about 1h completion of dropping；Continue the 30min that flows back at 40 DEG C；Room temperature is cooled to, to distilled water (50mL) is added in reaction system, acutely Stirring 10min, stratification；Lower floor's liquid is collected, the liquid that will be collected into is washed 2 times with distilled water (100mL)；By organic phase In moving to the flask equipped with silica gel (10g) and activated carbon (1g), boiling reflux 30min；Suction filtration, collects filtrate and vacuum distillation, During the solid obtained after distillation moved into the flask equipped with n-hexane (45mL), flow back 2h at 80 DEG C；Filter while hot, by gained Solid product is weighed after drying, and obtains pale yellow solid 18.15g, i.e. the bromo- 2- Aminopyrazines of 3,5- bis-, and yield is 47.8%.

¹H NMR(400MHz,DMSO)δ:8.14(s,1H),7.01(s,2H).

The preparation of the bromo- 2- Aminopyrazines (10) of step G 3- morpholines -5-

Morpholine (50mL) is added and is equipped with the three-neck flask of the bromo- 2- Aminopyrazines of 3,5- bis- of 12.50g, kept for 80 DEG C Lower back flow reaction 1h, TLC detection reaction are completed.System after backflow is cooled to room temperature, the burning equipped with frozen water (300mL) is added to It is stirred continuously in cup, separates out solid；Yellow solid 12.30g, i.e. 3- morpholines -5- with metallic luster are obtained after suction filtration, drying Bromo- 2- Aminopyrazines, yield is 96.1%.

¹H NMR(400MHz,DMSO)δ:7.70 (s, 1H), 6.28 (s, 2H), 3.85-3.62 (m, 4H), 3.04 (d, J= 4.0Hz,4H)。

The preparation of step H 4- (6- bromines imidazo [1,2-a] pyrazine -8- bases) morpholine (11)

4.4g chloroacetaldehydes are mixed with 15mL isopropanols, 10mL mixed solutions is taken and is added the round bottom equipped with 1.7g intermediates 10 In flask, it is warming up to 45 DEG C and starts to react 1h；During the mixed solution of remaining chloroacetaldehyde and isopropanol added into flask, 65 are warming up to DEG C reaction, TLC detection react, stop react.Room temperature is cooled to, is added in the beaker equipped with 300mL frozen water, separate out brown color Solid, suction filtration, drying obtain yellow-brown solid product, i.e. 4- (6- bromines imidazo [1,2-a] pyrazine -8- bases) morpholine.

The preparation of step I 4- (8- morpholinoes imidazo [1,2-a] pyrazine -6- bases) aniline (13)

With reference to step D methods, reacted by 4- (6- bromines imidazo [1,2-a] pyrazine -8- bases) morpholines and para-bromoaniline, be obtained 4- (8- morpholinoes imidazo [1,2-a] pyrazine -6- bases) aniline.

Step J N- (4- (8- morpholinoes imidazo [1,2-a] pyrazine -6- bases) phenyl) -4- phenylpyridine acid amides

Pyridine acids side chain compound is dissolved in 8mL dichloromethane, the 1 dry DMF of drop is added, oxalyl chloride is dropwise added dropwise (1.5 times of equivalents), the DIPEA mixed solutions of 4- (8- morpholinoes imidazo [1,2-a] pyrazine -6- bases) aniline and 1mL are added dropwise Into the dichloromethane solution of side chain, reaction is stopped after finishing reaction 10min, TLC detection reaction completely.

Reaction solution is washed 2 times with saturated sodium carbonate solution, takes organic layer, and solvent is evaporated off, and adds isopropanol stirring 30min, Suction filtration, filter residue is dissolved in 15mL dichloromethane, adds a small amount of silica gel, stirs 10min, and suction filtration takes filtrate, solvent is evaporated off, and is added 10mL isopropanols are washed, and suction filtration obtains neat solid product, i.e. N- (4- (8- morpholinoes imidazo [1,2-a] pyrazine -6- bases) benzene Base) -4- phenylpyridine acid amides.

Embodiment 11

N- (4- (- morpholino imidazo [1,2-a] pyrazine -6- bases) phenyl) -4- phenylpyridine acid amides

First according to step F synthetic intermediates 9, intermediate 9 and morpholine by step G synthetic intermediates 10, intermediate 10 with Chloroethene aldehyde reaction passes through step I synthetic intermediates by step H synthetic intermediates 11, then by intermediate 11 and para-bromoaniline reaction 13, last intermediate 13 is reacted by step J with pyridine acids side chain compound, obtains N- (4- (- morpholino imidazos [1,2- A] pyrazine -6- bases) phenyl) -4- phenylpyridine acid amides.

m.p.261-262℃；ESI-MS[M+H]⁺m/z:477.2；¹H NMR(400MHz,DMSO)δ9.55(s,1H), 7.59 (d, J=5.0Hz, 1H), 7.34 (s, 1H), 7.20 (s, 1H), 6.79 (d, J=7.8Hz, 5H), 6.74-6.65 (m, 3H), (m, the 4H) of 6.34 (dd, J=8.4,4.3Hz, 4H), 3.07 (s, 4H), 2.61-2.51

According to the method for embodiment 11, first according to step F synthetic intermediates 9, intermediate 9 and R²Substituted morpholine is by step Rapid G synthetic intermediates 10, intermediate 10 with chloroethene aldehyde reaction by step H synthetic intermediates 11, then by intermediate 11 with to bromine , by step I synthetic intermediates 13, last intermediate 13 passes through step J different substituents for aniline or the reaction of the amine of 5- bromopyridines -2 The side chain reaction of (e.g., pyridine acids or pyrimidine acids), is obtained the compound of embodiment 12~20 respectively.

Embodiment 12

4- (2,4 difluorobenzene base)-N- (4- (8- morpholinoes imidazo [1,2-a] pyrazine -6- bases) phenyl) picolinamide

m.p.266-267℃；ESI-MS[M+H]⁺m/z:513.2；¹H NMR(400MHz,DMSO)δ10.76(s,1H), 8.79 (d, J=5.1Hz, 1H), 8.52 (s, 1H), 8.26 (s, 1H), 7.96 (d, J=7.3Hz, 4H), 7.89 (s, 1H), 7.85-7.74 (m, 2H), 7.52 (s, 1H), 7.43 (d, J=9.6Hz, 1H), 7.26 (t, J=8.3Hz, 1H), 4.23 (s, 4H), 3.74 (d, J=4.3Hz, 4H)

Embodiment 13

4- (4- ethylphenyls)-N- (4- (8- morpholinoes imidazo [1,2-a] pyrazine -6- bases) phenyl) picolinamide

m.p.261-262℃；ESI-MS[M+H]⁺m/z:505.2；¹H NMR(400MHz,DMSO)δ10.88(s,1H), 8.78 (d, J=4.7Hz, 1H), 8.40 (d, J=7.2Hz, 3H), 8.08 (d, J=8.4Hz, 2H), 8.03 (s, 1H), 7.74 (d, J=7.6Hz, 2H), 7.54 (dd, J=18.7,6.0Hz, 2H), 7.50 (d, J=7.5Hz, 1H, 7.45 (d, J=7.5Hz, 2H), (d, J=7.7Hz, the 3H) of 3.91 (s, 4H), 3.77 (s, 4H), 2.68 (d, J=7.4Hz, 2H), 1.23

Embodiment 14

4- (2,4- 3,5-dimethylphenyls)-N- (4- (8- morpholinoes imidazo [1,2-a] pyrazine -6- bases) phenyl) picolinamide

m.p.234-235℃；ESI-MS[M+H]⁺m/z:505.2；¹H NMR(400MHz,DMSO)δ10.83(s,1H), 8.77 (d, J=5.0Hz, 1H), 8.62 (s, 1H), 8.00 (dd, J=17.9,9.6Hz, 6H), 7.69-7.63 (m, 2H), (d, J=29.3Hz, the 6H) of 7.25-7.11 (m, 3H), 4.20 (s, 4H), 3.77 (s, 4H), 2.28

Embodiment 15

N- (4- (8- morpholinoes imidazo [1,2-a] pyrazine -6- bases) phenyl) -4- (p-methylphenyl) picolinamide

m.p.271-273℃；ESI-MS[M+H]⁺m/z:491.2；¹H NMR(400MHz,DMSO)δ9.87(s,1H), 7.90 (d, J=5.2Hz, 1H), 7.56 (s, 1H), 7.44 (s, 1H), 7.22-7.15 (m, 3H), 6.95 (d, J=8.5Hz, 3H),6.73(s,1H),6.54(s,4H),3.45(s,4H),3.15–2.68(m,4H),0.68–0.16(m,3H).

Embodiment 16

5- (3- fluorophenyls)-N- (4- (8- morpholinoes imidazo [1,2-a] pyrazine -6- bases) phenyl) picolinamide

m.p.261-264℃；ESI-MS[M+H]⁺m/z:495.2；¹H NMR(400MHz,CDCl₃)δ10.18(s,1H), 8.85 (s, 2H), 8.40 (d, J=8.1Hz, 2H), 8.29 (d, J=8.1Hz, 1H), 8.09 (s, 3H), 7.88 (s, 1H), 7.75 (s, 1H), 7.52 (d, J=5.1Hz, 1H), 7.45 (s, 1H), 7.36 (d, J=9.2Hz, 1H), 7.20 (d, J=9.0Hz, 1H),4.17(s,4H),4.03(s,4H).

Embodiment 17

5- (4- methoxyphenyls)-N- (4- (8- morpholinoes imidazo [1,2-a] pyrazine -6- bases) phenyl) picolinamide

m.p.255-257℃；ESI-MS[M+H]⁺m/z:507.2；¹H NMR(400MHz,DMSO)δ11.05(s,1H), 8.74 (d, J=5.1Hz, 1H), 8.57 (s, 1H), 8.47 (s, 1H), 8.03 (d, J=8.6Hz, 2H), 8.02 (s, 2H), 7.87 (d, J=9.8Hz, 2H), 7.86 (d, J=8.6Hz, 2H), 7.51 (s, 1H), 7.14 (d, J=8.6Hz, 2H), 4.32 (s, 4H), 4.19 (s, 2H), 3.85 (d, J=3.9Hz, 3H)

Embodiment 18

6- (4- methoxyphenyls)-N- (4- (8- morpholines imidazo [1,2-a] pyrazine -6- bases) phenyl) pyrimidine -4- formyls Amine

m.p.269-271℃；ESI-MS[M+H]⁺m/z:508.2；¹H NMR(400MHz,DMSO)δ10.96(s,1H), 9.38 (s, 1H), 8.57 (d, J=22.1Hz, 2H), 8.46 (s, 1H), 8.32 (d, J=8.0Hz, 2H), 8.03 (dd, J= 14.7,8.4Hz, 4H), 7.96 (s, 1H), 7.59 (s, 1H), 7.14 (d, J=8.1Hz, 2H), 4.29 (s, 4H), 3.87 (s, 3H),3.80(s,4H).

Embodiment 19

6- (4- bromophenyls)-N- (4- (8- morpholines imidazo [1,2-a] pyrazine -6- bases) phenyl) pyrimidine -4- formamides

m.p.221-223℃；ESI-MS[M+H]⁺m/z:557.1；¹H NMR(400MHz,DMSO)δ10.91(s,1H), 9.49 (s, 1H), 8.65 (s, 1H), 8.59 (d, J=11.9Hz, 2H), 8.38 (d, J=8.3Hz, 2H), 8.00-7.94 (m, 3H), 7.85 (d, J=8.3Hz, 2H), 7.59 (d, J=2.7Hz, 2H), 4.30 (s, 4H), 3.79 (d, J=4.0Hz, 4H)

Embodiment 20

N- (4- (8- morpholinoes imidazo [1,2-a] pyrazine -6- bases) phenyl) -6- phenyl pyrimidine -4- formamides

m.p.272-275℃；ESI-MS[M+H]⁺m/z:478.1；¹H NMR(400MHz,DMSO)δ10.79(s,1H), 8.75 (d, J=5.1Hz, 1H), 8.58 (s, 1H), 8.39 (s, 1H), 8.04 (d, J=8.7Hz, 2H), 8.01 (s, 1H), 7.98 (d, J=6.3Hz, 2H), 7.95 (s, 1H), 7.89 (d, J=8.5Hz, 2H), 7.58 (s, 1H), 7.13 (d, J=8.5Hz, 2H), 4.29 (s, 4H), 3.80 (d, J=4.3Hz, 4H)

Embodiment 21

N- (5- (4- morpholinoes thieno [3,2-d] pyrimidine -2-base) pyridine -2- bases) -4- phenylpyridine acid amides

ESI-MS[M+H]+m/z:494.2

Embodiment 22

N- (5- (4- (3- methyl morpholines) thieno [3,2-d] pyrimidine -2-base) pyridine -2- bases) -4- phenylpyridine acid amides

ESI-MS[M+H]+m/z:509.2

Embodiment 23

N- (5- (6- ((dimethylamino) methyl) -4- (3- methyl morpholines) thieno [3,2-d] pyrimidine -2-base) pyridine - 2- yls) -4- (thiophene -2- bases) picolinamide

ESI-MS[M+H]⁺m/z:572.2

Embodiment 24

N- (5- (8- (3- methyl morpholines base) imidazo [1,2-a] pyrazine -6- bases) pyridine -2- bases) -4- (thiophene -2- bases) Picolinamide

ESI-MS[M+H]⁺m/z:498.2

Embodiment 25

N- (5- (8- (3- methyl morpholines base) imidazo [1,2-a] pyrazine -6- bases) pyridine -2- bases) -4- phenylpyridine acyls Amine

ESI-MS[M+H]+m/z:492.2

Embodiment 26

N- (5- (6- (2- hydroxyl propyl- 2- yls) -4- morpholinoes thieno [3,2-d] pyrimidine -2-base) pyridine -2- bases) -4- Phenylpyridine acid amides

ESI-MS[M+H]⁺m/z:553.2

Embodiment 27

N- (5- (4- morpholinoes -6- (the fluoro- 2- hydroxyls propyl- 2- yls of 1,1,1- tri-) thieno [2,3-d] pyrimidine -2-base) pyrroles Pyridine -2- bases) -4- phenylpyridine acid amides

ESI-MS[M+H]⁺m/z:607.2

Embodiment 28

N- (5- (4,6- dimorpholinos thieno [2,3-d] pyrimidine -2-base) pyridine -2- bases) -4- phenylpyridine acid amides

ESI-MS[M+H]⁺m/z:580.2

In vitro cytotoxic effect

To the heterocycle containing biaryl amide structure according to formula I or II of the present invention, simultaneously pyrimidine or pyrazine compounds are carried out External suppression lung carcinoma cell H460, colon cancer cell HT-29, human breast cancer cell MDA-MB-231, people glioblastoma are female thin Born of the same parents' oncocyte U87MG, human lung carcinoma cell H1975 and Liver cancer cell SMMC-7721 screening active ingredients, reference substance GDC-0941 according to Document (J.Med.Chem., 2008,51 (18), pp 5522-5532) methods described is prepared.

1) after 2~3 stabilizations of cell recovery and passage, it is made to disappear from blake bottle bottom with trypsin solution (0.25%) Change is got off.After cell dissociation buffer is poured into centrifuge tube, nutrient solution is added to terminate digestion afterwards.By centrifuge tube in 800r/min Lower centrifugation 10min, adds 5mL nutrient solutions after abandoning supernatant, piping and druming mixes cell, draws 10 μ L cell suspensions and adds cell Counted in tally, adjustment cell concentration is 10⁴Individual/hole.Except A1 holes for blank well is not added with extracellular in 96 orifice plates, remaining all adds Enter 100 μ L cell suspensions.96 orifice plates are put into incubator and cultivate 24h.

2) with 50 μ L dmso solution given the test agent, appropriate nutrient solution is subsequently adding, sample is dissolved into 2mg/mL Liquid, then in 24 orifice plates by Sample Dilution be 20,4,0.8,0.16,0.032 μ g/mL.

Each concentration add 3 holes, wherein around the row cell growing way of two row two it is affected by environment larger, only and be blanc cell Hole uses.96 orifice plates are put into incubator and cultivate 72h.

3) band medicine nutrient solution in 96 orifice plates is discarded, is rinsed cell twice with phosphate buffer solution (PBS), in every hole Add MTT (tetrazole) (0.5mg/mL) 100 μ L to be put into incubator after 4h, discard MTT solution, add the μ of dimethyl sulfoxide (DMSO) 100 L.Vibration makes survivaling cell fully be dissolved with MTT product on magnetic force oscillator, is put into measurement result in ELIASA.Pass through Bliss methods can obtain medicine IC₅₀Value.

The suppression lung carcinoma cell H460 of compound, colon cancer cell HT-29, human breast cancer cell MDA-MB-231, people dislike Property glioblastoma cells U87MG, human lung carcinoma cell H1975 and Liver cancer cell SMMC-7721 Activity Results are shown in Table 2.

PI3K α enzymatic activitys are tested

1st, solution is prepared

1) testing compound adds 1mL DMSO, is made into 10mM storage solutions.Positive compound GDC-0941 storing liquid concentration It is 10mM (being dissolved in DMSO) that the storing liquid concentration of positive compound cis-platinum is 2mM (being dissolved in DMSO).

2) DMSO diluted compounds storing liquids are used, 2mM solution (100X) is made into.

3) 2 μ L 2mM solution are taken, 18 μ L reaction solutions diluted compounds to 200 μM of (10X) solution are added.

4) the 2 above-mentioned solution of μ L and 18 μ L reaction solutions are added in working plate, 10X solution is made into.

5) take with the μ L of solution in upper plate 1 to detection plate.

6) 1 μ L kinase reaction liquid is added in the full suppression control of detection plate and null suppression control wells so that the concentration of DMSO It is 10%.

2nd, experimental procedure

1) layout of orifice plate

384 orifice plates are needed to arrange according to experiment, wherein：

A) HPE (complete to suppress control)：It is not added with kinases and compound, plus ATP, substrate and 1%DMSO.

B) ZPE (null suppression control)：It is not added with compound, plus kinases, ATP, substrate and 1%DMSO.

C) positive reference compound hole：Plus kinases, ATP, substrate and various concentrations positive compound.

D) testing compound hole：Plus kinases, ATP, substrate and testing compound.

2) agents useful for same is prepared

4XATP：ATP is diluted to 4X with reaction solution.

4X substrate solutions：Substrate is diluted to 4X with reaction solution.

2.5X kinase solutions：With reaction solution by kinase dilution to 2.5X.

3) kinase reaction

A) 1 μ L10X compounds (positive control of testing compound or various kinases) solution is added per hole according to arrangement, It is complete to suppress control and null suppression control wells 1 μ L reaction solutions of addition.

B) 4 μ L2.5X kinase solutions are added per hole according to arrangement.The full control wells that suppress add 4 μ L reaction solutions.

C) detection plate 1000rpm is centrifuged to mix.

D) 4XATP solution is mixed in equal volume with 4X substrate solutions, obtains 2XATP- substrate solutions.

E) the above-mentioned 2X ATP- substrate solutions of 5 μ L are added per hole according to arrangement.

F) detection plate 1000rpm is centrifuged to mix.

G) detection plate is placed in into 30 DEG C to react 1 hour.

H) 10 μ L Kinase glo plus or ADP-Glo reaction reagents are added per hole, 27 DEG C are placed 20 minutes.

I) 20 μ L Kinase Detection reagents are added per hole, 27 DEG C are placed 30 minutes.

J) Envision reads fluorescence values.

Note:Preset room temperature is needed before Kinase glo plus, ADP-Glo and Kinase Detection reagent uses Half an hour.

4) primary data analysis

Prism5.0 analyzes initial data.

The IC of compound is calculated according to Bliss methods₅₀

Experimental result is as shown in table 2.IC in table 1₅₀>=80%, represented with " +++ ", 80%>IC₅₀>=60%, with " ++ " Represent, 60%>IC₅₀>=40%, represented with "+", IC₅₀<=40%, represented with "-", " NA " represents inactive, and " ND " is represented Do not test.

The target compound anti tumor activity in vitro of table 2 and enzymatic activity

From above-mentioned result of the test it can be clearly seen that the acid amides containing biaryl of claimed formula I or II of the invention The heterocycle and pyrimidine or pyrazine compounds of structure, with good anti tumor activity in vitro.

The heterocycle containing biaryl amide structure and pyrimidine or pyrazine compounds of formula of I or II of the present invention can be applied individually With, but typically given with pharmaceutical carrier mixture, the selection of the pharmaceutical carrier will be according to required route of administration and standard medicine Thing put into practice, separately below with the various pharmaceutical dosage forms of such compound, for example tablet, capsule, injection, aerosol, suppository, The preparation method of film, pill, externally-applied liniment and ointment, illustrates its new opplication in pharmaceutical field.

Application examples 1：Tablet

With the compound 6g of embodiment 2, after adding auxiliary material 12g to mix according to the general pressed disc method of pharmacy, 100 are pressed into, often Piece weight 180mg.

Application examples 2：Capsule

With the compound 10g of embodiment 8, after auxiliary material 20g is mixed according to the requirement of pharmacy capsule, load hollow glue Capsule, each capsule weight 150mg.

Application examples 3：Injection

With the compound 10g of embodiment 9, according to pharmacy conventional method, charcoal absorption is carried out, through 0.65 μm of miillpore filter After filtering, insert nitrogen pot and be made hydro-acupuncture preparation, every dress 2mL, altogether filling 100 bottles.

Application examples 4：Aerosol

With the compound 20g of embodiment 12, after being dissolved with appropriate propane diols, after adding distilled water and other spoke material, it is made The settled solution of 100mL is obtained final product.

Application examples 5：Suppository

With the compound 5g of embodiment 15, by finely ground addition glycerine it is appropriate, the glycerin gelatine for having melted is added after grinding well, grind Mill is uniform, is poured into the model for having applied lubricant, and suppository 25 is obtained

Application examples 6：Film

With the compound 5g of embodiment 17, by heating for dissolving, 80 mesh sieves after the stirring expansion such as polyvinyl alcohol, medicinal glycerin, water Net filtration, then the compound of embodiment 12 is added to stirring and dissolving, film applicator masking 50 in filtrate.

Application examples 7：Pill

With the compound 15g of embodiment 18, after being mixed with the matrix 50g heating fusings such as gelatin, in instilling cryogenic liquid paraffin, The ball of dripping pill 1000 is obtained altogether.

Application examples 8：Externally-applied liniment

With the compound 20g of embodiment 20, according to auxiliary material 5g mixed grindings such as conventional dose method and emulsifying agents, then add steaming Distilled water is prepared to 400mL.

Application examples 9：Ointment

With the compound 5g of embodiment 24, it is finely ground after ground well with the oleaginous base 250g such as vaseline it is prepared.

Although describing the present invention by particular, modification and equivalent variations are for being proficient in this field Be will be apparent from for technical staff, and they are included within the scope of the invention.

Claims

1. a kind of heterocycle containing biaryl amide structure and pyrimidine or pyrazine compounds, it is characterised in that structure is such as following to lead to Shown in Formulas I or II：

Wherein：

X=S, Y=CH or X=CH, Y=S；

Z is phenyl ring, five yuan or hexa-atomic heteroaromatic；

A, B, D, E, G are respectively CH or N；

R₁Selected from H,

R²Selected from H or-CH₃；

Ar for phenyl, naphthyl, 5~10 unit's heteroaryls, 5~10 yuan of saturations or fractional saturation heterocyclic radical, the phenyl, naphthyl, Heteroaryl and heterocyclic radical contain 1~3 hetero atom selected from O, N and S, and optional 1~4 identical or different R of Ar³Substitution；

R³Selected from hydrogen, halogen, hydroxyl, trifluoromethyl, trifluoromethoxy, amino, azido, nitro, cyano group, sulfydryl, (C₁~C₄) Alkyl, (C₃~C₆) cycloalkyl, (C₂~C₄) alkenyl, (C₂~C₄) alkynyl, (C₁~C₄) alkoxy, (C₁~C₄) alkylthio group, allyl Base, (2- methyl) pi-allyl, (C₁~C₄) alkoxy methyl, (C₁~C₄) alkyl acyl or (C₁~C₃) alkylenedioxy group takes Dai Ji.

R⁴、R⁵It is identical or different, separately selected from (C₁~C₆) alkyl or (C₃~C₆) cycloalkyl；Or R⁴And R⁵With and it The nitrogen-atoms that is connected form 5~10 yuan of saturated heterocyclyls together, the saturated heterocyclyl except with R⁴And R⁵The nitrogen of connection is former It is sub outer, optionally containing 1~3 hetero atom selected from O, N and S.

2. the heterocycle and pyrimidine or pyrazine compounds of biaryl amide structure are contained according to claim 1, it is characterised in that：

Wherein：

X=S, Y=CH or X=CH, Y=S；

Z is phenyl ring or pyridine ring；

A, B, D, E, G are respectively C or N；

R₁Selected from H,

R²Selected from H or-CH₃；

Ar is phenyl, naphthyl, pyridine radicals, pyrimidine radicals or thienyl, and the phenyl, naphthyl, pyridine radicals, pyrimidine radicals, thienyl are appointed Select 1~4 identical or different R³Substitution；

R³Selected from hydrogen, halogen, hydroxyl, trifluoromethyl, trifluoromethoxy, amino, azido, nitro, cyano group, sulfydryl, (C₁~C₄) Alkyl, (C₃~C₆) cycloalkyl, (C₁~C₄) alkenyl, (C₁~C₄) alkynyl, (C₁~C₄) alkoxy, (C₁~C₄) alkylthio group, allyl Base, (2- methyl) pi-allyl, (C₁~C₄) alkoxy methyl, (C₁~C₄) alkyl acyl or (C₁~C₃) alkylenedioxy group takes Dai Ji.

It is selected from：

3. the heterocycle and pyrimidine or pyrazine compounds of biaryl amide structure are contained according to claim 1, it is characterised in that： The compounds of formula I is selected from the one kind in following compounds：

4- (2,4- 3,5-dimethylphenyls)-N- (4- (4- morpholinoes thieno [2,3-d] pyrimidine -2-base) phenyl) picolinamide,

6- (4- methoxyphenyls)-N- (4- (4- morpholinoes thieno [2,3-d] pyrimidine -2-base) phenyl) pyrimidine -4- formamides,

N- (4- (4- morpholinoes thieno [2,3-d] pyrimidine -2-base) phenyl) -6- (4- (trifluoromethyl) pyrimidine -4- formyls Amine,

4- (2,4- 3,5-dimethylphenyls)-N- (4- (8- morpholinoes imidazo [1,2-a] pyrazine -6- bases) phenyl) picolinamide,

6- (4- methoxyphenyls)-N- (4- (8- morpholines imidazo [1,2-a] pyrazine -6- bases) phenyl) pyrimidine -4- formamides,

N- (5- (6- ((dimethylamino) methyl) -4- (3- methyl morpholines) thieno [3,2-d] pyrimidine -2-base) pyridine -2- Base) -4- (thiophene -2- bases) picolinamide,

N- (5- (8- (3- methyl morpholines base) imidazo [1,2-a] pyrazine -6- bases) pyridine -2- bases) -4- (thiophene -2- bases) pyridine Acid amides,

N- (5- (8- (3- methyl morpholines base) imidazo [1,2-a] pyrazine -6- bases) pyridine -2- bases) -4- phenylpyridines acid amides,

N- (5- (6- (2- hydroxyl propyl- 2- yls) -4- morpholinoes thieno [3,2-d] pyrimidine -2-base) pyridine -2- bases) -4- phenyl Picolinamide,

N- (5- (4- morpholinoes -6- (the fluoro- 2- hydroxyls propyl- 2- yls of 1,1,1- tri-) thieno [2,3-d] pyrimidine -2-base) pyridine -2- Base) -4- phenylpyridines acid amides,

4. a kind of one of the claims 1 to 3 of Pharmaceutical composition, including therapeutically effective amount is described containing the miscellaneous of biaryl amide structure Ring and pyrimidine or pyrazine compounds, its geometric isomer and its pharmaceutically acceptable salt, hydrate, solvate or preceding Medicine is used as active component and pharmaceutically acceptable excipients.

5. a kind of heterocycle and pyrimidine or pyrazine compounds containing biaryl amide structure as claimed in claim 1, control in preparation Treat and/or prevent the application in proliferative disease medicine.

6. a kind of heterocycle and pyrimidine or pyrazine compounds containing biaryl amide structure as claimed in claim 1, control in preparation Application in the medicine for the treatment of and/or pre- anti-cancer.

7. a kind of heterocycle and pyrimidine or pyrazine compounds containing biaryl amide structure as claimed in claim 1, control in preparation Treat and/or prevent the application in the medicine of prostate cancer, lung cancer and cervical carcinoma.