CN104230954A - 2,4-diaminopyrimidine compounds and medical applications thereof - Google Patents

2,4-diaminopyrimidine compounds and medical applications thereof Download PDF

Info

Publication number
CN104230954A
CN104230954A CN201310228635.2A CN201310228635A CN104230954A CN 104230954 A CN104230954 A CN 104230954A CN 201310228635 A CN201310228635 A CN 201310228635A CN 104230954 A CN104230954 A CN 104230954A
Authority
CN
China
Prior art keywords
halogen
alkyl
replaced
hydrogen
compound
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.)
Pending
Application number
CN201310228635.2A
Other languages
Chinese (zh)
Inventor
张翱
耿美玉
宋子兰
艾菁
刘志清
陈奕
彭霞
丁健
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 CN201310228635.2A priority Critical patent/CN104230954A/en
Publication of CN104230954A publication Critical patent/CN104230954A/en
Pending legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D498/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D498/08—Bridged systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
    • C07D239/24—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members
    • C07D239/28—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
    • C07D239/46—Two or more oxygen, sulphur or nitrogen atoms
    • C07D239/48—Two nitrogen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/08—Bridged systems

Abstract

The invention relates to a 2,4-diaminopyrimidine compounds shown as a general formula I, a pharmaceutically acceptable salt or a pharmaceutically acceptable solvate thereof, a preparation method thereof, a pharmaceutical composition comprising the compound, and applications of the compounds in drugs for preventing or treating diseases related to cell abnormal proliferation, morphologic changes and hyperkinesia which are related to anaplastic lymphoma kinase in biological bodies, and drugs related to angiogenesis and cancer metastasis, particularly drugs for treating or preventing tumor growth and metastasis. The general formula I is shown in the description.

Description

2,4-diaminopyrimidines and medicinal use thereof
Technical field
The present invention relates to that a class has SRCA LK selective inhibitory activity 2, 4-diaminopyrimidines and pharmacy acceptable salt thereof or pharmaceutically acceptable solvate, its preparation method, comprise the pharmaceutical composition of this compound, and these compounds are for the preparation of the abnormal cell proliferation relevant to gradually changeable lymphoma enzyme in prevention or treatment organism, the disease that metamorphosis and hypoerkinesia etc. are relevant, and the purposes in the medicine of the disease relevant to angiogenesis or metastasis of cancer, especially the purposes in the medicine for the preparation for the treatment of or prophylaxis of tumours growth and transfer.
Background technology
Gradually changeable lymphoma enzyme (ALK) is a kind of receptor tyrosine kinase, is under the jurisdiction of insulin receptor superfamily.Be found in the earliest in gradually changeable large celllymphoma (ALCL), in the ALCL of about 60%-85%, have the expression of ALK, and normal ALK is single-minded is expressed in neural system, in especially neonatal brain.In human body, ALK gene expression level declines along with reaching maturity of brain, and the amount in ripe cerebral tissue is very low, expresses and there is certain regionality; Other system does not especially find the expression of ALK in hemopoietic system.ALK gene lacks expression in the non-hematopoietic system cancer of the overwhelming majority and healthy tissues, shows that the distribution range of alk protein is extremely narrow.
ALK gene is positioned at karyomit(e) 2p23 site, and the mRNA of the transcribed generation size 6222bp of ALK in normal circumstances servant source, is made up of 29 exons, the I type membrane-spanning protein ALK of coding 1620 aminoacid sequence 200KDa.ALK gene is in dormant state usually, causes cell deterioration to develop into malignant tumour owing to occurring to merge with other genes.But the gene that can occur to merge with its has a lot, at nonsmall-cell lung cancer (non-small cell lung cancer, NSCLC) mainly merge with EML4 gene (echinoderms microtubule-associated protein sample 4) in, becoming lymphom kinase (EML4-ALK) fusion gene between echinoderms microtubule-associated protein sample 4-in the incidence of NSCLC is 2% ~ 7%.
Along with deepening continuously of nonsmall-cell lung cancer (NSCLC) molecular biology research, individualized treatment based on molecular marked compound has been gone to clinical from laboratory, and the treatment of NSCLC patient late achieves significant clinical progress.It is also important that, except traditional Pathological cassification, the difference that NSCLC can also express according to various molecular marked compound, carry out molecular phenotype classification, and with the driven nature gene relevant to tumorigenesis for target spot, research and develop new medicine, carry out individuation molecular targeted therapy targetedly, improve patient's prognosis.In the ideal situation, all NSCLC patients should carry out the detection of related molecular marker thing before the treatment, implementing and treating targetedly, improving result for the treatment of when fully understanding patient tumors molecular expression signature.In this context, Tyrosylprotein kinase has become in recent years very powerful and exceedingly arrogant molecular target, its selective depressant, or has become the focus of antitumor drug research around the Mutiple Targets micromolecular inhibitor of ALK.
At present, the micromolecular inhibitor PF02341066 of Pfizer company exploitation is ratified listing by U.S. FDA on August 26th, 2011, and this is also the micromolecular inhibitor of a unique ALK gone on the market.But existing clinical study shows to have occurred resistance to PF02341066, PF02341066 bioavailability in vivo has much room for improvement simultaneously.Now the inhibitor of a kind of single target ALK does not go on the market, but the AF-802 being in clinical 2 phases is all ALK selective depressants with being in LDK-378 and ASP-3026 of clinical 1 phase, has good Prospect of R & D.
The present invention is directed to tradition 2, 4-diaminopyrimidines causes target compound suppress potassium-channel hERG generation is obvious thus has the shortcomings such as cardiac toxic owing to having very strong alkalescence, the phenyl ring of pyrimidine-2-phenylamino substituting group part is introduced the amide group of replacement, by reducing, cardiac toxic is alleviated to the suppression of potassium-channel, thus it is novel to obtain a class formation, security is high, to multiple Tyrosylprotein kinase (EGFR, PDGFR, FGFR, c-Met, Flt3 etc.), especially ALK is had to the poly-substituted miazines compound of higher activity, acquisition one class new type antineoplastic medicine is expected to its further investigation.
Summary of the invention
An object of the present invention is to provide a class with 2,4-di-amino-pyrimidine is the compound of parent nucleus, its pharmacy acceptable salt or pharmaceutically acceptable solvate, and described compound is a class tyrosine kinase inhibitor, has ALK selective inhibitory.
Shown in the following general formula I of structure of 2,4-diaminopyrimidines of the present invention:
Wherein:
R 1for the heterocyclic radical of substituted or unsubstituted 4-10 unit or the heteroaryl of substituted or unsubstituted 5-10 unit,
Wherein, the substituting group of described replacement is halogen; The C1-C6 alkyl not replacing or replaced by halogen or 4-7 unit heterocyclic radical; C3-C6 cycloalkyl; C6-C10 aryl; 4-7 unit heterocyclic radical; 5-8 unit heteroaryl; Or-C (O) R '; Wherein, R ' is the C1-C6 alkyl not replacing or replaced by halogen, hydroxyl, amino or dimethylamino; The C2-C6 alkenyl not replacing or replaced by halogen, hydroxyl, amino or dimethylamino; Or C3-C6 cycloalkyl;
R 2for hydrogen, halogen, trifluoromethyl, C1-C6 alkoxyl group or C1-C6 alkyl;
The C1-C4 alkylidene group that A is direct key or does not replace or replaced by amino;
X is hydrogen, halogen or trifluoromethyl;
L is-S (=O) 2-,-C (=O) N (R c)-or-S (=O) 2n (R c)-; Wherein, R cfor hydrogen or C1-C4 alkyl;
M is hydrogen, C1-C8 alkyl or C3-C8 cycloalkyl.
Preferably, in the compound shown in general formula I,
Wherein, R 1for the heterocyclic radical of substituted or unsubstituted 4-10 unit or the heteroaryl of substituted or unsubstituted 5-10 unit; Wherein, the substituting group of described replacement is halogen; The C1-C4 alkyl not replacing or replaced by halogen or 4-7 unit heterocyclic radical; C3-C6 cycloalkyl; C6-C10 aryl; 4-7 unit heterocyclic radical; 5-8 unit's heteroaryl or-C (O) R '; Wherein, R ' is the C1-C4 alkyl not replacing or replaced by halogen, hydroxyl, amino or dimethylamino; The C2-C4 alkenyl not replacing or replaced by halogen, hydroxyl, amino or dimethylamino; C3-C6 cycloalkyl;
R 2for hydrogen, halogen, trifluoromethyl, C1-C4 alkoxyl group or C1-C4 alkyl;
The C1-C2 alkylidene group that A is direct key or does not replace or replaced by amino;
X is hydrogen, halogen or trifluoromethyl;
L is-S (=O) 2-or-C (=O) N (R c)-; Wherein, R cfor hydrogen or C1-C2 alkyl;
M is hydrogen, C1-C4 alkyl or C3-C6 cycloalkyl.
More preferably, in the compound shown in general formula I,
Wherein, R 1for:
Wherein, R 0for H; The C1-C4 alkyl not replacing or replaced by halogen, 4-7 unit heterocyclic radical; C3-C6 cycloalkyl; C6-C10 aryl; 4-7 unit heterocyclic radical; 5-8 unit's heteroaryl or-C (O) R '; Wherein, R ' is the C1-C4 alkyl not replacing or replaced by halogen, hydroxyl, amino or dimethylamino; The C2-C4 alkenyl not replacing or replaced by halogen, hydroxyl, amino or dimethylamino; C3-C6 cycloalkyl; Preferably, R 0for H; Methyl; Trifluoromethyl; Oxetanylmethoxy; 2-morpholinyl ethyl; Pyrimidyl; Cyclopropyl formyl radical; 2-hydroxyacetyl; 4-dimethylamino-2-alkene-1-butyryl radicals;
R 2for hydrogen, halogen, trifluoromethyl, C1-C4 alkoxyl group or C1-C4 alkyl; Preferably, R 2for hydrogen or methoxyl group;
The C1-C2 alkylidene group that A is direct key or does not replace or replaced by amino;
X is hydrogen, halogen or trifluoromethyl; Preferably, X is halogen; More preferably, X is chlorine;
L is-S (=O) 2-or-C (=O) NH-;
M is hydrogen or C1-C4 alkyl; Preferably, M is C1-C4 alkyl; More preferably, M is methyl or sec.-propyl.
Most preferably, typical compound of the present invention includes but not limited to the compound shown in table one:
Table one
The pharmacy acceptable salt of the compound that described general formula I represents comprises without limitation: inorganic acid salt, example hydrochloric acid salt, hydrobromate, nitrate, vitriol, phosphoric acid salt etc.; Organic acid salt, as formate, acetate, propionic salt, benzoate, maleate, fumarate, succinate, tartrate, Citrate trianion etc.; Alkylsulfonate, as metilsulfate, ethyl sulfonate etc.; Arylsulphonate, as benzene sulfonate, tosilate etc.
The pharmaceutically acceptable solvate of the compound that described general formula I represents comprises the solvate of compound that general formula I represents and water, ethanol, Virahol, ether, acetone etc. without limitation.
Define above-mentioned group:
Described halogen can be fluorine, chlorine, bromine, and iodine, the halogen number simultaneously occurred and kind are not limit.
Described alkyl can be the straight or branched alkyl with 1-8 carbon atom; Comprise without limitation: methyl, ethyl, n-propyl, sec.-propyl, normal-butyl, isobutyl-, the tertiary butyl etc.
Described alkenyl comprises and has 2-6 carbon atom and the straight or branched thiazolinyl comprising at least one double bond.
Described aryl be have 6-10 carbon atom aromatic group.
Described heteroaryl can be five yuan to ten yuan containing one or more heteroatomic aromatic group be selected from nitrogen, oxygen and sulphur, such as, described heteroaryl can be selected from the heteroatoms in nitrogen, oxygen and sulphur containing 1-5, preferably containing 1 ~ 3 heteroatoms be selected from nitrogen, oxygen and sulphur, more preferably containing 1 ~ 2 heteroatoms be selected from nitrogen and oxygen; The example of described heteroaryl can comprise imidazolyl, pyrimidyl etc.
Described heterocyclic radical can be quaternary to ten yuan be selected from heteroatomic saturated or undersaturated nonaromatic monocycle in nitrogen, oxygen and sulphur and ring, volution, condensed ring, bridged ring etc. containing one or more, such as, described heterocyclic radical can be selected from the heteroatoms in nitrogen, oxygen and sulphur containing 1-5, preferably containing 1 ~ 3 heteroatoms be selected from nitrogen, oxygen and sulphur, more preferably containing 1 ~ 2 heteroatoms be selected from nitrogen and oxygen; Such as; described heterocyclic radical is including, but not limited to homopiperazine base, Pyrrolidine base, 7H-purine radicals, 8-oxygen-3-azabicyclic [3.2.1] octyl, 2 of the piperazinyl of morpholinyl, piperidyl, piperazinyl, N-alkyl or acyl substituted, homopiperazine base, N-alkyl or acyl substituted; 5-diaza-bicyclo [2.2.1] heptane base, 5; 6; 7,8-tetrahydrochysene-[1,2; 4] triazolo [4,3-a] pyrazinyl etc.
Another object of the present invention is to provide the preparation method of 2, the 4-diaminopyrimidines that above-mentioned general formula I represents.
The compound that general formula I represents can be obtained by the method for replacement or coupling by compound ii and compound III.
Such as, compound ii and compound III are carried out substitution reaction and are obtained the compound that general formula I represents under camphorsulfonic acid catalysis:
Particularly, compound ii and compound III being dissolved in the solvent as Virahol, adding D-(+) after camphorsulfonic acid, microwave heating is obtained by reacting compound of Formula I;
Or compound ii and compound III obtain through linked reaction the compound that general formula I represents in the presence of a catalyst:
Particularly, compound ii, compound III, palladium, two diphenylphosphine-9,9-dimethyl xanthene (xantphos) of part 4,5-and mineral alkali cesium carbonate are dissolved in as 1, in the solvent of 4-dioxane, microwave heating is obtained by reacting compound of Formula I;
Wherein, R 1, R 2, A, X, L and M definition the same.
2, the 4-diaminopyrimidines that general formula I of the present invention represents, its pharmacy acceptable salt or pharmaceutically acceptable solvate are a class tyrosine kinase inhibitor, have ALK and c-Met selective inhibitory.Compound of the present invention plays the effect of inhibition tumor cell growth by abnormal cell proliferation, metamorphosis and hypoerkinesia etc. that suppression gradually changeable lymphoma enzyme is correlated with, and these compounds also have the effect of angiogenesis inhibiting or anticancer transfer.
Therefore, another object of the present invention be to provide that above-mentioned general formula I represents 2,4-diaminopyrimidines, its pharmacy acceptable salt or pharmaceutically acceptable solvate, for the preparation of preventing or treating the purposes in the medicine of the disease that in organism, abnormal cell proliferation, metamorphosis and the hypoerkinesia relevant to gradually changeable lymphoma enzyme is correlated with and the disease of being correlated with angiogenesis or metastasis of cancer, are especially growing and the purposes in the medicine shifted for the preparation for the treatment of or prophylaxis of tumours.
Another object of the present invention is to provide a kind of pharmaceutical composition, its comprise treatment significant quantity one or more general formula Is represent 2,4-diaminopyrimidines, its pharmacy acceptable salt or pharmaceutically acceptable solvate, and pharmaceutically acceptable carrier or vehicle.
Another object of the present invention is to provide a kind of method for the treatment of the disease that in organism, abnormal cell proliferation, metamorphosis and the hypoerkinesia relevant to the overexpression of gradually changeable lymphoma enzyme is correlated with and the disease of being correlated with angiogenesis or metastasis of cancer, described method comprises and comprises 2,4-diaminopyrimidines, its pharmacy acceptable salt or pharmaceutically acceptable solvate that general formula I represents or its mixture pharmaceutical composition as activeconstituents to patient therapeuticallv's significant quantity.
Embodiment
Below in conjunction with specific embodiment, the present invention is further elaborated.These embodiments are only for purpose of explanation, and do not limit the scope of the invention and essence.
Preparation embodiment
1h-NMR Varian MercuryAMX300 type instrument measures; 2; 4,5-trichloropyrimidine, isatoic anhydride; m-nitraniline; 2-(isopropelsulfonyl) aniline, palladium, methoxyacetic acid; 4-methyl isophthalic acid-piperazine acetic acid; 4-methoxyl group-3-N-methyl-p-nitroaniline, the two diphenylphosphine-9 of 4,5-; 9-dimethyl xanthene; 2-(7-azo benzotriazole)-N, N, N'; N'-tetramethyl-urea phosphofluoric acid ester; DIPEA and N-hydroxyl-7-azepine benzotriazole are purchased from J & K Chemica lark prestige chemical reagents corporation, and all the other reagent are produced by Chinese Medicine reagent company limited.All solvents are before use all through re-distillation, and the anhydrous solvent used is all obtain by standard method drying treatment; Except explanation, it is all carry out under nitrogen protection and TLC tracking that institute responds, all through saturated sodium-chloride water solution washing and anhydrous sodium sulfate drying process during aftertreatment; The purifying of product all uses silica gel (200 ~ 300 order) column chromatography except explanation; Wherein silica gel (200 ~ 300 order) is produced by Haiyang Chemical Plant, Qingdao, and GF-254 thin-layer silicon offset plate is produced by Yantai Jiang You silica gel development corporation, Ltd..
The synthesis of preparation embodiment 1 compound S 1
The synthesis of intermediate 1-3:
M-nitraniline (1-1) and triethylamine are dissolved in dry DCM, slowly drip the dry DCM solution of chloroacetyl chloride under ice bath, after temperature slowly being risen to room temperature after finishing, 30min reacts completely.After reaction solution dchloromethane, with the washing of water, saturated sodium bicarbonate solution and saturated common salt, organic phase anhydrous sodium sulfate drying, concentrates to obtain intermediate 1-3.
The synthesis of intermediate 1-4:
Intermediate 1-3 is dissolved in acetonitrile, adds 2eq8-oxygen-3-azabicyclic [3.2.1] octane and 2eq salt of wormwood, heated overnight at reflux, after reacting completely, reaction solution silica-gel plate sample upper prop is obtained intermediate 1-4.
The synthesis of intermediate 1-5:
Be dissolved in methyl alcohol by intermediate 1-4, add 0.2eq palladium carbon (palladium content 10%), after hydrogen exchange, room temperature reaction spends the night.Filtered by palladium carbon after reacting completely, filtrate directly removes under reduced pressure crude product 1-5 is for subsequent use.
The synthesized reference document of intermediate 1-6: WO2009143389.
The synthesis of compound S 1:
By 1.4eq intermediate 1-5,1eq intermediate 1-6 and 2eq D-(+) camphorsulfonic acid are dissolved in Virahol, 80 DEG C, microwave reaction one hour, after reacting completely, reaction solvent is removed under reduced pressure, add chloroform and saturated sodium bicarbonate extraction, organic phase is washed with saturated common salt again, upper prop CHCl after anhydrous sodium sulfate drying 3: MeOH=80:1 ~ 30:1 obtains compound S 1. 1H?NMR(300MHz,CDCl 3)δ9.65(s,1H),9.16(s,1H),8.60(d,J=8.5Hz,1H),8.16(s,1H),7.90(dd,J=8.1,1.7Hz,2H),7.60(t,J=7.9Hz,1H),7.48(d,J=7.5Hz,1H),7.36–7.18(m,2H),7.08(d,J=7.2Hz,2H),4.37(s,2H),3.35–3.14(m,1H),3.08(s,2H),2.64(d,J=1.8Hz,4H),2.02(s,4H),1.32(d,J=6.9Hz,6H)。
The synthesis of preparation embodiment 2 compound S 2
Except using (1S, 4S)-2-BOC-2,5-diaza-bicyclo [2.2.1] heptane to replace outside 8-oxygen-3-azabicyclic [3.2.1] octane, the same S1 of synthesis of compound S 2. 1HNMR(300MHz,CDCl 3)δ9.63(s,1H),9.18(s,1H),8.60(dd,J=8.4,0.9Hz,1H),8.17(s,1H),7.90(dd,J=8.0,1.5Hz,1H),7.84(t,J=2.0Hz,1H),7.60(ddd,J=8.9,7.4,1.7Hz,1H),7.52–7.40(m,1H),7.32–7.18(m,2H),7.12(dd,J=3.9,3.0Hz,2H),3.63(s,1H),3.48(s,1H),3.39(d,J=2.6Hz,1H),3.31(d,J=12.5Hz,2H),3.24(dd,J=8.5,5.2Hz,1H),3.09(d,J=10.0Hz,1H),2.99–2.89(m,2H),2.66(d,J=9.7Hz,1H),1.81(d,J=9.7Hz,1H),1.32(d,J=6.9Hz,6H).
The synthesis of preparation embodiment 3 compound S 3
The synthesis of intermediate 3-2:
1eq m-nitraniline (1-1) and 1.2eq Boc-L-proline(Pro) (3-1) are dissolved in dry methylene dichloride 50ml, 2eq2-(7-azo benzotriazole)-N is added under ice bath, N, N', N'-tetramethyl-urea phosphofluoric acid ester (HATU), 5eq N, N-diisopropylethylamine (DIPEA) and 1eq N-hydroxyl-7-azepine benzotriazole (HOAt), rise to stirred overnight at room temperature, be spin-dried for methylene dichloride, add ethyl acetate, 1N HCl extracts, and merges organic layer, use saturated sodium bicarbonate and NaCl successively, anhydrous sodium sulfate drying.Filtration is spin-dried for upper prop, sherwood oil: ethyl acetate=2:1 obtains product 1-3, productive rate equivalent. 1H?NMR(300MHz,CDCl 3)δ10.04(s,1H),8.44(t,J=2.1Hz,1H),7.69(d,J=41.2Hz,2H),7.31(s,1H),4.52(s,1H),3.52(dd,J=10.0,6.8Hz,1H),2.38(s,1H),1.96(dd,J=20.4,9.7Hz,4H),1.51(s,9H).
The synthesis of intermediate 3-3:
Intermediate 3-2 is dissolved in methyl alcohol, adds the palladium carbon of 10%, hydrogen exchange, stirred at ambient temperature 2 hours, filter palladium carbon, be spin-dried for filtrate, obtain intermediate 1-4, be directly used in next step reaction.
The synthesized reference document of intermediate 3-4: Bioorganic & Medicinal Chemistry Letters, 21 (2), 660-663; 2011.
The synthesis of intermediate 3-5:
1eq intermediate 3-3,1.4eq intermediate 3-4,20%mmol palladium, two diphenylphosphine-9,9-dimethyl xanthene (xantphos) of 0.5eq part 4,5-and 3eq cesium carbonate are mixed in microwave tube, add 2ml1,4-dioxane, microwave 110oC, 1 hour.Be chilled to room temperature, add ethyl acetate washing, merge organic layer, saturated sodium bicarbonate is washed, anhydrous sodium sulfate drying, and filtration is spin-dried for, upper silicagel column, methylene dichloride: methyl alcohol: ammoniacal liquor=30:1:0.1 obtains intermediate 3-5. 1H?NMR(300MHz,DMSO)δ11.69(s,1H),9.91(s,1H),9.45(s,1H),8.78(dd,J=25.7,6.6Hz,2H),8.18(s,1H),7.90–7.66(m,2H),7.41(dd,J=17.7,10.1Hz,2H),7.28–7.03(m,3H),4.21(d,J=20.2Hz,1H),3.33(s,2H),2.79(d,J=4.4Hz,3H),2.15(s,1H),1.84(d,J=8.8Hz,3H),1.32(d,J=33.3Hz,9H).
Compound S 3 synthesizes:
Be dissolved in methylene dichloride by intermediate 3-5, add trifluoroacetic acid, stirred overnight at room temperature, be spin-dried for solvent, extraction into ethyl acetate, washing, obtains compound S 3. 1h NMR (300MHz, CDCl 3) δ 11.07 (s, 1H), 9.69 (s, 1H), 8.67 (d, J=8.3Hz, 1H), 8.11 (s, 1H), 7.77 (s, 1H), 7.44 (dd, J=15.3,7.6Hz, 3H), 7.10 – 6.99 (m, 2H), 6.28 (s, 1H), 3.88 (dd, J=9.1,5.3Hz, 1H), 3.03 (d, J=4.8Hz, 3H), 2.20 (d, J=8.1Hz, 1H), 2.04 (dd, J=12.4,6.2Hz, 2H), 1.80 – 1.74 (m, 4H).
The synthesis of preparation embodiment 4 compound S 4
Except use 5-BOC-hexahydropyrrolo also [3,4-B] pyrroles replace outside 8-oxygen-3-azabicyclic [3.2.1] octane, the same S1 of synthesis of compound S 4. 1HNMR(300MHz,CDCl 3)δ9.62(s,1H),9.13(s,1H),8.59(d,J=8.4Hz,1H),8.16(s,1H),7.88(d,J=8.0Hz,1H),7.81(s,1H),7.58(t,J=7.9Hz,1H),7.49(d,J=7.5Hz,1H),7.33–7.18(m,3H),7.10(d,J=8.0Hz,1H),3.44(d,J=16.8Hz,1H),3.30–3.15(m,3H),3.06(t,J=7.8Hz,1H),2.99–2.89(m,1H),2.83(d,J=4.8Hz,2H),2.73(m,1H),2.59(dd,J=11.8,4.5Hz,1H),2.45–2.33(m,1H),2.26–2.07(m,2H),1.52(m,1H),1.31(d,J=6.8Hz,6H).
The synthesis of preparation embodiment 5 compound S 5
Except using 1-cyclopropane carbonyl piperazine to replace 8-oxygen-3-azabicyclic [3.2.1] octane, 2-methoxyl group-5-N-methyl-p-nitroaniline is used to replace outside m-nitraniline, the same S1 of synthesis of compound S 5. 1HNMR(300MHz,CDCl 3)δ9.73(s,1H),9.58(s,1H),8.60(d,J=8.4Hz,1H),8.36(d,J=2.6Hz,1H),8.13(s,1H),7.87(dd,J=7.9,1.6Hz,1H),7.55(t,J=8.9Hz,1H),7.46(dd,J=8.8,2.6Hz,1H),7.23–7.16(m,1H),7.14(s,1H),6.84(d,J=8.9Hz,1H),3.90(s,3H),3.75(s,4H),3.29–3.21(m,1H),3.18(s,2H),2.63(s,4H),1.75(m,1H),1.31(d,J=6.9Hz,6H),1.00(dt,J=6.1,3.0Hz,2H),0.84–0.72(m,2H).
The synthesis of preparation embodiment 6 compound s 6
Except use cis-2-Boc-hexahydropyrrolo also [3,4-c] pyrroles replace outside 8-oxygen-3-azabicyclic [3.2.1] octane, the same S1 of synthesis of compound s 6. 1HNMR(300MHz,CDCl 3)δ9.60(s,1H),9.04(s,1H),8.59(d,J=7.7Hz,1H),8.17(s,1H),7.89(dd,J=7.9,1.6Hz,1H),7.85(s,1H),7.60(t,J=7.9Hz,1H),7.53(d,J=8.1Hz,1H),7.29–7.19(m,2H),7.14(d,J=9.9Hz,2H),3.54(s,2H),3.42(d,J=11.2Hz,2H),3.23(m,3H),2.85(s,2H),2.74(s,2H),1.70(s,2H),1.31(d,J=6.9Hz,6H).
The synthesis of preparation embodiment 7 compound S 7
Except using N-methylhomopiperazin to replace except 8-oxygen-3-azabicyclic [3.2.1] octane, the same S1 of synthesis of compound s 6. 1HNMR(300MHz,CDCl 3)δ9.62(s,1H),9.33(s,1H),8.60(d,J=8.4Hz,1H),8.16(s,1H),7.89(dd,J=8.0,1.6Hz,1H),7.86(s,1H),7.60(t,J=7.9Hz,1H),7.48(d,J=8.2Hz,1H),7.23(dd,J=9.0,7.4Hz,2H),7.14(d,J=6.7Hz,2H),3.30–3.20(m,3H),2.85(dd,J=8.0,3.9Hz,4H),2.69(dd,J=12.8,6.8Hz,4H),2.42(d,J=2.1Hz,3H),1.93–1.82(m,2H),1.32(d,J=6.9Hz,6H).
The synthesis of preparation embodiment 8 compound S 8
Except using Boc-L-Histidine to replace except Boc-L-proline(Pro), the same S3 of synthesis of compound S 8. 1HNMR(300MHz,CDCl 3)δ9.64(s,1H),9.61(s,1H),8.57(d,J=8.4Hz,1H),8.14(s,1H),7.88(d,J=8.0Hz,1H),7.83(s,1H),7.55(s,2H),7.38(d,J=7.8Hz,1H),7.24–7.12(m,4H),6.86(s,1H),3.75(s,1H),3.49(s,1H),3.25(dd,J=13.8,6.8Hz,2H),3.07(s,1H),1.29(t,J=9.1Hz,6H).
The synthesis of preparation embodiment 9 compound S 9
Except using 4-(2-(piperazine-1-base) ethyl) morpholino for except 8-oxygen-3-azabicyclic [3.2.1] octane, the same S1 of synthesis of compound S 9. 1HNMR(300MHz,CDCl 3)δ9.63(s,1H),9.09(s,1H),8.59(d,J=8.3Hz,1H),8.17(s,1H),7.90(d,J=6.6Hz,1H),7.84(s,1H),7.60(dt,J=8.8,4.3Hz,1H),7.49(d,J=8.3Hz,1H),7.33–7.18(m,2H),7.08(d,J=6.5Hz,2H),3.84–3.64(m,4H),3.33–3.18(m,1H),3.12(s,2H),2.57(m,16H),1.32(d,J=6.8Hz,6H).
The synthesis of preparation embodiment 10 compound S 10
Except using Boc-piperazine to replace except 8-oxygen-3-azabicyclic [3.2.1] octane, the same S1 of synthesis of compound S 10. 1HNMR(300MHz,CDCl 3)δ9.63(s,1H),9.13(s,1H),8.60(d,J=8.5Hz,1H),8.17(s,1H),7.90(dd,J=7.9,1.6Hz,1H),7.84(s,1H),7.60(t,J=7.9Hz,1H),7.48(d,J=8.3Hz,1H),7.27(d,J=9.8Hz,1H),7.22(d,J=8.6Hz,1H),7.11(d,J=6.9Hz,1H),7.06(s,1H),3.31–3.17(m,1H),3.11(d,J=2.9Hz,2H),3.00–2.89(m,4H),2.59(d,J=4.7Hz,4H),1.36–1.27(d,J=2.9Hz,6H).
The synthesis of preparation embodiment 11 compound S 11
The oxyacetic acid of S10 and 1.3eq is dissolved in DMF, adds 1.2eq O-benzotriazole-N under ice bath, N, N', N'-tetramethyl-urea Tetrafluoroboric acid (TBTU) and 2eq DIPEA.Pour in mixture of ice and water by reaction solution after question response is complete, be extracted with ethyl acetate twice, organic phase concentrates upper prop and obtains S11. 1H?NMR(300MHz,CDCl 3)δ9.65(s,1H),8.86(s,1H),8.60(d,J=8.5Hz,1H),8.17(s,1H),7.90(dt,J=8.5,1.6Hz,1H),7.86(s,1H),7.60(t,J=8.7Hz,1H),7.53–7.44(m,1H),7.28(d,J=8.0Hz,1H),7.23(d,J=8.1Hz,1H),7.14(s,1H),7.11–7.03(m,1H),4.18(d,J=4.5Hz,2H),3.75(s,2H),3.60(d,J=4.4Hz,1H),3.36(s,2H),3.31–3.21(m,1H),3.18(s,2H),2.64(s,4H),1.32(d,J=6.9Hz,6H).
The synthesis of preparation embodiment 12 compound S 12
Except using 1-methylpiperazine to replace except 8-oxygen-3-azabicyclic [3.2.1] octane, the same S1 of synthesis of compound S 12. 1H?NMR(300MHz,CDCl 3)δ9.62(s,1H),9.09(s,1H),8.59(dd,J=8.5,0.9Hz,1H),8.16(s,1H),7.92–7.79(m,2H),7.63–7.52(m,1H),7.46(ddd,J=8.1,2.1,0.9Hz,1H),7.37(s,1H),7.25–7.23(m,1H),7.22–7.20(m,1H),7.08(ddd,J=8.0,2.0,0.9Hz,1H),3.30–3.17(m,1H),3.12(s,2H),2.64(s,4H),2.49(s,4H),2.31(s,3H),1.30(d,J=6.9Hz,6H).
The synthesis of preparation embodiment 13 compound S 13
Except using (2E)-4-dimethylamino-2-butylene acid to replace except oxyacetic acid, the same S11 of synthesis of compound S 13. 1HNMR(400MHz,CDCl 3)δ9.65(s,0H),8.98(s,0H),8.62(d,J=8.3Hz,0H),8.18(s,0H),7.90(dd,J=8.0,1.5Hz,0H),7.87(s,0H),7.62–7.56(m,1H),7.49(dd,J=8.0,1.6Hz,1H),7.41(s,1H),7.28(t,J=4.0Hz,1H),7.24(dd,J=10.4,4.8Hz,1H),7.14–7.07(m,1H),6.88(dt,J=15.1,5.9Hz,1H),6.42(d,J=15.2Hz,1H),3.69(d,J=40.3Hz,4H),3.30–3.20(m,1H),3.17(s,2H),3.09(dd,J=5.9,1.2Hz,2H),2.66–2.60(m,4H),2.27(s,6H),1.32(d,J=6.9Hz,6H).
The synthesis of preparation embodiment 14 compound S 14
The synthesis of compound 14-3:
Except using 1-N-Boc-3-azetidinecarboxylic acid to replace except Boc-L-proline(Pro), the synthesis of compound 14-3 is with compound 3-3.
The synthesis of compound S 14:
Except using compound 14-3 to replace except compound 1-5, the same S1 of synthesis of compound S 14. 1H?NMR(300MHz,CDCl3)δ9.60(s,1H),8.57(d,J=8.3Hz,1H),8.36(s,1H),8.13(s,1H),7.96–7.75(m,2H),7.56(t,J 1=7.7Hz,1H),7.45(s,1H),7.36(s,1H),7.30–7.07(m,3H),3.87(m,4H),3.47(s,1H),3.32–3.11(m,1H),1.30(d,J=6.8Hz,6H)。
The synthesis of preparation embodiment 15 compound S 15
The synthesis of compound 15-3:
Except using morpholine-4-guanidine-acetic acid to replace except Boc-L-proline(Pro), the synthesis of compound 15-3 is with compound 3-3.
The synthesis of compound S 15:
Except using compound 15-3 to replace except compound 1-5, the same S1 of synthesis of compound S 15. 1H?NMR(300MHz,CDCl 3)δ9.63(s,1H),9.03(s,1H),8.59(d,J=8.5Hz,1H),8.17(s,1H),7.89(d,J=6.5Hz,1H),7.84(s,1H),7.58(t,J=8.6,7.3,1H),7.47(d,J=8.2Hz,1H),7.36–7.16(m,2H),7.10(d,J=8.0,1H),3.84–3.68(m,4H),3.31–3.17(m,1H),3.13(s,2H),2.68–2.54(m,4H),1.31(d,J=6.9Hz,6H)。
The synthesis of preparation embodiment 16 compound S 16
The synthesis of compound 16-3:
Except using N-Boc-AzeOH to replace except Boc-L-proline(Pro), the synthesis of compound 16-3 is with compound 3-3.
The synthesis of compound S 16:
Except using compound 16-3 to replace except compound 1-5, the same S1 of synthesis of compound S 16. 1H?NMR(300MHz,CDCl 3)δ9.61(s,1H),9.53(s,1H),8.59(d,J=8.3Hz,1H),8.16(s,1H),7.87(dd,J=7.9,1.5Hz,2H),7.65–7.45(m,2H),7.44–7.33(m,1H),7.22(dd,J=16.5,6.8Hz,3H),4.42(t,J=8.5Hz,1H),3.81(q,J=8.4Hz,1H),3.42–3.08(m,2H),2.67(s,2H),2.54–2.20(m,1H),1.28(t,J=9.9Hz,7H)。
The synthesis of preparation embodiment 17 compound S 17
Except using 1-cyclopropane carbonyl piperazine to replace 8-oxygen-3-azabicyclic [3.2.1] octane, 4-methoxyl group-3-N-methyl-p-nitroaniline is used to replace outside m-nitraniline, the same S1 of synthesis of compound S 17. 1H?NMR(300MHz,CDCl 3)δ9.64(s,1H),8.77(s,1H),8.63(d,J=8.5Hz,1H),8.34(s,1H),8.18(s,1H),7.87(d,J=7.9Hz,1H),7.59(t,J=7.9Hz,1H),7.50(s,1H),7.38–7.29(m,1H),7.19(t,J=7.6Hz,1H),6.84(d,J=8.7Hz,1H),3.88(s,3H),3.67(s,4H),3.31–3.18(m,1H),3.12(s,2H),2.59(s,4H),1.77–1.62(m,1H),1.30(d,J=6.8Hz,6H),1.03–0.91(m,2H),0.77(dd,J=7.3,3.5Hz,2H).
The synthesis of preparation embodiment 18 compound S 18
S10 and 1.5eq oxetanone is dissolved in THF, stirring at room temperature 2 hours, adds 1.6eq NaHB (AcO) 3, in room temperature for overnight.After reacting completely, direct silica gel mixed sample concentrates upper prop and obtains S18. 1h NMR (400MHz, CDCl 3) δ 9.66 (s, 1H), 9.06 (s, 1H), 8.62 (d, J=8.4Hz, 1H), 8.19 (s, 1H), 7.91 (dd, J=8.0, 1.4Hz, 1H), 7.87 (s, 1H), 7.61 (t, J=7.9Hz, 1H), 7.51 (d, J=8.1Hz, 1H), 7.30 (d, J=3.1Hz, 1H), 7.28 (d, J=3.2Hz, 1H), 7.24 (t, J=5.7Hz, 1H), 7.09 (d, J=8.0Hz, 1H), 4.69 (t, J=6.6Hz, 2H), 4.63 (t, J=6.2Hz, 2H), 3.59 – 3.51 (m, 1H), 3.31 – 3.20 (m, 1H), 3.16 (s, 2H), 2.70 (s, 4H), 2.43 (s, 4H), 1.33 (d, J=6.9Hz, 6H).
The synthesis of preparation embodiment 19 compound S 19
Except using 1-cyclopropane carbonyl piperazine to replace except 8-oxygen-3-azabicyclic [3.2.1] octane, the same S1 of synthesis of compound S 19. 1H?NMR(300MHz,CDCl 3)δ9.63(s,1H),8.98(s,1H),8.60(dd,J=8.4,0.9Hz,1H),8.16(d,J=2.6Hz,1H),7.87(dt,J=2.8,1.4Hz,1H),7.84(t,J=2.0Hz,1H),7.61–7.52(m,1H),7.48(s,1H),7.48–7.41(m,1H),7.28–7.17(m,2H),7.13–7.04(m,1H),3.70(s,4H),3.29–3.17(m,1H),3.14(d,J=6.4Hz,2H),2.61(s,4H),1.72(tt,J=8.0,4.7Hz,1H),1.30(d,J=6.9Hz,6H),1.02–0.92(m,2H),0.83–0.71(m,2H).
The synthesis of preparation embodiment 20 compound S 20
Except using 3-(trifluoromethyl)-5,6,7,8-tetrahydrochysene-[1,2,4] triazolo [4,3-a] pyrazine to replace outside 8-oxygen-3-azabicyclic [3.2.1] octane, the same S1 of synthesis of compound S 20. 1H?NMR(300MHz,CDCl 3)δ9.60(s,1H),8.80(s,1H),8.58(d,J=8.5Hz,1H),8.11(s,1H),7.84(d,J=7.9Hz,1H),7.80(s,1H),7.55(t,J=8.0Hz,1H),7.46(s,2H),7.24–7.15(m,2H),7.11(d,J=7.8Hz,1H),4.21(d,J=5.8Hz,2H),4.02(s,2H),3.42(s,2H),3.22(dt,J=13.6,6.8Hz,1H),3.12(t,J=5.2Hz,2H),1.27(t,J=7.2Hz,6H)。
The synthesis of preparation embodiment 21 compound S 21
Except using 1-(2-pyrimidyl) piperazine to replace except 8-oxygen-3-azabicyclic [3.2.1] octane, the same S1 of synthesis of compound S 21. 1H?NMR(300MHz,CDCl 3)δ9.63(s,1H),9.12(s,1H),8.60(d,J=8.4Hz,1H),8.30(d,J=4.7Hz,2H),8.16(s,1H),7.87(d,J=8.2Hz,2H),7.57(dd,J=15.1,6.6Hz,2H),7.44(d,J=8.1Hz,1H),7.31–7.08(m,3H),6.51(t,J=4.7Hz,1H),3.86(s,4H),3.31–3.19(m,1H),3.16(s,2H),2.76–2.57(m,4H),1.27(t,J=8.0Hz,6H)。
Two, test example
EXPERIMENTAL EXAMPLE one: molecular receptor SRCA LK molecular level activity rating
1. receptor tyrosine kinase ALK molecular level enzyme is lived and is suppressed preliminary assessment experiment
(1) the PBS(10mM sodium phosphate buffer of enzyme reaction substrate Poly (Glu, Tyr) 4:1 without potassium ion, 150mM NaCl, pH7.2-7.4) be diluted to 20 μ g/mL, 125 μ L/ hole coated elisa plates, put 37 ° of C and react 12-16 hour.Discard liquid in hole.Wash plate, contain the PBS without potassium ion of 0.1%Tween-20 with the T-PBS(in 200 μ L/ holes) wash plate three times, each 5 minutes.Dry enzyme plate 1-2 hour in 37 ° of C baking ovens.
(2) every hole adds with reaction buffer (50mM HEPES pH7.4,50mM MgCl 2, 0.5mM MnCl 2, 0.2mM Na 3vO 4, 1mM DTT) and the ATP solution 49 μ L that dilutes, add 1 μ L compound in every hole, add compound to be tested, the more each kinases territory recombinant protein adding 50 μ L reaction buffers dilutions starts reaction, each experiment need be established without ATP control wells holes.Put 37 ° of C shaking tables (100rpm) and react 1 hour.Discard liquid in hole, T-PBS washes plate three times.
(3) add antibody PY99100 μ L/ hole (T-PBS1:500 of antibody containing BSA5mg/mL dilutes), 37 ° of C shaking tables react 0.5 hour.Discard liquid in hole, T-PBS washes plate three times.
(4) add the anti-100 μ L/ holes of sheep anti mouse two (T-PBS1:2000 of antibody containing BSA5mg/ml dilutes) of horseradish peroxidase-labeled, 37 ° of C shaking tables react 0.5 hour.Discard 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%H 2o 20.1M citric acid-sodium citrate damping fluid (pH=5.4) dilution), 25 ° of C lucifuges reaction 1-10 minute.
(6) 2M H is added 2sO 450 μ L/ hole stopped reactions, to decline orifice plate microplate reader VERSAmax reading with wavelengthtunable, wavelength is 490nm.
(7) interpretation of result
2. receptor tyrosine kinase ALK enzyme is lived and is suppressed IC 50evaluation experimental
Table two: compound is to receptor tyrosine kinase ALK enzyme suppression level alive
The test result display of representative compound, the compound of this type is under the concentration of 1 μM, all more than 90% is reached to the inhibiting rate of ALK, under the concentration of 0.1 μM, inhibiting rate is more than 80%, the activity of some compounds and positive control quite or more excellent, are therefore very potential ALK inhibitor.
EXPERIMENTAL EXAMPLE two: molecular receptor Tyrosylprotein kinase c-Met molecular level activity rating
1. receptor tyrosine kinase c-Met molecular level enzyme is lived and is suppressed preliminary assessment experiment
(1) PBS (10mM sodium phosphate buffer, 150mM NaCl, pH7.2-7.4) of enzyme reaction substrate Poly (Glu, Tyr) 4:1 without potassium ion is diluted to 20g/ml, 125l/ hole coated elisa plate, puts 37 DEG C of reaction 12-16 hour.Discard liquid in hole.Wash plate, wash plate three times, each 5 minutes with the T-PBS (PBS without potassium ion containing 0.1%Tween-20) in 200l/ hole.Dry enzyme plate 1-2 hour in 37 DEG C of baking ovens.
(2) every hole adds the ATP solution 50L diluted with reaction buffer (50mM HEPES pH7.4,50mM MgCl2,0.5mM MnCl2,0.2mM Na3VO4,1mM DTT), final concentration 5 μMs.Add 1 μ l compound (1%DMSO dissolves, and final concentration is 10 μMs) in every hole, then add 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.Each experiment need be established without ATP control wells holes and corresponding DMSO solvent control hole (negative control hole).Discard liquid in hole, T-PBS washes plate three times.
(3) add antibody PY99100l/ hole (T-PBS of antibody containing BSA5mg/ml dilutes, and concentration is 0.4g/ml), 37 DEG C of shaking tables react 0.5 hour.Discard liquid in hole, T-PBS washes plate three times.
(4) add the anti-100l/ hole of sheep anti mouse two (T-PBS of antibody containing BSA5mg/ml dilutes, and concentration is 0.5g/ml) of horseradish peroxidase-labeled, 37 DEG C of shaking tables react 0.5 hour.Discard liquid in hole, T-PBS washes plate three times.
(5) the OPD nitrite ion 100l/ hole (diluting with the 0.1M citric acid-sodium citrate damping fluid (pH=5.4) containing 0.03%H2O2) of 2mg/ml is added, 25 DEG C of lucifuge reaction 1-10 minute.(need with ultrasonic when OPD dissolves, nitrite ion needs now with the current).
(6) add 2M H2SO450l/ hole stopped reaction, to decline orifice plate microplate reader VERSAmax reading with wavelengthtunable, wavelength is 490nm.
(7) inhibiting rate of sample is tried to achieve by following formula:
2, receptor tyrosine kinase c-Met enzyme is lived and is suppressed IC50 evaluation experimental
The c-Met enzyme inhibiting compound (compound is at the inhibiting rate >50% of 10-5M to receptor tyrosine kinase c-Met) alive that clearly has above-mentioned screening obtained is made into gradient concentration, carries out IC50 evaluation.Calculate the IC50 value of the horizontal arrestin Tyrosylprotein kinase of each compound molecule by four parameter methods, result is listed in table 1.
Table four: compound is to receptor tyrosine kinase c-Met enzyme suppression level alive
The test result display of representative compound, the compound of this type, under the concentration of 1 μM, all has obvious restraining effect to c-Met enzyme, has restraining effect at the Concentration portion compound of 0.1 μM to c-Met enzyme.

Claims (9)

1. 2, the 4-diaminopyrimidines, its pharmacy acceptable salt or the pharmaceutically acceptable solvate that represent of the following general formula I of a class,
Wherein:
R 1for the heterocyclic radical of substituted or unsubstituted 4-10 unit or the heteroaryl of substituted or unsubstituted 5-10 unit, wherein, the substituting group of described replacement is halogen; The C1-C6 alkyl not replacing or replaced by halogen or 4-7 unit heterocyclic radical; C3-C6 cycloalkyl; C6-C10 aryl; 4-7 unit heterocyclic radical; 5-8 unit heteroaryl; Or-C (O) R '; Wherein, R ' is the C1-C6 alkyl not replacing or replaced by halogen, hydroxyl, amino or dimethylamino; The C2-C6 alkenyl not replacing or replaced by halogen, hydroxyl, amino or dimethylamino; Or C3-C6 cycloalkyl;
R 2for hydrogen, halogen, trifluoromethyl, C1-C6 alkoxyl group or C1-C6 alkyl;
The C1-C4 alkylidene group that A is direct key or does not replace or replaced by amino;
X is hydrogen, halogen or trifluoromethyl;
L is-S (=O) 2-,-C (=O) N (R c)-or-S (=O) 2n (R c)-; Wherein, R cfor hydrogen or C1-C4 alkyl;
M is hydrogen, C1-C8 alkyl or C3-C8 cycloalkyl.
2. 2,4-diaminopyrimidines according to claim 1, its pharmacy acceptable salt or pharmaceutically acceptable solvate, wherein,
R 1for the heterocyclic radical of substituted or unsubstituted 4-10 unit or the heteroaryl of substituted or unsubstituted 5-10 unit, wherein, the substituting group of described replacement is halogen; The C1-C4 alkyl not replacing or replaced by halogen or 4-7 unit heterocyclic radical; C3-C6 cycloalkyl; C6-C10 aryl; 4-7 unit heterocyclic radical; 5-8 unit's heteroaryl or-C (O) R '; Wherein, R ' is the C1-C4 alkyl not replacing or replaced by halogen, hydroxyl, amino or dimethylamino; The C2-C4 alkenyl not replacing or replaced by halogen, hydroxyl, amino or dimethylamino; C3-C6 cycloalkyl;
R 2for hydrogen, halogen, trifluoromethyl, C1-C4 alkoxyl group or C1-C4 alkyl;
The C1-C2 alkylidene group that A is direct key or does not replace or replaced by amino;
X is hydrogen, halogen or trifluoromethyl;
L is-S (=O) 2-or-C (=O) N (R c)-; Wherein, R cfor hydrogen or C1-C2 alkyl;
M is hydrogen, C1-C4 alkyl or C3-C6 cycloalkyl.
3. 2,4-diaminopyrimidines according to claim 2, its pharmacy acceptable salt or pharmaceutically acceptable solvate, wherein,
R 1for:
Wherein, R 0for H; The C1-C4 alkyl not replacing or replaced by halogen, 4-7 unit heterocyclic radical; C3-C6 cycloalkyl; C6-C10 aryl; 4-7 unit heterocyclic radical; 5-8 unit's heteroaryl or-C (O) R '; Wherein, R ' is the C1-C4 alkyl not replacing or replaced by halogen, hydroxyl, amino or dimethylamino; The C2-C4 alkenyl not replacing or replaced by halogen, hydroxyl, amino or dimethylamino; Or C3-C6 cycloalkyl;
R 2for hydrogen, halogen, trifluoromethyl, C1-C4 alkoxyl group or C1-C4 alkyl;
The C1-C2 alkylidene group that A is direct key or does not replace or replaced by amino;
X is hydrogen, halogen or trifluoromethyl;
L is-S (=O) 2-or-C (=O) NH-;
M is hydrogen or C1-C4 alkyl.
4. 2,4-diaminopyrimidines according to claim 3, its pharmacy acceptable salt or pharmaceutically acceptable solvate, wherein,
R 0for H; Methyl; Trifluoromethyl; Oxetanylmethoxy; 2-morpholinyl ethyl; Pyrimidyl; Cyclopropyl formyl radical; 2-hydroxyacetyl; 4-dimethylamino-2-alkene-1-butyryl radicals;
R 2for hydrogen or methoxyl group;
The C1-C2 alkylidene group that A is direct key or does not replace or replaced by amino;
X is halogen;
L is-S (=O) 2-or-C (=O) NH-;
M is methyl or sec.-propyl.
5. 2,4-diaminopyrimidines according to claim 1, its pharmacy acceptable salt or pharmaceutically acceptable solvate, wherein, described 2,4-diaminopyrimidines are selected from following compound:
。
6. 2,4-diaminopyrimidines according to any one of claim 1-5, its pharmacy acceptable salt or the pharmaceutically acceptable solvate purposes in the medicine of the disease of being correlated with for the preparation of abnormal cell proliferation, metamorphosis and the hypoerkinesia relevant to gradually changeable lymphoma enzyme in prevention or treatment organism.
7. 2,4-diaminopyrimidines according to any one of claim 1-5, its pharmacy acceptable salt or the pharmaceutically acceptable solvate purposes in the medicine for the preparation of prevention or the treatment disease relevant to angiogenesis or metastasis of cancer.
8. 2,4-diaminopyrimidines according to any one of claim 1-5, its pharmacy acceptable salt or the pharmaceutically acceptable solvate purposes in the medicine for the preparation for the treatment of or prophylaxis of tumours growth and transfer.
9. a pharmaceutical composition, it comprises according to any one of claim 1-5 2 of one or more treatment significant quantities, 4-diaminopyrimidines, its pharmacy acceptable salt or pharmaceutically acceptable solvate, and pharmaceutically acceptable carrier or vehicle.
CN201310228635.2A 2013-06-08 2013-06-08 2,4-diaminopyrimidine compounds and medical applications thereof Pending CN104230954A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201310228635.2A CN104230954A (en) 2013-06-08 2013-06-08 2,4-diaminopyrimidine compounds and medical applications thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201310228635.2A CN104230954A (en) 2013-06-08 2013-06-08 2,4-diaminopyrimidine compounds and medical applications thereof

Publications (1)

Publication Number Publication Date
CN104230954A true CN104230954A (en) 2014-12-24

Family

ID=52219948

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201310228635.2A Pending CN104230954A (en) 2013-06-08 2013-06-08 2,4-diaminopyrimidine compounds and medical applications thereof

Country Status (1)

Country Link
CN (1) CN104230954A (en)

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104987324A (en) * 2015-06-04 2015-10-21 湖北生物医药产业技术研究院有限公司 Pyrimidine derivative used as anaplastic lymphoma kinase (ALK) inhibitor
CN105085483A (en) * 2015-06-04 2015-11-25 湖北生物医药产业技术研究院有限公司 Kinase inhibitor and application thereof
CN105579457A (en) * 2013-03-20 2016-05-11 拜耳制药股份公司 Substituted N-biphenyl-3-acetylamino-benzamides and N-[3-(acetylamino)phenyl]-biphenyl-carboxamides and their use as inhibitors of the WNT signalling pathway
CN107586278A (en) * 2016-07-08 2018-01-16 中国科学院上海药物研究所 2,4 2 amine pyrimidine compounds, its preparation method, pharmaceutical composition and the purposes of a kind of fragment containing phenol
CN109836415A (en) * 2017-11-29 2019-06-04 北京博远精准医疗科技有限公司 Carbamide compounds as ALK inhibitor
CN114907288A (en) * 2022-05-13 2022-08-16 中国医学科学院医药生物技术研究所 Application of nitrobenzene compounds in preparation of pseudomonas aeruginosa quorum sensing inhibitor

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004074244A2 (en) * 2003-02-20 2004-09-02 Smithkline Beecham Corporation Pyrimidine compounds
CN101506177A (en) * 2006-07-21 2009-08-12 诺瓦提斯公司 2, 4 -di (arylaminio) -pyrimidine-5-carboxamide compounds as JAK kinases inhibitors
CN101563327A (en) * 2006-12-19 2009-10-21 健泰科生物技术公司 Pyrimidine kinase inhibitors

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004074244A2 (en) * 2003-02-20 2004-09-02 Smithkline Beecham Corporation Pyrimidine compounds
CN101506177A (en) * 2006-07-21 2009-08-12 诺瓦提斯公司 2, 4 -di (arylaminio) -pyrimidine-5-carboxamide compounds as JAK kinases inhibitors
CN101563327A (en) * 2006-12-19 2009-10-21 健泰科生物技术公司 Pyrimidine kinase inhibitors

Cited By (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105579457A (en) * 2013-03-20 2016-05-11 拜耳制药股份公司 Substituted N-biphenyl-3-acetylamino-benzamides and N-[3-(acetylamino)phenyl]-biphenyl-carboxamides and their use as inhibitors of the WNT signalling pathway
CN105579457B (en) * 2013-03-20 2017-08-04 拜耳制药股份公司 The substituted acetyl-amino benzamide of N biphenyl 3 and N [3 (acetyl-amino) phenyl] dibenzoyl amine and its it is used as WNT signal pathway inhibitor purposes
CN105085483B (en) * 2015-06-04 2019-01-01 湖北生物医药产业技术研究院有限公司 Kinase inhibitor and its application
CN105085483A (en) * 2015-06-04 2015-11-25 湖北生物医药产业技术研究院有限公司 Kinase inhibitor and application thereof
WO2016192131A1 (en) * 2015-06-04 2016-12-08 湖北生物医药产业技术研究院有限公司 Kinase inhibitor and application thereof
WO2016192132A1 (en) * 2015-06-04 2016-12-08 湖北生物医药产业技术研究院有限公司 Pyrimidine derivative serving as alk inhibitor
CN104987324A (en) * 2015-06-04 2015-10-21 湖北生物医药产业技术研究院有限公司 Pyrimidine derivative used as anaplastic lymphoma kinase (ALK) inhibitor
CN107586278A (en) * 2016-07-08 2018-01-16 中国科学院上海药物研究所 2,4 2 amine pyrimidine compounds, its preparation method, pharmaceutical composition and the purposes of a kind of fragment containing phenol
CN107586278B (en) * 2016-07-08 2020-01-10 中国科学院上海药物研究所 2, 4-diamino pyrimidine compound, preparation method, pharmaceutical composition and application thereof
CN109836415A (en) * 2017-11-29 2019-06-04 北京博远精准医疗科技有限公司 Carbamide compounds as ALK inhibitor
CN109836415B (en) * 2017-11-29 2020-11-06 北京博远精准医疗科技有限公司 Urea compounds as ALK inhibitors
CN114907288A (en) * 2022-05-13 2022-08-16 中国医学科学院医药生物技术研究所 Application of nitrobenzene compounds in preparation of pseudomonas aeruginosa quorum sensing inhibitor
CN114907288B (en) * 2022-05-13 2023-12-08 中国医学科学院医药生物技术研究所 Application of nitrobenzene compounds in preparation of pseudomonas aeruginosa quorum sensing inhibitor

Similar Documents

Publication Publication Date Title
AU2013269317B2 (en) New compounds
CN105461695B (en) Pyrimidine or pyrrolotriazine derivatives and its production and use
CA2853390C (en) Anticancer pyridopyrazines via the inhibition of fgfr kinases
KR101889565B1 (en) Pyrazolyl quinoxaline kinase inhibitors
CN104230954A (en) 2,4-diaminopyrimidine compounds and medical applications thereof
JP6386467B2 (en) Novel heterocyclic derivatives as modulators of kinase activity
CA2874967A1 (en) Pteridines as fgfr inhibitors
WO2022132914A1 (en) Prmts inhibitors
CN105143228A (en) [1,2,4]triazol[4,3-a]pyridine derivate, preparation method therefor or medical application thereof
CN102875570A (en) Quinazoline derivative, and preparation method and application thereof
CN104837844A (en) Pyrazole substituted imidazopyrzines as casein kinase 1D/E inhibitors
EA033655B1 (en) Azaquinazoline carboxamide derivatives
CN108239071A (en) Amide and thioamides analog derivative and its preparation method and application
CN107573327A (en) Indazolecarboxamides Pyridione derivatives and its production and use
CN102108078A (en) 1,4-substituted phthalazine compound and preparation method and applications thereof
CN113853375B (en) N-containing heteroaryl derivatives and pharmaceutical compositions for preventing or treating cancer comprising the same as active ingredient
CN109422760A (en) FGFR4 inhibitor and its application
CN104177342B (en) The indoles and naphthalene ketone derivant and its medical usage of heterocyclic radical substitution
CN107793397B (en) Optical isomer of substituted pyrimidine PI3K inhibitor and application thereof
CN112480109A (en) Pyrido [2,3-b ] pyrazine-3 (4H) -ketone derivatives and application thereof
CN105566307A (en) Heterocyclically substituted indolonaphthalenone derivative, and preparation method, medicinal composition and use thereof
CN103664903A (en) Benzoazepine compounds and preparation method and application thereof
KR20180100705A (en) The novel compounds and their pharmaceutically acceptable salts
CN104211687B (en) The indole of heterocyclic substituted naphthalene ketone derivant, its preparation method, medical composition and its use
CA3188077A1 (en) Egfr inhibitor, preparation method therefor and application 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
RJ01 Rejection of invention patent application after publication
RJ01 Rejection of invention patent application after publication

Application publication date: 20141224