CN105481858B - A kind of nitrogenous fused heterocyclic compound, preparation method, composition and application - Google Patents

A kind of nitrogenous fused heterocyclic compound, preparation method, composition and application Download PDF

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CN105481858B
CN105481858B CN201410534512.6A CN201410534512A CN105481858B CN 105481858 B CN105481858 B CN 105481858B CN 201410534512 A CN201410534512 A CN 201410534512A CN 105481858 B CN105481858 B CN 105481858B
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CN105481858A (en
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万惠新
许志勇
石辰
李春丽
许振民
夏广新
马珂
李玉峰
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Shanghai Pharmaceutical Group (Benxi) north pharmaceutical Co.,Ltd.
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Shanghai Pharmaceuticals Holding Co Ltd
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    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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    • Y02P20/00—Technologies relating to chemical industry
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    • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Abstract

The invention discloses a kind of nitrogenous fused heterocyclic compounds shown in formula I, its pharmaceutically acceptable salt, or their enantiomter, diastereoisomer, tautomer, solvate, metabolic precursor thereof or prodrug, or pharmaceutical composition and its application containing it.Nitrogenous fused heterocyclic compound of the invention has preferable anti-tumor activity.

Description

A kind of nitrogenous fused heterocyclic compound, preparation method, composition and application
Technical field
The present invention is specifically related to a kind of nitrogenous fused heterocyclic compound, preparation method, composition and application.
Background technique
Cyclin-dependent kinase (cyclin-dependent kinase, CDK) is a kind of serine (Ser)/Soviet Union's ammonia Sour (Thr) kinases, the CDK-cyclin formed as intracellular important signal transducers and period plain (cyclin) are compound Object participates in the growth of cell, proliferation, suspend mode or enters apoptosis.Cyclin plays in regulation of cell proliferation Important function, in tumour cell, G1 period phase element and CDK it is not normal be it is most frequent, these changes may be participated in there are many mechanism Change.These variations cause tumour often through the activation of oncogene and the silencing of tumor suppressor gene.Malignant cell is logical It crosses heredity and gradually life system influences the expression of cell cycle regulating protein, cause the overexpression and the expression of CDK inhibitor of period element Loss, consequent is CDK active out of control.
Cell cycle checkpoint (cell cycle checkpoint) refers to the speed limit position in control cell generation cycle Point is responsible for determining the integrality of DNA synthesis, monitoring DNA replication dna, DNA damage reparation and resistance before DNA replication dna and mitosis Disconnected cell enters m period, the accurate progress for adjusting the cell cycle, to prevent that mistake occurs in proliferating cycle.Cell response DNA damage makes Cell cycle checkpoint be activated, and leads to Cell-Cycle Blockade, to repair the DNA of damage, or passes through cell Apoptosis or the mode inducing cell death for terminating growth.Cell-Cycle Blockade frequently occurs in G1/S or G2/M intersection, cell Cycle detection point regulatory molecule is also the novel targets of anti-tumor drug.
Early stage Cycle Regulation agent such as CDK inhibitor, Auro inhibitor etc. generally lead to poison because specificity is not high Side effect is very big, and in recent years, selective CDK inhibitor etc. is reported in succession and enters clinical trial.Therefore, it finds and finds and is high The specific cell periodic adjustment agent of effect, low toxicity is a popular domain of anti-tumor drug research.
Summary of the invention
The present invention is to overcome existing Cycle Regulation agent in the prior art due to specific not high, poor selectivity Etc. reasons and show the defects of biggish clinical side effects, although the bad defect of the high activity of selectivity, and mention Supplied a kind of nitrogenous fused heterocyclic compound, its pharmaceutically acceptable salt and their enantiomter, diastereoisomer, Tautomer, solvate, metabolic precursor thereof or prodrug, preparation method, pharmaceutical composition and application containing it.Of the invention Nitrogenous fused heterocyclic compound has preferable anti-tumor activity.
The present invention provides a kind of shown in formula I nitrogenous fused heterocyclic compound, its pharmaceutically acceptable salt or it Enantiomter, diastereoisomer, tautomer, solvate, metabolic precursor thereof or prodrug；
Wherein, R1For hydrogen, halogen (preferably chlorine), C1~C6Alkyl (preferably methyl), C1~C6Alkoxy (preferably methoxyl group) Or C3~C6Naphthenic base (preferably cyclopropyl)；
R2For hydrogen, halogen (preferably bromine), C1~C6The C that alkyl, hydroxyl replace1~C6Alkyl (preferably CH3-CH (OH) -), C1 ~C6Alkyl oxygen-carbonyl-(preferably ethoxy-carbonyl), carboxyl, cyano, aldehyde radical (preferably carboxaldehyde radicals), C1~C6Acyl group (preferably second Acyl group),Or substituted or unsubstituted C3~C12Heteroaryl；DescribedIn R5And R6Independently be hydrogen, C1~C6Alkyl (preferably methyl) or C1~C6Alkoxy (preferably methoxyl group), alternatively, the R5And R6Atom connect cyclization, WithIn N atom form unitary azepine C2~C6Naphthenic base (preferably unitary aziridinyl)；The substitution or not Substituted C3~C12Hetero atom in heteroaryl is one of O, N and S or a variety of (preferably N)；The heteroatomic number For 1~4 (preferably 2)；The substituted C3~C12Being substituted by by halogen (preferably F), C in heteroaryl1~C6Alkoxy (preferably methoxyl group), trifluoromethyl, hydroxyl and C1~C6One of alkyl (preferably methyl) is a variety of replaced；
R3For hydrogen or C1~C6Alkyl (preferably methyl)；
R4For substituted or unsubstituted C1~C6Alkyl (preferably substituted or unsubstituted methyl, substituted or unsubstituted positive third Base, substituted or unsubstituted tert-butyl, substituted or unsubstituted isopropyl or substituted or unsubstituted sec-butyl), replace or not ReplaceSubstituted or unsubstituted C3~C6Naphthenic base (preferably substituted or unsubstituted C4、C5Or C6 Naphthenic base), C1~C6Alkyl oxygen-carbonyl-(preferably ethoxy-carbonyl -), substituted or unsubstituted C3~C12Aryl (preferably benzene Base), substituted or unsubstituted C3~C12Heteroaryl or substituted or unsubstituted C2~C6Heterocyclylalkyl；Described h, i and the j is respectively It is independently (the h preferably 2 or 3, the i preferably 1 or 2, the j preferably 2 or 3) of any integer in 1-5；Described Substituted or unsubstituted C3~C12Hetero atom in heteroaryl is one of O, N and S or a variety of (preferably N), the substitution Or unsubstituted C3~C12Heteroatomic number in heteroaryl is 1~4 (preferably 1)；Described is substituted or unsubstituted C2~C6Hetero atom in Heterocyclylalkyl is one of O, N and S or a variety of (preferably N or O), and described is substituted or unsubstituted C2~C6Heteroatomic number in Heterocyclylalkyl is 1~4 (preferably 1)；Described is substituted by by halogen (preferably F), C1~ C6Alkoxy (preferably methoxyl group), trifluoromethyl, hydroxyl and C1~C6One of alkyl (preferably methyl) is a variety of replaced；
Ar is W-L1-L2-Y-；
Wherein:
Y is selected from substituted or unsubstituted C3~C12Aryl, substituted or unsubstituted C3~C12Heteroaryl or substitution do not take " the C in generation3~C6Heterocycloalkenyl and C3~C12Aryl " or substituted or unsubstituted " C3~C6Heterocycle heterocycloalkenyl and C3~C12It is miscellaneous Aryl "；Substituted or unsubstituted " the C3~C6Heterocycloalkenyl and C3~C12Aryl " and substituted or unsubstituted " C3~C6 Heterocycle heterocycloalkenyl and C3~C12The hetero atom in heterocycloalkenyl in heteroaryl " it is independent for one of N, O or S or A variety of (preferred N independent), heteroatomic number independent is 1~4 (preferably 1) independent；Described takes Generation or unsubstituted " C3~C6Heterocycloalkenyl and C3~C12Aryl " and substituted or unsubstituted " C3~C6Heterocycle heterocycloalkenyl and C3 ~C12The number of the alkenyl in heterocycloalkenyl in heteroaryl " independent is 1；The substituted or unsubstituted C3~C12 Heteroaryl and substituted or unsubstituted " C3~C6Heterocycle heterocycloalkenyl and C3~C12The hetero atom in heteroaryl in heteroaryl " is each From being independently one of O, N and S or a variety of (preferred N independent), heteroatomic number independent is 1~4 (preferably 1) independent；Described is substituted by by halogen (preferably fluorine) or C1~C6Alkyl (preferably methyl) is replaced；
L2Selected from-(CH2)r,-O- ,-S- ,-C (=O)-or substituted or unsubstituted amino, r is any between 0~5 Integer (preferably 0 or 1)；Described is substituted by by C1~C3Replaced alkyl；
L1Selected from-(CH2)n, substituted or unsubstituted C3~C12Naphthenic base (preferably substituted or unsubstituted cyclohexyl) takes Generation or unsubstituted C3~C12Heterocyclylalkyl；N is the arbitrary integer (preferably 0 or 1) between 0~5；Described is substituted by by C1~ C6Replaced alkyl (preferably methyl) or halogen；The substituted or unsubstituted C3~C12Hetero atom in Heterocyclylalkyl be O, One of N and S or a variety of (preferably N), heteroatomic number are any integer (preferably 1 or 2) in 1~4；
W be hydrogen,Halogen (preferably bromine), hydroxyl, aldehyde radical,It is substituted or unsubstituted Amino, substituted or unsubstituted C1~C6Alkyl (preferably substituted or unsubstituted methyl, alternatively, substituted or unsubstituted ethyl) Or substituted or unsubstituted C1~C6Acyl group；DescribedIn X2For O, S orDescribedIn s be any integer (preferably s is 1 or 2) in 1~5, it is describedIn T be any integer (preferably t is 0 or 1) in 0~5, it is describedIn m be 0~5 in any Integer (preferably m is 1), it is describedIn X1For N, O ,-C (=O)-NH- orThe substitution Amino refer on nitrogen only one hydrogen be substituted or nitrogen on two hydrogen be substituted, when substituted amino refers to that two hydrogen are substituted on nitrogen When, two substituent groups are identical or different；Described is substituted by by C1~C6Alkyl (preferably methyl), C1~C6Alkyl sulphonyl (preferably mesyl), C1~C6Alkoxy (preferably methoxyl group), C1~C6Alkylthio group (preferably methyl mercapto), C3~C6Naphthenic base, The C that halogen, hydroxyl, amino, hydroxyl replace1~C6(ethyl, the hydroxyl that methyl, the hydroxyl that preferably hydroxyl replaces replace replace alkyl Propyl), the N of 2-12 carbon atom, the N of N- dialkyl amido, 2-12 carbon atom, N- dialkyl amido-alkyl-, 2-12 Alkyl-, the C of the alkoxy substitution of a carbon atom3~C6Naphthenic base (preferably cyclopropyl) and C2~C6One of Heterocyclylalkyl or It is a variety of replaced；The C2~C6Hetero atom in Heterocyclylalkyl is one of N, O and S or a variety of, and heteroatomic number is Any integer in 1~4.
As the R2For substituted or unsubstituted C3~C12When heteroaryl, the substituted or unsubstituted C3~C12It is miscellaneous Aryl is preferredDescribedIn A be oxygen, NH or substituted amino, taking in the substituted amino On behalf of by C1~C6Alkyl replaces (preferably being replaced by methyl).
As the R4For substituted or unsubstituted C3~C12When heteroaryl, the substituted or unsubstituted C3~C12It is miscellaneous Aryl is preferred
As the R4For substituted or unsubstituted C2~C6When Heterocyclylalkyl, the substituted or unsubstituted C2~C6It is miscellaneous C in naphthenic base2~C6Heterocyclylalkyl is preferred
As the R2It is substituted or unsubstitutedWhen, described is substituted or unsubstitutedIt is preferred that
The R1It is preferred that hydrogen or methyl.
The R2It is preferred that acetyl group, cyano, bromine ,-CH (OH) CH3、C1~C6Alkyl oxygen-carbonyl-or
The R3It is preferred that hydrogen or methyl.
The R4It is preferred that methyl, tert-butyl, isopropyl, cyclopenta, cyclobutyl, phenyl, n-propyl or cyclohexyl.
The Y is preferred The Y preferably connects L in left end2, such as when Y isWhen Connection type be
The L1It is preferred that OrThe L1Group preferably left end connect W, right end connect L2, such as work as L1ForWhen Connection type is
The Ar is preferred
Preferably, in the Formulas I compound represented, R1For hydrogen or methyl, R2For acetyl group, cyano, bromine ,-CH (OH) CH3、C1~C6Alkyl oxygen-carbonyl-orR3For hydrogen, R4For methyl, tert-butyl, isopropyl, cyclopenta, cyclobutyl, Phenyl, n-propyl or cyclohexyl, Ar areMore preferably, in the Formulas I compound represented, R1For Methyl, R2For acetyl group or cyano, R3For hydrogen, R4For methyl, cyclopenta, isopropyl or tert-butyl, Ar isIt is optimal, in the Formulas I compound represented, R1For methyl, R2For acetyl group or cyano, R3 For hydrogen, R4For cyclopenta or isopropyl, Ar is
In the present invention, the pharmaceutically acceptable salt of the Formulas I compound represented is preferably chemical combination shown in Formulas I Object and organic acid or inorganic acid are formed by salt or Formulas I compound represented and organic base or inorganic base is formed by salt.This In invention, the solvate of the Formulas I compound represented be preferably hydrate or Formulas I compound represented with it is organic molten Dosage form at solvate.
The organic acid can for this field it is conventional can at the various organic acids of salt, preferably methanesulfonic acid, p-methyl benzenesulfonic acid, Maleic acid, fumaric acid, citric acid, tartaric acid, malic acid, lactic acid, formic acid, acetic acid, propionic acid, trifluoroacetic acid, oxalic acid, succinic acid, Benzoic acid, isethionic acid, naphthalene sulfonic acids or salicylic acid etc..
The inorganic acid can for this field it is conventional can be at the various inorganic acids of salt, preferably hydrochloric acid, sulfuric acid or phosphoric acid etc..
The organic base can for this field it is conventional can be at the various organic bases of salt, preferably pyridines, imidazoles, pyrazine One of organic bases such as class, indoles, fast quinoline class, tertiary amines, phenyl amines are a variety of.The tertiary amines organic base preferably three Ethamine and/or N, N- diisopropylethylamine.The preferred N of phenyl amines organic base, accelerine.The pyridines have One of the preferred pyridine of machine alkali, picoline, 4-dimethylaminopyridine and 2- methyl -5- ethylpyridine are a variety of.
The inorganic base can for this field it is conventional can be at the various inorganic bases of salt, preferred as alkali hydride, alkali gold The hydroxide of category, the alkoxide of alkali metal, potassium carbonate, sodium carbonate, lithium carbonate, cesium carbonate, saleratus and sodium bicarbonate One of or it is a variety of.The preferred sodium hydride of the alkali metal hydride and/or hydrofining.The hydroxide of the alkali metal It is preferred that one of sodium hydroxide, potassium hydroxide and lithium hydroxide or a variety of.The preferred methanol of the alkoxide of the alkali metal One of sodium, sodium ethoxide, potassium tert-butoxide and sodium tert-butoxide are a variety of.
In the present invention, the nitrogenous fused heterocyclic compound shown in formula I is preferably following any compound:
In the present invention, " x~y carbon atom ... base " (wherein x and y refer to number) being previously mentioned refers to the total of the group Carbon number is that " x~y " is a.
For Formulas I compound represented of the present invention, organic synthesis and field of medicinal chemistry and technology people can be used A variety of method preparations known to member.Methodologies described hereinafter can be used, with synthetic method known to organic chemistry filed Together or variation on it understood by one of ordinary skill in the art synthesizes the compounds of this invention.
The preparation method of Formulas I compound represented of the present invention can use following from the starting material being easily obtained General approach and process prepare the compound of the present invention.It will be appreciated that when providing typical or preferred technological operation When condition (that is, reaction temperature, time, the molar ratio of reactant, solvent, pressure, etc.)；Other technological operations can also be used Condition, unless otherwise indicated.Optimum reaction condition can change with specific reactant used or solvent, but these conditions can To be determined by those skilled in the art by routine optimization process.
The method of Formulas I compound represented of the present invention described herein can be according to known in the art any suitable Suitable method is monitored.For example, nuclear magnetic resonance, infrared spectroscopy, spectrophotometric or mass spectral analysis, HPLC or thin-layer chromatography are supervised Control the generation of product.
The preparation of compound can be related to the protection and deprotection of multiple chemical groups.For the need protected and be deprotected It wants, and the selection of protecting group appropriate can readily be determined by those skilled in the art, the chemical mistake of protecting group Journey is in such as Greene et al., Protective Groups in Organic Synthesis, the second edition, Wiley&Sons, 1991, in find, be incorporated herein by reference herein in whole form.
Method reaction described herein can be carried out in suitable solvent, organic synthesis field technical staff is easily Select the solvent.Suitable solvent substantially not with raw material, intermediate or product it is described react carry out at a temperature of from it is anti- It answers, the temperature that the temperature that the reaction carries out can change in the boiling point temperature range from the freezing point of solvent to solvent, Given reaction can carry out in the mixture of a kind of solvent or multi-solvents.According to specific reaction step, Ke Yixuan Select the solvent for being suitable for specific reaction step.
In general, three kinds of reaction routes and technique at least as described below, which can be used, prepares the compounds of this invention, but not The reagent and solvent being limited in reaction condition.
Therefore, the present invention also provides the preparation methods of the Formulas I compound represented, are following either method:
Method one:
The preparation method of the Formulas I compound represented includes the following steps: to take off Formula II compound represented Except the reaction of protecting group, Formulas I compound represented is obtained, the PG in Formula II compound represented is the protecting group；
Wherein, described Ar, R1、R2、R3、R4As described above.
PG in the Formula II compound represented can be hydroxyl protection base, amino protecting group, carboxyl-protecting group or carbonyl The various conventional protecting groups in this fields such as base protecting group, in order on the Ar group and Formula II compound represented Amino can play the role of protecting certain reactive groups on Ar not react when being reacted and being linked together.
PG in the Formula II compound represented can for tertiary butyl oxycarbonyl, 9-fluorenylmethyloxycarbonyl, benzyloxycarbonyl group, Acetyl group, allyl, allyl carbonyl etc..
The condition of the reaction of the deprotection base can be the conventional removing condition of the various protecting groups in this field, such as hydrolyze Condition, the condition of aminolysis reaction, condition of hydrogenation of reaction etc..
When the W isAnd the X2When for NH, the PG is tertiary butyl oxycarbonyl, i.e. Formula II -1 Boc group on shown compound.
When the W isThe X2When for NH, and when the PG is tertiary butyl oxycarbonyl, preferably, The preparation method of the Formulas I compound represented includes the following steps: -1 compound represented of Formula II carrying out deprotection The Formulas I compound represented is made in the reaction of base；The L1、L2、Y、R1、R2、R3、R4As described above；
In the reaction of the deprotection base when the PG is tertiary butyl oxycarbonyl, the Y is preferred
In the reaction of the deprotection base when the PG is tertiary butyl oxycarbonyl, the L2It is preferred that- (CH2) r-, and r is 0.
In the reaction of the deprotection base when the PG is tertiary butyl oxycarbonyl, the L1It is preferred that- (CH2) n-, and n is 0.
In the reaction of the deprotection base when the PG is tertiary butyl oxycarbonyl, the R1It is preferred that methyl Or hydrogen.
In the reaction of the deprotection base when the PG is tertiary butyl oxycarbonyl, the R2It is preferred that hydrogen, Methyl, halogen, cyano, acetyl group ,-CO2CH2CH3、-CO2-N(CH3)-OCH3、-CO2NH2Or-CO2-N(CH3)2。
In the reaction of the deprotection base when the PG is tertiary butyl oxycarbonyl, the R3It is preferred that H or Methyl.
In the reaction of the deprotection base when the PG is tertiary butyl oxycarbonyl, the R4It is preferred that first Base, phenyl, cyclobutane base, pyridyl group ,-CO2CH2CH3, propyl, cyclopenta, butyl or cyclohexyl.
The process of the reaction of the deprotection base when the PG is tertiary butyl oxycarbonyl can use ability Traditional test methods (such as TLC, HPLC, GC or NMR) in domain are monitored, generally with the disappearance of such as -1 compound represented of Formula II Shi Zuowei reaction end.
The reaction of the described deprotection base when the PG is tertiary butyl oxycarbonyl, after, preferably, also It can further include the operation of post-processing.The method and condition of the post-processing can be conventional for the such post-reaction treatment in this field Method and condition, preferably: reaction system is washed, is dried, is filtered, solvent evaporated, then column chromatograph； Alternatively, solvent, washing, filtering is evaporated off in reaction system；Alternatively, solvent, thin-layer chromatography is evaporated off in reaction system.
In the reaction of the deprotection base when the PG is tertiary butyl oxycarbonyl, preferably, also further Include the following steps: in organic solvent, -1 compound represented of Formula V and 3 compound represented of formula are subjected to annulation, generated - 2 compound represented of formula III；Then -2 compound represented of formula III is subjected to oxidation reaction, changed shown in production III-1 Close object；- 1 compound represented of formula III is reacted with 2 compound represented of formula, is changed shown in the obtained Formula II -1 Close object；
The L1、L2、Y、R1、R2、R3、R4As described above；
The X is halogen, preferably bromine, fluorine, chlorine, iodine, more preferable bromine.
Wherein, the condition of the method for each step reaction in reaction route can be according to the method for these reactions of this field Normal condition carry out.
Organic solvent in the annulation can be the various conventional organic solvents of the such reaction in this field, preferably Methanol, ethyl alcohol, isopropanol, N,N-dimethylformamide, DMAC N,N' dimethyl acetamide, N-Methyl pyrrolidone, dioxane, four One of hydrogen furans and methylene chloride are a variety of.
Method two:
The preparation method of the Formulas I compound represented includes the following steps: -3 compound represented of formula III and formula 4 compounds represented carry out substitution reaction or transition metal-catalyzed coupling reaction, generate chemical combination shown in the Formulas I Object；LG group in -3 compound represented of formula III is leaving group；
The R1、R2、R3、R4As described above with Ar.
The leaving group can be the such various conventional leaving groups of reaction in this field, preferably halogen, methyl mercapto, first Sulfuryl, OH, amino or sulphonic acid ester.
The condition of the substitution reaction or the transition metal-catalyzed coupling reaction can be such anti-for this field The various normal conditions answered.The substitution reaction can by heat or pressurize or acid-base catalysis etc. under the conditions of carry out.
In the substitution reaction of the method two, the R1It is preferred that methyl or hydrogen.The R2It is preferred that hydrogen, methyl, halogen Element, cyano, acetyl group ,-CO2CH2CH3、-CO2-N(CH3)-OCH3、-CO2NH2Or-CO2-N(CH3)2.The R3It is preferred that H or Methyl.The R4It is preferred that methyl, phenyl, cyclobutane base, pyridyl group ,-CO2CH2CH3, propyl, cyclopenta, butyl or hexamethylene Base.
In the method two of the preparation method of the Formulas I compound represented, preferably, still further comprising following step It is rapid: -2 compound represented of Formula V and 3 compound represented of formula are subjected to annulation, production III-3 compound represented； LG group in compound shown in Formula V -2 is leaving group；The leaving group is as described above；
The R1、R2、R3、R4As described above with Ar.
The condition of the annulation is as described above.
In method two, as the R2When for halogen, -2 compound represented of Formula V is to change shown in Formula V -2-1 Close object.As the R2When for acetyl group, -2 compound represented of Formula V is Formula V -2-2 compound represented.When described R2When for halogen, -3 compound represented of formula III is formula III -3-1 compound represented.As the R2For acetyl When base, -3 compound represented of formula III is formula III -3-2 compound represented.
In the method two of the preparation method of the Formulas I compound represented, preferably, still further comprising following step It is rapid: the Formula V -2-1 compound represented can be generated by transition metal-catalyzed lower acylation reaction described in V-2-2 Compound represented.Formula III -3-1 the compound represented can give birth to by transition metal-catalyzed lower acylation reaction At the III-3-2 compound represented.As the R2When for halogen, compound described in the Formulas I can pass through Acylation reaction under transition metal-catalyzed, which generates, works as the R2Compound described in Formulas I when for acetyl group.
The R1、R3、R4, X, LG and Ar as described above.
Wherein, the condition of the method for each step reaction in reaction route can be according to the method for these reactions of this field Normal condition carry out.
In the method two of the preparation method of the Formulas I compound represented, preferably, still further comprising following step It is rapid: as the R2For C1~C6Alkyl oxygen-carbonyl-when, the Formulas I compound represented, in water and organic solvent (such as first Alcohol, ethyl alcohol, isopropanol, tetrahydrofuran, dioxane) in the mixed solvent, (preferably under alkaline condition by hydrolysis Hydrolysis, more preferably in sodium hydroxide, potassium hydroxide, the hydrolysis under the alkaline condition of lithium hydroxide) it can be generated and work as The R2Formulas I compound represented when for carboxyl.As the R2When for carboxyl, the Formulas I compound represented and N, It is available as the R that O- dimethyl hydroxylamine carries out condensation reaction2ForWhen Formulas I compound represented.Work as institute The R stated2ForWhen, the Formulas I compound represented and grignard reagent or lithium reagent, in anhydrous organic solvent In (such as tetrahydrofuran, methylene chloride), generation can be reacted as the R2Formulas I compound represented when for acetyl group.
The R1、R3、R4, X and Ar as described above.
Wherein, the condition of the method for each step reaction in reaction route can be according to the method for these reactions of this field Normal condition carry out.
In the method two of the preparation method of the Formulas I compound represented, preferably, still further comprising following step It is rapid: as the R2When for cyano, the Formulas I compound represented, in anhydrous solvent, with grignard reagent or lithium metal Addition reaction occurs for reagent, and hydrolysis occurs again for obtained product, last available R2For C1~C6The formula of acyl group I compound represented；
The R1、R3、R4, X and Ar as described above；The R14For C1~C6Alkyl.
Wherein, the condition of the method for each step reaction in reaction route can be according to the method for these reactions of this field Normal condition carry out.
In the method two of the preparation method of the Formulas I compound represented, preferably, still further comprising following step It is rapid: as the R2When for cyano, the Formulas I compound represented is in organic solvent, anti-with hydrazine, azanol or amine reagent With acyl chlorides, acid or acid anhydrides the available R of ring closure reaction occurs for Ying Hou, obtained product again2For substituted or unsubstituted C3~C12 Formulas I compound represented when heteroaryl；
The R1、R3、R4, X and Ar as described above；The R15For substituted or unsubstituted C3~C12Heteroaryl；Institute The substituted C stated3~C12Being substituted by by halogen (preferably F), C in heteroaryl1~C6Alkoxy (preferably methoxyl group), fluoroform Base, hydroxyl and C1~C6One of alkyl (preferably methyl) is a variety of replaced.
Wherein, the condition of the method for each step reaction in reaction route can be according to the method for these reactions of this field Normal condition carry out.
In the method two of the preparation method of the Formulas I compound represented, preferably, still further comprising following step It is rapid: by -1 compound represented of Formula IV, (preferably tetrahydrofuran, methylene chloride or N, N- dimethyl formyl in anhydrous solvent Amine), under the action of alkali (preferably sodium hydride, potassium hydroxide, sodium tert-butoxide or sodium methoxide), with cyano phosphite ester or its etc. Effect object is reacted, production V-2-4 compound represented；
The R1And LG is as described above.
The condition of the method for the reaction of the production V-2-4 compound represented can be according to the such reaction in this field Method normal condition carry out.
In the method two of the preparation method of the Formulas I compound represented, preferably, still further comprising following step Rapid: by Formula V -2-4 compound represented, (preferably bromine, N- bromo-succinimide, C5H6Br2N2O2 cross bromine pyrrole with halogenating agent Pyridine salt etc.) reaction, available Formula V -2-5 compound represented；
The R1, X and LG as described above.
The condition of the method for the reaction of the production V-2-5 compound represented can be according to the such reaction in this field Method normal condition carry out.
In the method two of the preparation method of the Formulas I compound represented, preferably, still further comprising following step It is rapid: by -2 compound represented of Formula IV, with the cyano compounds such as cyan-acetic ester, cyano propanone or the third dicyan in alkalinity or nothing Under solvent condition, condensation reaction is carried out, Formula V -2-6 compound represented is obtained；
The R1And LG is as described above.
The condition of the method for the reaction of the production V-2-6 compound represented can be according to the such reaction in this field Method normal condition carry out.
In the method two of the preparation method of the Formulas I compound represented, preferably, still further comprising following step It is rapid: in organic solvent (preferably n,N-Dimethylformamide, N-Methyl pyrrolidone or alcohols solvent), will to change shown in Formula IV -3 Object is closed, is performed under heating conditions and reacts with amino-compound, available -4 compound represented of Formula IV；It will be shown in Formula IV -4 Compound in a solvent with halogenating agent (preferably bromine, N- bromo-succinimide, C5H6Br2N2O2 cross bromopyridine salt etc.) It is reacted, available -5 compound represented of Formula IV；Functional group conversions by -5 compound represented of Formula IV by routine, Available -6 compound represented of Formula IV；In a heated condition by -6 compound represented of Formula IV, with urea, guanidine, thiocarbamide etc. Ring closure reaction occurs for reagent, carries out the operation such as functional group conversions or deprotection base again in the case of necessary, available Formula V -2-7 compound represented；
The R1, X and LG as described above；The R16For C1~C6Alkyl.
Wherein, the condition of the method for each step reaction in reaction route can be according to the method for these reactions of this field Normal condition carry out.
Method three:
As the R2When for halogen or acetyl group, the preparation method of the Formulas I compound represented includes following step It is rapid: by formula III -3-4 compound represented and 5 compound represented of formula in transition metal-catalyzed lower generation coupling reaction, to generate The Formulas I compound represented；
The R1、R3、R4, X and Ar as described above.
The condition of coupling reaction in the method three can be the various normal conditions of the such reaction in this field.
The present invention also provides containing nitrogenous fused heterocyclic compound, its pharmaceutically acceptable salt shown in above-mentioned Formulas I, or Person their enantiomter, diastereoisomer, tautomer, solvate, metabolic precursor thereof or prodrug pharmaceutical composition Object.
Wherein, the pharmaceutical composition nitrogenous fused heterocyclic compound as shown in one or more Formulas I, it pharmaceutically may be used The salt of receiving or their enantiomter, diastereoisomer, tautomer, solvate, metabolic precursor thereof or prodrug It is formed at least one pharmaceutic adjuvant.The selection of pharmaceutic adjuvant is different because of administration method and action character, generally can be filler, Diluent, adhesive, wetting agent, disintegrating agent, lubricant, emulsifier or suspending agent etc..
Pharmaceutical composition of the invention can by oral, injection (vein, muscle, in subcutaneous and coronary artery), it is sublingual, Buccal, per rectum, per urethra, Via vagina, intranasal, sucking or topic route application, optimization approach is oral.
The present invention also provides above-mentioned nitrogenous fused heterocyclic compounds shown in formula I, its pharmaceutically acceptable salt, or Their enantiomter, diastereoisomer, tautomer, solvate, metabolic precursor thereof or prodrug preparation prevention or Treat the application on the drug of the extremely relevant disease of cell cycle regulating.
The extremely relevant disease of cell cycle regulating of the disease preferred cycle protein dependent kinase regulation.
The cyclin-dependent kinase preferred CDK4 and/or CDK6.
The extremely relevant tumour of disease preferred cell cycle regulating.
The present invention also provides above-mentioned nitrogenous fused heterocyclic compounds shown in formula I, its pharmaceutically acceptable salt, or Their enantiomter, diastereoisomer, tautomer, solvate, metabolic precursor thereof or prodrug is in manufacturing cycle egg Application on white dependant kinase inhibitors.The cyclin-dependent kinase is as described above.
The present invention also provides above-mentioned nitrogenous fused heterocyclic compounds shown in formula I, its pharmaceutically acceptable salt, or Their enantiomter, diastereoisomer, tautomer, solvate, metabolic precursor thereof or prodrug has swollen in preparation Application on the drug of oncocyte inhibitory activity.
The preferred cancer cell of the tumour cell.The preferred breast cancer cell of the cancer cell.The breast cancer cell It is preferred that breast cancer cell MCF-7.
On the basis of common knowledge of the art, above-mentioned each optimum condition, can any combination to get each preferable reality of the present invention Example.
The reagents and materials used in the present invention are commercially available.
The positive effect of the present invention is that: the present invention provides a kind of new structural nitrogenous fused heterocyclic compound, Its pharmacodynamics and pharmacokinetics embodiment test data are shown: it has choosing to cyclin-dependent kinase in molecular level Selecting property inhibiting effect, and specificity is high, activity is strong；In cellular level and animal level to active with cyclin-dependent kinase Relevant tumour cell also has significant Inhibit proliferaton effect；The compounds of this invention stablizes the hepatomicrosome of people, mouse etc. Property it is good, metabolic enzyme inhibits without obvious, and rats and mice body absorption property is good, and bioavilability is high, has preferable druggability.
Specific embodiment
The present invention is further illustrated below by the mode of embodiment, but does not therefore limit the present invention to the reality It applies among a range.In the following examples, the experimental methods for specific conditions are not specified, according to conventional methods and conditions, or according to quotient The selection of product specification.
The structure of all compounds of the present invention can by nuclear magnetic resonance (1H NMR) and/or Mass Spectrometer Method (MS) identification.1H Nmr chemical is displaced (δ) with PPM record (10-6).NMR is carried out by Bruker AVANCE-400 spectrometer.
LC-MS measures by Agilent 1200HPLC/6120 mass spectrograph
Thin layer silica gel plate is Yantai Huanghai Sea HSGF254 or Qingdao GF254 silica gel plate.Column chromatography generally uses the Yantai Huanghai Sea 200-300 mesh silica gel is as carrier.
Embodiment 1 ' (preparation of intermediate):
Step 1:
Bis- chloro- 4- methyl 3-cyanopyridine (5g, 26.7mmol) (such as formula 1-a compound represented) of 2,6- is placed in seal pipe In, tert-butylamine (3.12mL, 29.4mmol) and DIPEA (5.3mL, 32.1mmol) are added thereto, is heated to 120 DEG C of reactions 18h.Reaction solution is cooled to room temperature after reaction, and reaction solution is diluted with ethyl acetate, water washing, and organic phase anhydrous sodium sulfate is dry Dry, silica gel column chromatography purifies (PE:EA=80:20 elution), obtains white solid 1.77g (such as formula 1-b compound represented).1H- NMR(400MHz,CDCl3): δ=6.15 (1H, s), 5.00 (1H, s), 2.37 (3H, s), 1.44 (9H, s) .LC-MS m/z:(M +H)+=224.10.
Step 2:
By (the change as shown in formula 1-b of the chloro- 4- methyl 3-cyanopyridine (1.77g, 7.94mmol) of 6- (tert-butyl amino) -2- Close object) it is dissolved in 40mL glacial acetic acid, the bromine water (0.49ml, 9.53mmol) being dissolved in 5mL glacial acetic acid, room temperature reaction are added dropwise thereto 1h, solvent evaporated, residue are dissolved in ethyl acetate, saturated sodium bicarbonate aqueous solution and saturated sodium-chloride water solution after reaction Washing, organic phase are dried, filtered with anhydrous sodium sulfate, solvent evaporated, and silica gel column chromatography purifies (PE:EA=90:10 elution), are obtained White solid 1.723g (such as formula 1-c compound represented).1H-NMR(400MHz,CDCl3): δ=5.66 (1H, s), 2.51 (3H,s),1.49(9H,s).LC-MS m/z:(M+H)+=302.00.
Step 3:
By the chloro- 4- methyl 3-cyanopyridine (1g, 3.3mmol) of the bromo- 6- of 5- (tert-butyl amino) -2- (as shown in formula 1-c Compound) be dissolved in 20mL anhydrous methylene chloride, be cooled to -78 DEG C under argon gas protection, be added dropwise thereto DIBAL-H (3.3ml, 3.3mmol, 1M in THF), -78 DEG C of reaction 2h rise back room temperature after reaction, and 1N hydrochloric acid solution is added thereto by pH value It is adjusted to 2-3,0.5h is stirred at room temperature in reaction solution, and following reaction liquid is adjusted to alkalinity with 1N NaOH solution, and rear ethyl acetate extraction has Machine is mutually dried, filtered with anhydrous sodium sulfate, solvent evaporated, and silica gel column chromatography purifies (PE:EA=90:10 elution), is obtained white solid Body 0.943g (such as formula 1-d compound represented).1H-NMR(400MHz,CDCl3): δ=10.31 (1H, s), 5.81 (1H, s), 2.69(3H,s),1.52(9H,s).LC-MS m/z:(M+H)+=305.00.
Step 4:
By the chloro- 4- methyl nicotine aldehyde (261mg, 0.86mmol) of the bromo- 6- of 5- (tert-butyl amino) -2- (such as formula 1-d institute The compound shown) and potassium tert-butoxide (290mg, 2.58mmol) be dissolved in 9ml NMP, thereto be added guanidine hydrochloride (411mg, 4.3mmol), microwave heating is cooled to room temperature after reaction to 180 DEG C of reaction 30min, 9ml water, reaction solution is added thereto 10min is stirred at room temperature, the solid filtering subsequently generated is washed with a small amount of methanol, and it is dry, yellow solid 86mg is obtained (such as formula 1-e institute The compound shown).1H-NMR (400MHz, d-DMSO): δ=8.91 (1H, s), 6.87 (2H, s), 5.90 (1H, s), 2.56 (3H,s),1.50(9H,s).LC-MS m/z:(M+H)+=310.10.
Step 5:
By the bromo- N of 6-7(tert-butyl) -5- picoline simultaneously [2,3-d] pyrimidine -2,7- diamines (86mg, 0.28mmol) (such as formula 1-e compound represented) is dissolved in 2.8ml trifluoroacetic acid and 0.3ml water, is heated to 80 DEG C of reaction 18h, and reaction terminates After be cooled to room temperature, 3ml water is added in solvent evaporated thereto, after with ammonium hydroxide be adjusted to alkalinity, 10min is stirred at room temperature in reaction solution, with The solid filtering generated afterwards, is washed with water, dry, obtains yellow solid 68mg (such as formula 1-f compound represented).LC-MS m/z: (M+H)+=254.00.
Embodiment 2 ' (preparation of intermediate):
Step 1:
Sodium hydride (1.092g, 27.3mmol) is dissolved in 25ml anhydrous tetrahydro furan, is added thereto in the case where 0 DEG C of argon gas is protected Enter cyanogen methyl acid phosphate diethylester (2.66ml, 16.38mmol), react at room temperature 0.5h, after be added be dissolved in 5ml THF thereto 1- (4- amino -2- (methyl mercapto) pyrimidine -5- base) ethyl ketone (1g, 5.46mmol) (such as Formulas I -16-e compound represented), room temperature Reaction is overnight.After reaction, Shui temper is slowly added into reaction solution to go out, after be extracted with ethyl acetate, the anhydrous sulphur of organic phase Sour sodium dries, filters, solvent evaporated, and silica gel column chromatography purifies (DCM:MeOH=97:3 elution), obtains yellow solid 635mg (such as Formula 2-a compound represented).1H-NMR (400MHz, d-DMSO): δ=8.93 (1H, s), 7.33 (2H, s), 6.54 (1H, d, J =1.2Hz), 2.51 (3H, s), 2.48 (3H, d, J=0.8Hz) .LC-MS m/z:(M+H)+=207.00.
Step 2:
By 5- methyl -2- (methyl mercapto) pyrido [2,3-d] pyrimidine -7- amine (650mg, 3.14mmol) (as shown in formula 2-a Compound) be dissolved in 15ml anhydrous tetrahydro furan, be added and be dissolved in 15ml anhydrous methylene chloride thereto in the case where 0 DEG C of argon gas is protected Sulfonic acid chloride (2.54ml, 31.4mmol), react at room temperature 1h.After reaction, unsaturated carbonate hydrogen is slowly added into reaction solution Sodium Shui Rong Ye temper goes out, after be extracted with dichloromethane, organic phase is dried, filtered with anhydrous sodium sulfate, solvent evaporated, silica gel column chromatography It purifies (DCM:MeOH=97:3 elution), obtains yellow solid 313mg (such as formula 2-b compound represented).1H-NMR(400MHz, D-DMSO): δ=9.04 (1H, s), 7.67 (2H, s), 6.67 (1H, d, J=1.2Hz), 2.52 (3H, s) .LC-MS m/z:(M+ H)+=195.10.
Step 3:
By chloro--methylpyridine simultaneously [2,3-d] pyrimidine -7- amine (224mg, the 1.16mmol) (chemical combination as shown in formula 2-b Object) it is dissolved in 10ml DMF, C5H6Br2N2O2 (363mg, 1.27mmol) is added thereto, room temperature reaction is overnight.After reaction, to Slowly add water 30ml in reaction solution, the solid filtering of generation is dry, obtains the yellow solid 180mg (chemical combination as shown in formula 2-c Object).LC-MS m/z:(M+H)+=272.90.
Embodiment 3 ' (preparation of intermediate):
Step 1:
By (the change as shown in Formulas I -16-e of 1- (4- amino -2- (methyl mercapto) pyrimidine -5- base) ethyl ketone (1g, 5.46mmol) Close object) it is dissolved in ethyl acetoacetate (3ml, 27.85mmol), it is heated to 180 DEG C of reaction 3h and is cooled to room temperature after reaction, Ethyl acetate 10ml is added into reaction solution, insoluble solid filtering is dry, obtains yellow solid 1.069g (as shown in formula 3-a Compound).1H-NMR (400MHz, d-DMSO): δ=12.60 (1H, s), 9.02 (1H, s), 2.58 (3H, s), 2.44 (3H, s),2.34(3H,s).LC-MS m/z:(M+H)+=250.05.
Step 2:
By 6- acetyl group -5- methyl -2- (methyl mercapto) pyrido [2,3-d] pyrimidine -7 (8H) -one (0.353g, It 1.42mmol) is dissolved in phosphorus oxychloride (3ml, 32.77mmol) (such as formula 3-a compound represented), 110 DEG C is heated to and reacted Night is cooled to room temperature after reaction, solvent evaporated, and residue is dissolved in saturated sodium bicarbonate solution, ethyl acetate extraction, nothing Aqueous sodium persulfate dries organic phase, filtering, and solvent evaporated obtains yellow solid 400mg (such as formula 3-b compound represented), without pure Change, direct plunges into next reaction.LC-MS m/z:(M+H)+=268.00.
Step 3:
By 1- (chloro- 5- methyl -2- (methyl mercapto) pyrido [2,3-d] pyrimidine -6- base of 7-) ethyl ketone (0.4g, 1.49mmol) (such as formula 3-b compound represented) is dissolved in ammonium hydroxide (5ml), is heated to 100 DEG C of reactions overnight, after reaction, is cooled to room temperature, Solvent evaporated, residue silica gel column chromatography purify (DCM:MeOH=95:5 elution), obtain yellow solid 200mg (as shown in formula 3-c Compound).LC-MS m/z:(M+H)+=249.10.
Embodiment 4 ' (preparation of intermediate):
Step 1:
Potassium tert-butoxide (4.3g) is dispersed in anhydrous THF (150mL), ice-water bath is cooling, is slowly added under nitrogen protection Phosphonoacetate (7mL), 0 DEG C is stirred to react 1h, and 2- methyl mercapto -4- amino-5-pyrimidine formaldehyde (5g) (such as Formulas I-is added 1-d compound represented), reaction 18h is stirred at room temperature.Water is added after reaction, ethyl acetate extraction merges organic layer, does It is dry, solvent is evaporated off and obtains white solid (7g, 99%) (such as formula 4-a compound represented):1H NMR(CDCl3,400MHz):δ 1.35(t,3H),2.54(s,3H),4.28(q,2H),5.28(br,2H),6.33(d,1H),7.56(d,1H),8.29(s, 1H).ESI-MS m/z 239.7[M+H]+。
Step 2:
By (E)-ethyl -3- (4- amino -2- (methyl mercapto) pyrimidine -5- base) acryloyl group (14g) (as shown in formula 4-a Compound) it is dissolved in methanol (200ml), it is added methanol solution of sodium methylate (21mL), 80 DEG C are stirred to react 6h.It is evaporated off after reaction Solvent is added water dissolution, is adjusted to pH=8 with 2N hydrochloric acid, product is largely precipitated, and white solid (8g, 73%) is obtained by filtration (such as formula 4-b compound represented):1H NMR(DMSO-d6,400MHz):δ2.56(s,3H),6.50(d,1H),7.89(d,1H),8.84 (s,1H),12.37(br,1H).ESI-MS m/z 193.9[M+H]+。
Step 3:
Simultaneously [2,3-d] pyrimidin-7-ones (300mg) (such as formula 4-b compound represented) are dispersed in ice vinegar to 2- methylsulfanyl pyridine Sour (10ml) is added bromine (0.16mL), and reaction 48h is stirred at room temperature.Methylene chloride is added after reaction, filters, uses methanol Washing, obtains white solid (390mg, 92%) (such as formula 4-c compound represented):1H NMR(DMSO-d6,400MHz):δ 2.57(s,3H),8.48(s,1H),8.85(s,1H),12.91(br,1H).ESI-MS m/z 271.9[M+H]+。
Step 4:
By 2- methyl mercapto -6- bromopyridine, simultaneously [2,3-d] pyrimidin-7-ones (300mg) (such as formula 4-c compound represented) disperse At phosphorus oxychloride (20mL), 110 DEG C are stirred to react 16h.Solvent is evaporated off after reaction, saturated sodium bicarbonate is added under ice-water bath Solution is adjusted to pH=8, filters to obtain white solid (290mg) (such as formula 4-d compound represented), does not purify, be directly used in next Step.
Step 5:
By the bromo- 7- chloropyridine of 2- methyl mercapto -6-, simultaneously [2,3-d] pyrimidine (290mg) (such as formula 4-d compound represented) is dissolved in Reaction 72h is stirred at room temperature in 7N methanolic ammonia solution (50mL).Solvent is evaporated off after reaction, obtains crude white solid (310mg) (such as Formulas I -6-c compound represented): ESI-MS m/z271.0 [M+H]+。
Embodiment 5 ' (preparation of intermediate):
Step 1:
Cyanogen methyl acid phosphate diethylester (12.5 grams, 70.6mmol) (such as Formulas I -16-e compound represented) is dissolved in tetrahydro furan In muttering, ice bath is cooling, and sodium hydride (3 equivalent) is added in solution, stirs half an hour, 4- amido -2- methylthiopyrimidine -5- formaldehyde (2.8 grams, 15mmol) are added at one time in solution, and the reaction solution that is stirred overnight at room temperature pours into extraction in ice water and goes out, ethyl acetate extraction Take organic layer anhydrous magnesium sulfate dry, filtering is concentrated to dryness, gained grease rapid column chromatography (ethyl acetate: petroleum ether =1:1) obtain target compound (such as formula 5-a compound represented).1H-NMR,DMSO,8.95(s,1H),7.40(brs, 2H),6.56(s,1H),2.50(s,3H)。LC-MS:C9H10N4S calcd[M+H]+207.065,found 207.0
Step 2:
By 5- methyl -7- amido -2- methylsulfanyl pyridine [2,3-d] pyrimidine (1.0 grams, 4.8mmol) (as shown in formula 5-a Compound) it is dissolved in 20 milliliters of tetrahydrofurans, room temperature is in batches for bis- bromo- 5,5- Dimethyl Hydan (1.6 grams, 5.6mmol) of 1,3- It is added, gained mixed liquor is stirred overnight at room temperature, and pours into extraction in ice water and goes out, ethyl acetate extraction, organic layer is done with anhydrous magnesium sulfate Dry, filtering is concentrated to dryness, gained grease rapid column chromatography (methylene chloride: methanol=10:1) obtains target yellow solid (300 milligrams) (such as Formulas I -5-d compound represented).1H-NMR,DMSO,9.19(s,1H),2.68(s,3H),2.57(s, 3H)。LC-MS:C9H9BrN4Scalcd[M+H]+283.97,found 284.1。
Embodiment 1:
Step 1:
By (the change as shown in Formulas I -16-e of 1- (4- amino -2- (methyl mercapto) pyrimidine -5- base) ethyl ketone (5.49g, 30mmol) Close object) be dissolved in 2-methyl cellosolve (20mL), be added thereto malononitrile (5.94g, 90mmol) and ammonium acetate (11.55g, 150mmol), 130 DEG C of reaction 2h are heated to.Reaction solution is cooled to room temperature after reaction, the solid filtering of generation, with water and second Alcohol washing, it is dry, 7- amino -5- methyl -2- (methyl mercapto) pyrido [2,3-d] pyrimidine -6- formonitrile HCN is obtained (such as Formulas I -16-d institute The compound shown), it is yellow solid 4.905g.1H-NMR (400MHz, d-DMSO): δ=9.14 (1H, s), 7.84 (2H, s), 2.75(3H,s),2.54(3H,s)。LC-MS m/z:(M+H)+=232.00.
Step 2:
(such as by 7- amino -5- methyl -2- (methyl mercapto) pyrido [2,3-d] pyrimidine -6- formonitrile HCN (171mg, 0.74mmol) Formulas I -16-d compound represented) it is dissolved in dry DMF (10mL), bromacetone (0.12mL, 1.47mmol) is added thereto, adds Heat is to 80 DEG C of reaction 48h.Reaction solution Jia Ru Shui temper goes out after reaction, ethyl acetate extraction, the dry organic phase of anhydrous sodium sulfate, Filtering, solvent evaporated, residue silica gel column chromatography purify to obtain 5,8- dimethyl -2- (methyl mercapto) imidazo [1', 2':1,6] Pyrido [2,3-d] pyrimidine -6- formonitrile HCN (such as Formulas I -16-c compound represented) is yellow solid 100mg.1H-NMR (400MHz,d-DMSO):δ9.44(1H,s),8.27(1H,s),2.83(3H,s),2.71(3H,s),2.40(3H,s)。LC-MS m/z:(M+H)+=269.95.
Step 3:
By 5,8- dimethyl -2- (methyl mercapto) imidazo [1', 2':1,6] pyrido [2,3-d] pyrimidine -6- formonitrile HCN (121mg, 0.45mmol) (such as Formulas I -16-c compound represented) be dissolved in chloroform (7mL) be added thereto m-CPBA (303mg, 1.35mmol, 77%), react at room temperature 2h.Reaction solution is washed with saturated sodium bicarbonate solution after reaction, and anhydrous sodium sulfate is dry Dry organic phase, filtering, solvent evaporated obtain 5,8- dimethyl -2- (methylsulfonyl) imidazo [1', 2':1,6] pyrido [2,3- D] pyrimidine -6- formonitrile HCN (such as Formulas I -16-b compound represented), it is yellow solid, direct plunges into and react in next step.1H-NMR (400MHz,CDCl3): δ=9.53 (1H, s), 8.27 (1H, s), 3.49 (3H, s), 3.00 (3H, s), 2.57 (3H, s).LC- MS m/z:(M+H)+=301.90.
Step 4:
By 5,8- dimethyl -2- (methylsulfonyl) imidazo [1', 2':1,6] pyrido [2,3-d] pyrimidine -6- formonitrile HCN (135mg, 0.45mmol) (such as Formulas I -16-b compound represented) is dissolved in chloroform (7mL), and tert-butyl 4- (4- ammonia is added thereto Base phenyl) piperazine -1- carboxylate (125mg, 0.45mmol), heated overnight at reflux.After reaction, solvent evaporated, residue Silica gel column chromatography purifies to obtain tert-butyl 4- (4- ((6- cyano -5,8- dimethyl-imidazo [1', 2':1,6] pyrido [2,3- D] pyrimidine -2-base) amino) phenyl) piperazine -1- carboxylate (such as Formulas I -16-a compound represented) is yellow solid 28mg.1H-NMR(400MHz,CDCl3):δ9.01(1H,s),7.89(1H,s),7.60(2H,d),7.01(2H,d),3.62(4H,t,J =4.8Hz), 3.16 (4H, t, J=4.8Hz), 2.83 (3H, s), 2.51 (3H, s), 1.50 (9H, s).LC-MSm/z:(M+H)+ =499.00.
Step 5:
By tert-butyl 4- (4- ((6- cyano -5,8- dimethyl-imidazo [1', 2':1,6] pyrido [2,3-d] pyrimidine - 2- yl) amino) phenyl) piperazine -1- carboxylate (28mg, 0.056mmol) (such as Formulas I -16-a compound represented) is dissolved in dichloro 4M HCl/dioxane (0.5mL) is added thereto and reacts at room temperature 1h for methane (5mL).Reaction solution saturated carbon after reaction Sour hydrogen sodium solution washing, the dry organic phase of anhydrous sodium sulfate, filtering, solvent evaporated, residue silica gel column chromatography purify to obtain formula I-16 compound represented is Chinese red solid 21mg.1H-NMR (400MHz, d-DMSO): δ=10.54 (1H, s), 9.26 (1H, s), 8.00 (1H, s), 7.79 (2H, s), 7.04 (2H, d), 3.26 (4H, t, J=4.4Hz), 3.13 (4H, t, J= 4.8Hz),2.78(3H,s),2.40(3H,s).LC-MS m/z:(M+H) +=399.00.
Embodiment 2:
By 5,8- dimethyl -2- (methylsulfonyl) imidazo [1', 2':1,6] pyrido [2,3-d] pyrimidine -6- formonitrile HCN (such as formula I-16-b compound represented) (135mg, 0.45mmol) be dissolved in chloroform (7mL), be added thereto 4- morpholine aniline (80mg, 0.45mmol), heated overnight at reflux.After reaction, solvent evaporated, residue silica gel column chromatography purify to obtain such as -17 institute of Formulas I The compound shown is yellow solid 61mg.1H-NMR (400MHz, d-DMSO): δ=10.51 (1H, s), 9.25 (1H, s), 7.99 (1H, s), 7.77 (2H, s), 7.01 (2H, d), 3.76 (4H, t, J=4.0Hz), 3.10 (4H, t, J=4.4Hz), 2.77 (3H,s),2.39(3H,s)。LC-MS m/z:(M+H)+=400.00.
Embodiment 3:
Step 1:
By the bromo- 7- aminopyridine of 2- methyl mercapto -6- simultaneously [2,3-d] pyrimidine (such as Formulas I -6-c compound represented) crude product (310mg) is dissolved in DMF (30mL), is added bromacetone (248 μ L), 80 DEG C are stirred to react 16h.Water, acetic acid are added after reaction Ethyl ester extraction merges organic layer, dry, and solvent is evaporated off and obtains white solid 70mg, is such as Formulas I -6-b compound represented: ESI-MS m/z 309.0[M+H]+。
Step 2:
Intermediate (such as Formulas I -6-b compound represented) (230mg) obtained by upper step is dissolved in chloroform (30mL), content is added 77% metachloroperbenzoic acid (340mg) is stirred at room temperature reaction 4h, 4- (4- tert-butoxycarbonyl-piperazine) aniline is added (140mg), 70 DEG C are stirred to react 18h.Solvent is evaporated off after reaction, water is added, ethyl acetate extraction merges organic layer, does It is dry, solvent is evaporated off, column chromatographic purifying (petroleum ether: ethyl acetate=10:1) obtains yellow solid (32mg) (as shown in Formulas I -6-a Compound):1H NMR(CDCl3,400MHz):δ1.52(s,9H),2.56(s,3H),3.17(t,4H),3.63(t,4H), 7.03(d,2H),7.63(d,2H),7.64(s,1H),8.06(s,1H),8.81(s,1H).ESI-MS m/z 537.8 [M+ H]+。
Step 3:
Intermediate (such as Formulas I -6-a compound represented) (8mg) obtained by upper step is dissolved in methylene chloride (1mL), 4N is added Reaction 5h is stirred at room temperature in the dioxane solution (1mL) of HCl.Solvent is evaporated off after reaction, is washed with methanol, is obtained by filtration Red solid (6mg) (such as -6 compound represented of Formulas I):1H NMR(D2O,400MHz):δ2.32(s,3H),3.34(t,4H), 3.37(t,4H),6.61(d,2H),6.78(d,2H),7.53(s,1H),7.79(s,1H),8.56(s,1H).ESI-MS m/ z437.9[M+H]+。
Embodiment 4:
Starting material (such as Formulas I -6-a compound represented) (10mg) is dissolved in anhydrous dioxane (10mL), is added three Butyl (1- ethoxy ethylene) tin (223 μ L), bi triphenyl phosphorus palladium chloride (6.5mg), lower 110 DEG C of nitrogen protection are stirred to react 14h.Potassium fluoride dihydrate (35mg) is added after reaction, 4N HCl dioxane (0.5ml) is stirred at room temperature 2h, is evaporated off Solvent, Thin Layer Chromatography (methylene chloride: methanol=10:1) obtain yellow solid (4mg) (such as -7 compound represented of Formulas I) :1H NMR(D2O,400MHz):δ2.31(s,3H),3.04(s,3H),3.34(t,4H),3.37(t,4H),6.60(d,2H), 6.79(d,2H),7.53(s,1H),7.79(s,1H),8.56(s,1H).ESI-MS m/z 402.0[M+H]+。
Embodiment 5:
Step 1:
Cyanogen methyl acid phosphate diethylester (2.5 grams, 14mmol) (such as Formulas I -1-d compound represented) is dissolved in tetrahydrofuran In, ice bath is cooling, and potassium tert-butoxide (3 equivalent) is added in solution, stirs half an hour, 4- amido -2- methylthiopyrimidine -5- formaldehyde (2.0 grams, 11.82mmol) are added at one time in solution, and the reaction solution that is stirred overnight at room temperature pours into extraction in ice water and goes out, acetic acid second Ester extracts the anhydrous magnesium sulfate drying of organic layer, and filtering is concentrated to dryness, gained grease rapid column chromatography (methylene chloride: first Alcohol=10:1) obtain target compound (such as Formulas I -1-c compound represented).1H-NMR,DMSO,8.82(s,1H),7.93(d, 1H, J=8.8Hz), 7.44 (brs, 1H), 6.73 (d, 1H, J=8.8Hz), 2.50 (s, 3H).LC-MS:C8H8N4S calcd [M+H]+193.05,found 193.0.
Step 2:
By (the change as shown in Formulas I -1-c of 2- methyl mercapto -7- amido-pyridine [2,3-d] pyrimidine (500 milligrams, 2.6mmol) Close object) it is dissolved in 2 milliliters of DMF, it is added 1- bromacetone (500 milligrams, 3.65mmol), gained reaction solution is heated to 50 DEG C of reactions It 12 hours, pouring into ice water, solution is extracted with (50 milliliter * 3) of ethyl acetate, and gained organic layer anhydrous sodium sulfate is dry, it filters, It is concentrated to dryness, gained crude product rapid column chromatography separation (ethyl acetate: petroleum ether=1:2) obtains target compound (120 milligrams) (such as Formulas I -1-b compound represented).1H-NMR, MeOD, 9.11 (s, 1H), 8.19 (s, 1H), 7.66 (d, 1H, J= 9.6Hz), 7.44 (d, 1H, J=9.6Hz), 2.73 (s, 3H), 2.49 (s, 3H).LC-MS:C11H10N4S calcd[M+H]+ 231.06,found 230.9.
Step 3:
Methyl mercapto compound (such as Formulas I -1-b compound represented) is dissolved in 5 milliliters of methylene chloride for 120 milligrams, Metachloroperbenzoic acid (1.1 equivalent) is added in room temperature, is stirred at room temperature 3 hours, and LC-MS detection reaction is completed, saturated sodium bicarbonate Solution extracts reaction of going out, and dichloromethane layer is separated, and anhydrous sodium sulfate is dry, filters, is concentrated to dryness, gained crude product (such as Formulas I- 1-a compound represented) it is directly used in next step
Step 4:
Crude product (such as Formulas I -1-a compound represented) obtained by upper step is dissolved in 5 milliliters of chloroform, 4- morpholine is added Base-aniline (100 milligrams, 0.56mmol), gained dark solution heated overnight at reflux, for revolving to doing, gained crude product is directly quick Column chromatography, obtains 30 milligrams of target compound (such as -1 compound represented of Formulas I).1H-NMR,DMSO,10.06(brs,1H), 9.07 (s, 1H), 8.02 (s, 1H), 7.77 (d, 2H, J=8.0Hz), 7.55 (d, 1H, J=9.2Hz), 7.24 (d, 1H, J= 9.6Hz), 7.01 (d, 2H, J=9.2Hz), 3.76 (t, 4H, J=4.8Hz), 3.10 (t, 4H, J=4.8Hz), 2.16 (s, 3H)。LC-MS:C20H20N6Ocalcd[M+H]+361.17,found 360.9
Embodiment 6:
Using the method for embodiment 5, compound I-1-a is first synthesized, is then closed using the method for the last two steps of embodiment 1 At obtaining such as Formulas I -2 compound represented 1H-NMR, DMSO, 9.18 (s, 1H), 8.90 (brs, 1H), 8.13 (s, 1H), 7.82- 7.76 (m, 3H), 7.37 (d, 1H, J=8.8Hz), 7.08-7.04 (m, 2H), 3.28-3.12 (m, 8H), 2.45 (s, 3H).LC- MS:C20H21N7calcd[M+H]+360.1,found 360.0。
Embodiment 7:
Step 1:
By the bromo- 5- methyl -2- methyl mercapto -7- amido of 6--pyridine [2,3-d] pyrimidine (500 milligrams, 2.6mmol) (such as Formulas I - 5-d compound represented) it is dissolved in 5 milliliters of DMF, it is added 1- bromacetone (400 milligrams, 2.62mmol), gained reaction solution adds Heat is reacted 12 hours to 50C, is poured into ice water, and solution is extracted with (50 milliliter * 3) of ethyl acetate, gained organic layer anhydrous slufuric acid Sodium is dry, filters, is concentrated to dryness, and gained crude product rapid column chromatography separation (ethyl acetate: petroleum ether=1:2) obtains target chemical combination Object (120 milligrams) (such as Formulas I -5-c compound represented).1H-NMR,DMSO,9.37(s,1H),8.29(s,1H),2.71(s, 3H),2.69(s,3H),2.39(s,3H)。LC-MS:C12H11BrN4Scalcd[M+H]+322.9,found322.8.
Step 2:
Above-mentioned 200 milligrams of methyl mercapto compound (such as Formulas I -5-c compound represented) is dissolved in 5 milliliters of chloroform In, metachloroperbenzoic acid (1.1 equivalent) is added in room temperature, is stirred at room temperature 3 hours, and LC-MS detection reaction is completed, unsaturated carbonate hydrogen Sodium solution extracts reaction of going out, and chloroform layer is separated, and anhydrous sodium sulfate is dry, filters, is concentrated to dryness, gained crude product is (such as formula I-5-b compound represented) it is directly used in next step
Step 3:
Crude product (such as Formulas I -5-b compound represented) obtained by upper step is dissolved in 5 milliliters of chloroform, 4- (4- amine is added Base phenyl) piperidines -1- carboxylic acid tert-butyl ester (200 milligrams, 0.72mmol), gained dark solution heated overnight at reflux, revolving to dry, The direct rapid column chromatography of gained crude product obtains 80 milligrams of target compound (such as Formulas I -5-a compound represented)
1H-NMR, DMSO, 1017 (s, 1H), 9.21 (s, 1H), 8.07 (s, 1H), 7.75 (d, 2H, J=7.6Hz), 7.02 (d, 2H, J=9.2Hz), 3.39 (m, 4H), 3.07 (m, 4H), 2.69 (s, 3H), 2.40 (s, 3H), 1.46 (s, 9H).LC-MS: C26H30BrN7O2calcd[M+H]+552.16,found 522.0
Step 4:
Above-mentioned 50 milligrams of boc-protected compound (such as Formulas I -5-a compound represented) is dissolved in 10 milliliters of dichloromethane In alkane, 6N hydrochloric acid/1,4- dioxane solution is added, is stirred overnight at room temperature, is spin-dried for, water dissolution is transferred to PH7.0 with ammonium hydroxide, is revolved Dry, rapid column chromatography separates (methylene chloride: methanol=10:1) and obtains 2.2 milligrams of (chemical combination as shown in Formulas I -5 of target compound Object)
1H-NMR,DMSO,10.15(s,1H),9.18(s,1H),8.04(s,1H),7.80-7.73(m,2H),6.99- 6.88(m,2H),3.08(brs,4H),3.00(brs,4H),2.67(s,3H),2.39(s,3H).
LC-MS:C21H22BrN7calcd[M+H]+452.1,found 452.0
Embodiment 8:
Step 1:
Starting material (80 milligrams, 0.1448mmol) (such as Formulas I -5-a compound represented) is dissolved in 5 milliliters of 1,4- bis- In six ring of oxygen, it is added tributyl ethoxy ethylene tin (200 milligrams, 0.5537mmol), Pd (PPh3) 2Cl2 (50 milligrams), argon gas Protection is heated to 110 and spends night reaction solution to be cooled to room temperature, and is added potassium fluoride (500 milligrams), 6N hydrochloric acid, room temperature is then added It is stirred overnight the rotation of reaction solution obtained by and removes organic solvent, aqueous solution is transferred to PH7.0 with ammonium hydroxide, and ethyl acetate extracts, and organic layer is anhydrous Sodium sulphate is dry, filtering, is concentrated to dryness crude product rapid column chromatography (methylene chloride: methanol=10:1) obtained by and provides target chemical combination Object (11 milligrams) (such as -3 compound represented of Formulas I).1H-NMR,DMSO,10.08(s,1H),9.19(s,1H),7.97(s, 1H), 7.80 (d, 2H), 7.04 (d, 2H, J=7.2Hz), 5.79 (brs, 1H), 5.55 (brs, 1H), 3.32-3.16 (m, 8H), 2.63(s,3H),2.40(s,3H),1.49(brs,3H)。LC-MS:C23H27N7Ocalcd[M+H]+418.2,found 418.0。
Embodiment 9:
Step 1:
By 2- methyl mercapto -5- methyl -7- amino -6- pyrido [2,3-d] pyrimidinecarboxylic acid ethyl ester (500mg) (such as Formulas I -20- G compound represented) it is dissolved in DMF (20mL), it is added bromoacetophenone (715mg), 80 DEG C are stirred to react 16h.After reaction Water is added, ethyl acetate extraction merges organic layer, dry, it is evaporated off solvent, column chromatographic purifying (petroleum ether: ethyl acetate=10: 1) yellow solid (90mg) (such as Formulas I -20-f compound represented) is obtained:1H NMR(CDCl3,400MHz):δ1.54(t,3H), 2.68(s,3H),2.77(s,3H),4.63(q,2H),7.28-7.38(m,1H),7.45-7.48(m,2H),8.04(d,2H), 8.58(s,1H),9.16(s,1H).ESI-MS m/z 378.9[M+H]+。
Second, three steps:
Raw material (180mg) (such as Formulas I -20-f compound represented) is dissolved in chloroform (20mL), is added between content 77% Chloroperoxybenzoic acid (220mg) is stirred at room temperature reaction 4h, 4- (4- tert-butoxycarbonyl-piperazine) aniline (131mg) is added, 70 DEG C are stirred Mix reaction 28h.Solvent is evaporated off after reaction, water is added, methylene chloride extraction merges organic layer, and it is dry, solvent, column is evaporated off Chromatographic purifying (petroleum ether: ethyl acetate=1:1) obtains yellow solid (63mg) (such as Formulas I -20-d compound represented):1H NMR(CDCl3,400MHz):δ1.52(s,9H),2.65(s,3H),3.19(t,4H),3.65(t,4H),4.61(q,2H), 7.05(d,2H),7.36(d,2H),7.44-7.48(m,3H),7.65(d,2H),8.03(d,2H),8.42(s,1H),9.04 (s,1H).ESI-MS m/z 608.0[M+H]+。
Step 4:
Raw material (55mg) (such as Formulas I -20-d compound represented) is dissolved in methanol (5mL), 4N NaOH solution is added (2mL), 70 DEG C are stirred to react 1h.Solvent is evaporated off after reaction, is adjusted to pH=8 with 2N hydrochloric acid, product is largely precipitated, and filters To yellow solid (50mg, 96%) (such as Formulas I -20-c compound represented): ESI-MS m/z 580.0 [M+H]+。
Step 5:
Raw material (50mg) (such as Formulas I -20-c compound represented) is dissolved in DMF (5mL), HATU (49mg) and DIPEA is added N is added after reaction 0.5h is stirred at room temperature in (22 μ L), and reaction 12h is stirred at room temperature in O- dimethyl hydroxylamine hydrochloride (12.6mg).Instead Water is added after answering, ethyl acetate extraction merges organic layer, and it is dry, solvent, column chromatographic purifying (petroleum ether: acetic acid second is evaporated off Ester=1:1) obtain yellow solid (25mg) (such as Formulas I -20-b compound represented): ESI-MS m/z 623.0 [M+H]+。
Step 6:
Raw material (25mg) (such as Formulas I -20-b compound represented) is dissolved in super dry THF (2mL), is delayed under 0 DEG C of nitrogen protection It is slow that 3N methyl-magnesium-bromide diethyl ether solution (0.1mL) is added dropwise.Solvent is evaporated off after reaction, water, ethyl acetate extraction is added, merges Organic layer, it is dry, solvent is evaporated off, column chromatographic purifying (petroleum ether: ethyl acetate=1:1) obtains yellow solid (10mg) (such as formula I-20-a compound represented): ESI-MS m/z 578.0 [M+H]+。
Step 7:
4N HCl dioxane solution (1mL) is added in raw material (10mg) (such as Formulas I -20-a compound represented), room temperature It is stirred to react 5h.Solvent is evaporated off after reaction, is washed with ethyl acetate, yellow solid (4mg) is obtained by filtration (such as -20 institute of Formulas I The compound shown):1H NMR(CD3OD,400MHz):δ2.84(s,3H),2.90(s,3H),3.44(s,4H),3.48(s,3H), 7.19(s,2H),7.62(d,3H),7.84(s,2H),7.97(d,2H),8.71(s,1H),9.52(s,1H).ESI-MS m/ z479.0[M+H]+。
Embodiment 10:
4NHCl dioxane solution (1mL) is added in raw material (20mg) (such as Formulas I -20-d compound represented), room temperature is stirred Mix reaction 5h.Solvent is evaporated off after reaction, is washed with methanol, red solid (15mg, 89%) is obtained by filtration (such as -14 institute of Formulas I The compound shown):1H NMR(DMSO-d6,400MHz):δ1.41(t,3H),2.58(s,3H),2.87(s,4H),3.05(s, 4H),4.48(q,2H),7.03(d,2H),7.35-7.38(m,1H),7.46-7.50(m,2H),7.81(s,2H),8.07(d, 2H),8.62(s,1H),9.23(s,1H).ESI-MS m/z 509.0[M+H]+。
Embodiment 11:
4N HCl dioxane solution (1mL) is added in raw material (10mg) (such as Formulas I -20-b compound represented), room temperature It is stirred to react 5h.Solvent is evaporated off after reaction, is washed with ethyl acetate, yellow solid (5mg, 59%) is obtained by filtration (such as formula I-19 compound represented):1H NMR(CD3OD,400MHz):δ2.82(s,3H),3.43(s,4H),3.51(s,4H),3.57 (s,3H),3.68(s,3H),7.22(s,2H),7.63(s,3H),7.85(s,2H),7.97(s,2H),8.74(s,1H),9.49 (s,1H).ESI-MS m/z 523.0[M+H]+。
Embodiment 12:
Step 1:
By 2- methyl mercapto -5- methyl -6- cyano -7- aminopyridine simultaneously [2,3-d] pyrimidine (500mg) (such as Formulas I -16-d institute The compound shown) it is dissolved in DMF (20mL), it is added bromoacetophenone (861mg), 80 DEG C are stirred to react 16h.It is added after reaction Water, ethyl acetate extraction merge organic layer, dry, and solvent is evaporated off, and column chromatographic purifying (petroleum ether: ethyl acetate=10:1) obtains To yellow solid (140mg) (such as Formulas I -95-d compound represented): ESI-MS m/z 331.9 [M+H]+。
Second, three steps:
Raw material (140mg) (such as Formulas I -95-d compound represented) is dissolved in chloroform (20mL), is added between content 77% Chloroperoxybenzoic acid (195mg) is stirred at room temperature reaction 4h, 4- (4- tert-butoxycarbonyl-piperazine) aniline (94mg) is added, 70 DEG C are stirred Mix reaction 28h.Solvent is evaporated off after reaction, water is added, methylene chloride extraction merges organic layer, and it is dry, solvent, column is evaporated off Chromatographic purifying (petroleum ether: ethyl acetate=1:1) obtains yellow solid (78mg) (such as Formulas I -95-b compound represented):1H NMR(CDCl3,400MHz):δ1.52(s,9H),2.85(s,3H),3.20(t,4H),3.65(t,4H),7.05(d,2H), 7.39(d,2H),7.46-7.50(m,3H),7.63(s,2H),8.04(d,2H),8.38(s,1H),9.03(s,1H).ESI-MS m/z 561.0[M+H]+。
Step 4:
Raw material (56mg) (such as Formulas I -95-b compound represented) is dissolved in methanol (5mL), 4N NaOH solution is added (0.2mL) and hydrogen peroxide (0.23mL), 30 DEG C are stirred to react 5h.Na is added after reaction2S2O3Solution washing, uses dichloro Methane extraction, is evaporated off solvent and obtains yellow solid (10mg) (such as Formulas I -95-a compound represented): ESI-MS m/z 579.0 [M+H]+。
Step 5:
4N HCl dioxane solution (1mL) is added in raw material (10mg) (such as Formulas I -95-a compound represented), room temperature It is stirred to react 5h.Solvent is evaporated off after reaction, is washed with ethyl acetate, yellow solid (5mg) is obtained by filtration (such as -95 institute of Formulas I The compound shown):1H NMR(DMSO-d6,400MHz):δ2.69(s,3H),3.26-3.40(m,8H),7.14(d,2H), 7.42-7.44(m,1H),7.50-7.54(s,2H),7.90(s,2H),8.10(d,2H),8.37(br,1H),8.71(s,1H), 9.35(s,1H),9.41(br,2H).ESI-MS m/z 479.0[M+H]+。
Embodiment 13:
4N HCl dioxane solution (1mL) is added in raw material (20mg) (such as Formulas I -95-b compound represented), room temperature It is stirred to react 5h.Solvent is evaporated off after reaction, is washed with ethyl acetate, yellow solid (15mg, 91%) is obtained by filtration (such as formula I-15 compound represented):1H NMR(DMSO-d6,400MHz):δ2.82(s,3H),2.86(s,3H),3.06(s,4H), 7.05(s,2H),7.37-7.41(m,1H),7.48-7.52(s,2H),7.85(s,2H),8.12(s,2H),8.64(s,1H), 9.28(s,1H).ESI-MS m/z 461.0[M+H]+。
Embodiment 14:
Step 1:
By 2- methyl mercapto -5- methyl -6- cyano -7- aminopyridine simultaneously [2,3-d] pyrimidine (500mg) (such as Formulas I -16-d institute The compound shown) it is dissolved in DMF (20mL), it is added bromo ring fourth ethyl ketone (765mg), 80 DEG C are stirred to react 16h.After reaction plus Entering water, ethyl acetate extraction merges organic layer, and it is dry, solvent, column chromatographic purifying (petroleum ether: ethyl acetate=10:1) is evaporated off Obtain yellow solid (160mg) (such as Formulas I -21-c compound represented):1H NMR(DMSO-d6,400MHz):δ1.99-2.05 (m,2H),2.28-2.35(m,4H),2.73(s,3H),2.84(s,3H),3.67-3.71(m,1H),8.32(s,1H),9.45 (s,1H).ESI-MS m/z 310.0[M+H]+。
Second, three steps:
Raw material (160mg) (such as Formulas I -21-c compound represented) is dissolved in chloroform (20mL), is added between content 77% Chloroperoxybenzoic acid (238mg) is stirred at room temperature reaction 4h, 4- (4- tert-butoxycarbonyl-piperazine) aniline (115mg) is added, 70 DEG C are stirred Mix reaction 28h.Solvent is evaporated off after reaction, water is added, methylene chloride extraction merges organic layer, and it is dry, solvent, column is evaporated off Chromatographic purifying (petroleum ether: ethyl acetate=1:1) obtains yellow solid (56mg) (such as Formulas I -21-a compound represented):1H NMR(CDCl3,400MHz):δ1.52(s,9H),2.01-2.14(m,2H),2.32-2.47(m,4H),2.85(s,3H),3.19 (s,4H),3.64(s,4H),3.75-3.79(m,1H),7.04(d,2H),7.62(s,2H),7.94(s,1H),9.03(s, 1H).ESI-MS m/z 539.0[M+H]+。
Step 4:
4N HCl dioxane solution (1mL) is added in raw material (20mg) (such as Formulas I -21-a compound represented), room temperature It is stirred to react 5h.Solvent is evaporated off after reaction, is washed with ethyl acetate, yellow solid (9mg) is obtained by filtration (such as -21 institute of Formulas I The compound shown):1H NMR(CD3OD,400MHz):δ2.07-2.29(m,2H),2.44-2.48(m,2H),2.54-2.59(m, 2H),3.03(s,3H),3.46-3.52(m,8H),3.88-3.92(m,1H),7.14-7.25(m,2H),7.70-7.84(m, 2H),8.28(s,1H),9.49(s,1H).ESI-MS m/z 439.0[M+H]+。
Embodiment 15:
Step 1:
By 2- methyl mercapto -5- methyl -6- cyano -7- aminopyridine simultaneously [2,3-d] pyrimidine (chemical combination as shown in Formulas I -16-d Object) (250mg) be dissolved in DMF (15mL), it is added pyridine bromide ethanone hydrobromide (550mg), 80 DEG C are stirred to react 46h.Reaction After be added water, ethyl acetate extraction merges organic layer, dry, and solvent, column chromatographic purifying (petroleum ether: ethyl acetate is evaporated off =10:1) obtain yellow solid (90mg) (such as Formulas I -22-c compound represented):1H NMR(DMSO-d6,400MHz):δ2.76 (s,3H),2.90(s,3H),7.40-7.44(m,1H),7.96-7.97(m,1H),8.17(d,1H),8.67(d,1H),8.91 (s,1H),9.16(s,1H).ESI-MS m/z 333.0[M+H]+。
Step 2:
Raw material (such as Formulas I -22-c compound represented) (120mg) is dissolved in chloroform (20mL), is added between content 77% Chloroperoxybenzoic acid (330mg) is stirred at room temperature reaction 4h, 4- (4- tert-butoxycarbonyl-piperazine) aniline (80mg) is added, 70 DEG C are stirred Mix reaction 28h.Solvent is evaporated off after reaction, water is added, methylene chloride extraction merges organic layer, and it is dry, solvent, column is evaporated off Chromatographic purifying (petroleum ether: ethyl acetate=1:1) obtains yellow solid (10mg) (such as Formulas I -22-a compound represented): ESI- MS m/z 562.0[M+H]+。
Step 3:
4N HCl dioxane solution (1mL) is added in raw material (10mg) (such as Formulas I -22-a compound represented), room temperature It is stirred to react 5h.Solvent is evaporated off after reaction, is washed with ethyl acetate, yellow solid (5mg) is obtained by filtration (such as -22 institute of Formulas I The compound shown):1H NMR(CDCl3,400MHz):δ3.03(s,3H),3.10(s,4H),3.22(s,4H),6.68(d,2H), 6.84(d,2H),7.07(s,1H),7.64(s,1H),7.85(s,1H),8.35(s,1H),8.69(s,1H),8.85(s,1H), 9.06(s,1H).ESI-MS m/z 461.0[M+H]+。
Embodiment 16:
Using the synthetic method in embodiment 7, after corresponding raw material is replaced, synthesis obtains boc-protected intermediate Compound (such as Formulas I -23-a compound represented), then the compound (100mg) is dissolved in methylene chloride, 4N HCl dioxy is added Reaction 5h is stirred at room temperature in six ring solution (3mL).Solvent is evaporated off after reaction, is washed with ethyl acetate, it is solid that yellow is obtained by filtration Body (42mg) (such as -23 compound represented of Formulas I):1H NMR(DMSO-d6,400MHz):δ1.37(t,3H),2.82(s,3H), 3.25(s,8H),4.37(q,2H),7.08(s,2H),7.75(s,2H),8.57(s,1H),9.12(br,2H),9.32(s, 1H).ESI-MS m/z 457.0[M+H]+。
Embodiment 17:
Using the synthetic method in embodiment 7, after corresponding raw material is replaced, synthesis obtains yellow solid (45mg) (such as -24 compound represented of Formulas I):1H NMR(DMSO-d6,400MHz):δ0.98(t,3H),1.76(q,2H),2.75-2.78 (m,2H),2.83(s,3H),3.25(s,4H),3.42(s,4H),7.10(d,2H),7.80(s,2H),8.11(s,1H),9.36 (s,1H),9.54(br,2H).ESI-MS m/z 427.0[M+H]+。
Embodiment 18:
First and second step:
It synthesizes to obtain raw material (such as Formulas I -27-f compound represented) using the similar method of embodiment 9, by the raw material (650mg) is dissolved in chloroform (20mL), and the metachloroperbenzoic acid (405mg) of content 77% is added, and reaction 4h is stirred at room temperature, is added 4- (4- tert-butoxycarbonyl-piperazine) aniline (389mg), 70 DEG C are stirred to react 28h.It is evaporated off solvent after reaction, is added water, two Chloromethanes extraction merges organic layer, dry, solvent is evaporated off, column chromatographic purifying (petroleum ether: ethyl acetate=1:1) obtains yellow Solid (130mg) (such as Formulas I -27-d compound represented): ESI-MS m/z 600.3 [M+H]+。
Step 3:
Raw material (130mg) (such as Formulas I -27-d compound represented) is dissolved in ethyl alcohol (5mL), 4N NaOH solution is added (2mL), 70 DEG C are stirred to react 1h.Solvent is evaporated off after reaction, is adjusted to pH=8 with 2N hydrochloric acid, product is largely precipitated, and filters To yellow solid (50mg) (such as Formulas I -27-c compound represented): ESI-MS m/z 572.3 [M+H]+。
Step 4:
Raw material (50mg) (such as Formulas I -27-c compound represented) is dissolved in DMF (5mL), HATU (50mg) and DIPEA is added N is added after reaction 0.5h is stirred at room temperature in (22 μ L), and reaction 12h is stirred at room temperature in O- dimethyl hydroxylamine hydrochloride (12.8mg).Instead Water is added after answering, ethyl acetate extraction merges organic layer, and it is dry, solvent, column chromatographic purifying (petroleum ether: acetic acid second is evaporated off Ester=1:1) obtain yellow solid (53mg) (such as Formulas I -27-b compound represented): ESI-MS m/z 615.3 [M+H]+。
Step 5:
Raw material (53mg) (such as Formulas I -27-b compound represented) is dissolved in super dry THF (2mL), in 0 DEG C under nitrogen protection 3N methyl-magnesium-bromide diethyl ether solution (0.3mL) is slowly added dropwise.Solvent is evaporated off after reaction, water, ethyl acetate extraction is added, closes And organic layer, it is dry, solvent is evaporated off, column chromatographic purifying (petroleum ether: ethyl acetate=1:1) obtains yellow solid (22mg) (such as Formulas I -27-a compound represented):1H NMR(CDCl3,400MHz):δ1.52(s,9H),1.72-1.74(m,2H),1.81- 1.86(m,4H),2.14-2.16(m,2H),2.58(s,3H),2.88(s,3H),3.17(t,4H),3.25-3.29(m,1H), 3.64(t,4H),7.03(d,2H),7.47(br,1H),7.64(d,2H),7.90(s,1H),9.04(s,1H).ESI-MS m/z 570.3[M+H]+。
Step 6:
4N HCl dioxane solution (1mL) is added in raw material (22mg) (such as Formulas I -27-a compound represented), room temperature It is stirred to react 5h.Solvent is evaporated off after reaction, is washed with ethyl acetate, yellow solid (15mg) (such as Formulas I -27 is obtained by filtration Compound represented):1H NMR(CDCl3,400MHz):δ1.73-1.75(m,2H),1.81-1.84(m,4H),2.12-2.18 (m,2H),2.58(s,3H),2.89(s,3H),3.11(s,4H),3.20(s,4H),3.25-3.30(m,1H),7.03(d, 2H),7.42(br,1H),7.63(d,2H),7.90(s,1H),9.04(s,1H).ESI-MS m/z 470.3[M+H]+。
Embodiment 19:
Using the synthetic method in embodiment 18, after corresponding raw material is replaced, synthesis is obtained as shown in Formulas I -13 Compound.1H-NMR,DMSO,10.22(s,1H),9.25(s,1H),7.93(s,1H),7.79(m,2H),7.05(m,2H), 3.33-3.29(m,5H),3.22-3.16(m,4H),2.76(s,3H),2.51(s,3H),1.40(d,6H).LC-MS: C25H29N7Ocalcd[M+H]+444.24,found 444.0.
Embodiment 20:
Using the synthetic method in embodiment 18, after corresponding raw material is replaced, synthesis is obtained as shown in Formulas I -12 Compound.1H-NMR,DMSO(δ),10.25(s,1H),9.25(s,1H),7.88(s,1H),7.77(m,2H),7.05(m, 2H),3.29(m,4H),3.19(m,4H),2.51(s,3H),1.46(s,9H)。LC-MS:C26H31N7Ocalcd[M+H]+ 458.26,found 458.0.
Embodiment 21:
Using the synthetic method in embodiment 18, after corresponding raw material is replaced, synthesis is obtained as shown in Formulas I -30 Compound.1H-NMR,DMSO(δ),10.22(s,1H),9.24(s,1H),7.91(s,1H),7.75(m,2H),7.03(m, 2H),3.15-3.09(m,4H),2.90-2.94(m,2H),2.75-2.72(m,4H),2.65-2.62(m,2H),2.51(s, 3H),2.07-2.04(m,2H),1.82-1.76(m,2H),1.52-1.24(m,6H).LC-MS:C28H33N7Ocalcd[M+ H]+484.27,found 484.2.
Embodiment 22:
Using the synthetic method in the first step of embodiment 18, second step and the 6th step, after corresponding raw material is replaced, Synthesis is obtained such as -25 compound represented of Formulas I.1H-NMR,DMSO(δ),9.19(s,1H),7.88(s,1H),7.73(m,2H), 7.00(m,2H),4.45(q,2H),3.05(m,4H),2.86(m,4H),2.70(m,1H),2.52(s,3H),2.04-1.89 (m,2H),1.81-1.69(m,3H),1.48-1.17(m,8H).LC-MS:C29H35N7O2calcd[M]+513.29,found 513.2.
Embodiment 23:
It synthesizes to obtain as shown in Formulas I -94 using the synthetic method in the first step of embodiment 18, second step and the 6th step Compound.1H-NMR,DMSO(δ),9.19(s,1H),7.88(s,1H),7.73(m,2H),7.00(m,2H),4.45(q,2H), 3.05(m,4H),2.86(m,4H),2.70(m,1H),2.52(s,3H),2.04-1.89(m,2H),1.81-1.69(m,3H), 1.48-1.17(m,8H).LC-MS:C28H34N8O2calcd[M]+515.28,found 515.3.
Embodiment 24:
Using the synthesis side of the synthetic method in embodiment 18 in the first step, second step, third step, the 4th step and the 6th step Method, after corresponding raw material is replaced, synthesis is obtained such as -28 compound represented of Formulas I.1H-NMR,DMSO(δ),10.19(s, 1H),9.18(s,1H),7.89(s,1H),7.79(m,2H),7.05(m,2H),3.48(s,3H),3.38(s,3H),3.31- 3.30(m,4H),3.18-3.17(m.4H),2.72-2.68(m,1H),2.45(s,3H),2.02-1.96(m,2H),1.81- 1.1.70(m,3H),1.54-1.35(m,5H).LC-MS:C29H36N8O2calcd[M+H]+529.30,found 529.3.
Embodiment 25:
It, will be corresponding using the synthetic method in embodiment 18 in the first step, second step, third step, the 4th step and the 6th step After raw material is replaced, synthesis is obtained such as -29 compound represented of Formulas I.1H-NMR,DMSO(δ),10.20(s,1H),9.19 (s,1H),7.89(s,1H),7.79(m,2H),7.06(m,2H),3.31(m,4H),3.22-3.17(m,5H),3.09(s, 3H),2.81(s,3H),2.43(s,3H),2.06-1.99(m,2H),1.82-1.79(m,3H),1.47-1.17(m,5H).LC- MS:C29H36N8Ocalcd[M+H]+513.30,found 513.3.
Effect example 1 (activity data of the compound of the present invention in kinases and cellular level)
Test method:
1, CDK kinase inhibiting activity experiment usesUltra technology, when ATP concentration is Km, respectively Test-compound is screened on CDK4/CycD3, CDK6/CycD3 kinases.In test process, test-compound is initially dense Degree is selected as 3333nM, and each test-compound selects 10 gradient dilution concentration, and gradient dilution multiple is 3 times, every concentration 1 A multiple holes are detected.
CDK4/CycD3 and CDK6/CycD3 is purchased from Carna Biosciences, Inc.；Dimethyl sulfoxide, ATP, DTT Solution is purchased from Sigma-Aldrich；EDTA solution is purchased from GIBCO；Detection Buffer, 10 × andUltra Europium-anti-phospho-eIF4E-binding protein 1 (Thr37/46) andUltra ULightTM- eIF4E-bindingprotein 1 (Thr37/46) Peptide, is purchased from Perkinelmer。
Testing procedure: 1, using 1 × reaction buffer kinases, substrate, ATP is respectively configured to 2.5 × enzyme/substrate it is mixed Close liquid and 2.5 × ATP solution.In experiment, CDK4/CycD3 kinases it is final concentration of: 0.76ng/ul, ATP are final concentration of: 80uM；CDK6/CycD3 kinases it is final concentration of: 0.5ng/ul, ATP are final concentration of: 50uM；Into 384 orifice plates be added 2.5 × Enzyme/Substrate cocktail, be incubated at room temperature 5 minutes；2.5 × ATP solution is added, is reacted at room temperature 30 minutes.2, it usesDetection Buffer, 1 × preparation 2 ×Ultra Europium-anti-phospho- EIF4E-binding protein 1 (Thr37/46) is spare.After enzyme reaction carries out 30 minutes, 10mM is added into 384 orifice plates EDTA, react at room temperature 5 minutes.It addsUltra Europium-anti-phospho-eIF4E- Binding protein 1 (Thr37/46) is reacted at room temperature 1 hour.3,384 orifice plates are put in HERAEUS Multifuge 2000rpm is centrifuged 2 minutes in X1R centrifuge；According to surveying and determination in the enterprising line number of EnVisionTM, the laser of 337nM wavelength is selected to make For exciting light, measure RFU665nM and RFU615nM, and using RFU665nM/RFU615nM × 10000 as final data into Row analysis.4, Log (inhibitor) vs.response-Variable is carried out to data using Graphpad Prism 5.0 Slope (four parameters) curve matching, calculates corresponding IC50 (half maximal inhibitory concentration)。
2, MCF-7 cell growth inhibition assay usesLuminescent Cell Viability The measurement of Assay method.Experiment inhibits Mcf- on human breast carcinoma Mcf-7 cell, through the method detection compound of CellTiter The effect of 7 cell Proliferations, in detection process, test-compound initial concentration is selected as 10 μM, and each embodiment compound selects 9 A gradient dilution concentration, gradient dilution multiple are 3 times, and 2 multiple holes of every concentration are detected.Mcf-7 cell is purchased from the triumphant base in Nanjing Biotechnology Development Co., Ltd；Luminescence method cell viability detection kit is purchased from Promega, article No.: G7573；Dimethyl sulfoxide is purchased from Sigma-Aldrich, article No.: D8418；384 porocyte culture plates, Corning, article No.: 3570。
Testing procedure: 1. normal culture human breast cancer cell Mcf-7, after digestion, by 675, every hole cell density kind plate in 384 orifice plates；2. after kind of plate 1d, one block of plate is used to survey background Celltiter value, it is denoted as Control D1.Remaining plate dosing: setting Cell controls group.Test-compound initial concentration is 10 μM, successively 9 gradient dilution concentration, and gradient dilution multiple is 3 times, often 2 multiple holes of concentration are detected.3. after handling 7 days 25ul CellTiter detection liquid, oscillation is added in every hole in test-compound 2min is mixed well, centrifugation, is stood balance and is detected after ten minutes, record fluorescence signal, medicine group is denoted as Drug D7, cell pair Control D7 is denoted as according to group.4. carrying out Log (inhibitor) to data using Graphpad Prism 5.0 Vs.response-Variable slope (four parameters) curve matching, calculates corresponding IC50(half maximal inhibitory concentration)。
Test result is shown in Table 1.
The active testing result of table 1, kinases and cellular level

Claims (10)

1. a kind of nitrogenous fused heterocyclic compound or its pharmaceutically acceptable salt shown in formula I；
Wherein, R1For hydrogen, halogen, C1~C6Alkyl, C1~C6Alkoxy or C3~C6Naphthenic base；
R2For hydrogen, halogen, C1~C6The C that alkyl, hydroxyl replace1~C6Alkyl, C1~C6Alkyl oxygen-carbonyl-, carboxyl, cyano, aldehyde Base, C1~C6Acyl group orDescribedIn R5And R6It independently is hydrogen, C1~C6Alkyl or C1~C6Alcoxyl Base, alternatively, the R5And R6Atom connect cyclization, withIn N atom form unitary azepine C2~C6Naphthenic base；
R3For hydrogen；
R4For substituted or unsubstituted C1~C6Alkyl, substituted or unsubstituted C3~C6Naphthenic base, C1~C6Alkyl oxygen-carbonyl-, Substituted or unsubstituted C3~C12Aryl, substituted or unsubstituted C3~C12Heteroaryl or substituted or unsubstituted C2~C6Heterocycle Alkyl；The substituted or unsubstituted C3~C12Hetero atom in heteroaryl is one of O, N and S or a variety of, described Substituted or unsubstituted C3~C12Heteroatomic number in heteroaryl is 1~4；The substituted or unsubstituted C2~C6 Hetero atom in Heterocyclylalkyl is one of O, N and S or a variety of, described substituted or unsubstituted C2~C6In Heterocyclylalkyl Heteroatomic number be 1~4；Described is substituted by by halogen, C1~C6Alkoxy, trifluoromethyl, hydroxyl and C1~C6Alkane One of base is a variety of replaced；
Ar is W-L1-L2-Y-；
Wherein:
Y is selected from
L2Selected from-(CH2)r,-O- ,-S- ,-C (=O)-or substituted or unsubstituted amino, r is 0 or 1；It is described be substituted by by C1~C3Replaced alkyl；
L1Selected from-(CH2)n, substituted or unsubstituted C3~C12Naphthenic base, substituted or unsubstituted C3~C12Heterocyclylalkyl；N is 0 or 1；Described is substituted by by C1~C6Replaced alkyl or halogen；The substituted or unsubstituted C3~C12In Heterocyclylalkyl Hetero atom be one of O, N and S or a variety of, heteroatomic number is any integer in 1~4；
W be hydrogen,Halogen, hydroxyl, aldehyde radical,Substituted or unsubstituted amino, substitution or Unsubstituted C1~C6Alkyl or substituted or unsubstituted C1~C6Acyl group；DescribedIn X2For O, S or DescribedIn s be 1~5 in any integer, it is describedIn t be 0 Any integer in~5, it is describedIn m be 0~5 in any integer, it is describedIn X1For N, O ,-C (=O)-NH- orThe substituted amino refers to only one hydrogen on nitrogen Be substituted or nitrogen on two hydrogen be substituted, when substituted amino refers to that two hydrogen are substituted on nitrogen, two substituent groups be it is identical or It is different；Described is substituted by by C1~C6Alkyl, C1~C6Alkyl sulphonyl, C1~C6Alkoxy, C1~C6Alkylthio group, C3~C6 The C that naphthenic base, halogen, hydroxyl, amino, hydroxyl replace1~C6The N of alkyl, 2-12 carbon atom, N- dialkyl amido, 2-12 The alkoxy alkyl-, the C that replace of the N of carbon atom, N- dialkyl amido-alkyl-, 2-12 carbon atom3~C6Naphthenic base and C2 ~C6One of Heterocyclylalkyl is a variety of replaced；The C2~C6Hetero atom in Heterocyclylalkyl is one in N, O and S Kind is a variety of, and heteroatomic number is any integer in 1~4.
2. nitrogenous fused heterocyclic compound or its pharmaceutically acceptable salt shown in formula I as described in claim 1, feature It is:
As the R1When for halogen, the halogen is chlorine；As the R1For C1~C6When alkyl, the C1~C6Alkyl For methyl；As the R1For C1~C6When alkoxy, the C1~C6Alkoxy is methoxyl group；As the R1For C3~C6 When naphthenic base, the C3~C6Naphthenic base is cyclopropyl；
And/or as the R2When for halogen, the halogen is bromine；As the R2The C replaced for hydroxyl1~C6When alkyl, The C that the hydroxyl replaces1~C6Alkyl is CH3-CH(OH)-；As the R2For C1~C6Alkyl oxygen-carbonyl-when, it is described C1~C6Alkyl oxygen-carbonyl-is ethoxy-carbonyl；As the R2When for aldehyde radical, the aldehyde radical is carboxaldehyde radicals；When described R2For C1~C6When acyl group, the C1~C6Acyl group is acetyl group；As the R2ForWhen, and it is describedIn R5And R6It independently is C1~C6When alkyl, the C1~C6Alkyl is methyl；As the R2ForWhen, and it is describedIn R5And R6It independently is C1~C6When alkoxy, the C1~C6Alkoxy is Methoxyl group；As the R2ForWhen, and the R5And R6Atom connect cyclization, withIn N atom Form unitary azepine C2~C6When naphthenic base, the unitary azepine C2~C6Naphthenic base is unitary aziridinyl；
And/or as the R4For substituted or unsubstituted C1~C6When alkyl, the substituted or unsubstituted C1~C6Alkyl For substituted or unsubstituted methyl, substituted or unsubstituted n-propyl, substituted or unsubstituted tert-butyl, substituted or unsubstituted Isopropyl or substituted or unsubstituted sec-butyl；As the R4For substituted or unsubstituted C3~C6When naphthenic base, described is taken Generation or unsubstituted C3~C6Naphthenic base is substituted or unsubstituted C4、C5Or C6Naphthenic base；As the R4For C1~C6Alkyl Oxygen-carbonyl-when, the C1~C6Alkyl oxygen-carbonyl-is ethoxy-carbonyl-；As the R4For substituted or unsubstituted C3 ~C12When aryl, the C3~C12Aryl is phenyl；As the R4For substituted or unsubstituted C3~C12When heteroaryl, institute The substituted or unsubstituted C stated3~C12Hetero atom in heteroaryl is O or N, and/or, the substituted or unsubstituted C3~ C12Heteroatomic number in heteroaryl is 1；As the R4For substituted or unsubstituted C2~C6It is described when Heterocyclylalkyl Substituted or unsubstituted C2~C6Hetero atom in Heterocyclylalkyl is N or O, and/or, the substituted or unsubstituted C2~ C6Heteroatomic number in Heterocyclylalkyl is 1；
And/or as the L1For substituted or unsubstituted C3~C12When naphthenic base, the substituted or unsubstituted C3~C12 Naphthenic base is substituted or unsubstituted cyclohexyl；As the L1For substituted or unsubstituted C3~C12It is described when Heterocyclylalkyl Substituted or unsubstituted C3~C12Hetero atom in Heterocyclylalkyl is N, and/or, heteroatomic number is 1 or 2；
And/or when the W is halogen, the halogen is bromine；When the W isWhen, it is described S be 1 or 2, and/or, the t be 0 or 1, and/or, the m be 1；When the W is substituted or unsubstituted C1~C6 When alkyl, the substituted or unsubstituted C1~C6Alkyl is substituted or unsubstituted methyl, alternatively, substituted or unsubstituted Ethyl.
3. nitrogenous fused heterocyclic compound or its pharmaceutically acceptable salt shown in formula I as claimed in claim 2, feature It is:
As the R4For substituted or unsubstituted C3~C12When heteroaryl, the substituted or unsubstituted C3~C12Heteroaryl ForAs the R4For substituted or unsubstituted C2~C6When Heterocyclylalkyl, the substituted or unsubstituted C2~ C6C in Heterocyclylalkyl2~C6Heterocyclylalkyl is
4. nitrogenous fused heterocyclic compound or its pharmaceutically acceptable salt shown in formula I as described in claim 1, feature It is:
The R1For hydrogen or methyl；
And/or the R2For acetyl group, cyano, bromine ,-CH (OH) CH3、C1~C6Alkyl oxygen-carbonyl-or
And/or the R4For methyl, tert-butyl, isopropyl, cyclopenta, cyclobutyl, phenyl, n-propyl or cyclohexyl；
And/or the L1For
And/or the Ar is
5. nitrogenous fused heterocyclic compound or its pharmaceutically acceptable salt shown in formula I as described in claim 1, feature It is: in the Formulas I compound represented, R1For hydrogen or methyl, R2For acetyl group, cyano, bromine ,-CH (OH) CH3、C1~C6 Alkyl oxygen-carbonyl-orR3For hydrogen, R4For methyl, tert-butyl, isopropyl, cyclopenta, cyclobutyl, phenyl, positive third Base or cyclohexyl, Ar are
6. nitrogenous fused heterocyclic compound or its pharmaceutically acceptable salt shown in formula I as described in claim 1, feature Be: the pharmaceutically acceptable salt of the Formulas I compound represented is Formulas I compound represented and organic acid or inorganic acid It is formed by salt.
7. nitrogenous fused heterocyclic compound or its pharmaceutically acceptable salt shown in formula I as described in claim 1, feature Be: the nitrogenous fused heterocyclic compound shown in formula I is following any compound:
8. a kind of preparation method such as Formulas I compound represented of any of claims 1-7, for it is following either one Method:
Method one:
The preparation method of the Formulas I compound represented includes the following steps: Formula II compound represented carrying out removing guarantor The reaction for protecting base, obtains Formulas I compound represented, and the PG in Formula II compound represented is the protecting group；
Wherein, described Ar, R1、R2、R3、R4With described in claim 1；
Method two:
The preparation method of the Formulas I compound represented includes the following steps: -3 compound represented of formula III and 4 institute of formula The compound shown carries out substitution reaction or transition metal-catalyzed coupling reaction, generates the Formulas I compound represented；Formula LG group in III-3 compound represented is leaving group；
The R1、R2、R3、R4With Ar with described in claim 1；
Method three:
As the R2When for halogen or acetyl group, the preparation method of the Formulas I compound represented include the following steps: by Formula III -3-4 compound represented and 5 compound represented of formula generate described in transition metal-catalyzed lower generation coupling reaction Formulas I compound represented；
The R1、R3、R4, X and Ar be the same as described in claim 1.
9. one kind containing the nitrogenous fused heterocyclic compound as shown in Formulas I of any of claims 1-7 or its pharmaceutically The pharmaceutical composition of acceptable salt.
10. a kind of fused heterocyclic compound nitrogenous as shown in Formulas I of any of claims 1-7 or its can pharmaceutically connect Application of the salt received on the drug of preparation prevention or the extremely relevant disease for the treatment of cell cycle regulating.
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