CN104072481B - A kind of Anti-angiogenic compounds - Google Patents

A kind of Anti-angiogenic compounds Download PDF

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CN104072481B
CN104072481B CN201410125752.0A CN201410125752A CN104072481B CN 104072481 B CN104072481 B CN 104072481B CN 201410125752 A CN201410125752 A CN 201410125752A CN 104072481 B CN104072481 B CN 104072481B
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compound
reactant mixture
dcm
acid
tlc
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CN104072481A (en
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侯睿
罗红蓉
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NINGBO FANGCHANG PHARMACEUTICAL CO., LTD.
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Ningbo Fangchang Pharmaceutical Co Ltd
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A61P27/06—Antiglaucoma agents or miotics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems

Abstract

The invention provides noval chemical compound of a kind of anti-angiogenic rebirth and its production and use.The compounds of this invention has good anti-angiogenic rebirth effect, it was initially believed that this compounds is by suppression VEGFR2(i.e. KDR) produce activity.This compounds can be used for the treatment to protein kinase caused by abnormal diseases such as new vessels, VEGFR2, PDGFR β, KIT, such as wet MD, malignant tumor etc..

Description

A kind of Anti-angiogenic compounds
Technical field
The present invention relates to a kind of noval chemical compound and purposes.
Background technology
Angiogenesis (angiogenesis), is to germinate from existing blood vessel to generate the process of neovascularity.This process and blood Endothelial cell migrates relevant with propagation.Angiogenesis is relevant, such as malignant tumor with multiple mankind's major disease.Have now been found that, Ocular angiogenesis disease, including age-related macular degeneration (AMD), diabetic retinopathy, neovascular green grass or young crops Light eye etc., the common feature of this kind of disease be all ocular angiogenesis paraplasm (gold dawn, etc., anti vegf agents is at eye Application in section's disease and Recent Advances in Mechanism, China and foreign countries' medical treatment, 2012).
Wherein, degeneration of macula mainly has dryness and moist two kinds, and the feature of wet MD (AMD) is, choroidal New vessels enter under retina and then occur hemorrhage, ooze out and the pathological change such as edema.Moist rapid DE, Even more serious compared with dryness.At present, the most preferably it is in progress in terms of the treatment of wet MD.Laser in early days burns bright hemostasis Replaced by blood vessel endothelial factor antagonist, but because of the latter's poor effect, quickly replaced by photodynamic therapy.Photodynamic therapy Although the curative effect of improve, but still undesirable.Occur in that the most again new blood vessel endothelial factor antagonist Lucentis(thunder pearl is single Anti-), it is the recombinant of a kind of humanized's VEGF hypotype monoclonal antibody fragment, new vessels can be reduced and generate.2006, this medicine Thing is approved by the fda in the United States for treating wet MD, and curative effect is good；Meanwhile, such anti vegf agents pair is currently also found Diabetic retinopathy, neovascular glaucoma have therapeutical effect.But owing to Lucentis is antibody medicine, price is high, It can't be popularized in the whole world.Therefore, the little molecule neovascularization inhibitor medicine that research curative effect is good, cheap is to work as The focus of modern international pharmaceutical industry keen competition.
Summary of the invention
Object of the present invention is to provide a kind of noval chemical compound, and its production and use.
The invention provides the compound as shown in formula A and pharmaceutically acceptable salt thereof or prodrug:
Wherein, Z0For O or S；X be C, Y be N；；Z1、Z2It is respectively and independently selected from N or-CR8；
r7Selected from hydrogen or C1-C6Alkyl；r8Selected from hydrogen or C1-C6Alkyl；r10Selected from H, amino, hydroxyl, sulfydryl, C1-C6Alkane Base or-(CH2)n1NHr1, wherein, n1=1-5, r1Alkyl for H or C1-3；
r9Selected from H, amino, hydroxyl, sulfydryl, (C-r11r12)nNr13r14、(C-r11r12)nOr15、(C-r11r12)nC(O) Er13、(C-r11r12)nS(O)mr17；Or r8And r9Saturated 4-7 unit heterocycle or replacement is formed together with the atom being connected with them Heterocycle, wherein, heterocycle or substituted heterocycle contain 1 or 2 hetero atom selected from N, O or S, and its substituent group is 0,1, or 2, respectively Independently selected from oxo group, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxyl, C1-C6 halogenated alkoxy, C1-C6 Alkanoyl, C1-C6 alkylamino radical, C3-C7 cycloalkyl, halogen, hydroxyl, amino, carbonyl, amino carbonyl, C1-C6 aminoalkyl carbonyl, C1-C4 acyl group, C1-C6 alkoxy carbonyl, C1-C6 sulfonyl, amino-sulfonyl；
Ar2Selected from phenyl, naphthyl, monocycle or bicyclic heteroaryl, bicyclo-or tricyclic heterocyclic base, the most each heteroaryl or miscellaneous Ring group has 1,2,3 or 4 hetero atom selected from N, O or S；Described phenyl, naphthyl, heteroaryl or heterocyclic group be unsubstituted or Substituted phenyl, naphthyl, heteroaryl or heterocyclic group, its substituent group is 1,2, or 3, separately selected from C1-C8 alkyl, C1-C8 haloalkyl, hydroxyl C1-C6 alkyl, C1-C8 alkoxyl, C1-C8 halogenated alkoxy, halogen, hydroxyl, amino, amino C- C6 alkyl, C1-C6 alkyl amino, phenyl, C3-C7 cycloalkyl, the C3-C7 cycloalkyl of volution, amino-sulfonyl, C1-C6 alkyl Amino-sulfonyl；
M is 0,1, or 2；N is 0,1,2, or 3；E is empty, oxygen or-Nr18；
r11,r12And r18It is identical or different, is respectively and independently selected from hydrogen or C1-C4 alkyl；r13,r14,r15,r16And r17 Be independently selected from hydrogen, C1-C6 alkyl, C1-C6 acyl group, phenyl and heterocycle, or substituted C1-C6 alkyl, C1-C6 acyl group, phenyl and Heterocycle, its substituent group for for 0,1 or 2, is respectively and independently selected from C1-C4 alkoxyl, C1-C4 alkyl, halogen, hydroxyl, amino, C1-C6 aminoalkyl.
Further, described structural formula of compound is as follows:
Wherein, X be C, Y be N；
R1Selected from H, amino, hydroxyl or sulfydryl；R2Selected from H, amino, hydroxyl, sulfydryl or-(CH2)nNHR8, wherein, n=1-5, R8Alkyl for H or C1-3；R3Selected from H or C1-6 alkyl；R4、R5、R6It is respectively and independently selected from H, halogen, the alkyl of C1-6 or halogen Replace alkyl；R7Alkyl or halogen selected from H, C1-6；
Or, R2、R3Coupled carbon atom collectively form substituted or non-substituted containing 1-2 heteroatomic five yuan or Hexatomic ring, described hetero atom is N, O or S, and its substituent group is the alkyl of C1-6.
Further, X, Y differ；R1Selected from H or amino；R2Selected from H, amino or-(CH2)nNHR8, wherein, n=1-3, R8Alkyl for H or C1-2；R3Selected from H or C1-2 alkyl；R4、R5、R6It is respectively and independently selected from halogen, the alkyl of C1-2 or halogen to take Substituted alkyl；R7Selected from H or halogen；
Or, R2、R3Coupled carbon atom collectively forms substituted or non-substituted five yuan containing 1 N or hexatomic ring, Substituent group is the alkyl of C1-3.
Further, R2Selected from amino or-(CH2)nNHR8；Or, R2、R3Coupled carbon atom collectively forms and takes Generation or non-substituted five yuan containing 1 N or hexatomic ring, substituent group is the alkyl of C1-3.
Further, described structural formula of compound is as follows:
When compound is Formula II, W1, W2 are respectively and independently selected from C or hetero atom, n=1 or 2；R9 is H, C1-C4 alkyl or halogen Element；R10 is halogen；Wherein, during n=1, it is C during W1, W2 difference；Preferably, during n=1, W1, W2 are hetero atom simultaneously, hetero atom It is preferably N；During n=2, W1, W2 are C simultaneously；
When compound is formula III, W3 is selected from C or hetero atom, and hetero atom is preferably N；R10 is halogen.
Further, described halogen is F or Cl.
Preferably, described compound is:
Present invention also offers above-claimed cpd and pharmaceutically acceptable salt thereof or prodrug and prepare angiogenesis Purposes in inhibitor.
Further, described angiogenesis inhibitors is VEGF R2 inhibitor.
Further, described inhibitor is the medicine of anti-ocular angiogenesis.
Further, described medicine is choroid neovascularization inhibitors.
Further, described medicine is treatment or prevention wet MD, diabetic retinopathy or new hemopoietic The medicine of pipe glaucoma.
Present invention also offers above-claimed cpd and pharmaceutically acceptable salt thereof or prodrug preparation VEGFR2, PDGFR-β is or/and purposes in KIT inhibitor class medicine.
Present invention also offers a kind of pharmaceutical composition, it is containing above-claimed cpd and pharmaceutically acceptable salt thereof Ophthalmic preparation.In addition to the above-claimed cpd that the present invention provides, preparation can also comprise other and known there is similar therapeutic The medicine of purposes.
Further, described ophthalmic preparation is eye drop, Eye ointments or ophthalmically acceptable injection.
The method being known in the art can prepare the salt of the compounds of this invention.With acid treatment, or with suitable anion Exchanger, can become salt with above-claimed cpd.The pharmaceutically acceptable salt of the compound of the present invention, can have from above-claimed cpd There is the organic or inorganic acid-addition salts of basic nitrogen atom.
Preferably, suitable mineral acid includes but not limited to, halogen acids, example hydrochloric acid, sulphuric acid, or phosphoric acid.
Preferably, suitable organic acid includes, but not limited to carboxylic acid, phosphoric acid, sulfonic acid or amino carboxylic acid, such as acetic acid, and third Acid, octanoic acid, capric acid, dodecylic acid, hydroxyacetic acid, lactic acid, fumaric acid, succinic acid, adipic acid, 1,5-pentanedicarboxylic acid., suberic acid, nonyl two Acid, malic acid, tartaric acid, citric acid, aminoacid, such as glutamic acid or aspartic acid, maleic acid, hydroxy acid, citraconic acid, ring Cyclohexane carboxylic-acid, adamantanecarboxylic acid, benzic acid, salicylic acid, 4 aminosallcylic acids, phthalic acid, phenylacetic acid, mandelic acid, Cortex Cinnamomi Acid, methane or ethane sulfonic acid sulfonic acid, 2-ethylenehydrinsulfonic acid, ethane-1,2-disulfonic acid, benzenesulfonic acid, 2-LOMAR PWA EINECS 246-676-2,1,5-naphthalene two sulphur Acid, 2-toluene sulfonic acide, p-methyl benzenesulfonic acid, ethyl sulfuric acid, the acid of lauryl sulphate acid, N Cyclohexylamino acetic acid, N-first Base-N-ethyl-N-propyl-amino sulfonic acid, or other organic acid, such as ascorbic acid.
It addition, it can also be used in isolated or purified by the most unacceptable salt, such as picrate or perchloric acid Salt.But, for therapeutic use, can only be pharmaceutically acceptable salt or free cpds, with applicable pharmaceutical preparation Form.
Pharmaceutically acceptable prodrug of the present invention, refers to that described compound obtains after modifying for chemical structure Conversion through enzyme or non-enzymatic in vivo discharge active component and play the compound of drug effect.One embodiment of the present invention In, further comprises isotope-labeled above-claimed cpd or its pharmaceutically acceptable salt, described compound isotopically labelled is Refer to herein listed by compound identical, but one or more atom is replaced by another atom, this atom former Protonatomic mass or mass number are different from atomic mass common in nature or mass number.The isotope bag in compound can be introduced Include hydrogen, carbon, nitrogen, oxygen, sulfur, i.e. 2H, 3H, 13C, 14C, 15N, 17O, 18O, 35S.Containing above-mentioned isotope and/or other atom Isotopic compound and stereoisomer thereof, and the pharmaceutically useful salt of this compound, stereoisomer should be included in this Within invention scope.
Key intermediate and compound in the present invention carry out separating and purification, and the mode used is normal in organic chemistry Isolation and purification method and the example of described method include filtering, extract, be dried, be spin-dried for and various types of chromatographs.Can Selectively, can make intermediate the most purified i.e. carry out next step reaction.
In some embodiments, one or more compounds of the present invention can be used in conjunction with one another.Also may select will The compound of the present invention is used in combination with other active agent any, for preparing regulating cell function or the medicine for the treatment of disease Thing or pharmaceutical composition.If using one group of compound, then can by these compounds simultaneously, respectively or in an orderly manner to tested Object is carried out.
The compounds of this invention has good anti-angiogenic rebirth effect, it was initially believed that this compounds is by suppression VEGFR2(i.e. KDR) produce activity.This compounds can be used for the protein kinase caused by abnormal disease such as new vessels, VEGFR2 Sick treatment, such as wet MD, malignant tumor etc..
Accompanying drawing explanation
Fig. 1 the compounds of this invention inhibitory action to Brachydanio rerio vascular development
Fig. 2 negative control result figure, wherein, Brachydanio rerio vascular development is normal
Fig. 3 medicine group result figure, wherein, Brachydanio rerio vascular development position is suppressed by the compounds of this invention part
Fig. 4 medicine group result figure, wherein, Brachydanio rerio vascular development position is completely inhibited by the compounds of this invention
Fig. 5 the compounds of this invention is to VEGFR2 In-vitro Inhibitory Effect
The inhibitory action of Fig. 6 the compounds of this invention KDR4 choroidal neovascularization, wherein, A: negative control, B:KDR4
The inhibitory action of Fig. 7 the compounds of this invention V01 choroidal neovascularization, wherein, A: negative control, B:V01
The nuclear-magnetism figure of Fig. 8 compound V01
The mass spectrum of Fig. 9 compound V3
The mass spectrum of Figure 10 compound V4
The mass spectrum of Figure 11 compound V5
The nuclear-magnetism figure of Figure 12 compound V5
The mass spectrum of Figure 13 compound V6
The mass spectrum of Figure 14 compound K DR3
Figure 15 KDR4 compound mass spectrum
The mass spectrum of Figure 16 KDR6
The mass spectrum of Figure 17 KDR6A
The mass spectrum of Figure 18 KDR8
The nuclear-magnetism figure of Figure 19 KDR8A
The mass spectrum of Figure 20 KDR8A
The mass spectrum of Figure 21 KDR8B
The dose effect curve of Figure 22 compound V01
Figure 23 compound V01 is on mice eye new vessels and hemorrhage impact, A: right eye V01 process, B: left eye PBS pair According to
Detailed description of the invention
The preparation of embodiment 1 intermediate of the present invention
Step is as follows:
1st step: mixing Michaelis acid (75.3g, 0.522mol) and triethyl orthoformate (92mL, 0.55mol), is heated to 55 DEG C, maintain 90 minutes, be then cooled to 45 DEG C.Compound 1 (92g, 0.5mol) is dissolved in methanol (200mL), and joins reaction Mixture reacts 45 minutes, keeps the temperature of reactant mixture less than 50 DEG C simultaneously, be then stirred at room temperature overnight, thin layer Chromatograph (PE/EA=3:1) monitoring is to having reacted.Reactant mixture is cooled to 0 DEG C, and precipitate filters, and obtains white after drying Purify compound 2 (155.9g, yield: 96%).
2nd step: compound 2 (155.9g, 0.441mol) (1.1L) in Dowtherm A heat pipe is heated to 100 DEG C, then it is slowly added to connect in the flask having Dowtherm A heat pipe in (500mL is preheating to 210 DEG C), keeps simultaneously Temperature is higher than 207 DEG C.Reactant stirs reaction one hour at 210 DEG C, is then cooled to room temperature.The precipitation being collected by filtration, After washing with ether, acetone, obtain the Purify compound 3 (67g, yield: 61%) of brown after drying.
3rd step: at room temperature, with stirring and toward phosphorus oxychloride P DEG C l3 (35mL) adds compound 3 (10g, 0.0398mol).After interpolation, reactant mixture return stirring 3 hours, thin layer chromatography (dichloromethane DCM/ methanol=10:1) with Track has reacted, then reactant mixture injects frozen water, and pH=8 is adjusted in sodium carbonate alkalization, then extracts with ethyl acetate, and collection has Machine phase, washs with saline, dried with Na2SO4, crude product again by silica gel column chromatography (petroleum ether PE/ ethyl acetate EA=20:1, 1:1), faint yellow solid pure compound 4 (6.0g, yield: 56%) is obtained.
4th step: by compound 4 (10.0g, 37mmol), zinc powder (0.36g, 5.55mmol), Zn (CN) 2 (2.69g, 22.9Zn), dppf (8.82g, 15.9mmol) and Pd2 (dba) 3 (7.8g, 8.51mmol) is mixed in DMA (50mL), 80 DEG C It is stirred overnight.Thin layer chromatography (PE/EA=5:1) tracks to almost all of compound 4 and is consumed.Reactant mixture is cooled to During room temperature is fallen back, filtering, filtrate is extracted with ethyl acetate, collects organic facies, washs with saline, is dried dense on na 2 so 4 Contracting obtains crude product, and silica gel chromatography (PE/EA=50:1,10:1) obtains faint yellow solid pure compound 5, and (6.0g produces Amount: 62.3%).
5th step: mixing cpd 5 (6.0g, 23.08mmol), sodium hydroxide (10g, 250mmol) are at ethylene glycol (70mL) Stir 3 hours at cohobation device with water (10mL).Thin layer chromatography (DCM/ methanol=10:1) tracks to reaction and completes.Will reaction Mixture is cooled to room temperature, adds aqueous citric acid solution and is acidified to pH=4, filters, and it is pure that precipitation obtains faint yellow solid after drying Compound 6 (6.0g, yield: 93.2%).
6th step: stirring mixing compound 6 (6.0g, 21.5mmol), in methanol (100mL), is added dropwise at 0 DEG C SOCl2(4.7mL,64.5mmol).After interpolation, reactant mixture is stirred at room temperature 1 hour, then heated overnight at reflux. Thin layer chromatography (DCM/ methanol=10:1) tracks to reaction and completes.Reactant mixture evaporation is removed major part solvent, and residue is noted Enter frozen water, add solid sodium carbonate regulation pH=8, and extract by ethyl acetate, collect organic facies, after washing with saline, Na2SO4 is upper to be dried, and concentrates and obtains bright-yellow solid compound 7 (3.9g, yield: 61.9%).
7th step: mixing compound 7 (3.9g, 13.3mmol) and 10%Pd/C (1.0g) are in methanol (150mL), in room temperature Lower H2 gas stirs 2 hours.Thin layer chromatography (PE/EA=2:1) tracks to reaction and completes.Being filtered by reactant mixture, filtrate concentrates After obtain the solid chemical compound 8 (2.6g, yield: 96.3%) of brown.
8th step: stirring mixing compound 8 (0.5g, 2.46mmol), in methanol (5mL), adds 2N NaOH under room temperature (2.5mL,5mmol.After interpolation, reactant mixture is stirred at room temperature night.Thin layer chromatography (DCM/ methanol=10:1) Indicator Reaction Complete.Being evaporated by reactant mixture, residue diluted with water, and extract by ethyl acetate, discard ethyl acetate layer, aqueous phase adds Aqueous citric acid solution also regulates pH=2, filters, and precipitation obtains pure compound 9 (0.4g, yield: 86%) after drying.
9th step: mixing compound 9 (0.4g, 2.11mmol), 3-5-trifluoromethylaniline (375mg, 2.32mmol), HATU (960mg, 2.53mmol) and DIPEA (820mg, 6.33mmol) are stirred overnight under DMF (5mL) room temperature in N2.Thin layer color Spectrum (DCM/ methanol=10:1) display reaction completes.Ethyl acetate after reactant mixture dilute with water is extracted, collects organic facies, Wash with saline, be dried on na 2 so 4 after concentrating and obtain crude product, pure by the solid obtaining yellow after silica gel chromatography Compound 10 (0.4g, yield: 57%).
The embodiment 2 compound V01(present invention also can be designated as V01) preparation
Step 1:
Compound 11A (20g, 0.15mol), DMAP (1.9g, 15.4mmol) and (BoC)2O (75g, 0.34mol), four Mixing in hydrogen furan (750mL), reactant mixture is stirred at room temperature overnight.Thin layer chromatography (PE/EA=3:1) display reaction completes. Be suspended in PE/EA (10:1,200mL) after being concentrated by reactant mixture, obtain after filtration white solid pure compound 11 (50g, 100%)。
Step 2:
Mixing cpd 10 (50mg, 0.15mmol) and Cs2CO3 (150mg, 0.45mmol) are in DMSO (1mL), at N2 In half an hour, is stirred at room temperature, be subsequently adding compound 11 (60mg).Consequent reactant mixture stirring half an hour, use Thin layer chromatography (DCM/ methanol=10:1) detection is to having reacted.After reactant mixture dilute, extracting with EA, collection has Machine phase, washs with saline, dried with Na2SO4, prepares the Purify compound 12 (30mg, 32%) of yellow after chromatogram purification.
Under room temperature, N2 stirs mixing cpd 12 (20mg, 0.032mmol) and TFA (0.1mL) half an hour.Thin layer color Spectrum (DCM/ methanol=10:1) display is to having reacted.After reactant mixture is alkalized by Na2CO3, extract with DCM, collect organic Phase, washs with saline, to be dried on Na2SO4, obtains crude product after concentration, by obtaining consolidating of yellow after silica gel chromatography Body pure compound V01 (4.5mg, 33%), Structural Identification sees Fig. 8.
The embodiment 3 compound V3(present invention also can be designated as V03) preparation
Step 1:
Stirring mixing compound 13 (5.0g, 45mmol) and pyridine (200mL), by (Boc) at 65 DEG C2O(14.7g, 67.5mmol) it is added drop-wise in said mixture.After interpolation, 85 DEG C of stirring reactions 4 hours.Thin layer chromatography (DCM/ methanol=10: 1) track to after reaction completes, reactant mixture be cooled to 0 DEG C, adds dense HCl (100mL) and H2O (50mL), EA and extract After, collect organic facies, use NaHCO3 solution washing, dried with Na2SO4, obtain the grease of yellow, be suspended in Et2O, filters, it is thus achieved that compound as white solid 14 (4.3g, yield: 45.2%).
Stirring mixing compound 14 (2.4g, 11.4mmol) and N, N dimethyl aniline (6.6mL) in DCM (84mL), 0 In DEG C N2, drip POCl3(3.2mL,34.2mmol).After interpolation, reactant mixture is stirred at room temperature 2 hours.Thin layer chromatography (PE/EA=2:1) track to after reaction completes, reactant mixture be injected frozen water.Organic facies salt is washed, and does on na 2 so 4 Crude product is obtained, by silica gel chromatography, it is thus achieved that white solid pure compound 15 (1.8g, 70%) after dry concentration.
Stirring mixing compound 15 (100mg, 0.47mmol) and DMAP (12mg, 0.09mmol), room in THF (1mL) Temperature is lower adds (BOC)2O (124mg, 0.57mmol), is then stirred overnight under room temperature.Thin layer chromatography (PE/EA=2:1) shows instead After should completing, by reactant mixture dilute with water, EA extracts, and collects organic facies, washes with salt, is dried on na 2 so 4 after concentrating Obtain crude product, by silica gel chromatography, it is thus achieved that white solid pure compound 16 (70mg, 44.8%).
Under room temperature, at N2Stirring mixing compound 10 (50mg, 0.15mmol) and Cs in middle DMSO (1mL)2CO3(150mg, 0.45mmol) half an hour, it is subsequently adding compound 16 (60mg, 0.18mmol), is stirred for half an hour.Thin layer chromatography (PE/EA= After 2:1) showing that reaction completes, by reactant mixture dilute with water, EA extracts, and collects organic facies, washes with salt, on na 2 so 4 It is dried after concentrating and obtains crude product, by silica gel chromatography, it is thus achieved that yellow solid pure compound 17 (50mg, 53.3%).
Step 2:
Under room temperature, in N2, in DMSO (1mL), stirring mixes compound 17 (50mg, 0.08mmol) and TFA (0.2mL) half Hour.After thin layer chromatography (DCM/ methanol=10:1) shows that reaction completes, after this reactant mixture is alkalized by Na2CO3, use DCM Extract.Organic facies is with brine wash, is dried on na 2 so 4 after concentrating and obtains crude product, by obtaining yellow after silica gel chromatography Purify compound V3 (15mg, 44%), its Structural Identification data see Fig. 9.
The embodiment 3 compound V4(present invention also can be designated as V04) preparation
Step 1:
Stirring mixing compound 18 (50g, 0.47mol), in DCM (600mL), adds TEA (94g, 0.93mol), then At 0 DEG C, in N2, dropping bromoacetate (94g, 0.56mol).Reactant mixture is stirred at room temperature overnight.Thin layer chromatography (PE/EA=1:1) after showing that reaction completes, reactant mixture salt is washed, and is dried on na 2 so 4 after concentrating and obtains crude product, By silica gel chromatography (using PE/EA=20:1to10:1to5:1 eluting), it is thus achieved that yellow oily pure compound 19 (46g, 51%)。
At 95 DEG C, toluene (800mL) stirring mixing compound 19 (38g, 196.65mmol) and TEA (29.9g, 295mmol), 4-bromobutyrate (72.9g, 373.65mmol) it is added dropwise over. add rear heated at reflux overnight.Thin layer chromatography (PE/EA=5:1), after showing that reaction completes, reactant mixture concentrates, by silica gel chromatography (with PE/EA=20:1to5: wash De-), it is thus achieved that yellow oily pure compound 20 (32g, yield:53%).
Stirring mixing compound 20 (32g, 104.3mmol) in toluene (800mL), at 0 DEG C, adds t-BuOK (51.2g,456.3mmol).After interpolation, reactant mixture at room temperature reacts 1 hour.Thin layer chromatography (PE/EA=5:1) shows After having reacted, adding 2N HCl and regulate pH=6, then EA extracts, and collects organic facies, washes with salt, is dried dense on na 2 so 4 Obtain crude product 21 (17g, yield:62.5%) after contracting, for dark yellow oil, can be directly used for next step reaction, be not required to further Purification.
((11g, 161.65mmol) is dissolved in methanol (280mL) to MeONa, and reactant mixture is cooled to 5 DEG C, adds second Acid carbonamidine (3.0g, 29.15mmol).Stirring reactant mixture half an hour, add compound 21 (17g, 65.1mmol).Will Reactant mixture 40 DEG C is stirred overnight.After thin layer chromatography (DCM/ methanol=10:1) shows that reaction completes, reactant mixture is cooled down To room temperature, evaporation removes most solvent.Residue water processes and EA extracts, and organic facies salt is washed, and does on na 2 so 4 Obtain crude product after dry concentration, silica gel chromatography obtain faint yellow solid pure compound 22 (2.5g, yield: 16%).
It is 0.5MPa H at pressure2In, it is stirred overnight mixing compound 22 (2.5g, 10.4mmol), 10%Pd/C (0.5g) (B DEG C)2O (2.7g, 12.4mmol) is in MeOH (40mL).After thin layer chromatography (DCM/ methanol=10:1) shows that reaction completes, Reactant mixture filtering and concentrating is obtained yellow solid pure compound 23 (2.6g, 100%).
Mixing compound 23 (1.6g, 6.3mmol), PPh in 1,2-DCE (64mL)3(3.33g, 12.6mmol) and CCl4 (2.93g, 18.9mmol), 70 DEG C are heated 1 hour.After thin layer chromatography (DCM/ methanol=10:1) shows that reaction completes, reaction is mixed After compound concentrates, silica gel chromatography obtain faint yellow solid pure compound 24 (1.38g, 81%).
In room temperature N2, stirring mixing compound 10 (100mg, 0.3mmol) and Cs in DMSO (2mL)2CO3(300mg, 0.9mmol) half an hour, add compound 24 (97mg, 0.36mmol).Reactant mixture stirring mixes half an hour.Thin layer After chromatograph (DCM/MeOH=10:1) shows that reaction completes, reactant dilute with water, then EA extracts.Organic facies salt is washed, Na2SO4 is upper is dried acquisition crude product after concentration, is obtained yellow solid by silica gel chromatography pure after being concentrated by reactant mixture Compound 25 (21mg, 12.4%).
Step 2:
In room temperature N2, middle stirring mixing compound 25 (21mg, 0.037mmol) and TFA (0.2mL) half an hour.Thin layer Chromatograph (DCM/MeOH=10:1) shows that reaction completes.After reactant mixture is alkalized by Na2CO3, extract with DCM.Organic facies is used Salt is washed, and is dried on na 2 so 4 after concentrating and obtains crude product, the Purify obtaining yellow after silica gel chromatography closes Thing V4 (10mg, 57.9%), its Structural Identification data see Figure 10.
The embodiment 4 compound V5(present invention also can be designated as V05) preparation
Step 1:
(S)-1-phenylethylamine (10g, 8.26mmol) and the 1,2-DCE of ethyl propanoic acid (11.9g, 8.26mmol) (200mL), in solution, it is dividedly in some parts NaBH (OAc)3(35g, 16.52mmol).After adding, reactant mixture is at room temperature stirred Mix 4 hours.Then reactant mixture is added NaHCO3Aqueous solution alkalizes, and extracts with DCM, separates organic facies, and with saturated Brine It, is dried on na 2 so 4, and is concentrated to give thick compound 34(20.5g, yield: 100%), can be without entering one Step purification is directly used in next step.
At 1,2-DCE(200mL) in stirring mixing compound 34 (20.5g, 82mmol) and glyoxylic acid ethyl ester ((33mL, 165mmol, the toluene solution of 50%), NaBH (OAc)3(35g, 165mmol), is stirred at room temperature 4 hours.Then will reaction Mixture NaHCO3Aqueous solution alkalizes, and extracts with DCM.Separate organic facies and wash with saturated aqueous common salt, doing at Na2SO4 Dry and be concentrated to give thick compound 35(26g, 100%), can be directly used in without further purification in next step.
Compound 35 (26g, 78mmol) and the t-BuOK (22g, 156mmol) mixing in toluene (600mL), stir back Flow through night.Then by reactant mixture NaHCO3Aqueous solution alkalization also extracts with DCM.Organic phases washed with brine, Na2SO4 is dried and concentrates, and obtains crude compound 36.
Take crude compound 36 and pass through silica gel chromatography (PE/EA=20:1 eluting) purification, obtain pure compound 37(3g).
Mixing cpd 37 (3.0g, 0.01mol) in ethanol (50mL) and formamidine acetate (3.24g, 0.03mol), mixture adds NaOEt (2.38g, 0.035mol) in solution.Stir this reactant mixture in 90 DEG C overnight, Then evaporate to remove most solvent.Residue H2O dilutes, and being acidified to pH with 2N HCl is 6, then extracts with DCM. Organic phases washed with brine, is dried at Na2SO4 and concentrates, obtain crude product, passed through Silica gel chromatography, obtains pure Compound 38(1.0g, yield: 35.8%), for yellow solid.
By compound 38 (1.0g, 10.4mmol), the Pd/C(0.1g of 10%) and (B DEG C)2O (2.7g, 12.4mmol) mixes Together in MeOH(40mL) in, it is stirred overnight in the hydrogen that pressure is 0.5 MPa.TLC(DCM/ methanol=10:1) show to have reacted Become.Reactant mixture is filtered, filtrate is concentrated, obtains crude product, passed through Silica gel chromatography, obtain pure chemical combination Thing 39(0.7g, productivity: 71%), for yellow solid.
Take compound 39 (0.7g, 6.3mmol) and triphenylphosphine PPh3(3.33g, 12.6mmol) is at carbon tetrachloride (10mL) mixing in, in 80 DEG C of stirred overnight.TLC(DCM/ methanol=15:1) show that reaction completes.Reactant mixture is concentrated, And by residue by Silica gel chromatography, obtain pure compound 40(0.4g, and 53.4%), for light yellow oil.
Under a nitrogen, compound 10 (150mg, 0.45mmol) and Cs2CO3(440mg, 1.35mmol) is in DMSO(2ml) in Mixing, is stirred at room temperature half an hour, is subsequently adding compound 40 (153mg, 0.54mmol).The reactant mixture of gained is stirred Mix half an hour, TLC(DCM/ methanol=10:1) show that reaction completes.By reactant mixture dilute with water and extract with EA.Organic facies Wash with saline, at Na2SO4It is dried and concentrates, obtain crude product, passed through the TLC purification of preparation, obtain pure compound 41(60mg, 23%), for yellow solid.
Step 2:
Take compound 41 (60mg, 0.104mmol) and mixture TFA(0.3mL) under a nitrogen, be stirred at room temperature half little Time.TLC(DCM/ methanol=10:1) show that reaction completes.By reactant mixture NaHCO3Aqueous solution alkalization also extracts with DCM.Have Machine saline washs, at Na2SO4Upper being dried concentrates, and obtains crude product, is passed through the TLC purification of preparation, obtains purification and closes Thing V5 (20mg, 40.3%), for yellow solid, its Structural Identification data see Figure 11,12.
The embodiment 5 compound V6(present invention also can be designated as V06) preparation
Step 1:
In flask equipped with mechanical agitator and the 3L of calcium chloride tube, load compound 26 (100g, 0.716mol) and Ethanol (1.0L).This mixture is stirred 20 minutes, then drips triethylamine (72.5g, 0.716mol).By the mixture of gained Stir 10 minutes, then, add ethyl acrylate (61.6g, 0.716mol).Reactant mixture is stirred at room temperature 17 little Time.(BOC)2O (234.5g, 1.08mol) is at room temperature added dropwise over.After completion of dropwise addition, reactant mixture is at room temperature stirred Mix overnight.Concentrate reactant mixture to remove in major part EtOH.Residue is dissolved in water (3L), and uses Et2O(1L × 3) extraction.The organic facies merged ammonium chloride (500L × 3) aqueous solution and saline (500L × 3) wash, and anhydrous Na 2SO4 is done Dry, and be concentrated to give thick compound 28 (300g, 92%), for yellow oil, its for next step without extra pure Change.
Sodium (27.6g, 0.765mol) adds in dehydrated alcohol (1.5L).When solid sodium is wholly absent, by compound 28 (300g, 1.04mol) adds in hot solution.Reactant mixture refluxes overnight.Thin layer chromatography (petrol ether/ethyl acetate=4:1) Show that initiation material is consumed completely.Being evaporated by reactant mixture, be dissolved in by residue in water (1L), then citric acid is water-soluble It is 6 that liquid is acidified to pH.Then by mixture EtOAc(1 liter × 3) extraction.Extract is merged, and washes with saline (1L × 3) Wash, at Na2SO4It is dried and evaporates, obtaining compound 29 (169g, 63.4%), for brown oil.This crude product is without further Purification is used for next step.
Feldalat NM (2.6g, 48.58mmol) is dissolved in MeOH(50mL) in, and this solution is cooled to 5 DEG C.Add acetic acid Carbonamidine (3.0g, 29.15mmol), stirs half an hour by reactant mixture.Add compound 29 (5.0g, 19.43mmol), will Reactant mixture is stirred at reflux overnight.TLC(DCM/ methanol=10:1) show that reaction completes.Reactant mixture is cooled to room temperature, And evaporate to remove major part solvent.Residue with water is processed and extracts with EA.Organic phases washed with brine, at Na2SO4Dry Dry and concentrate, obtain crude product, passed through Silica gel chromatography, obtain pure compound 30(680mg, yield: 14.7%), for light yellow solid.
Under 0 DEG C of nitrogen, phosphorus oxychloride (P DEG C of l3,174mg, 1.26mmol) joins stirring and is mixed with compound 30 (100mg, 0.42mmol) and the DCM(4mL of DMA (0.28mL)) in, after adding, reactant mixture is poured into In frozen water, alkalize by adding solid sodium carbonate, then extract with DCM.Organic phases washed with brine, at Na2SO4It is dried and dense Contracting, obtains crude product, by it by preparation TLC purification, obtains pure compound 31 (60mg, 55.6%), for white solid.
Under a nitrogen, compound 10 (50mg, 0.15mmol) and Cs2CO3(150mg, 0.45mmol) is mixed in DMSO (1mL) in, mixture is stirred at room temperature half an hour, is subsequently adding compound 31 (46mg, 0.18mmol).By the reaction of gained Mixture stirs half an hour, TLC(DCM/ methanol=10:1) show that reaction completes.By reactant mixture dilute with water and extract with EA Take.Organic phases washed with brine, is dried at Na2SO4 and concentrates, obtain crude product, is passed through the TLC purification of preparation, obtains Pure compound 32 (13mg, 15.7%), for yellow solid.
Step 2:
Under nitrogen, stir half an hour under compound 32 (13mg, 0.038mmol) and mixture room temperature TFA(0.1mL). TLC(DCM/ methanol=10:1) show that reaction completes.Reaction the mixture alkalization of Na2CO3 aqueous solution and DCM are extracted.Organic Wash with saline, be dried at Na2SO4 and concentrate, obtain crude product, by it by preparation TLC purification, obtain pure chemical combination Thing, V6 (6mg, yield: 56.3%), for yellow solid, its Structural Identification data see Figure 13.
The preparation method of embodiment 6KDR3
1st step: by compound 1(1.4 gram, 0.01mmol), 4-(benzyloxymethyl)-6-chloropyrimide (4.7 grams, 0.02) and K2CO3(4.2 gram, 0.03 mole) at DMF(50mL) in mixing, 100 DEG C, under N2 stirring be 5 hours.TLC(PE/EA=4:1) Show that reaction completes.Reactant mixture is cooled to room temperature dilute with water, extracts with EA.Collect organic facies, wash with saline, It is dried with Na2SO4 and concentrates, obtaining crude product, being passed through column chromatography purification, obtaining pure compound 2(3.0 gram, productivity: 90%), for yellow solid.
Step 2: under N2 atmosphere, compound 2(1.2 gram, 3.56 mMs), hydrazine hydrate (0.36 gram, 7.12 mMs) With Raney Ni(0.2g) THF(20mL) in mixture return stirring 1 hour.TLC(PE/EA=2:1) show to have reacted Become.Reactant mixture is filtered, filtering and concentrating, obtain crude product, passed through column chromatography purification, obtain pure compound 3 (0.8 gram, productivity: 73%), for yellow solid.
Step 3: at 0 DEG C under a nitrogen, by TCDI(0.78 gram, 4.4mmol) and compound 3(0.9 gram, 2.93mmol) Be dried DCM(10mL) in be slowly mixed together.After adding, reactant mixture is stirred at room temperature half an hour.TLC(PE/EA=1: 1) show reactant mixture completes.Reactant mixture is concentrated, obtains crude product, purify with flash chromatography, obtain Crude compound 4(1.0 gram, productivity: 100%), for pale solid.
Step 4: by compound 4(1.1 gram, 3.15 mMs), 4-methyl-5-(trifluoromethyl) and benzene 1,2-diamidogen (0.6 Gram, 3.15 mMs) and the THF(50 milliliter of carbodiimide (1.2 grams, 6.30 mMs)) under a nitrogen, stirred at reflux 2 is little Time.TLC(DCM/ methanol=10:1) show, reactant mixture completes.This reactant mixture, and filtering precipitate is diluted with EA. Filtrate is evaporated, obtains crude product, used column chromatography (PE/EA=10:1 to 4:1) purification, obtain pure compound 5(0.8 Gram, productivity: 50%), for brown oil.
Step 5: compound 5(0.7 gram, 1.38 mMs) and TFA(15ml) in mixture in 60 DEG C of stirred overnight.Will Reactant mixture is warming up to 70 DEG C, and stirring 1 hour at this temperature.TLC(DCM/ methanol=10:1) show, reactant mixture In complete.Reactant mixture being cooled to room temperature and is poured in frozen water, alkalizing to pH with the NaOH of 2N is 10, then extracts with EA Take.Organic phases washed with brine, is dried with Na2SO4 and concentrates, and obtains the crude product (0.4g, yield: 70%) of compound 6, for Yellow oil, it is directly used in next step without further purification.
Step 6: at the compound 6(60mg that stirred, every day 0.1445mmol) DCM(5ml) in, add TEA(30 milli Gram, 0.289mmol) and MSCL(25 milligram, 0.2168mmol) under 0 DEG C of nitrogen protection.After completion of dropwise addition, reaction terminates.Will This mixture DCM dilutes, and washs with saline, is dried at Na2SO4 and concentrates, obtain the crude product of compound 7, and it is without entering One step purification is directly used in next step.
7th step: compound 7(60 milligram, 0.12 mM) and MeNH2(0.6 milliliter, 1.2 mMs, in MeOH Anhydrous THF(5ml 2N)) in mixture in stirred overnight at room temperature.TLC(DCM/ methanol=10:1) show, in reactant mixture Complete.Reactant mixture is concentrated and purification residue, TLC purification three times, obtains pure KDR3(6 milligram, productivity: 12%), for White solid, its Structural Identification data see Figure 14.
Prepared by embodiment 7KDR4 compound
The first step: by compound 1(1.0 gram, 4.5 mMs) at ACN(20 milliliter) in mixing 2-chloro-5-nitropyridine 2- Chloro-5-nitropyridine (0.8 gram, 5 mMs) and cesium carbonate (3.3 grams, 10 mMs), be stirred at room temperature overnight.TLC(PE/ EA=1:1) show reactant mixture completes.By reactant mixture dilute with water, extract with EA.Organic phases washed with brine, Na2SO4 is dried and concentrates, and obtains crude product, is passed through purification by flash chromatography, obtains pure compound 2(0.4 gram, yield: 24.5%), for yellow solid.
Second step: to compound 2(0.3 gram, 0.83 mM) THF(5mL) in stirring mixture in add in Ruan Nickel (0.1 gram), is then heated to backflow.H2N-NH2H2O(1.66 mM) THF(5mL) in solution be added dropwise over.Will The reactant mixture of gained stirs 10 minutes.TLC(DCM/ methanol=15:1) show, reactant mixture completes.Reaction is mixed Thing filters, and filtering and concentrating obtains crude product, passed through purification by flash chromatography, obtains pure compound 3(0.16 gram, productivity: 50%), for yellow solid.
3rd step: at compound 3(0.16 gram, 0.483mmol) agitating solution in, under the protection of 0 DEG C of nitrogen, at DCM (10mL) TCDI(95mg, 0.531mmol are added in).After completion of dropwise addition, reactant mixture is stirred at room temperature.Overnight.TLC (DCM/ methanol=10:1) shows, completes in reactant mixture.Reactant mixture is concentrated, obtains crude product, passed through quickly Chromatogram purification, obtains pure compound 4(50 milligram, productivity: 28%) it is pale solid.
4th step: by compound 4(50 milligram, 0.134mmol), 4-methyl-5-(trifluoromethyl)-benzene-1,2-diamidogen (25.5mg, 0.134 mM) and EDCI(51 Mick, 0.268 mM) at anhydrous THF(10mL) in, refluxed under nitrogen is stirred Mix 1 hour.TLC(DCM/MEOH=15:1) show, reactant mixture completes.Diluted reaction mixture, by EA and filtration precipitation Thing, obtains crude product, by it by pre-TLC purification, obtains pure compound 5(40 milligram, yield: 56%), for white solid.
5th step: compound 5(20 milligram, 0.038 mM) and the mixture of trifluoroacetic acid (0.2 milliliter), in room temperature Lower stirring half an hour.TLC(DCM/ methanol=5:1) show reactant mixture completes.Reactant mixture is evaporated to remove TFA, Residue is dissolved in THF, solid K2CO3 alkalizes, filter and concentrated filtrate, obtain pure compound KDR4(10 milligram, produce Rate: 62.5%), for white solid, its Structural Identification data see Figure 15.
Prepared by embodiment 10KDR6 compound
The first step: 7-methoxy quinoline-4-alcohol (10.0 grams, 57.1mmol) is dissolved in 50ml phosphorus oxychloride, then will be anti- Mixture is answered to heat 3 hours at 120 DEG C.Then mixture is cooled to room temperature, and pours frozen water (50 milliliters) into.Extract with EtOAc Take this mixture (100mLx3), organic layer saturated aqueous common salt is washed, is dried with anhydrous sodium sulfate and concentrates, slightly being produced Thing, is used flash-chromatography (with PE/EA=30:1 eluting) purification, and the product obtained (8.5 grams, yield: 76.9%), in vain Color solid.
Second step: 4-chloro-7-methoxy quinoline (5.5G, 28.4mmol), zinc (276.9mg, 4.26mmol), Zn(CN) 2 (2.07 grams, 17.6mmol), DPPF(6.65 gram, 12.21mmol), PD2(DBA) and 3(5.98 gram, 6.53mmol) it is dissolved in In the DMAc of 100mL, mixture argon covered and heats at 160 DEG C overnight, then mixture being cooled to room temperature, and leads to Cross kieselguhr to filter, by filtrate with H2O process, then with EA extraction, organic layer saturated aqueous common salt is washed, use anhydrous slufuric acid Sodium is dried, and is concentrated under vacuum.By flash-chromatography (with PE/EA=10:1 eluting) purification, obtain product 7-methoxyl group quinoline Quinoline-4-formonitrile HCN (4.38 grams, yield: 83.7%), for brown solid.
3rd step: 7-methoxy quinoline-4-formonitrile HCN (4.38 grams, 23.78mmol) is dissolved in the H2O of 30mL, adds hydrogen Sodium oxide (2.38 grams, 59.45mmol).Then 100 DEG C are heated the mixture to overnight.Reactant mixture is cooled to room temperature, And extract with EA, to remove impurity.Aqueous phase is acidified, is 2 by adding 6N hydrochloric acid to pH.By the precipitate of yellow by filtering Collecting and be dried, obtain the 7-quininic acid of 2.5g, productivity 52%, for brown solid.
4th step: 7-quininic acid (2.0 grams, 9.84mmol) and DIPEA(3.82 gram, 29.52mmol) molten Solution is in DMF.Then reactant mixture is cooled to 0 DEG C, adds HATU(4.49 gram, 11.81mmol) and 3-(trifluoromethyl) Aniline (1.74 grams, 10.82mmol).This mixture argon covers, and is then stirred at room temperature overnight.This mixture water is dilute Release and use EtOAc(3 × 50 milliliter) extraction.Washing organic layer, washs with saline, is dried with anhydrous sodium sulfate, and under vacuo Concentrate, obtain crude product, passed through purification by flash chromatography, and PE/EA=1:1 eluting, obtain product (2.5g, yield: 79.6%) it is light yellow solid.
5th step: 7-methoxyl group-N-(3-(trifluoromethyl) phenyl) quinoline-4-Methanamide (0.6 gram, 1.733mmol) in DCM(50 milliliter) in solution, 10 DEG C of drop wise under nitrogen add BBr3(1.73mL, 6.93 mMs.After adding, reaction is mixed Thing is stirred at room temperature 100 hours.TLC(DCM/ methanol=15:1) show that reaction completes.Reactant mixture is poured in ice-water And extract with DCM.Washing organic layer, with saline wash, anhydrous Na 2SO4 is dried, the product of concentration, for brown solid (0.5 gram, Productivity: 87%), use it for next step, it is not necessary to be further purified.
6th step: by 7-hydroxy-n-(3-(trifluoromethyl) phenyl) quinoline-4-Methanamide (50 milligrams, 0.151mmol), The tert-butyl group (6-chloropyrimide-4-base) methyl (methyl) t-butyl carbamate (46.6mg, 0.181mmol) and K2CO3 (41.7mg, 0.302mmol) is at DMSO(2ml) solution mixes, it is stirred overnight at 90 DEG C.TLC(PE/EA=1:2) reaction is shown Complete.By reactant mixture dilute with water and extract with EA.Washing organic layer, washs with saline, and anhydrous Na 2SO4 is dried, dense Contracting, crude product, this is by pre--TLC purification, obtains pure product (25 milligrams, productivity: 30%), for white solid.
7th step: tertbutyl methyl ((6-(4-(3-(trifluoromethyl) phenylcarbamoyl) quinoline-7-base epoxide) phonetic Pyridine-4-base) methyl) t-butyl carbamate (25 milligrams, 0.045mmol) and TFA(0.25 milliliter) and mixture, at 0 DEG C Stir half an hour.TLC(DCM/ methanol=5:1) show that reaction completes.Evaporation TFA, and residue is alkalized by aqueous solution. Na2CO3 and extracting with DCM.Washing organic facies, washs with saline, is dried with Na2SO4 and concentrates, obtaining crude product, being led to Cross pre--TLC purification, obtain pure compound 06(7 milligram, yield: 21%), for white solid, its Structural Identification data see Figure 16.
Prepared by embodiment 11KDR6A compound
The first step: 7-methoxy quinoline-4-alcohol (10.0 grams, 57.1mmol) is dissolved in 50ml phosphorus oxychloride, then will be anti- Mixture is answered to heat 3 hours at 120 DEG C.Then mixture is cooled to room temperature, and pours frozen water (50 milliliters) into.Extract with EtOAc Take this mixture (100mLx3), organic layer saturated aqueous common salt is washed, is dried with anhydrous sodium sulfate and concentrates, slightly being produced Thing, is used flash-chromatography (with PE/EA=30:1 eluting) purification, and the product obtained (8.5 grams, yield: 76.9%), in vain Color solid.
Second step: 4-chloro-7-methoxy quinoline (5.5G, 28.4mmol), zinc (276.9mg, 4.26mmol), zinc (CN) 2 (2.07 grams, 17.6mmol), DPPF(6.65 gram, 12.21mmol), PD2(DBA) and 3(5.98 gram, 6.53mmol) it is dissolved in In the DMAc of 100mL, mixture argon covered and heats at 160oC overnight, then mixture being cooled to room temperature, and leads to Cross kieselguhr to filter, by filtrate with H2O process, then with EA extraction, organic layer saturated aqueous common salt is washed, use anhydrous slufuric acid Sodium is dried, and is concentrated under vacuum.By flash-chromatography (with PE/EA=10:1 eluting) purification, obtain product 7-methoxyl group quinoline Quinoline-4-formonitrile HCN (4.38 grams, yield: 83.7%), for brown solid.
3rd step: 7-methoxy quinoline-4-formonitrile HCN (4.38 grams, 23.78mmol) is dissolved in the H2O of 30mL, adds hydrogen Sodium oxide (2.38 grams, 59.45mmol).Then 100 DEG C are heated the mixture to overnight.Reactant mixture is cooled to room temperature, And extract with EA, to remove impurity.Aqueous phase is acidified, is 2 by adding 6N hydrochloric acid to pH.By the precipitate of yellow by filtering Collecting and be dried, obtain the 7-quininic acid of 2.5g, productivity 52%, for brown solid.
4th step: 7-quininic acid (406mg, 2 mMs) and DIPEA(775mg, 6 mMs) it is dissolved in DMF(5mL) in.Reactant mixture is cooled to 5 DEG C, is subsequently adding HATU(912mg, 2.4mmol) and 4-fluoro-3-(fluoroform Base) aniline (394mg, 2.2 mMs).This mixture uses argon and covering at ambient temperature, overnight.TLC(PE/EA=1:2) Show that reaction completes.By reactant mixture dilute with water and extract with EA.Washing organic layer, washs with saline, anhydrous Na 2SO4 It is dried, and concentrates, crude product, by it by CCTO purification, obtain pure product (220 milligrams, yield: 30%), for brown solid.
5th step: at N-(4-fluoro-3-(trifluoromethyl) phenyl)-7-methoxy quinoline-4-Methanamide (0.2 gram, DCM(10ml 0.549mmol)) in solution, at 0 DEG C, N2 stirred under argon adds 4N BBr3(0.68 milliliter, 2.75mmol) DCM solution.After adding, being stirred at room temperature by reactant mixture is 20 hours.Then reactant mixture is stirred 4 at-30 DEG C Hour.TLC(DCM/ methanol=10:1) show that initiation material is still.Reactant mixture is poured in ice-water and extracts with DCM.Will Organic layer saturated aqueous common salt washs, and is dried with anhydrous sodium sulfate and concentrates, obtaining crude product, by it by pre-TLC purification, obtaining Raw material (120 milligrams) and pure product N-(4-fluoro-3-(trifluoromethyl base) phenyl)-7-hydroxyquinoline-4-Methanamide (60 milli Gram, 78%), for white solid.
6th step: by fluoro-for N-(4-3-(trifluoromethyl) phenyl)-7-hydroxyquinoline-4-Methanamide (30 milligrams, 0.086mmol), the tert-butyl group (6-chloropyrimide-4-base) methyl (methyl) t-butyl carbamate (26.5mg, 0.103mmol) and K2CO3(23.8mg, 0.172mmol) at DMF(1mL) in, stir 15 hours at 50 DEG C.TLC(DCM/ methanol=10:1) show instead Should complete.By reactant mixture dilute with water.And extract EA.Organic layer saturated aqueous common salt is washed, does with anhydrous sodium sulfate Dry and concentrate, obtain crude product, passed through pre--TLC purification, obtain pure product (6-(4-(4-fluoro-3-(trifluoromethyl uncle Butyl) phenylcarbamoyl) quinoline-7-base epoxide) pyrimidine-4-yl) methyl (methyl) t-butyl carbamate (20 milligrams, Yield: 41%), for yellow solid.
7th step: mixture (the 6-(4-(4-fluoro-3-(trifluoromethyl) phenylcarbamoyl of the first tert-butyl ester) quinoline-7- Base epoxide) pyrimidine-4-yl) methyl (methyl) t-butyl carbamate (20 milligrams, 0.035mmol) and trifluoroacetic acid (0.3 milli Rise) it is stirred at room temperature one hour.TLC(DCM/MEOH=5:1) show that reaction completes.Evaporation TFA, and by residue by Na2CO3 alkalization also extracts with DCM.Washing organic facies, washs with saline, is dried with Na2SO4 and concentrates, obtaining crude product, will It, by pre--TLC purification, obtains pure compound 06A(5 milligram, yield: 30%), for yellow solid, its Structural Identification data See Figure 17.
Prepared by embodiment 12KDR8 compound
1st step: the mixture of ethyl oxalate (70 milliliters) is heated to 120 DEG C, adds compound 1(50 gram, 0.4mol), And heat the mixture to 180 DEG C, maintain 5 minutes.By mixture refrigerated overnight, white depositions is collected by filtration, is dried, from And obtain compound 2(75.0 gram of white powder, yield: 59.11%).
Step 2: by compound 2(25 gram, 0.11 mole) it is dissolved in the dimethylbenzene (1L) of boiling, it is slowly added into P2S5 (9.5 grams, 0.0385 mole), backflow, until having reacted (about 5 hours), it is further continued for backflow.Amide TLC(PE/EA=5/1) table After this reaction bright completes, being cooled down by reactant mixture, and extract with 1N NaOH, aqueous phase hydrochloric acid is acidified, and collects yellow mercury oxide Thing, is then dissolved in EtOAc, washs with H2O, is dried with Na2SO4, and concentrates, and the compound obtained 3(19 gram is received Rate: 70.9%) it is a kind of red oil.
Step 3: compound 3(30 gram, 0.142 mole is dissolved in 1N sodium hydroxide (500mL), with the potassium ferricyanide (135 grams, 0.416mol, in 340ml water) oxidation, carry out to ferricyanide solution by being slowly added to thioamides, make temperature keep Less than 10 DEG C.After 1h, in TLC(DCM/MeOH=3/1) show that reaction is completely.Reactant mixture is being diluted in water, Acidifying (being 6 by the HCl to pH of 2N), then filters, the compound 4(15g obtained, yield: 50.48%).
Step 4: compound 4(1 gram, 4.34 mMs) at DMF(20mL) in stirring and dissolving, add 1-methyl-5- (trifluoromethyl)-1H-pyrazol-3-amine (0.717g, 4.35mmol), is subsequently added and adds in room temperature under nitrogen Enter HATU (2.47g, 6.5mmol) and DIEA (1.12g, 8.6mmol).After being stirred overnight under at a temperature of identical, TLC (PE/EA=2/1) show that this reaction completes.This reactant mixture is poured in water, and extracts with EtOAc.Washing will organically Phase, washs with saline, is dried on na 2 so 4 and concentrates, and obtains thick product, by silica gel chromatography (PE/EA eluting=20/ 1-5/1) it is purified, obtain pure compound 5(900 milligram, yield: 75.5%).
In step 5:-10 DEG C nitrogen, at compound 5(0.4g, 1.12mmol) DCM(10mL) in solution, add BBr3 (10mL, 4N in DCM).After 4 hours, TLC(PE/EA=1/1) show that reaction completes.Then reaction add ice, crosses and filter Remove undissolved material.Separating filtrate, extracts with DCM.By organic phases washed with brine, it is dried with Na2SO4 and concentrates, obtaining Thick product, this is purified, TLC obtain the compound (100mg, yield: 26.02%) of pure compound 6.
Step 6: the DMF(10mL of the solution (50mg, 0.146mmol) of compound 6) solution adds K2CO3 (70mg, 0.500mmol) with tert-butyl (6-chloropyrimidin-4-yl) methyl (methyl) carbamate (41.41mg,0.161mmol).By reactant mixture at 50 DEG C of stirred under nitrogen overnight.TLC(PE/EA=1/1) show to have reacted Become.Reactant mixture is washed with water, extracts with EA.Organic facies salt washs 4 times, is dried with Na2SO4 and concentrates, and obtains thick Product, TLC purification obtain pure compound 7(40mg, yield: 48.59%).
7th step: compound 7(40mg, 0.071mmol) and TFA(0.4mL) mixed at room temperature under a nitrogen, stir half an hour. TLC(DCM/ methanol=10/1) show that reaction completes.Reactant mixture is evaporated to remove TFA.Residue is alkalized, uses Na2CO3 Alkalization, to pH9, then extracts with DCM.By organic phases washed with brine, sodium sulfate is dried, and concentrates, obtains crude product, led to Cross TLC purification, obtain pure compound 8(20mg, yield: 44%).Its Structural Identification data see Figure 18.
Prepared by embodiment 13KDR8A compound
1st step: ethyl oxalate (70 milliliters) is heated to 120 DEG C, adds compound 1(50 gram, 0.4mol), and will mixing Thing is heated to 180 DEG C, and heat time heating time is: 5 minutes.After cooling, mixture, in refrigerator overnight, adds 50 milliliters of water, then shapes Become white depositions.Collect this solid, be dried, obtain 2(75.0 gram of compound of white powder, yield: 59.11%).
Step 2: by compound 2(25 gram, 0.11 mole) it is dissolved in the dimethylbenzene (1L) of boiling, it is slowly added to P2S5 (9.5 grams, 0.0385 mole), backflow, until having reacted (about 5 hours), it is further continued for backflow.TLC(PE/EA=5/1) represent anti- After should completing, being cooled down by reactant mixture, and extract with 1N NaOH, aqueous phase hydrochloric acid is acidified, and collects yellow mercury oxide, then It is dissolved in EtOAc, washs with H2O, be dried with Na2SO4, and concentrate, the compound obtained 3(19 gram, yield: 70.9%), for red oil.
Step 3: crude compound 3(30g, 0.142mol) it is dissolved in 1N sodium hydroxide (500mL), with the potassium ferricyanide (135 Gram, 0.416mol, in 340ml water) oxidation, carry out to ferricyanide solution by being slowly added to thioamides, make temperature Keep below 10 DEG C.After 1h, in TLC(DCM/MeOH=3/1) show that reaction is completely.By reactant mixture in water dilute Release, the HCl of 2N regulating to pH is 6, then filters, the compound 4(15g obtained, yield: 50.48%)
Step 4: compound 4(1 gram, 4.78 mMs) at DMF(40 milliliter) solution adds 3-(trifluoromethyl) aniline, It is subsequently added HATU(2.73 gram, 7.17 mMs) and DIEA(1.24 gram, 9.56 mMs), stir under nitrogen at room temperature.? Be stirred overnight rear TLC(PE/EA=2/1 at a temperature of identical) show that reaction completes after, reactant mixture is poured in water, and uses EtOAc extracts.Washing organic facies, washs with saline, is dried at Na2SO4 and concentrates, obtain crude product, passed through silica gel color Spectrum (with PE/EA=20/1-5/1 eluting) purification, obtains pure compound 5(1.68 gram, yield: 99.0%).
Step 5:-10 DEG C under a nitrogen, compound 5(0.1 gram, 0.283 mM) DCM(10mL) solution adds 4N BBr3(0.5mL).After 4 hours, TLC(PE/EA=1/1) show that reaction completes after, add ice, then filter to remove described Undissolved material.Filtrate extracts with DCM.Wash organic facies with saline, be dried with Na2SO4 and concentrate, obtain crude product, will Its logical TLC purification, obtains pure compound 6(64mg, yield: 66.65%).
Step 6: compound 6(64 milligram, 0.189 mM) DMF(10 milliliter) in solution, add K2CO3(78 milligram, 0.567 mM) and the tert-butyl group (6-chloropyrimide-4-base) methyl (methyl) t-butyl carbamate (tert-butyl (6- Chloropyrimidin-4-yl) methyl (methyl) carbamate, 53 milligrams, 0.208 mM).By reactant mixture 50 DEG C are stirred overnight under a nitrogen, TLC(PE/EA=1/1) show that reaction completes.Being washed with water by reactant mixture, EA extracts. Washing organic facies, washs four times with saline, is dried with Na2SO4 and concentrates, obtaining crude product, is passed through pre--TLC purification, To pure compound 7(20mg, yield: 19.68%).
7th step: compound 7(20 milligram, 0.035 mM) and mixture TFA(0.2mL), room temperature is stirred under a nitrogen Mix half an hour.In TLC(DCM/MeOH=10/1) show that reaction completes.Reactant mixture evaporation is removed TFA.By residue by SNa2CO3 regulation pH is 9, then extracts with DCM.Washing organic facies, washs with saline, is dried with Na2SO4 and concentrates, obtaining Crude product, is passed through pre--TLC purification, is obtained pure compound K DR08A(12mg, yield: 73.07%), its Structural Identification Data see Figure 19,22.
Prepared by embodiment 14KDR8B compound
1st step: the mixture of ethyl oxalate (70 milliliters) is heated to 120 DEG C, adds compound 1(50 gram, 0.4mol), And heat the mixture to 180 DEG C, maintain 5 minutes.By refrigerated overnight after mixture, form white depositions.Collect this solid In by filtering, and it is dried, thus obtains 2(75.0 gram of the compound for white powder, yield: 59.11%).
Step 2: by compound 2(25 gram, 0.11 mole) it is dissolved in the dimethylbenzene (1L) of boiling, it is slowly added into P2S5 (9.5 grams, 0.0385 mole), backflow, until completing (about 5 hours) .TLC(PE/EA=5/1 by reactant) show that reaction completes. This reactant mixture cools down, and with the extraction of 1N NaOH, aqueous phase hydrochloric acid acidifying, collects yellow mercury oxide, then dissolved In EtOAc, wash with H2O, be dried with Na2SO4, and concentrate, the compound obtained 3(19 gram, yield: 70.9%) it is red Grease.
Step 3: compound 3(30 gram, 0.142mol) it is dissolved in 1N sodium hydroxide (500 milliliters), use the potassium ferricyanide (135 grams, 0.416mol, in 340ml water) aoxidize, and carry out to ferricyanide solution by being slowly added to thioamides, make Temperature keeps below 10 DEG C.After 1h, in TLC(DCM/MeOH=3/1) show that reaction completes.By reactant mixture in water Dilution, is acidified (being 6 by 2N HCl to pH), then filters, the compound 4(15g obtained, yield: 50.48%).
Step 4: compound 4(1 gram, 4.78 mMs) DMF(20mL) in solution, under room temperature under nitrogen, stirring adds 4- Fluoro-3-(trifluoromethyl) anilinechloride (4-fluoro-3-(trifluoromethyl) aniline Hydrochloride, 1.24 grams, 5.74 mMs), it is subsequently added HATU(2.73 gram, 7.17 mMs) and DIEA(1.85 gram, 14.34 mMs). After being stirred overnight at a temperature of identical, TLC(PE/EA=2/1) show that reaction completes.Reactant mixture is poured in water, and Extract with EtOAc.Wash organic facies with saline, be dried at Na2SO4 and concentrate, obtain crude product, passed through silica gel chromatography (with PE/EA=20/1～5/1 eluting) purification, obtains pure compound 5(1.3 gram, yield: 73.4%).
Step 5: compound 5(0.5 gram, 1.35 mMs) DCM(10mL) in solution, under-10 DEG C of nitrogen, add 4N BBr3(2.5mL), after reacting 4 hours, TLC(PE/EA=1/1) show that reaction completes.In reaction add ice, then filter with Remove undissolved material.Separating filtrate, extracts with DCM.Saline washing organic facies, is dried with Na2SO4 and concentrates, obtaining Crude product, is passed through pre--TLC purification, is obtained pure compound 6(250mg, yield: 51.97%).
Step 6: compound 6(70 milligram, 0.196 mM) DMF(10mL) in solution, add K2CO3(81.46 milli Gram, 0.589 mM) and the tert-butyl group (6-chloropyrimide-4-base) methyl (methyl) t-butyl carbamate (tert-butyl (6- Chloropyrimidin-4-yl) methyl (methyl) carbamate, 55.70 milligrams, 0.216 mM), reaction is mixed Compound is at 50 DEG C of stirred under nitrogen overnight.TLC(PE/EA=1/1) show that reaction completes.Reactant mixture is washed with water, with EA Extraction.Washing organic facies, washs 4 times with saline, is dried at Na2SO4 and concentrates, obtain crude product, passed through TLC purification, Obtain pure compound 7(40 milligram, 39.51%).
7th step: compound 7(38 milligram, 0.071 mM) and TFA(0.4 milliliter) mixture, under nitrogen room temperature Stir half an hour.In TLC(DCM/MeOH=10/1) show that reaction completes.Reactant mixture evaporation is removed TFA.By residue Being regulated to pH by Na2CO3 is 9, then extracts with DCM.Washing organic facies, washs with saline, is dried with Na2SO4 and concentrates, To crude product, passed through TLC purification, obtained pure compound K DR8B(15 milligram, 47.75%), its Structural Identification data are joined See Figure 21.
Beneficial effects of the present invention is illustrated below by way of test example.
Test example 1 Brachydanio rerio vascular development inhibition test
Brachydanio rerio (Danio rerio) is a kind of bony fish that Cyprinidae (Cyprinidae) short hundredweight Buddhist nun fish belongs to (Danio).Its Gene is up to 85% with the similarity of human gene, and raun once can lay eggs 200～300 pieces, its fertilization and embryo development procedure Carry out the most in vitro, just can grow shaping in 24 hours, and embryo's entire body is transparent, it is simple to observe the change of intracorporeal organ tissue. Various features becomes one of 5 kinds of Fish as Laboratory Animals of Liao Shi International Organization for Standardization accreditation.At present, Brachydanio rerio is the mankind Being used widely in disease research, the research in terms of cardiovascular system is the most.
Experimental technique: this experiment uses the Brachydanio rerio usually used as examination compound on vascular formation function influence to be animal Model.After selected for postnatal zebrafish embryo, it be put in culture dish and be placed in incubator fostering 3-5 days, the present invention Compound is pressed variable concentrations (1 μM, 10 μMs, 30 μMs, 100 μMs) after Brachydanio rerio is born and is directly added in culture fluid.After 24 hours Check the developmental state of vertebra blood vessel and take a picture with fluorescence microscope.130B is pazopanib (Pazopanib), right as the positive According to, DMSO is as negative control.
Result of the test sees Fig. 1-4, table 1a, 1b.
The table 1a:KDR series little molecule inhibitory action to FLK-1 transgenic zebrafish vascular development
Concentration (uM) KDR3 KDR4 KDR6 KDR8 KDR8A
1 0% 0% 0% 0% 0%
10 0% 0% 0% 0% 0%
30 0% 10% 0% 0% 0%
100 50% 100% 10% 5% 10%
The table 1b KDR series little molecule inhibitory action to FLK-1 transgenic zebrafish vascular development
AI: vascular study rate
From Fig. 1-4, the compounds of this invention KDR4 and V01-v6 all can suppress Brachydanio rerio vascular development, especially V01, V3, v6, its drug activity is notable.
From table 1b, the activity of compound DKR4 is substantially better than KDR3, KDR6, KDR8 and KDR8A, compound V01, The activity of V03 is substantially better than KDR4.
The effect experiment of test example 2 the compounds of this invention
1. pair VEGFR2 body outer suppressioning test
The experimental technique of detection the compounds of this invention suppression VEGFR2 tyrosine phosphorylation is as follows:
1) cultivation to P3-P5HUVEC cell is transferred to 6 orifice plates, every hole about 2*105Cell
2) treat that cell grows to 70-80%, add candidate small molecule inhibitor (10nM, 100nM or1 μM), hatch 30min
3) add VEGF50ng/ml to stimulate, continue 10min
4) add the termination of non denatured lysate to react, collect cell pyrolysis liquid, carry out protein quantification
5) SDS-PAGE electrophoresis, transferring film, with the TTBS buffer (Tris-of 5% defatted milk preparation after cutting film Buffered saline/0.01%Tween20) close 2 hours
6) wash film, close overnight with anti-phosphorylation VEGFR2 antibody (1:1000 dilution) 4 DEG C
7) 1 hour is hatched with the goat antirabbit two of horseradish peroxidase-labeled is anti-altogether with film after within the 2nd day, washing film
8) develop the color with chemical luminescence reagent kit (Millipore) after washing film, photograph.
Result of the test sees Fig. 5.
As shown in Figure 5, the compounds of this invention KDR4 and V01, V3 all can suppress VEGFR2, and wherein, compound V01, V3 are imitated Fruit is more preferable compared with KDR4.
Test example 3 the compounds of this invention inhibitory action to choroidal neovascularization
Mice is c57/BL, uses the choroidal neovascularization (Choroidal of induced with laser Neovascularization, CNV) animal model tests.The one that this model is currently also most widely used is used to grind Study carefully medicine and CNV is formed the animal model of impact.Use 532 laser C57/bl6 mice (4 laser spots of each retina) OK Laser retinal light coagulates the CNV induced and is formed.Laser is injected after implementing immediately: a mice right eye injection concentration The KDR4 of 150uM, the V01 of another mice right eye injection concentration 1uM, two mice left eye injection all injections and medicine same volume PBS as comparison.Inject latter 5 days and animal is put to death and obtains eyeball.After removing anterior ocular segment, vitreous body and retina, make Choroid tile Parallel I solectin dyes.Zeiss fluorescence microscope system is used to carry out taking pictures and the measurement of CNV area.Knot Fruit sees Fig. 6, Fig. 7.
According to Fig. 6, compound K DR4(150uM), V01(1uM) all can significantly inhibit choroidal neovascularization formed, Can effectively treat or alleviate wet MD.
Test example 4 the compounds of this invention is on kinase whose impact
Protein kinase (protein kinases) is also known as protein phosphorylation enzyme (protein phosphakinase).One The enzyme of class catalytic proteins phosphorylation reaction.It can transfer to protein molecule the γ-phosphoric acid on adenosine triphosphate (ATP) On amino acid residue on the hydroxyl of some serine, threonine or tyrosine residue, thus change protein, the conformation of enzyme and work Property.The phosphorylation of protein is the important step of multi-signal pathway, and the important life of intracellular major part lives through journey all It is unable to do without protein phosphorylation.
Protein kinase falls into 5 types: Protein Serine/threonine kinase, protein tyrosine kinase, protein groups histidine kinase, Albumen tryptophan kinases and albumen aspartyl/glutamyl kinases.Protein kinase is in the regulation of cell processes and maintains Having arrived important function, all observed abnormal kinase in numerous disease state, described morbid state includes: pernicious swollen Tumor, immune disease, cardiovascular disease, diabetes, infectious disease, arthritis and other immunologic derangement, nervous system are as old Year dementia, A Mohaici disease AD etc., it has now been found that to relevant with protein kinase more than 400 kinds of human diseasess.
Wherein, VEGFR(vascular endothelial growth factor receptor) family member is receptor tyrosine kinase, as VEGFR1, VEGFR2 etc., this receptoroid malignant tumor growth, transfer in and vascular proliferative disease (as degeneration of macula, Tumor) etc. have a major impact during advancing of disease.
PDGFR(platelet derived growth factor receptor) family member is receptor tyrosine kinase, such as PDGFR α and PDGFR β and colony-stimulating factor 1 receptor, stem cell factor receptor KIT etc., currently also find sending out of such kinases and tumor Raw, development has close ties.As PDGFR unconventionality expression melanoma, meningioma, neuroendocrine tumor, ovarian cancer, Being found in carcinoma of prostate, pulmonary carcinoma and cancer of pancreas, the abnormal activation of KIT is then the direct inducement of many tumorigenesis.
1) suppression dose response studies
Compound V01 is dissolved in 100%DMSO, is diluted to 3 groups of series concentration, and DMSO all keeps 1% in last test. Test maximum concentration is 50uM.Make with the activity inhibitor D-82041 DEISENHOFEN (Staurosporine) of non-selective protein kinase For reference, maximum concentration is 1uM.The results are shown in Table 3, Figure 22.
Table 2
Target Supplier [enzyme], nM [ATP],μM Incubation time, hr
KDR Invitrogen 0.25 80 3
The table 3 inhibitory activity to KDR
Compound IC50(μM) 95% confidence rate Hill
Staurosporine 0.00942 0.00121 1.002392
V01 0.0207 0.00287 1.007953
2) suppression specificity research
To compound VO1, testing 22 kinds of kinase activity inhibition tests, test concentrations is 5mM, is repeated twice, at ATP Km tests.First V01 is dissolved in 100%DMSO, and concentration of ordinary dissolution is 100 times of final test concentration, during all final tests DMSO is 1%.The activity inhibitor D-82041 DEISENHOFEN (Staurosporine) of non-selective protein kinase is as comparison, test Time use Cmax 10mM.
Concrete test method and reagent are as shown in the table:
Table 4
Kinases is tested average inhibition such as following table for twice by V01:
Table 5
Knowable to result of the test, V01 is up to 100% to the suppression ratio of protein kinase K DR, relevant to tumor to other two kinds Protein kinase also have certain inhibitory action, wherein, PDGFR-β suppression ratio is 52.7%, and KIT suppression ratio is 68%, but V01 is to it The remaining protein kinase inhibiting activity overwhelming majority is less than 5%.As can be seen here, compare with the small molecule kinase inhibitors of existing research and development, this Invention compound has high specific and high efficiency inhibitory action to KDR, and has PDGFR-β and KIT both with tumor The protein kinase of close association also has certain inhibitory action, this indicate that compound V01 to KDR, PDGFR-β and KIT this three Plant the diseases such as the relevant various tumors of protein kinase abnormal activation and old wet MD and all have potential therapeutic activity.
3) research of new vessels suppression
Test 5 Mus altogether. in cornea central authorities, burn with 75% silver nitrate solution and 25% potassium nitrate rod induced chemical Modeling.Immediately at conjunctiva of right eye hemostasis 0.2ml compound V01 after chemical burn, then drip 0.2ml compound V012 every day Secondary.After 7 days, cornea is taken pictures and is compared.
Shown in Figure 23, compound V01 can significantly reduce new vessels and hemorrhage.
In sum, the compounds of this invention has good anti-angiogenic rebirth effect, it was initially believed that this compounds is logical Cross suppression VEGFR2(i.e. KDR) produce activity.This compounds can be used for new vessels, VEGFR2, PDGFR-β, KIT etc. The treatment of protein kinase caused by abnormal disease, such as wet MD, malignant tumor etc..

Claims (8)

1. the compound of anti-angiogenic rebirth and pharmaceutically acceptable salt thereof, it is characterised in that: described compound is:
2. compound described in claim 1 and pharmaceutically acceptable salt use in preparing angiogenesis inhibitors class medicine thereof On the way.
Purposes the most according to claim 2, it is characterised in that: described angiogenesis inhibitors is VEGF Receptor inhibitor 2.
Purposes the most according to claim 2, it is characterised in that: described inhibitor is the medicine of anti-ocular angiogenesis.
Purposes the most according to claim 4, it is characterised in that: described medicine is choroid neovascularization inhibitors.
6. according to the purposes described in claim 4 or 5, it is characterised in that: described medicine be treatment or prevention wet MD, Diabetic retinopathy or the medicine of neovascular glaucoma.
7. compound described in claim 1 and pharmaceutically acceptable salt thereof are preparing VEGFR2, PDGFR-β or/and KIT suppresses Purposes in agent class medicine.
8. a pharmaceutical composition, it is characterised in that: it is containing compound described in claim 1 and pharmaceutically acceptable The ophthalmic preparation of salt.
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CN105906553B (en) * 2016-04-21 2019-01-29 重庆威尔德·浩瑞医药化工有限公司 A kind of synthetic method of N- benzyl -4- methyl piperidine -3- keto hydrochloride
CN108912116A (en) * 2018-08-15 2018-11-30 翟学旭 A kind of nitrogen-containing heterocycle analog derivative and its application in retinal disease
CN112194598B (en) * 2020-10-15 2021-12-21 郑州猫眼农业科技有限公司 Process for the preparation of 3- (tert-butoxycarbonyl-R-oxycarbonylmethyl-amino) -propanoate
CN115572238B (en) * 2022-09-27 2023-10-17 常州永和精细化学有限公司 Preparation method of N- (2-ethoxyphenyl) -N' - (2-ethylphenyl) -oxamide

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1933839A (en) * 2004-01-23 2007-03-21 安进公司 Compounds and methods of use

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE10349587A1 (en) * 2003-10-24 2005-05-25 Merck Patent Gmbh Benzimidazolylderivate
JO3265B1 (en) * 2008-12-09 2018-09-16 Novartis Ag Pyridyloxyindoles Inhibitors of VEGF-R2 and Use Thereof for Treatment of Disease

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1933839A (en) * 2004-01-23 2007-03-21 安进公司 Compounds and methods of use

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