CN103709162B - Tri-substituted imidazole benzodiazine ketonic compound and its production and use - Google Patents

Tri-substituted imidazole benzodiazine ketonic compound and its production and use Download PDF

Info

Publication number
CN103709162B
CN103709162B CN201210378551.2A CN201210378551A CN103709162B CN 103709162 B CN103709162 B CN 103709162B CN 201210378551 A CN201210378551 A CN 201210378551A CN 103709162 B CN103709162 B CN 103709162B
Authority
CN
China
Prior art keywords
compound
benzodiazine
substituted
under condition
imidazo
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN201210378551.2A
Other languages
Chinese (zh)
Other versions
CN103709162A (en
Inventor
龙亚秋
耿美玉
许忠良
王勇
艾菁
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Shanghai Institute of Materia Medica of CAS
Original Assignee
Shanghai Institute of Materia Medica of CAS
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Shanghai Institute of Materia Medica of CAS filed Critical Shanghai Institute of Materia Medica of CAS
Priority to CN201210378551.2A priority Critical patent/CN103709162B/en
Publication of CN103709162A publication Critical patent/CN103709162A/en
Application granted granted Critical
Publication of CN103709162B publication Critical patent/CN103709162B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/12—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains three hetero rings
    • C07D471/14—Ortho-condensed systems

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Nitrogen Condensed Heterocyclic Rings (AREA)

Abstract

The present invention relates to 1 shown in formula I, 3,5 three replacement 1H imidazos [4,5 h] 1,6 benzodiazine 2 (3H) ketonic compounds, its isomer and pharmaceutically acceptable salt, Preparation Method And The Use and comprise the compositions of described compound.The invention still further relates to described compound, its isomer and pharmaceutically acceptable salt and the compositions containing it is preparing the purposes in the medicine treating the diseases relevant with protein kinase particularly c Met such as tumor disease as Mutiple Targets kinases inhibitor.

Description

Tri-substituted imidazole benzodiazine ketonic compound and its production and use
Technical field
The present invention relates to medicinal chemistry art, be specifically related to 1,3,5-tri-replacements-1H-imidazo [4,5-h] 1,6-phenodiazine Miscellaneous naphthalene-2 (3H)-one compound, its isomer and pharmaceutically acceptable salt, Preparation Method And The Use and comprise describedization The compositions of compound.The invention still further relates to described compound, its isomer and pharmaceutically acceptable salt and the combination containing it Thing is relevant with protein kinase particularly c-Met for treating tumor disease etc. in preparation as Mutiple Targets kinases inhibitor Purposes in the medicine of disease.
Background technology
Malignant tumor is commonly encountered diseases and the frequently-occurring disease of a kind of serious threat human health, increases so that cell or mutant are abnormal Growing and shifting to surrounding tissue is feature, and the death that the mankind cause because of malignant tumor occupy the second of all mortalities Position, is only second to cardiovascular and cerebrovascular disease.The Therapeutic Method of tumor has operative treatment, radiotherapy and Drug therapy (chemotherapy) Deng, but the most still based on chemotherapy, the targeting designed in particular for the key enzyme in tumor mechanism The appearance of property antitumor drug so that chemotherapy can successfully heal the sick or extend significantly the life-span of patient.
In recent years, tyrosine protein kinase (Protein tyrosine kinase, PTK) is as targeting antineoplastic medicine The target of thing, has caused medicine scholar interest widely.Tyrosine protein kinase is the important factor in signal transduction process, ginseng With a series of cell functions, grow with cell, break up, breed closely related [Microsc Res Tech.2003Jan 1；60 (1): 70-75], it can be catalyzed the γ phosphate of ATP and transfers to, on the tyrosine residue of many key proteins, make phenolic hydroxyl group Phosphorylation.The expression product of many cancer genes the most all has PTK activity, and the PTK activity in a lot of malignant conversioning cells is far away Higher than normal cell.If PTK activity therefore can be suppressed, it is possible to block the growth of tumor cell.PTK has become as the note that induces one Purpose antitumor drug novel targets [Curr Drug Targets.2003, Feb；4(2):113-121].
The antineoplastic agent of targeting PTK achieves rapid progress, Gleevec, Iressa and Crizotinib at last decade Successively being approved by the FDA in the United States listing, fully demonstrating receptor tyrosine kinase is an effective antitumour drug target, and Mutiple Targets tyrosine-kinase enzyme level (multiple targeted tyrosine kinase inhibition) also becomes very Rich promising oncotherapy New Policy [European J.Cancer.2006, Jun；42,1351-1356].Because the biggest portion Point tumor does not all rely on a bars pathway to maintain its growth and survival, so, Mutiple Targets medicine can suppress many Weight signal transduction path or the multi-step of a bars pathway, not only have synergism, and it be thin to be difficult to induced tumor Born of the same parents produce drug resistance.The new ideas of this molecular targeted tumor pharmacother also obtain successful clinical evidence, as recently by It is multiple that the medicine Crizotinib of the treatment advanced Non-small cell lung of FDA approval listing is simultaneously targeting c-Met and ALK etc. exactly The Mutiple Targets medicine of protein kinase.
Mostly the tyrosine kinase inhibitor of document report is the derivant of pyrimidine or quinazoline structure, the 1H-miaow of the present invention Azoles also [4,5-h] 1,6-benzodiazine-2 (3H)-one compounds is the multiple inhibitors of kinases of a class new construction, and it is right to show The kinase whose inhibitory activity such as c-Met, FGFR, Abl, Lck, KDR, IGF-1 α, ALK, and can effectively suppress related neoplasms thin The cell of born of the same parents system such as BaF3-TPR-Met grows.
Patent documentation related to the present invention is listed below:
WO98/13350 discloses a series of quinolines vascular endothelial growth factor receptor inhibitors.The most also 1,8-bis-is included Embodiment 53:2-acetylaminohydroxyphenylarsonic acid 5-(2-fluoro-5-hydroxy-4-methyl aniline) in azanaphthalene analog derivative, such as this patent- 1,8-benzodiazine.
WO99/32450 discloses 4-hydroxyquinoline-2-carboxamide derivatives for treating herpesvirus infection.
WO98/11073 discloses Bioquin-7CA's amine derivative for treating herpesvirus infection.
WO02/30931 discloses 8-hydroxyl-1,6-benzodiazine-7-carbonyl amines compound sick for treating HIV-1 Poison infects.
CN2008102000645.4 discloses 5,8-bis-replacement-1,6-benzodiazine-7-amidocarbonylation compound for Treatment breast carcinoma, colon cancer, ovarian cancer, carcinoma of prostate.
CN201010615647.7 discloses 5,8-bis-replacement-1,6-benzodiazine-7-amidocarbonylation compound and Dimer compound is used for treating breast carcinoma, ovarian cancer, malignant melanoma, Skin Squamous Cell Carcinoma, colon cancer, pulmonary carcinoma, hepatocarcinoma, pancreas Cancer, acute myeloid leukaemia and carcinoma of prostate.
Summary of the invention
It is an object of the present invention to provide a kind of 1,3,5-tri-replacement-1H-imidazo by shown in following formula I [4, 5-h] 1,6-benzodiazine-2 (3H)-one compound, its isomer and pharmaceutically acceptable salt,
Wherein,
R1For hydrogen, C1-C6Alkyl, C3-C6Cycloalkyl, amino C1-C6Alkyl, amino C3-C6Cycloalkyl, C1-C6Alkyl amino C1-C6Alkyl, C1-C6Alkoxycarbonyl amido C3-C6Cycloalkyl, C6-C10Aryl C1-C6Alkyl, adamantyl C1-C6Alkyl or C5- C10Heteroaryl C1-C6Alkyl；
R2For hydrogen, C1-C6Alkyl, substituted or unsubstituted C6-C10Aryl C1-C6Alkyl, wherein, described substituent group has 1-2 Individual and be each independently halogen ,-CN ,-CF3、-NO2, hydroxyl, amino, C1-C6Alkyl, C1-C6Alkoxyl, halogen substiuted C1-C6Alkoxyl, C5-C10Heteroaryl C1-C6Alkyl, C1-C6Alkyl sulphonyl, C6-C10Phenyl sulfonyl, adamantyl C1-C6Alkane Base or C5-C10Heteroarylsulfonyl；
R3For hydrogen, halogen, hydroxyl, C1-C6Alkyl oxy, substituted or unsubstituted C6-C10Aryloxy, wherein, described in take Dai Ji has 1-2 and is each independently halogen, aldehyde radical, C1-C6Hydroxyalkyl, 1-hexahydropyridine base carbonyl, N-morpholinyl carbonyl, (R)-1-PhenethyIamino carbonyl or 4-methylpiperazine-1-yl-carbonyl, or C5-C10Heteroaryl epoxide；
Wherein, the hetero atom in described heteroaryl is one or more in N, S and O；
Preferably,
R1For hydrogen, C1-C6Alkyl, C3-C6Cycloalkyl, amino C1-C6Alkyl, amino C3-C6Cycloalkyl, C1-C6Alkyl amino C1-C6Alkyl, C1-C4Alkoxycarbonyl amido C3-C6Cycloalkyl, phenyl C1-C3Alkyl, adamantyl C1-C3Alkyl or C5-C10Miscellaneous Aromatic ring yl C1-C6Alkyl；
R3For hydrogen, F, Cl, Br, hydroxyl, C1-C6Alkyl oxy, substituted or unsubstituted phenyl epoxide, wherein, described replacement Base has 1-2 and is each independently Cl, carboxaldehyde radicals, C1-C3Hydroxyalkyl, 1-hexahydropyridine base carbonyl, N-morpholinyl carbonyl, (R)-1-PhenethyIamino carbonyl or 4-methylpiperazine-1-yl-carbonyl, or pyridine radicals epoxide；
R2For hydrogen,Wherein,
L is C1-C6Alkyl or-SO2-；
R4And R5It is each independently hydrogen, halogen ,-CN ,-CF3、-NO2, hydroxyl, amino, C1-C3Alkyl, C1-C6Alkoxyl Or the substituted C of fluorine1-C6Alkoxyl；
Hetero atom in described heteroaryl is one or more in N and O；
It is highly preferred that
R1For C3-C6Cycloalkyl, amino C3-C6Cycloalkyl, tertbutyloxycarbonylamino C3-C6Cycloalkyl,Benzene first Base or 1-phenylethyl；
R3For Br, hydroxyl, C1-C6Alkyl oxy, substituted or unsubstituted phenyl epoxide, wherein, described substituent group has 1-2 Individual and be each independently Cl, carboxaldehyde radicals, C1-C3Hydroxyalkyl, 1-hexahydropyridine base carbonyl, N-morpholinyl carbonyl, (R)-1-benzene second Base amino carbonyl or 4-methylpiperazine-1-yl-carbonyl, 3-pyridine radicals epoxide or 2-pyridine radicals epoxide；
R2For hydrogen,Wherein,
L is C1-C3Alkyl or-SO2-；
R4And R5It is each independently hydrogen, F, Cl ,-CN ,-CF3, methyl, methoxyl group or-OCF3；
Most preferably, described compound, its isomer and pharmaceutically acceptable salt are following compound:
It is a further object of the present invention to provide 1,3,5-tri-replacement-1H-imidazo [4,5-h] shown in a kind of above-mentioned formula I 1,6-benzodiazine-2 (3H)-one compound, its isomer and the preparation method of pharmaceutically acceptable salt, the method is following Shown in reaction scheme:
Wherein, R1、R2、R3Definition same as described above, and the preferably group shown in table；
Reaction reagent and condition: (a) isopropanol, backflow；(b) thionyl chloride, backflow；(c) sodium borohydride, oxolane, 0 ℃；(d)TsNHCH2COOCH3, DEAD (diethyl azodiformate), triphenylphosphine, oxolane, 0 DEG C；(e) Feldalat NM, first Alcohol, 0 DEG C～room temperature；(f) NBS (N-bromo-succinimide), dichloromethane, room temperature；(g) TsCl, triethylamine, dichloromethane； (h)R1NH2, oxolane, backflow；(i) LiOH solution/methanol, or NaOH solution/THF, backflow；(j) DPPA (nitrine di(2-ethylhexyl)phosphate Phenyl ester), triethylamine, toluene, backflow；(k) trifluoracetic acid/dichloromethane, room temperature；(l)R2X, K2CO3, DMF, room temperature；X be Cl or Br；M () substituted phenol, described substituent group has 1-2 and is each independently carboxaldehyde radicals or C1Hydroxyalkyl, DMF, 110 DEG C； N () substituted benzene sulfonyl chloride, described substituent group has 1-2 and is each independently Cl or methyl, K2CO3, THF, backflow；(o) boron Sodium hydride, THF, room temperature；
The method specifically includes following steps:
(1) compound 1 pyridine 2,3-dicarboxylic anhydride selective esterification under condition a is obtained compound 2；Compound 2 is at bar Under part b, chloro obtains compound 3；Compound 3 obtains compound 4 with the selective reduction under condition c of acyl chlorides form；Compound 4 Compound 5 is obtained by Mitsunobo reaction under condition d；Compound 5 cyclization under condition e obtains 1,6-benzodiazine carbonyl Acid methyl compound 6；Compound 6 and N-bromo-succinimide react under condition f and obtain the acid of 5-Br-1,6-benzodiazine carbonyl Methyl compound 7；8 hydroxyls of compound 7 obtain compound 8 with Ts protection under condition g, and (synthetic route of compound 8 is shown in List of references WO2002030426 and WO2002030930)；Compound 8 under condition h with amine (R1NH2) occur nucleophilic displacement of fluorine anti- 8 bit amino substituted compounds 9 should be obtained；Compound 9 is hydrolyzed to compound 10 under alkalescence condition i further；Compound 10 exists Occur Cutis to reset in the presence of condition j DPPA, then occur molecule nucleophilic substitution to generate 1H-imidazo [4,5-h] 1,6- Benzodiazine-2 (3H)-one compound 11；With
(2)R1Available for the compound 11 direct Deprotection under condition k during the aminocyclohexyl of end Boc protection Compound S1；Compound 11 under the conditions of l 3 introduce corresponding substituent groups obtain as described in compound S10 in formula I, S13-S20；And/or
(3)R1For end Boc protection aminocyclohexyl time compound 11 under condition l with R2X reaction obtains compound 12；Compound 12 removes Boc under condition k and generates 1,3,5-tri-replacement-1H-imidazo [4,5-h] 1,6-bis-of described formula I Azanaphthalene-2 (3H)-one compound S2-S9；And/or
(4)R1For end Boc protection aminocyclohexyl time compound 11 be heated to reflux under condition n 8 hours with take The benzene sulfonyl chloride reaction in generation obtains substituted 3-benzenesulfonyl substituted-1H-imidazo [4,5-h] 1,6-benzodiazine-2 (3H)-one compound, sloughs Boc through 20% trifluoracetic acid/dichloromethane under condition k and obtains described compounds of formula I S11 or S12；And/or
(5) 5 bromine substituted 1H-imidazo [4,5-h] 1,6-benzodiazine-2 (3H)-one compounds are in the basic conditions I.e. can get described compound S22 with substituted phenol reactant, by compound S22 further under condition o in sodium borohydride Reduction obtains described compounds of formula I S23；And/or
(6) compound 12 obtain with substituted phenol reactant in the basic conditions 1,3,5-tri-replacement-1H-imidazo [4, 5-h] 1,6-benzodiazine-2 (3H)-one compound 13, compound 13 is sloughed Boc through 20% trifluoracetic acid/dichloromethane and is obtained Described compounds of formula I S26-S27.
Another object of the present invention be to provide a kind of comprise therapeutically effective amount selected from compound shown in formula I, its isomery The compositions of one or more in body and pharmaceutically acceptable salt.
The a further object of the present invention is to provide compound shown in formula I, its isomer and pharmaceutically acceptable salt and bag Compositions containing it is being prepared as the application in the medicine of Mutiple Targets kinases inhibitor.
Described kinases includes c-Met, FGFR, Abl, Lck, KDR, IGF-1 α and ALK.
Described medicine is for treatment and/or prevents the disease relevant with protein kinase particularly c-Met, such as tumor etc..
By to c-Met kinase activity screen, it is found by the applicant that: the compound that above-mentioned formula I represents under 10 μMs to c- Met kinases has efficient inhibitory activity.Find simultaneously, the compound that formula I represents under 10 μMs to BaF3-TPR-Met cell There is efficient inhibitory activity.
Therefore, the compound that formula I represents can effectively targeting c-Met mediation signal path, it is possible to for and c- The treatment of the relevant diseases such as the tumor that Met kinases overexpression causes.
The pharmaceutical composition of the compound represented containing formula I allowed on galenic pharmacy can play effective targeting c-equally The signal path of Met mediation, it is possible to for the treatment of the relevant diseases such as the tumor that causes with c-Met kinases overexpression.
Accompanying drawing explanation
Fig. 1 shows the compounds of this invention inhibitory action to multiple protein kinases.
Detailed description of the invention
Preparation embodiment:
Below in conjunction with preparation embodiment, the invention will be further described, but is not intended to the present invention.
Compound1H-NMR spectroscopic data is measured and is used Varian Mercury-300MHz or Varian Mercury- 400MHz nuclear magnetic resonance, NMR, elementary analysis uses Vario EL analyzer, and fusing point Buchi-510 capillary tube method measures, and temperature is not Calibrated.Infrared spectrum is by Bio-Rad FTS-185 determination of infrared spectroscopy；Mass spectrum EI-MS Finnigan MAT 95 mass spectrum Instrument, ESI-MS uses Finnigan LCQ Deca mass spectrograph to measure.Specific rotation is by P-1030 (A012360639) automatic polarimeter Measure.Rapid column chromatography is carried out on silica gel H (10-40 μM).Reagent purification is with reference to Purification of laboratory Chemicals；D.D.Perrin；W.L.F.Armarego and D.R.Perrin Eds.,Pergamon Press: Oxiford,1980。
As being not specifically noted, reagent of the present invention and method etc. are reagent well known in the art and method.
Preparation embodiment 1 pyridine-2,3-dioctyl phthalate-2-isopropyl ester (2)
Compound 1 pyridine 2,3-dicarboxylic anhydride (48.1g, 0.32mol) is dissolved in the 2-isopropanol of 100mL and is heated to reflux 16 hours, then solution is cooled to-20 DEG C and obtains compound as white solid 2 (44.4g, 68%).Fusing point: 140-141 DEG C；1H NMR(CDCl3, 300MHz): δ 8.87 (d, J=4.2Hz, 1H), 8.35 (d, J=7.8Hz, 1H), 7.55 (dd, J=5.1, 8.1Hz, 1H), 5.36 (septet (septet), J=6.3Hz, 1H), 1.41 (d, J=6.3Hz, 6H).
Preparation embodiment 2 3-hydroxymethyl-pyridine-2-isopropyl formate (4)
Compound 2 (52.7g, 0.25mol) is dissolved in the thionyl chloride of 400mL and is heated to reflux to solution becoming homogeneous, Then solvent it is spin-dried for.Adding 2 × the anhydrous THF of 50mL revolves steaming, removes the thionyl chloride of residual.Obtained red liquid is dissolved in The anhydrous THF of 400mL is cooled to 0 DEG C, is dividedly in some parts sodium borohydride (28.6g, 0.76mol), stir 4 hours at 0 DEG C, will be anti- Answer solution careful pour in frozen water, 3 × 200mL dichloromethane extract, add anhydrous Na2SO4It is dried.Column chromatography (petroleum ether: second Acetoacetic ester=3:2) obtain yellow solid compound 4 (18.8g, 38%).1H NMR(CDC13, 300MHz): δ 8.69 (dd, J= 1.5,4.7Hz, 1H), 7.88 (dd, J=1.5,7.7Hz, 1H), 7.46 (dd, J=4.7,7.8Hz, 1H), 5.35 (septet, J =6.4Hz, 1H), 4.81 (m, 2H), 1.45 (d, J=6.3Hz, 6H)；EI-MS m/z:195(M)+。
Preparation embodiment 3 3-{ [methoxycarbonyl-methyl-(toluene-4-sulfonyl)-amino]-methyl }-pyridine-2-carboxylic acids Isopropyl ester (5)
By compound 4 (1.734g, 8.89mmol), 2-para toluene sulfonamide acetic acid methyl ester (TsNHCH2COOCH3) (2.163g, 8.89mmol), and triphenylphosphine (3.499g, 13.338mmol) is dissolved in the anhydrous THF of 100mL, is cooled to 0 DEG C, inflated with nitrogen is protected.DEAD (2.165mL, 13.338mmol) is dissolved in the anhydrous THF of 10mL, is added dropwise over DEAD.Remove ice Bath, is spin-dried for obtaining red oil compound 5 after stirring two hours and is directly used in the next step.
8-hydroxyl-1,6-benzodiazine-7-carboxylate methyl ester (6)
The compound 5 (8.89mmol) that the reaction of upper step obtains is dissolved in 50mL absolute methanol, is cooled to 0 DEG C.Slowly Add Feldalat NM (1.681g, 31.123mmol).Remove ice bath, stir 3 hours.Spin off solvent, add 20mL water, 20mL acetic acid Ethyl ester, organic facies saturated sodium carbonate back extraction.Merge aqueous phase, adjust pH to 7, maintain aqueous phase pH to 7, extract 5 times with dichloromethane. Organic facies anhydrous sodium sulfate is dried, and column chromatography (petroleum ether: ethyl acetate=2:1) obtains pale solid compound 6 (0.62g, two step productivity 65%).Fusing point: 179-180 DEG C；1H NMR(CDC13,300MHz):δ11.79(s,1H),9.20(s, 1H), 8.85 (s, 1H), 8.32 (d, J=8.2Hz, 1H), 7.71 (dd, J=4.1,8.2Hz, 1H), 4.12 (s, 3H).
Preparation embodiment 4 5-bromo-8-hydroxyl-1,6-benzodiazine-7-carboxylate methyl ester (7)
Under room temperature, NBS (30mg, 0.167mmol) is joined the 1mL CH of compound 6 (34mg, 0.167mmol)2Cl2Molten In liquid, stir 1 hour.Filter, be dried to obtain compound as white solid 7 (30mg, productivity 85%)；1H NMR(d6-DMSO, 300MHz): δ 9.26 (dd, J=1.5,4.2Hz, 1H), 8.59 (dd, J=1.6,8.4Hz, 1H), 8.00 (dd, J=4.2, 8.4Hz,1H),3.94(s,3H)。
Preparation embodiment 5 methyl 5-bromo-8-tolysulfonyl Oxy-1,6-benzodiazine-7-carboxylate (8)
By paratoluensulfonyl chloride, compound 7, triethylamine in chloroform after 50 DEG C of stirrings 5 hours successively with saturated ammonium chloride, Saturated common salt is washed, and is dried.Column chromatography obtains compound as white solid 8, productivity: 76%.1H NMR(300MHz,CDCl3):δ 9.05 (d, J=4.2Hz, 1H), 8.59 (d, J=7.8Hz, 1H), 7.85 (d, J=8.1Hz, 2H), 7.70 (dd, J=4.2, 7.8Hz, 1H), 7.34 (d, J=8.1Hz, 2H), 3.83 (s, 3H), 2.47 (s, 3H).
Preparation embodiment 6 tert-butyl group (1r, 4r)-4-(7-(methoxy carbonic acyl radical)-5-bromo-1,6-benzodiazine-8-amino) Cyclohexyl carbamate (9-1)
By compound 8, triethylamine and single Boc protection that Isosorbide-5-Nitrae-trans cvclohexvl diamidogen is heated to reflux 8 in oxolane is little Time, it is spin-dried for THF, adds dichloromethane, use saturated Na successively2CO3Aqueous solution, saturated aqueous ammonium chloride, saturated common salt are washed, Anhydrous Na2SO4It is dried.Column chromatography obtains yellow solid compound 9-1, productivity: 78%.1H NMR(300MHz,CDCl3):δ 8.95 (dd, 1H, J=1.5,4.2Hz), 8.88 (d, 1H, J=7.5Hz), 8.47 (dd, 1H, J=1.5,8.4Hz), 7.64 (dd, 1H, J=4.2,8.4Hz), 4.92 (m, 1H), 4.42 (m, 1H), 3.97 (s, 3H), 3.47 (m, 1H), 2.20 (m, 2H), 2.05(m,2H),1.44(s,9H),1.43-1.23(m,4H)；EI-MS m/z:478(M)+,480(M+2)+。
Preparation embodiment 7
The bromo-8-of 5-((1r, 4r)-4-t-butoxycarbonyl amino Cyclohexylamino)-1,6-benzodiazine-7-carboxylic acid (10-1)
Compound 10-1 is by compound 9-1 in 1N NaOH solution/THF, and under 60 degree, 10h hydrolysis prepares.Yellow green is solid Body, productivity: 85%.Fusing point: 122-126 DEG C；1H NMR(300MHz,CDCl3): δ 8.96 (dd, 1H, J=1.5,4.2Hz), 8.88 (d, 1H, J=7.5Hz), 8.45 (dd, 1H, J=1.5,8.4Hz), 7.64 (dd, 1H, J=4.2,8.4Hz), 4.96 (m, 1H),4.41(m,1H),3.47(m,1H),2.21(m,2H),2.08(m,2H),1.44(s,9H),1.43-1.23(m,4H)； EI-MS m/z:464(M)+,466(M+2)+。
Preparation embodiment 8 tert-butyl group ((1r, 4r)-4-(5-Br-2-ketone-2,3-dihydro-1H-imidazo [4,5-h] 1,6- Benzodiazine-1-base)-hexamethylene)-carboxylate (11-1)
Compound 10-1, triethylamine and DPPA are heated to reflux in toluene 8 hours, are spin-dried for solvent, add dichloromethane, Use saturated Na successively2CO3Aqueous solution, saturated aqueous ammonium chloride, saturated common salt are washed, anhydrous Na2SO4It is dried.Column chromatography obtains Yellow solid compound 11-1, productivity: 83%.1H NMR(300MHz,CDCl3): δ 9.05 (d, J=3.9Hz, 1H), 8.58 (d, J=8.7Hz, 1H), 7.47 (dd, J=3.9,8.7Hz, 1H), 4.47 (br s, 1H), 2.19 (d, J=12.3Hz, 2H), 1.87 (d, J=11.7Hz, 2H), 1.48 (s, 9H), 1.30-1.42 (m, 4H)；13C NMR(100MHz,d6-DMSO):δ154.9 152.8 139.6 137.4 133.3 121.3 119.3 115.7 77.5 48.3 32.0 28.3 28.0；MS-EI m/z: 461(M)+,463(M+2)+。
Preparation embodiment 9 1-((1r, 4r)-4-aminocyclohexane-)-5-Br-1-H-imidazo [4,5-h] 1,6-phenodiazine Miscellaneous naphthalene-2 (3H)-one (S1)
Compound 11-1 removes Boc in the trifluoracetic acid/dichloromethane of 20% and generates compound S1.Greenish yellow solid, Productivity: 78%.1H NMR(300MHz,d6-DMSO): δ 9.07 (d, J=3.0Hz, 1H), 8.56 (d, J=6.9Hz, 1H), 7.54 (dd, J=3.0,6.9Hz, 1H), 5.25 (br s, 1H), 2.85 (t, 1H), 2.66 (d, J=12Hz, 2H), 1.96 (d, J= 11.1Hz, 2H), 1.74 (d, J=10.2Hz, 2H), 1.31 (m, 2H)；13C NMR(100MHz,d6-DMSO):δ154.1140.1 135.2 120.2 116.7 49.2 34.4 29.9；MS-ESI m/z:362(M+H)+；HR-ESI MSC15H16BrN5O3(M+ H)+Value of calculation: 362.0616, measured value: 362.0620.
Preparation embodiment 10 1-((1r, 4r)-4-aminocyclohexane-)-3-benzyl-5-Br-1-H-imidazo [4,5-h] 1,6-benzodiazine-2 (3H)-one (S2)
1, the tert-butyl group ((1r, 4r)-4-(3-benzyl-5-Br-2-ketone-2,3-dihydro-1H-imidazo [4,5-h] 1,6-bis- Azanaphthalene-1-base)-hexamethylene) preparation of-carboxylate (12-1)
By compound 11-1, Anhydrous potassium carbonate and bromobenzyl room temperature reaction 12 hours in DMF, it is spin-dried for solvent, adds dichloro Methane, uses saturated Na successively2CO3Aqueous solution, saturated aqueous ammonium chloride, saturated common salt are washed, anhydrous Na2SO4It is dried.Post layer Analysis obtain the yellow solid compound tert-butyl group ((1r, 4r)-4-(3-benzyl-5-Br-2-ketone-2,3-dihydro-1H-imidazo [4, 5-h] 1,6-benzodiazine-1-base)-hexamethylene)-carboxylate (12-1), productivity: 60%.1H NMR(300MHz,CDCl3):δ 9.00 (d, J=3.6Hz, 1H), 8.56 (d, J=8.7Hz, 1H), 7.54 (d, J=6.9Hz, 2H), 7.42 (dd, J=3.6, 8.7Hz, 1H), 7.31 (m, 3H), 5.25 (s, 2H), 4.45 (br s, 1H), 2.16 (d, J=12.3Hz, 2H), 1.84 (d, J= 10.8Hz,2H),1.47(s,9H),1.26-1.40(m,4H)。
2,1-((1r, 4r)-4-aminocyclohexane-)-3-benzyl-5-Br-1-H-imidazo [4,5-h] 1,6-diaza The preparation of naphthalene-2 (3H)-one (S2)
Compound 12-1 removes Boc in the trifluoracetic acid/dichloromethane of 20% and generates compound S2.Yellow solid, produces Rate: 45%.1H NMR(300MHz,CD3OD): δ 9.06 (d, J=3.6Hz, 1H), 8.68 (d, J=8.4Hz, 1H), 7.60 (m, 1H), 7.43 (d, J=6.9Hz, 2H), 7.32 (m, 3H), 5.24 (s, 2H), 2.84 (m, 2H), 2.21 (d, J=14.7Hz, 2H), 2.02 (d, J=8.1Hz, 2H), 1.59-1.72 (m, 2H)；MS-EI m/z:451(M)+,453(M+2)+。
Preparation embodiment 11 1-((1r, 4r)-4-aminocyclohexane-)-3-(the fluoro-benzyl of 4-)-5-Br-1-H-imidazo [4,5-h] 1,6-benzodiazine-2 (3H)-one (S3)
In addition to replacing bromobenzyl with 4-fluorine bromobenzyl, the preparation method of compound S3 is identical with the preparation method of compound S2. Yellow solid, productivity: 61%.1H NMR(300MHz,CDCl3): δ 8.96 (d, J=3.9Hz, 1H), 8.56 (d, J=7.5Hz, 1H), 7.53 (t, J=9.0Hz, 2H), 7.42 (dd, J=3.9,7.5Hz, 1H), 6.99 (t, J=8.4Hz, 2H), 5.20 (s, 2H), 2.96 (t, 1H), 2.75 (m, 2H), 2.03 (d, J=13.2Hz, 2H), 1.87 (d, J=9.9Hz, 2H), 1.34-1.46 (m,2H)；MS-ESI m/z:470(M+H)+；C22H21BrFN5O·1/4CF3COOH value of calculation: C 54.17, H 4.29, N14.04, measured value: C 54.20, H 4.32, N 14.03.
Preparation embodiment 12 1-((1r, 4r)-4-aminocyclohexane-)-3-(3-fluoro-4-methoxy-benzyl)-5-Br-1- H-imidazo [4,5-h] 1,6-benzodiazine-2 (3H)-one (S4)
In addition to replacing bromobenzyl with 3-fluoro-4-methoxyl group bromobenzyl, the preparation method of compound S4 and the preparation of compound S2 Method is identical.Yellow solid, productivity: 83%.1H NMR(300MHz,CDCl3): δ 8.96 (d, J=3.9Hz, 1H), 8.55 (d, J =8.7Hz, 1H), 7.42 (dd, J=3.9,8.7Hz, 1H), 7.29 (d, J=3.9Hz, 1H), 7.26 (m, 1H), 6.90 (t, J =8.7Hz, 1H), 5.15 (s, 2H), 3.84 (s, 3H), 2.94 (t, J=11.1Hz, 1H), 2.74 (m, 2H), 2.00 (d, J= 12.0Hz, 2H), 1.86 (d, J=11.1Hz, 2H), 1.31-1.43 (m, 2H)；13C NMR(100MHz,CDCl3):δ153.7 153.3 150.8 147.3 147.2 139.2 138.4 137.9 134.6 129.4 129.3 124.7 120.6 120.5 116.6116.4 113.2 56.1 49.5 42.9 35.8 28.3；MS-ESI m/z:500(M+H)+；C23H23BrFN5O2· 3/2H2O value of calculation: C 52.38, H 4.97, N 13.28, measured value: C 52.42, H 4.73, N 13.06.
Preparation embodiment 13 1-((1r, 4r)-4-aminocyclohexane-)-3-(3-trifluoromethyl benzyl)-5-Br-1-H-miaow Azoles also [4,5-h] 1,6-benzodiazine-2 (3H)-one (S5)
In addition to replacing bromobenzyl with 3-trifluoromethyl bromobenzyl, the preparation method of compound S5 and the preparation method of compound S2 Identical.Yellow solid, productivity: 85%.1H NMR(300MHz,CDCl3): δ 8.98 (d, J=2.7Hz, 1H), 8.57 (d, J= 8.7Hz, 1H), 7.82 (s, 1H), 7.70 (d, J=7.5Hz, 1H), 7.53 (d, J=8.1Hz, 1H), 7.46 (m, 2H), 5.28 (s, 2H), 2.98 (t, J=10.2Hz, 1H), 2.76 (m, 2H), 2.04 (d, J=11.7Hz, 2H), 1.88 (d, J=13.8Hz, 2H),1.36-1.48(m,2H)；MS-EI m/z:519(M)+,521(M+2)+；HR-EI MS C23H21BrF3N5O(M)+Calculate Value: 519.0882, measured value: 519.0878；C23H21BrF3N5O·1/6CF3COOH value of calculation: C 51.96, H3.96, N 12.98, measured value: C 51.98, H 3.88, N 12.93.
Preparation embodiment 14 1-((1r, 4r)-4-aminocyclohexane-)-3-(4-chlorobenzyl)-5-Br-1-H-imidazo [4,5-h] 1,6-benzodiazine-2 (3H)-one (S6)
In addition to replacing bromobenzyl with 4-chlorine bromobenzyl, the preparation method of compound s 6 is identical with the preparation method of compound S2. Yellow solid, productivity: 60%.1H NMR(300MHz,CDCl3): δ 8.97 (d, J=3.9Hz, 1H), 8.57 (d, J=8.7Hz, 1H), 7.49 (d, J=8.4Hz, 2H), 7.44 (dd, J=3.9,8.7Hz, 1H), 7.29 (d, J=8.4Hz, 2H), 5.20 (s, 2H), 3.00 (t, J=11.7Hz, 1H), 2.75 (m, 2H), 2.05 (d, J=13.2Hz, 2H), 1.88 (d, J=12.9Hz, 2H),1.38-1.50(m,2H)；MS-ESI m/z:488(M+H)+；HR-ESI MS C22H21BrClN5O(M+Na)+Value of calculation: 508.0516, measured value: 508.0503；C22H21BrClN5O·1/6CF3COOH value of calculation: C 53.03, H 4.22, N13.85, Measured value: C 53.19, H 4.25, N 13.76.
Preparation embodiment 15 1-((1r, 4r)-4-aminocyclohexane-)-3-(2-chlorobenzyl)-5-Br-1-H-imidazo [4,5-h] 1,6-benzodiazine-2 (3H)-one (S7)
In addition to replacing bromobenzyl with 2-chlorine bromobenzyl, the preparation method of compound S7 is identical with the preparation method of compound S2. Yellow solid, productivity: 49%.1H NMR(300MHz,CDCl3): δ 9.00 (d, J=2.7Hz, 1H), 8.57 (d, J=8.7Hz, 1H), 7.38-7.47 (m, 2H), 7.13-7.23 (m, 2H), 7.06 (d, J=6.0Hz, 1H), 5.39 (s, 2H), 3.03 (t, J= 12.6Hz, 1H), 2.81 (m, 2H), 2.09 (d, J=10.8Hz, 2H), 1.93 (d, J=10.5Hz, 2H), 1.42-1.54 (m, 2H)；MS-ESI m/z:488(M+H)+；C22H21BrClN5O·1/3CF3COOH value of calculation: C 51.88, H 4.10, N13.34, Measured value: C 52.10, H 4.36, N 13.17.
Preparation embodiment 16 1-((1r, 4r)-4-aminocyclohexane-)-3-(3-trifluoro-methoxybenzyl)-5-Br-1-H- Imidazo [4,5-h] 1,6-benzodiazine-2 (3H)-one (S8)
In addition to replacing bromobenzyl with 3-trifluoromethoxy bromobenzyl, the preparation method of compound S8 and the preparation side of compound S2 Method is identical.Yellow solid, productivity: 83%.1H NMR(300 MHz,CDCl3): δ 8.99 (s, 1H), 8.57 (d, J=9.0 Hz, 1H), 7.58 (d, J=7.8 Hz, 2H), 7.43 (dd, J=9.0 Hz, 1H), 7.17 (d, J=7.8 Hz, 2H), 5.23 (s, 2H), 3.10 (m, 1H), 2.78 (m, 2H), 2.13 (d, J=13.2 Hz, 2H), 1.91 (d, J=13.5 Hz, 2H), 1.47- 1.59(m,2H)；13CNMR(100 MHz,CDCl3):δ153.8 152.6 148.8 137.9 135.1 130.3 121.1 120.7 49.6 43.0 34.528.1；MS-EI m/z:535(M)+,537(M+2)+；HR-EI MS C23H21BrF3N5O2(M)+ Value of calculation: 535.0831, measured value: 535.0837.
Preparation embodiment 17 1-((1r, 4r)-4-aminocyclohexane-)-3-(3,4-dichloro benzyl)-5-Br-1-H-imidazoles And [4,5-h] 1,6-benzodiazine-2 (3H)-one (S9)
Except with 3,4-dichloro bromobenzyl is replaced outside bromobenzyl, the preparation method phase of the preparation method of compound S9 and compound S2 With.Yellow solid, productivity: 71%.1H NMR(300 MHz,CDCl3): δ 8.97 (d, J=2.7Hz, 1H), 8.56 (d, J= 10.5Hz, 1H), 7.61 (s, 1H), 7.43 (dd, J=2.7,10.5 Hz, 1H), 7.38 (s, 2H), 5.17 (s, 2H), 2.97 (t, 1H), 2.74 (m, 2H), 2.03 (d, J=12.0 Hz, 2H), 1.89 (d, J=16.2 Hz, 2H), 1.35-1.48 (m, 2H)；13C NMR(100 MHz,CDCl3):δ153.8 152.4 138.9 137.9 136.4 134.7 132.5 131.9 130.5 128.1 120.7120.6 49.4 42.6 35.5 28.2；MS-EI m/z:519(M)+,521(M+2)+；HR-EI MS C22H20BrCl2N5O(M)+Value of calculation: 519.0228, measured value: 519.0229；C22H21BrFN5O·1/4CF3COOH calculates Value: C 49.16, H 3.71, N 12.74, measured value: C 49.28, H 3.75, N 12.77.
Preparation embodiment 18 tert-butyl group ((1r, 4r)-4-(3-(3,4-dichloro benzyl)-5-Br-2-ketone-2,3-dihydro- 1H-imidazo [4,5-h] 1,6-benzodiazine-1-base)-hexamethylene)-carboxylate (S10)
Except with 3,4-dichloro bromobenzyl is replaced outside bromobenzyl, the preparation method of compound S10 and the preparation side of compound 12-1 Method is identical.Yellow solid, productivity: 91%.1H NMR(300MHz,CDCl3): δ 9.02 (d, J=3.9Hz, 1H), 8.57 (d, J= 9.0Hz, 1H), 7.62 (s, 1H), 7.44 (dd, J=3.9,9.0Hz, 1H), 7.38 (s, 2H), 5.18 (s, 2H), 4.43 (br S, 1H), 3.71 (br s, 1H), 2.88 (br s, 1H), 2.17 (d, J=12.6Hz, 2H), 1.85 (d, J=11.4Hz, 2H), 1.47(s,9H),1.21-1.40(m,4H)；MS-ESI m/z:622(M+H)+。
Preparation embodiment 19 1-((1r, 4r)-4-aminocyclohexane-)-3-(4-Methyl benzenesulfonyl epoxide)-5-Br-1-H- Imidazo [4,5-h] 1,6-benzodiazine-2 (3H)-one (S11)
By compound 11-1, Anhydrous potassium carbonate and paratoluensulfonyl chloride back flow reaction 12 hours in THF, it is spin-dried for solvent, Add dichloromethane, use saturated Na successively2CO3Aqueous solution, saturated aqueous ammonium chloride, saturated common salt are washed, anhydrous Na2SO4Dry Dry.Column chromatography obtains yellow solid compound and directly removes Boc generation compound in the trifluoracetic acid/dichloromethane of 20% S11.Greenish yellow solid, productivity: 78%.1H NMR(300MHz,CDCl3): δ 8.99 (d, J=2.4Hz, 1H), 8.58 (d, J= 8.4Hz, 1H), 8.20 (d, J=8.1Hz, 2H), 7.52 (dd, J=2.4,8.4Hz, 1H), 7.35 (d, J=7.8Hz, 1H), 2.97 (t, J=12.3Hz, 1H), 2.66 (m, 2H), 2.05 (d, J=10.5Hz, 2H), 1.83 (d, J=11.4Hz, 2H), 1.35-1.48(m,2H)；13C NMR(100MHz,CDCl3):δ154.0 146.0 137.8 135.1 129.8 128.6 122.149.4 34.6 27.6 21.7；MS-ESI m/z:518(M+H)+；C22H22BrFN5O3S·1/4CF3COOH value of calculation: C49.59, H 4.12, N 12.85, measured value: C 49.48, H 4.10, N 12.84.
Preparation embodiment 20 1-((1r, 4r)-4-aminocyclohexane-)-3-(3,4-dichloro-benzenes sulfonyloxy)-5-Br-1- H-imidazo [4,5-h] 1,6-benzodiazine-2 (3H)-one (S12)
Except paratoluensulfonyl chloride being replaced with 3, outside the substituted benzene sulfonyl chloride of 4-dichloro, the preparation side of compound S12 Method is identical with the preparation method of compound S11.Yellow solid, productivity: 45%.1H NMR(300MHz,CDCl3):δ9.01(d,J =4.2Hz, 1H), 8.61 (d, J=8.7Hz, 1H), 8.46 (d, J=2.1Hz, 1H), 8.18 (d, J=8.4Hz, 1H), 7.65 (d, J=8.7Hz, 1H), 7.55 (dd, J=4.2,8.7Hz, 1H), 2.94 (m, 1H), 2.64 (m, 2H), 2.03 (d, J= 12.3Hz,2H),1.88(m,2H),1.33-1.45(m,2H)；MS-ESI m/z:572(M+H)+；C21H18BrCl2N5O3S·2/ H2O·1/10CF3COOH value of calculation: C 43.17, H 3.23, N 11.87, measured value: C 43.43, H 3.52, N 11.61.
Preparation embodiment 21 5-Br-1-cyclohexyl-3-(3,4-dichloro benzyl)-1H-imidazo [4,5-h] [1,6]-two Azanaphthalene-2 (3H)-one (S13)
Compound 8, triethylamine and cyclohexylamine are heated to reflux in oxolane 8 hours, are spin-dried for THF, add dichloromethane Alkane, uses saturated Na successively2CO3Aqueous solution, saturated aqueous ammonium chloride, saturated common salt are washed, anhydrous Na2SO4It is dried, reduces pressure dense Contracting, the compound that column chromatography obtains is directly used in the next step.
By compound obtained in the previous step in 1N NaOH solution/THF, under 60 degree, 10h hydrolyzes, and the compound prepared is straight Connect for the next step.
Compound obtained in the previous step, triethylamine and DPPA are heated to reflux in toluene 8 hours, are spin-dried for solvent, add Dichloromethane, uses saturated Na successively2CO3Aqueous solution, saturated aqueous ammonium chloride, saturated common salt are washed, anhydrous Na2SO4It is dried, Concentrating under reduced pressure, the compound that column chromatography obtains is directly used in the next step.
By compound obtained in the previous step, Anhydrous potassium carbonate and 3, the substituted bromobenzyl of 4-dichloro room temperature reaction 12 in THF Hour, it is spin-dried for solvent, adds dichloromethane, use saturated Na successively2CO3Aqueous solution, saturated aqueous ammonium chloride, saturated aqueous common salt Wash, anhydrous Na2SO4It is dried.Column chromatography obtains yellow solid compound S13.Productivity 82.3%:1H NMR(300MHz,CDCl3): δ 8.98 (d, J=4.2Hz, 1H), 8.54 (d, J=8.7Hz, 1H), 7.62 (s, 1H), 7.42 (dd, J=4.2,8.7Hz, 1H), 7.38 (d, J=0.9Hz, 2H), 5.17 (s, 2H), 2.64 (m, 1H), 1.89 (m, 4H), 1.71 (m, 2H), 1.46 (m, 4H)；13C NMR(100MHz,CDCl3):δ153.7,152.5,139.0,138.6,137.8,136.6,134.5,132.6,131.9, 130.6,130.6,128.1,120.6,42.7,31.2,29.9,26.1,25.2；MS-EI m/z 505(M)+；HR-EI MSC22H19BrCl2N4O(M)+Value of calculation: 505.2225, measured value: 505.2221.
Preparation embodiment 22 2-((5-Br-1-cyclohexyl-2-ketone-1H-imidazo [4,5-h] [1,6]-benzodiazine-3 (2H)-methyl) phenethyl cyanogen (S14)
Except by 3, the substituted bromobenzyl of 4-dichloro replaces with outside 2-cyano-benzyl bromide, the preparation method of compound S14 and change The preparation method of compound S13 is identical.Yellow solid, productivity 93%,1H NMR(CDCl3) 9.011 (d, J=4.2,1H), 8.573 (d, J=8.7,1H), 7.698 (d, J=7.5,1H), 7.468 (m, 2H), 7.365 (m, 2H), 5.497 (s, 2H), 2.628 (m, 1H),1.898(m,4H),1.721(m,2H),1.433(m,4H).13C NMR(CDCl3)153.72,152.62,139.87, 139.10,138.61,137.89,134.70,133.08,133.02,129.71,127.99,120.77,120.69,117.13, 111.76,55.55,41.82,29.91,26.09,25.13；MSEI m/z 461[M+1]+.
Preparation embodiment 23 5-Br-1-cyclohexyl-3-(4-luorobenzyl)-1H-imidazo [4,5-h] [1,6]-diaza Naphthalene-2 (3H)-one (S15)
Except by 3, the substituted bromobenzyl of 4-dichloro replaces with outside 4-cyanogen bromobenzyl, the preparation method of compound S15 and chemical combination The preparation method of thing S13 is identical.Yellow solid, productivity 89%,1H NMR(CDCl3) 8.979 (d, J=3.9,1H), 8.556 (d, J=8.7,1H), 7.553 (d, J=6.6,1H), 7.411 (dd, J=3.9,8.7Hz, 1H), 7.006 (d, J=6.6Hz, 2H), 5.204(s,2H),2.649(m,1H),1.886(m,4H),1.587(m,2H),1.431(m,4H).13C NMR(CDCl3) 163.57,161.13,153.58,152.65,139.23,138.49,137.81,135.58,134.35,132.40,132.37, 130.66,130.58,120.47,115.55,115.33,55.37,43.04,29.92,26.11,25.17；MSEI m/z454 [M+1]+.
Preparation embodiment 24 1-benzyl-5-Br-3-(3,4-dichloro benzyl)-1H-imidazo [4,5-h] [1,6]-phenodiazine Miscellaneous naphthalene-2 (3H)-one (S16)
In addition to cyclohexylamine is replaced with benzylamine, the preparation method phase of the preparation method of compound S16 and compound S13 With.Yellow solid, productivity 72.3%:1H NMR(300MHz,CDCl3): δ 8.98 (d, J=4.2Hz, 1H), 8.52 (d, J= 8.7Hz, 1H), 7.56 (m, 3H), 7.42 (dd, J=4.2,8.7Hz, 1H), 7.37 (m, 2H), 5.78 (s, 2H), 5.21 (s, 2H)；13C NMR(100MHz,CDCl3):δ154.1,153.1,138.7,138.6,137.7,137.4,136.4,135.5, 134.7,132.6,132.1,130.6,130.5,128.4,128.0,127.7,121.0,120.7,115.5,46.3,42.9； MS-EI m/z 513(M)+；HR-EI MS C23H15BrCl2N4O(M)+Value of calculation: 513.2015, measured value: 513.2010.
Preparation embodiment 25 (S)-5-Br-3-(3,4-dichloro benzyl)-1-(1-phenethyl)-1H-imidazo [4,5-h] [1,6]-benzodiazine-2 (3H)-one (S17)
In addition to cyclohexylamine is replaced with (S)-1-phenethylamine, the preparation method of compound S17 and the system of compound S13 Preparation Method is identical.Yellow solid, productivity 84%,1H NMR(CDCl3) 8.958 (d, J=3.9,1H), 8.557 (d, J=8.4, 1H), 7.584 (m, 3H), 7.425 (dd, J=3.9,8.4,1H), 7.368 (m, 2H), 7.132 (m, 1H), 5.183 (s, 2H), 2.142 (d, J=7.2,3H)；MSEI m/z 528[M+1]+.
Preparation embodiment 26 (R)-5-Br-3-(3,4-dichloro benzyl)-1-(1-phenethyl)-1H-imidazo [4,5-h] [1,6]-benzodiazine-2 (3H)-one (S18)
In addition to cyclohexylamine is replaced with (R)-1-phenethylamine, the preparation method of compound S18 and the system of compound S13 Preparation Method is identical.Yellow solid, productivity 86%,1H NMR(CDCl3) 8.958 (d, J=4.2,1H), 8.554 (d, J=8.7, 1H), 7.584 (m, 3H), 7.423 (dd, J=4.2,8.7,1H), 7.367 (m, 2H), 7.156 (m, 1H), 5.168 (s, 2H), 2.144 (d, J=7.2,3H)；MSEI m/z 528[M+1]+.
Preparation embodiment 27 5-Br-1-cyclopropyl-3-(3,4-dichloro benzyl)-1H-imidazo [4,5-h] [1,6]-two Azanaphthalene-2 (3H)-one (S19)
In addition to cyclohexylamine is replaced with cyclopropylamine, the preparation method of compound S19 and the preparation method of compound S13 Identical.Yellow solid, productivity 86.3%:1H NMR(300MHz,CDCl3): δ 9.02 (d, J=4.2Hz, 1H), 8.57 (d, J= 8.4Hz, 1H), 7.64 (s, 1H), 7.45 (dd, J=4.2,8.4Hz, 1H), 7.37 (d, J=1.2Hz, 2H), 5.16 (s, 2H), 3.49(m,1H),1.56(m,2H),1.25(m,2H)；MS-EI m/z:464(M)+；HR-EI MSC19H13BrCl2N4O(M)+Meter Calculation value: 463.9645, measured value: 463.9640.
Preparation embodiment 28 N-((3r, 5r, 7r)-diamantane (obsolete)-1-methyl)-8-(((3r, 5r, 7r)-diamantane (obsolete)-1-first Base) amino)-5-Br-1,6-benzodiazine-7-Methanamide (S20)
Except cyclohexylamine is replaced with Adamantanemethylamine, 3,4-dichloro bromobenzyls replace with outside diamantane (obsolete) methylene bromide, The preparation method of compound S20 is identical with the preparation method of compound S13.Yellow solid, productivity 76%,1H NMR(CDCl3) 9.016 (d, J=4.2,1H), 8.453 (d, J=8.4,1H), 8.123 (t, J=6.6,1H), 7.621 (dd, J=4.2,8.4, 1H), 3.849 (s, 2H), 3.167 (d, J=6.6,2H), 2.007 (m, 6H), 1.684 (m, 24H)；MSEI m/z 562[M+1 ]+.
Preparation embodiment 29 1-cyclohexyl-3-(3,4-dichloro benzyl)-5-(pyridine-3-epoxide)-1H-imidazo [4,5- H] [1,6]-benzodiazine-2 (3H)-one (S21)
By compound S13, Anhydrous potassium carbonate and 3-pyridone in DMF 110 DEG C react 6 hours, be spin-dried for solvent, add Dichloromethane, uses saturated Na successively2CO3Aqueous solution, saturated aqueous ammonium chloride, saturated common salt are washed, anhydrous Na2SO4It is dried. Column chromatography obtains yellow solid compound S21.Productivity 93%,1H NMR(CDCl3) 9.025 (d, J=4.2,1H), 8.696 (d, J =8.7,1H), 8.633 (d, J=8.1,2H), 7.609 (m, 1H), 7.525 (s, 1H), 7.504 (dd, J=4.2,8.4,1H), 7.392 (dd, J=4.2,8.7,1H), 7.334 (d, J=4.5,1H), 7.118 (d, J=8.1,1H), 4.899 (s, 2H), 2.667(m,1H),1.894(m,4H),1.758(m,2H),1.447(m,4H).13C NMR(CDCl3)154.42,154.04, 152.58,150.72,144.46,142.23,140.15,136.79,136.14,133.26,132.26,131.79,130.92, 130.49,128.26,126.75,124.65,119.23,110.48,55.11,42.62,30.01,26.11,25.15；MSEI m/z519[M+1]+.
Preparation embodiment 30 4-((1-cyclohexyl-3-(3,4-dichloro benzyl)-2-ketone-2,3-dihydro-1H-imidazo [4, 5-h] [1,6] benzodiazine-5-epoxide) benzaldehyde (S22)
In addition to 3-pyridone is replaced with 4-hydroxy benzaldehyde, the preparation method of compound S22 and compound S21 Preparation method identical.Yellow solid, productivity 92%,1H NMR(CDCl3) 10.059 (s, 1H), 9.052 (d, J=4.2,1H), 8.6960 (d, J=8.4,1H), 8.015 (d, J=8.7,2H), 7.874 (d, J=8.4,2H), 7.380 (dd, J=4.2, 8.4,1H), 7.346 (d, J=1.8,2H), 4.919 (s, 2H), 2.670 (m, 1H), 1.899 (m, 4H), 1.720 (m, 2H), 1.571(m,4H)；MSEI m/z 546[M+1]+.
Preparation embodiment 31 1-cyclohexyl-3-(3,4-dichloro benzyl)-5-(4-(methylol) benzene oxo)-1H-imidazo [4,5-h] [1,6] benzodiazine-2 (3H)-one (S23)
Compound S22 50mg is dissolved in dry THF, adds the NaBH of 4eq4, under room temperature, stirring reaction 6h, adds ice Water terminates reaction, rear adds EA extraction, and organic layer is with saturated NH4Cl solution washs, and saturated NaCl washs, anhydrous Na2SO4Dry Dry, concentrating under reduced pressure, column chromatography obtains target compound S23, yellow solid, productivity 96%,1H NMR(CDCl3) 9.021 (d, J= 4.2,1H), 8.729 (d, J=8.4,1H), 7.504 (d, J=8.4,2H), 7.360 (m, 2H), 7.286 (d, J=7.8,2H), 7.196 (d, J=8.1,2H), 4.880 (s, 2H), 4.798 (s, 2H), 2.625 (m, 1H), 1.885 (m, 4H), 1.752 (m, 2H),1.496(m,4H)；MSEI m/z 548[M+1]+.
Preparation embodiment 32 1-((1r, 4r)-4-aminocyclohexyl)-3-(3,4-dichloro benzyl)-5-hydroxyl-1H-imidazoles And [4,5-h] [1,6]-benzodiazine-2 (3H)-one (S24)
By compound S10, potassium carbonate in DMF 110 DEG C react 6 hours, be spin-dried for solvent, add dichloromethane, use successively Saturated Na2CO3Aqueous solution, saturated aqueous ammonium chloride, saturated common salt are washed, anhydrous Na2SO4It is dried.It is solid that column chromatography obtains yellow Body compound, after in the trifluoracetic acid/dichloromethane of 20% remove Boc generate compound S24.Yellow solid, productivity: 56.7%.1H NMR(300MHz,CDCl3): δ 8.94 (d, J=2.5Hz, 1H), 8.52 (d, J=8.8Hz, 1H), 7.52 (d, J =1.9Hz, 1H), 7.41 (dd, J=8.7,4.2Hz, 1H), 7.33 7.26 (m, 2H), 5.09 (s, 2H), 3.31 3.17 (m, 1H),2.86–2.62(m,2H),2.29–2.01(m,2H),1.95–1.79(m,2H),1.72–1.47(m,2H)；MS-EI m/ z:457(M)+；HR-EI MS C22H21Cl2N5O2(M)+Value of calculation: 457.1072, measured value: 457.1040.
Preparation embodiment 33 1-((1r, 4r)-4-aminocyclohexyl)-3-(3,4-dichloro benzyl)-5-(3-hydroxyl pyrrole Pyridine)-1H-imidazo [4,5-h] [1,6]-benzodiazine-2 (3H)-one (S25)
By compound S10,3-pyridone, Anhydrous potassium carbonate in DMF 110 DEG C react 6 hours, be spin-dried for solvent, add Dichloromethane, uses saturated Na successively2CO3Aqueous solution, saturated aqueous ammonium chloride, saturated common salt are washed, anhydrous Na2SO4It is dried. Column chromatography obtains yellow solid compound (productivity: 56.7%.), in the trifluoracetic acid/dichloromethane of 20%, then remove Boc Generate compound S25.Yellow solid, productivity 77.6%,1H NMR(300MHz,CDCl3): δ 9.07 (d, J=4.2Hz, 1H), 8.82 (d, J=8.6Hz, 1H), 8.57 (d, J=2.5Hz, 1H), 8.53 (d, J=4.8Hz, 1H), 7.78 (d, J=7.0Hz, 1H), 7.63 7.51 (m, 2H), 7.38 (d, J=8.3Hz, 1H), 7.35 (d, J=1.9Hz, 1H), 7.07 (d, J=8.3Hz, 1H),4.91(s,2H),2.96–2.80(m,2H),2.31–2.18(m,2H),2.07–1.96(m,2H),1.78–1.58(m, 2H)；MS-EI m/z:534(M)+；HR-EI MS C27H24Cl2N6O2(M)+Value of calculation: 534.1342, measured value: 534.1356.
Preparation embodiment 34 4-((1-((1r, 4r)-4-aminocyclohexyl)-3-(3,4-dichloro benzyl)-2-oxo-2, 3-dihydro-1H-imidazo [4,5-h] [1,6]-benzodiazine-5-oxo) benzaldehyde (S26)
In addition to 3-pyridone is replaced with 4-hydroxy benzaldehyde, the preparation method of compound S26 and compound S25 Preparation method identical.Yellow solid, productivity 63.4%,1H NMR(300MHz,CDCl3):δ10.04(s,1H),9.07(d,J =4.3Hz, 1H), 8.80 (d, J=8.7Hz, 1H), 8.05 (d, J=8.4Hz, 2H), 7.90 (s, 1H), 7.57 7.53 (m, 1H), 7.42 (d, J=8.8Hz, 2H), 7.37 (s, 1H), 7.13 (d, J=11.3Hz, 1H), 4.94 (s, 2H), 3.00 2.80 (m,2H),2.30–2.16(m,2H),2.12–1.96(m,2H),1.77–1.59(m,2H)；MS-EI m/z:561(M)+；HR- EI MS C29H25Cl2N5O3(M)+Value of calculation: 561.2353, measured value: 561.2359.
Preparation embodiment 35 1-((1r, 4r)-4-aminocyclohexyl)-3-(3,4-dichloro benzyl)-5-(4-(methylol) Phenoxy group)-1H-imidazo [4,5-h] [1,6]-benzodiazine-2 (3H)-one (S27)
Compound S26 50mg is dissolved in dry THF, adds the NaBH of 4eq4, under room temperature, stirring reaction 6h, adds ice Water terminates reaction, rear adds EA extraction, and organic layer is with saturated NH4Cl solution washs, and saturated NaCl washs, anhydrous Na2SO4Dry Dry, concentrating under reduced pressure, after in the trifluoracetic acid/dichloromethane of 20% remove Boc generate compound S27.It is solid that column chromatography obtains yellow Body, productivity: 72.6%.1H NMR(300MHz,CDCl3): δ 9.03 (d, J=4.0Hz, 1H), 8.78 (d, J=8.5Hz, 1H), 7.86 (s, 1H), 7.50 (d, J=7.8Hz, 3H), 7.42 7.35 (m, 1H), 7.21 (d, J=8.1Hz, 2H), 7.12 (d, J= 8.2Hz,1H),4.88(s,2H),4.70(s,2H),3.03–2.94(m,1H),2.84–2.75(m,1H),2.11–2.04(m, 2H),1.93–1.88(m,2H),1.55–1.36(m,4H)；MS-EI m/z:563(M)+；HR-EIMS C29H27Cl2N5O3(M)+ Value of calculation: 563.1491, measured value: 563.1495.
Preparation embodiment 36 4-((1-((1r, 4r)-4-aminocyclohexyl)-3-(3,4-dichloro benzyl)-2-oxo-2, 3-dihydro-1H-imidazo [4,5-h] [1,6]-benzodiazine-5-oxo)-2-chlorobenzaldehyde (S28)
In addition to 3-pyridone is replaced with 2-chlorine 4-hydroxy benzaldehyde, the preparation method of compound S28 and chemical combination The preparation method of thing S25 is identical.Yellow solid, productivity 65.5%:1H NMR(300MHz,CDCl3):δ10.04(s,1H), 9.12 9.03 (m, 1H), 8.74 (d, J=8.5Hz, 1H), 8.15 8.05 (m, 1H), 7.24 7.20 (m, 1H), 7.92 (d, J =8.4Hz, 1H), 7.43 (d, J=8.3Hz, 2H), 7.30 7.27 (m, 1H), 7.14 7.03 (m, 1H), 4.86 (s, 2H), 4.57–4.34(m,2H),3.90–3.60(m,2H),3.02–2.85(m,2H),2.26–2.06(m,2H),1.43–1.37(m, 2H)；MS-EI m/z:595(M)+；HR-EI MS C29H24Cl3N5O3(M)+Value of calculation: 595.1047, measured value: 595.1042.
Preparation embodiment 37 1-((1r, 4r)-4-aminocyclohexyl)-3-(3,4-dichloro benzyl)-5-(4-(piperidines-1- Carbonyl) benzene oxo)-1H-imidazo [4,5-h] [1,6]-benzodiazine-2 (3H)-one (S29)
The synthesis of ether: be dissolved in dry DMF by the S10 of 0.15mmol, adds the methyl parahydroxybenzoate of 1.5eq, after Add 5eq and be now baked the anhydrous K milled2CO3, 110 DEG C of stirring reactions, TLC follows the tracks of, and adds EA and rinse, with saturated after having reacted NH4Cl solution washs, and saturated NaCl washs, anhydrous Na2SO4Being dried, concentrating under reduced pressure, the compound that column chromatography obtains is directly used in The next step.
Saponification: be dissolved in methanol aqueous solution by the compound obtained in the previous step of 1eq, the rear NaOH adding 4eq, under room temperature Stirring reaction, TLC follows the tracks of reaction, adds EA and rinse, with saturated NH after having reacted4Cl solution washs, and saturated NaCl washs, Anhydrous Na2SO4Being dried, concentrating under reduced pressure, the compound being recrystallized to give is directly used in the next step.
The synthesis of amide: the compound obtained in the previous step of 1eq is dissolved in dry THF solution, rear addition 1.5eq's Piperidines, adds the triethylamine of 3-5eq, the Bop of 2eq, stirring reaction under room temperature, and TLC follows the tracks of, and adds EA and rinse after having reacted, With saturated NH4Cl solution washs, and saturated NaCl washs, anhydrous Na2SO4It is dried, concentrating under reduced pressure, the compound that column chromatography obtains It is directly used in the next step.
The protectant removing of Boc: the compound that Boc protects is dissolved in DCM, adds TFA afterwards, be allowed to the dense of last TFA Degree is about about 30%, and stirring reaction under room temperature, TLC follows the tracks of, reacted rear concentrating under reduced pressure, and column chromatography obtains compound S29.
1H NMR(300MHz,CDCl3): δ 9.12 (d, J=5.5Hz, 1H), 8.87 (d, J=8.6Hz, 1H), 7.60 (d, J =8.5Hz, 3H), 7.47 (d, J=5.8Hz, 2H), 7.39 (d, J=8.0Hz, 2H), 7.20 (d, J=9.5Hz, 1H), 4.98 (s,2H),3.80–3.75(m,1H),3.56–3.46(m,1H),3.03–2.82(m,3H),2.31–2.22(m,2H),2.11– 2.00(m,2H),1.84–1.68(m,6H),1.68–1.50(m,4H),1.40–1.26(m,4H)；MS-EI m/z:644(M)+； HR-EI MS C34H34Cl2N6O3(M)+Value of calculation: 644.2069, measured value: 644.2072.
Preparation embodiment 38 1-((1r, 4r)-4-aminocyclohexyl)-3-(3,4-dichloro benzyl)-5-(4-(morpholine- 4-carbonyl) phenoxy group)-1H-imidazo [4,5-h] [1,6]-benzodiazine-2 (3H)-one (S30)
In addition to piperidines is replaced morpholine, the preparation method of compound S30 is identical with the preparation method of compound S29. Yellow solid, productivity 92%,1H NMR(300MHz,CDCl3): δ 8.96 (d, J=5.7Hz, 1H), 8.61 (d, J=8.2Hz, 1H), 7.64 (s, 1H), 7.56 7.49 (m, 1H), 7.45 (d, J=8.3Hz, 1H), 7.40 7.35 (m, 1H), 5.15 (s, 2H), 4.63 (t, J=6.2Hz, 2H), 3.78 (t, J=6.3Hz, 2H), 3.69 3.50 (m, 4H), 3.18 (dd, J=14.6, 7.3Hz,4H),2.19(s,1H),2.12–2.05(m,2H),1.98–1.94(m,1H),1.32–1.25(m,4H),1.20– 1.11(m,4H)；MS-EI m/z:646(M)+；HR-EI MS C33H32Cl2N6O4(M)+Value of calculation: 646.1860, measured value: 646.1862.
Preparation embodiment 39 4-((1-((1r, 4r)-4-aminocyclohexyl)-3-(3,4-dichloro benzyl)-2-oxo-2, 3-dihydro-1H-imidazo [4,5-h] [1,6]-benzodiazine-5-oxo)-N-((R)-1-phenethyl) Benzoylamide (S31)
In addition to piperidines is replaced with (R)-1-phenethylamine, the preparation method of compound S31 and the preparation of compound S29 Method is identical.Yellow solid, productivity 82%,1H NMR(300MHz,CDCl3): δ 8.958 (d, J=3.9,1H), 8.557 (d, J =8.4,1H), 7.584 (m, 3H), 7.425 (dd, J=3.9,8.4,1H), 7.368 (m, 2H), 7.132 (m, 1H), 5.183 (s, 2H), 2.142 (d, J=7.2,3H)；MS-EI m/z:680(M)+；HR-EI MS C37H34Cl2N6O3(M)+Value of calculation: 680.2069, measured value: 680.2071.
Preparation embodiment 40 1-((1r, 4r)-4-aminocyclohexyl)-3-(3,4-dichloro benzyl)-5-(4-(4-methyl piperazine Piperazine-1-carbonyl) phenoxy group)-1H-imidazo [4,5-h] [1,6]-benzodiazine-2 (3H)-one (S32)
In addition to piperidines is replaced with N methyl piperazine, the preparation method of compound S32 and the preparation side of compound S29 Method is identical.Yellow solid, productivity 92%, NMR (400MHz) δ 9.07 (dd, J=4.2,1.6Hz, 1H), 8.78 (dd, J= 8.5,1.7Hz, 1H), 7.67 (d, J=8.6Hz, 2H), 7.53 (dd, J=8.5,4.3Hz, 1H), 7.43 (d, J=4.9Hz, 1H), 7.41 (s, 1H), 7.41 7.38 (m, 2H), 7.14 (dd, J=8.3,2.0Hz, 1H), 5.60 5.31 (m, 1H), 4.93 (s, 2H), 4.17 3.73 (m, 1H), 3.42 3.34 (m, 4H), 2.96 (s, 3H), 2.94 2.76 (m, 2H), 2.25 (d, J= 12.0Hz, 2H), 2.03 (d, J=11.2Hz, 2H), 1.79 1.63 (m, 2H) .MS-EI m/z:660 (M)+；
EXPERIMENTAL EXAMPLE 1:1,3,5-tri-replacement-1H-imidazo [4,5-h] 1,6-benzodiazine-2 (3H)-one compound Inhibitory action kinase whose to c-Met
Receptor tyrosine kinase c-Met molecular level enzyme is lived and is suppressed preliminary assessment experiment
(1) enzyme reaction substrate Poly (Glu, Tyr)4:1With without potassium ion PBS (10mM sodium phosphate buffer, 150mMNaCl, pH7.2-7.4) it is diluted to 20 μ g/mL, 125 μ L/ hole coated elisa plates, put 37 DEG C and react 12-16 hour.Discard Liquid in hole.Wash plate, wash plate three times, every time with the T-PBS (PBS without potassium ion containing 0.1%Tween-20) in 200 μ L/ holes 5 minutes.ELISA Plate 1-2 hour it is dried in 37 DEG C of baking ovens.
(2) every hole adds with reaction buffer (50mM HEPES pH 7.4,50mM MgCl2,0.5mM MnCl2,0.2mM Na3VO4, 1mM DTT) and ATP (adenosine triphosphate) the solution 50 μ L that dilutes, final concentration 5 μMs.Every hole adds 1 μ L chemical combination of the present invention Thing (1%DMSO dissolves, final concentration of 10 μMs), adds the c-Met protein tyrosine kinase of 50 μ L reaction buffer dilutions. Put 37 DEG C of shaking tables (100rpm) and react 1 hour.Experiment need to set without ATP control wells holes and corresponding DMSO solvent control hole every time (negative control hole).Discarding liquid in hole, T-PBS washes plate three times.
(3) (antibody T-PBS containing BSA5mg/mL dilutes, and concentration is 0.4 μ g/ to add antibody PY99 100 μ L/ hole ML), 37 DEG C of shaking tables react 0.5 hour.Discarding liquid in hole, T-PBS washes plate three times.
(4) sheep anti mouse two adding horseradish peroxidase-labeled resists the 100 μ L/ hole (antibody T-containing BSA5mg/mL PBS dilutes, and concentration is 0.5 μ g/mL), 37 DEG C of shaking tables react 0.5 hour.Discarding liquid in hole, T-PBS washes plate three times.
(5) the OPD nitrite ion 100 μ L/ hole of 2mg/mL is added (with containing 0.03%H2O20.1M citric acid-citric acid Sodium buffer (pH=5.4) dilutes), 25 DEG C of lucifuges are reacted 1-10 minute.(need to be with ultrasonic when OPD dissolves, nitrite ion need to now be joined existing With).
(6) 2M H is added2SO450 μ L/ hole stopped reactions, decline orifice plate microplate reader VERSAmax reading with wavelengthtunable, Wavelength is 490nm.
(7) suppression ratio of sample is tried to achieve by following equation:
Result is listed in table 1.
(compound is 10 for the compound clearly with c-Met enzyme inhibitory action alive above-mentioned screening obtained-5M is to receptor The suppression ratio of tyrosine kinase c-Met > 50%) it is made into gradient concentration, carry out IC50Evaluate.Each compound is calculated by four parametric methods The IC of molecular level suppression protein tyrosine kinase50Value, result is listed in table 1.
Result: research finds that multiple the compounds of this invention have inhibitory activity in various degree, part chemical combination to c-Met kinases Thing is up to 80% to c-Met kinase inhibition rate under 10 μMs of concentration.The compound useful effect of the prompting present invention swashs in c-Met Enzyme, is the c-Met kinase inhibitor of novel structure.Detailed data is shown in Table 1.In table, space represents and does not carries out dependence test, without phase Close data.
Table 1 compound inhibition kinase whose to c-Met
From the experimental result of table 1 it can be seen that the compound useful effect of the present invention is in c-Met kinases, it it is novel structure C-Met kinase inhibitor.
EXPERIMENTAL EXAMPLE 2:1,3,5-tri-replacement-1H-imidazo [4,5-h] 1,6-benzodiazine-2 (3H)-one compound Impact on the cell strain multiplication capacity of c-Met mediation
Inoculation is in the BaF3/TPR-Met cell (8000/ hole) in 96 well culture plates of exponential phase, every hole 100 μ L, is separately added into the compounds of this invention of 10 μ L variable concentrations after overnight incubation, arrange 3 concentration, each multiple hole of concentration 3.Chemical combination After thing effect 72 hours, every hole adds 20 μ L MTT (5mg/mL), adds 100 μ L tri-liquid (10% after 37 DEG C of cultivation 4hr SDS-5% isobutanol-0.01M HCl), 37 DEG C are overnight, measure OD value under 570nm wavelength.Medicine is calculated by with following equation Suppression ratio to growth of tumour cell: suppression ratio (%)=(OD control wells-OD dosing holes)/OD control wells × 100%.Experiment weight Multiple twice.
Result: multiple compounds of the present invention have obvious inhibitory action to BaF3-TPR-Met cell proliferation, show this Compound can suppress by the cell-proliferation activity of c-Met activation mediation.Concrete data are shown in Table 2.
The suppression ratio % of table 2 compound on tumor cell proliferation
Multiple compounds of the present invention have obvious inhibitory action to BaF3-TPR-Met cell proliferation, show this compound Can suppress by the cell-proliferation activity of c-Met activation mediation.
EXPERIMENTAL EXAMPLE 3: the compound S9 inhibitory action to multiple protein kinases
Experimentation is shown in Millpore`s Kinaseprofile assay services.www.millipore.com
Concrete data are shown in Fig. 1.Result: the compound S9 of the present invention show to c-Met, FGFR, Abl, Lck, KDR, The kinase whose inhibitory activity such as IGF-1 α, ALK, show that this compounds is the multiple inhibitors of kinases of a class new construction.
1H-imidazo [4,5-h] 1,6-benzodiazine-2 (3H) the-one compound of the present invention show to c-Met, The kinase whose inhibitory activity such as FGFR, Abl, Lck, KDR, IGF-1 α, ALK, show that this compounds is the multiple of a class new construction Inhibitors of kinases.

Claims (9)

1. one kind by 1,3,5-tri-replacement-1H-imidazo [4,5-h] 1,6-benzodiazine-2 (3H)-one shown in following formula I Compound and pharmaceutically acceptable salt thereof,
Wherein,
R1For C3-C6Cycloalkyl, amino C3-C6Cycloalkyl, C1-C4Alkoxycarbonyl amido C3-C6Cycloalkyl, phenyl C1-C3Alkyl or Adamantyl C1-C3Alkyl；
R3For F, Cl, Br, hydroxyl, pyridine radicals epoxide or substituted or unsubstituted phenyl epoxide, wherein, described substituent group has 1-2 Individual and be each independently Cl, carboxaldehyde radicals, C1-C3Hydroxyalkyl, 1-hexahydropyridine base carbonyl, N-morpholinyl carbonyl, (R)-1-benzene second Base amino carbonyl or 4-methylpiperazine-1-yl-carbonyl；
R2For hydrogen,Wherein,
L is C1-C6Alkyl or-SO2-；
R4And R5It is each independently hydrogen, halogen ,-CN ,-CF3、C1-C3Alkyl, C1-C6Alkoxyl or the substituted C of fluorine1-C6Alcoxyl Base.
Compound the most according to claim 1 and pharmaceutically acceptable salt thereof, wherein,
R1For C3-C6Cycloalkyl, amino C3-C6Cycloalkyl, tertbutyloxycarbonylamino C3-C6Cycloalkyl,Benzyl or 1-phenylethyl；
R3For Br, hydroxyl, 3-pyridine radicals epoxide, 2-pyridine radicals epoxide or substituted or unsubstituted phenyl epoxide, wherein, described Substituent group has 1-2 and is each independently Cl, carboxaldehyde radicals, C1-C3Hydroxyalkyl, 1-hexahydropyridine base carbonyl, N-morpholinyl carbonyl Base, (R)-1-PhenethyIamino carbonyl or 4-methylpiperazine-1-yl-carbonyl；
R2For hydrogen,Wherein,
L is C1-C3Alkyl or-SO2-；
R4And R5It is each independently hydrogen, F, Cl ,-CN ,-CF3, methyl, methoxyl group or-OCF3。
Compound the most according to claim 2 and pharmaceutically acceptable salt thereof, it is following compound:
4. one kind comprise therapeutically effective amount selected from compound according to any one of claim 1 to 3 and pharmaceutically The compositions of one or more in acceptable salt.
Compound the most according to any one of claim 1 to 3, its pharmaceutically acceptable salt and comprise its compositions In preparation as the application in the medicine of Mutiple Targets kinases inhibitor.
Application the most according to claim 5, wherein, described kinases includes c-Met, FGFR, Abl, Lck, KDR, IGF-1 α And ALK.
7. according to the application described in claim 5 or 6, wherein, described medicine is relevant with protein kinase for treatment and/or prevention Disease.
Application the most according to claim 7, wherein, described disease is tumor.
9. the preparation method of compound a S1-S20, S22-S23 and S26-S27 as claimed in claim 3, the method as with Shown in lower reaction scheme:
Wherein, R1、R2、R3Definition compound corresponding with claim 3 respectively described in identical；
Reaction reagent and condition: (a) isopropanol, backflow；(b) thionyl chloride, backflow；(c) sodium borohydride, oxolane, 0 DEG C； (d)TsNHCH2COOCH3, DEAD, triphenylphosphine, oxolane, 0 DEG C；(e) Feldalat NM, methanol, 0 DEG C～room temperature；(f) NBS, two Chloromethanes, room temperature；(g) TsCl, triethylamine, dichloromethane；(h)R1NH2, oxolane, backflow；(i) LiOH solution/methanol, or NaOH solution/THF, backflow；(j) DPPA, triethylamine, toluene, backflow；(k) trifluoracetic acid/dichloromethane, room temperature；(l)R2X, K2CO3, DMF, room temperature；X is Cl or Br；(m) substituted phenol, described substituent group have 1-2 and be each independently carboxaldehyde radicals or C1Hydroxyalkyl, DMF, 110 DEG C；(n) substituted benzene sulfonyl chloride, described substituent group has 1-2 and is each independently Cl or methyl, K2CO3, THF, backflow；(o) sodium borohydride, THF, room temperature；
The method specifically includes following steps:
(1) compound 1 pyridine 2,3-dicarboxylic anhydride selective esterification under condition a is obtained compound 2；Compound 2 is under condition b Chloro obtains compound 3；Compound 3 obtains compound 4 with the selective reduction under condition c of acyl chlorides form；Compound 4 is in condition Compound 5 is obtained by Mitsunobo reaction under d；Compound 5 cyclization under condition e obtains 1,6-benzodiazine carbonyl acid methyl ester Compound 6；Compound 6 and N-bromo-succinimide react under condition f and obtain 5-Br-1,6-benzodiazine carbonyl acid esterification Compound 7；8 hydroxyls of compound 7 obtain compound 8 with Ts protection under condition g；Compound 8 under condition h with R1NH2Occur Nucleophilic substitution obtains 8 bit amino substituted compounds 9；Compound 9 is hydrolyzed to compound 10 under alkalescence condition i further； Compound 10 occurs Cutis to reset in the presence of condition j DPPA, then occurs molecule nucleophilic substitution to generate 1H-imidazo [4,5-h] 1,6-benzodiazine-2 (3H)-one compound 11；With
(2)R1For end Boc protection aminocyclohexyl time compound 11 direct Deprotection under condition k can get chemical combination Thing S1；Compound S10, S13-in compound 11 formula I as described in obtain 3 corresponding substituent groups of introducing under the conditions of l S20；And/or
(3)R1For end Boc protection aminocyclohexyl time compound 11 under condition l with R2X reaction obtains compound 12； Compound 12 removes Boc under condition k and generates 1,3,5-tri-replacement-1H-imidazo [4,5-h] the 1,6-phenodiazine of described formula I Miscellaneous naphthalene-2 (3H)-one compound S2-S9；And/or
(4)R1For end Boc protection aminocyclohexyl time compound 11 be heated to reflux under condition n 8 hours with substituted benzene Sulfonic acid chloride reaction obtains substituted 3-benzenesulfonyl substituted-1H-imidazo [4,5-h] 1,6-benzodiazine-2 (3H)-one Compound, sloughs Boc through 20% trifluoracetic acid/dichloromethane under condition k and obtains described compounds of formula I S11 or S12； And/or
(5) 5 bromine substituted 1H-imidazo [4,5-h] 1,6-benzodiazine-2 (3H)-one compounds in the basic conditions with take The phenol reactant in generation i.e. can get described compound S22, is reduced further by compound S22 under condition o in sodium borohydride Obtain described compounds of formula I S23；And/or
(6) compound 12 obtains 1,3,5-tri-replacement-1H-imidazo [4,5-h] in the basic conditions with substituted phenol reactant 1,6-benzodiazine-2 (3H)-one compound 13, compound 13 is sloughed Boc through 20% trifluoracetic acid/dichloromethane and is obtained described Compounds of formula I S26-S27.
CN201210378551.2A 2012-09-29 2012-09-29 Tri-substituted imidazole benzodiazine ketonic compound and its production and use Active CN103709162B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201210378551.2A CN103709162B (en) 2012-09-29 2012-09-29 Tri-substituted imidazole benzodiazine ketonic compound and its production and use

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201210378551.2A CN103709162B (en) 2012-09-29 2012-09-29 Tri-substituted imidazole benzodiazine ketonic compound and its production and use

Publications (2)

Publication Number Publication Date
CN103709162A CN103709162A (en) 2014-04-09
CN103709162B true CN103709162B (en) 2016-12-07

Family

ID=50402551

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201210378551.2A Active CN103709162B (en) 2012-09-29 2012-09-29 Tri-substituted imidazole benzodiazine ketonic compound and its production and use

Country Status (1)

Country Link
CN (1) CN103709162B (en)

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6211376B1 (en) * 1996-09-10 2001-04-03 Pharmacia & Upjohn Company 8-hydroxy-7-substituted quinolines as anti-viral agents
CN1469878A (en) * 2000-10-12 2004-01-21 Aza-and polyaza-naphthalenyl carboxamides useful as HIV integrase inhibitors
US6809097B1 (en) * 1996-09-25 2004-10-26 Zeneca Limited Quinoline derivatives inhibiting the effect of growth factors such as VEGF
CN101684119A (en) * 2008-09-27 2010-03-31 中国科学院上海药物研究所 5,8-disubstituted-1,6-quinazoline-7-amidocarbonylation compound, preparing method, composite and application thereof
CN102372711A (en) * 2010-08-18 2012-03-14 山东轩竹医药科技有限公司 Imidazo quinoline PI3K and mTOR (mammalian target of rapamycin) dual inhibitor
CN102558172A (en) * 2010-12-30 2012-07-11 中国科学院上海药物研究所 5,8-disubstituted-1,6-naphthyridine-7-carbonyl amide compounds, dimer compounds of 5,8-disubstituted-1,6-naphthyridine-7-carbonyl amide compounds, and preparation method and use of 5,8-disubstituted-1,6-naphthyridine-7-carbonyl amide compounds and dimer compounds of 5,8-disubstituted-1,6-naphthyridine-7-carbonyl amide compounds

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6211376B1 (en) * 1996-09-10 2001-04-03 Pharmacia & Upjohn Company 8-hydroxy-7-substituted quinolines as anti-viral agents
US6809097B1 (en) * 1996-09-25 2004-10-26 Zeneca Limited Quinoline derivatives inhibiting the effect of growth factors such as VEGF
CN1469878A (en) * 2000-10-12 2004-01-21 Aza-and polyaza-naphthalenyl carboxamides useful as HIV integrase inhibitors
CN101684119A (en) * 2008-09-27 2010-03-31 中国科学院上海药物研究所 5,8-disubstituted-1,6-quinazoline-7-amidocarbonylation compound, preparing method, composite and application thereof
CN102372711A (en) * 2010-08-18 2012-03-14 山东轩竹医药科技有限公司 Imidazo quinoline PI3K and mTOR (mammalian target of rapamycin) dual inhibitor
CN102558172A (en) * 2010-12-30 2012-07-11 中国科学院上海药物研究所 5,8-disubstituted-1,6-naphthyridine-7-carbonyl amide compounds, dimer compounds of 5,8-disubstituted-1,6-naphthyridine-7-carbonyl amide compounds, and preparation method and use of 5,8-disubstituted-1,6-naphthyridine-7-carbonyl amide compounds and dimer compounds of 5,8-disubstituted-1,6-naphthyridine-7-carbonyl amide compounds

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
Synthesis and c-Met kinase inhibition of 3,5-disubstituted and 3,5,7-trisubstituted quinolines: identification of 3-(4-acetylpiperazine-1-yl)-5-(3-nitrobenzylamino)-7-(trifluoromethyl)quinoline as a novel anticancer agent;Wang YX et al.;《Journal of Medicinal Chemistry》;20110315;第54卷(第7期);第2127-2142页 *
蛋白酪氨酸激酶小分子抑制剂的研究新进展;王勇等;《有机化学》;20111231;第31卷(第10期);第1595-1606页 *

Also Published As

Publication number Publication date
CN103709162A (en) 2014-04-09

Similar Documents

Publication Publication Date Title
Hanan et al. Discovery of potent and selective pyrazolopyrimidine janus kinase 2 inhibitors
Huang et al. Discovery of novel 2-aminobenzamide inhibitors of heat shock protein 90 as potent, selective and orally active antitumor agents
CN102898386B (en) Quinazoline derivant, its preparation method, intermediate, composition and application thereof
CA2971872C (en) Mutant idh1 inhibitors useful for treating cancer
TWI573790B (en) Substituted indazole derivatives active as kinase inhibitors
US11370770B2 (en) 3-arylindazoles as selective MEK4 inhibitors
CN114269735A (en) Dihydro or tetrahydro quinazoline compound and intermediate, preparation method and application thereof
CN108026101A (en) Condensed pyramidine compounds or its salt
CN109641887A (en) It can be used as the thiazole of the mutation IDH1 inhibitor for treating cancer
CN104341425B (en) Deuterated acetylene-derivative, its pharmaceutical composition and application
CN103965120A (en) Quinoline and quinazoline derivative, preparation method, intermediate, composition and application
TW200417546A (en) New compounds
WO2005085252A1 (en) Imidazo ‘1,2-a’ pyrazine compounds which interact with protein kinases
CA2901267C (en) Novel n-(2,3-dihydro-1h-pyrrolo[2,3-b]pyridin-5-yl)-4-quinazolinamine and n-(2,3-dihydro-1h-indol-5-yl)-4-quinazolinamine derivatives as perk inhibitors
Wang et al. Design, synthesis, biological evaluation and molecular modeling of novel 1H-pyrrolo [2, 3-b] pyridine derivatives as potential anti-tumor agents
TW201008938A (en) Novel triazolo[4,3-a]pyridine derivatives, process for the preparation thereof, use thereof as medicaments, pharmaceutical compositions and novel use, in particular as MET inhibitors
Crawford et al. Structure-guided design of group I selective p21-activated kinase inhibitors
TWI760005B (en) Fluorinated heterocyclic derivatives with macrocyclic structure and uses thereof
CN105732616A (en) Pyrrolopyridine compounds containing biaryl amide structure, preparation method and applications thereof
Zhao et al. Synthesis and evaluation of a series of pyridine and pyrimidine derivatives as type II c-Met inhibitors
CN107089968B (en) 1- (2-aminopyridine -4- base) -3- piperidine formamide derivative and its synthetic method and application
Wang et al. Bioisosteric replacement of an acylureido moiety attached to an indolin-2-one scaffold with a malonamido or a 2/4-pyridinoylamido moiety produces a selectively potent Aurora-B inhibitor
CN107151233A (en) Pyridine derivatives containing hydrazone and application thereof
CN102558172B (en) 5,8-bis-replaces-1,6-naphthyridine-7-amidocarbonylation compound and dimer compound thereof, Preparation Method And The Use
CN104804001B9 (en) 4-substituted pyrrolo [2,3-d ] pyrimidine compounds and uses thereof

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant