CN103524409A - Quinoline compound as well as preparation method and application thereof - Google Patents

Quinoline compound as well as preparation method and application thereof Download PDF

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CN103524409A
CN103524409A CN201310446924.XA CN201310446924A CN103524409A CN 103524409 A CN103524409 A CN 103524409A CN 201310446924 A CN201310446924 A CN 201310446924A CN 103524409 A CN103524409 A CN 103524409A
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quinoline
dimethoxy
oxygen base
dihydroquinoline
methane amide
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CN103524409B (en
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不公告发明人
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Shanghai Yifeng Biotechnology Co.,Ltd.
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Shanghai Ren Li Pharmaceutical Technology Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D215/00—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems
    • C07D215/02—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom
    • C07D215/16—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D215/20—Oxygen atoms
    • C07D215/22—Oxygen atoms attached in position 2 or 4
    • C07D215/233—Oxygen atoms attached in position 2 or 4 only one oxygen atom which is attached in position 4
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings

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Abstract

The invention relates to the technical field of medicine, in particular to a quinoline tyrosine kinase inhibitor as well as a preparation and an application of the quinoline tyrosine kinase inhibitor in preparation of antineoplastic drugs. The structure formula (I) of a compound is as follows: the compound has good tyrosine kinase inhibitory activity, particularly, VEGFR2 and VEGFR3 inhibitory activities, and partial compound is subjected to liver microsome metabolic rate study, absorption characteristic study in a Caco-2 model and drug metabolism kinetic study.

Description

Quinolines and preparation method thereof and application
Technical field
The present invention relates to medical technical field, particularly a class quinoline tyrosine kinase inhibitor, its preparation method and the application in preparing antitumor drug thereof.
Background technology
Malignant tumour is one of principal disease having a strong impact on human health, threat human life, and the World Health Organization and hygiene department of national governments all classify capture cancer as a top priority as.At present, conventional cancer therapy drug is mainly cytotoxic drug clinically, and poor selectivity, toxic side effect that this class medicine is difficult to avoid because its cytotoxic inherent nature has by force, easily produce the shortcomings such as resistance.Therefore, find that specificity is high, toxicity is little, the new target drone of patient's better tolerance just becomes cancer therapy drug research in the urgent need to.In recent years, along with the develop rapidly of life science, numerous specificity target spots based on cancer cells generation, development mechanism are out identified, as: suppress the vascular endothelial growth factor (VEGFR1, VEGFR2, VEGFR3) of tumor-blood-vessel growth etc.Vasculogenesis (angiogenesis) refers to from already present blood vessel grows the vascular system making new advances.Normal vasculogenesis only exists in some short-term, specific physiological process, as reproduction, wound healing etc.Abnormal vasculogenesis is one of pathological manifestations of the malignant diseases such as tumour, rheumatic arthritis, diabetic retinopathy.Since Folkman propose vasculogenesis and tumour develop closely-related hypothesis since, a large amount of clinical practices and experimental study have confirmed that vasculogenesis that inhibition tumour mediates can suppress growth and the transfer of tumour effectively.
Vegf receptor is the important target of angiogenesis inhibitor.In recent years; the research of the micromolecular inhibitor of target vegf receptor is very active; the inhibitor of a large amount of configurations is in the news; but still there are some problems in these inhibitor at present; for example they are all the competitive inhibitors of ATP; and more than in cell, especially the ATP concentration in cancer cells can reach 5mmol/L, so inhibitor activity should at least reach nmole level level and just can show effective restraining effect.In addition, vegf receptor belongs to Tyrosylprotein kinase superfamily, the conduction of bio signal in this family member's wide participation body.Due to the homology of sequence, the three-dimensional structure of their ATP-binding site is high conservative, therefore how to improve the selectivity of inhibitor in these family members of crucial importance.The present invention design, synthesized novel VEGFR1, VEGFR2, VEGFR3 acceptor inhibitor series compound, has found efficient, the VEGFR2 of low toxicity, VEGFR3 acceptor inhibitor.
Summary of the invention
The object of the present invention is to provide the tyrosine kinase inhibitor of a class formation novelty, particularly VEGFR inhibitor; Another object of the present invention is to provide the preparation method of this compounds; A further object of the present invention is to provide the application of this compounds in preparation prevention or medicine for treating tumor thing.
Concrete technical scheme of the present invention is as follows:
In a first aspect of the present invention, contriver is in the process of antitumor drug research, found that a class is suc as formula the compound shown in I, or its each optical isomer, each crystal formation, pharmaceutically acceptable inorganic or organic salt, hydrate, solvate or prodrug, wherein structural formula I is:
Figure BDA0000388073950000021
Wherein R is selected from H, C 1-10alkyl, C 2-6thiazolinyl, C 2-6alkynyl, C 1-6alkoxyl group, C 1-6alkylthio, C 1-6carbonyl, C 1-6the carbonylamino of carbalkoxy, carbonylamino, replacement, sulfuryl amino, phenyl, substituted-phenyl, cycloalkyl, substituted cycloalkyl, Heterocyclylalkyl, substituted heterocycle alkyl, heteroaryl or substituted heteroaryl.
Described " alkyl ", except as otherwise noted, refers to and contains 1-10 carbon atom.Alkyl includes but not limited to methyl, ethyl, n-propyl, sec.-propyl, normal-butyl, isobutyl-, the tertiary butyl.Described " alkoxyl group " refers to oxygen containing alkyl.The alkyl definition relating in described alkoxyl group, alkylthio, alkyl carbonyl, alkoxy carbonyl etc. as above-mentioned.
Described " thiazolinyl " refers to the straight or branched alkyl of 2-6 the carbon atom that contains one or more carbon-carbon double bonds.Include but not limited to vinyl, propenyl and butenyl.
Described " aryl ", except as otherwise noted, refers to the mononuclear aromatics that contains 6 carbon atoms, the double ring arene of 10 carbon atoms, and the thrcylic aromatic hydrocarbon of 14 carbon atoms, and can have 1-4 substituting group on each ring.Aryl includes but not limited to phenyl, naphthyl, anthryl.
Described " cycloalkyl ", except as otherwise noted, refers to the undersaturated cyclic hydrocarbon of saturated or part that contains 3-8 carbon atom.Cycloalkyl includes but not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl.
Described " heteroaryl ", refers to mononuclear aromatics, the double ring arene of a 8-12 atom or the thrcylic aromatic hydrocarbon of 11-14 atom of 5-8 atom, and contains one or more heteroatomss (for example N, O, S).Heteroaryl includes but not limited to pyridyl, furyl, imidazolyl, benzimidazolyl-, pyrimidyl, thienyl, quinolyl, indyl.
Described " Heterocyclylalkyl ", refers to the monocycle non-aromatics alkyl, the dicyclo of a 8-12 atom or the tricyclic hydrocarbon base of 11-14 atom that contain 3-8 atom, and contains one or more heteroatomss (for example N, O, S).Heterocyclylalkyl includes but not limited to piperazinyl, pyrrolidyl, alkyl dioxin, morpholinyl, tetrahydrofuran base.
The present invention also comprises corresponding all pharmaceutically acceptable salt, hydrate or the prodrug of above-claimed cpd.These salt can be positively charged in compound part with there is contrary electrical electronegative formation; Or part electronegative in compound forms with positive charge.
The compounds of this invention can contain a nonaromatic pair of key, has one or more asymmetric centers.So these compounds can be used as racemic mixture, independent enantiomer, independent diastereomer, non-enantiomer mixture, cis or trans-isomer(ide) existence.All these isomer are all expected.
Described " prodrug of structural formula I " is often referred to a kind of material, after using by appropriate means, can in subject, carry out metabolism or chemical reaction and be transformed at least one compound or its salt of structural formula I.
Preferably, the R group in formula I is selected from H, C 1-10alkyl, C 1-6alkoxyl group, or be selected from following structural unit:
Figure BDA0000388073950000031
Wherein, R 1, R 2independently be selected from separately H, halogen, C 1-6alkyl, C 1-6thiazolinyl, cyano group, C 1-6alkynyl, C 1-6alkoxyl group, nitro, trifluoromethyl, trifluoromethoxy or C 1-6alkyl-carbonyl.
Preferred, formula I compound of the present invention, is selected from following compound:
N-(2,4,4-trimethylpentane-2-yl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide (compound number KL-01)
N-(2-ethylphenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide (compound number KL-02)
N-(2,4-3,5-dimethylphenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide (compound number KL-03)
N-(2-isopropyl phenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide (compound number KL-04)
N-(4-n-butylphenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide (compound number KL-05)
N-(4-p-methoxy-phenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide (compound number KL-06)
N-(3-p-methoxy-phenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide (compound number KL-07)
N-(2-p-methoxy-phenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide (compound number KL-08)
N-(2-methoxyl group-5-aminomethyl phenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide (compound number KL-10)
N-(4-acetylphenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide (compound number KL-11)
N-(4-fluorophenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide (compound number KL-12)
N-(3-fluorophenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide (compound number KL-13)
N-(2-fluorophenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide (compound number KL-14)
N-(2,4 difluorobenzene base)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide (compound number KL-15)
N-(4-chloro-phenyl-)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide (compound number KL-16)
N-(3-chloro-phenyl-)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide (compound number KL-17)
N-(2-chloro-phenyl-)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide (compound number KL-18)
N-(4-chloro-2-methyl phenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide (compound number KL-19)
N-(4-(trifluoromethyl) phenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide (compound number KL-20)
N-(2-(trifluoromethyl) phenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide (compound number KL-21)
N-(4-cyano-phenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide (compound number KL-22)
N-(2-chloro-4 nitrophenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H-methane amide (compound number KL-23)
N-(the chloro-3-nitrophenyl of 4-)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide (compound number KL-24)
N-(2-methyl-4-nitrophenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide (compound number KL-25)
N-cyclopentyl-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide (compound number KL-26)
N-cyclohexyl-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide (compound number KL-27)
N-(benzo ［ d ］ ［ 1,3 ］ dioxolane-5-yl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide (compound number KL-28).
The compound that the present invention is listed, as afoul with name in structural formula, with chemical structural formula, be as the criterion.
In table 1, listed preferred structure of the present invention:
Table 1
Figure BDA0000388073950000041
Figure BDA0000388073950000061
Compound of the present invention can be prepared as according to ordinary method the form of pharmaceutical salts; Comprise its organic acid salt and inorganic acid salt: mineral acid includes, but is not limited to hydrochloric acid, sulfuric acid, phosphoric acid, bisphosphate, Hydrogen bromide, nitric acid etc., organic acid includes, but is not limited to acetic acid, toxilic acid, fumaric acid, tartrate, succsinic acid, lactic acid, tosic acid, Whitfield's ointment, oxalic acid etc.
As a second aspect of the present invention, compound of the present invention can prepare with following methods:
Synthetic route is as follows:
Figure BDA0000388073950000072
(1) 4-is chloro-6,7-dimethoxy-quinoline and 6-hydroxyl-1, and 2,3,4-tetrahydroquinoline reacts and generates 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline (II) under the existence of alkali, catalyzer;
(2) 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline (II) reacts with the isocyanate compound of various replacements and generates corresponding N-replacement-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-Carbox amide (I);
(3) according to ordinary method, be prepared as the form of pharmacy acceptable salt.
The present invention also provides a kind of pharmaceutical composition, and it contains pharmaceutically acceptable vehicle or carrier, and above-mentioned compound or its each optical isomer, each crystal formation, pharmaceutically acceptable inorganic or organic salt, hydrate, solvate or prodrug.
The present invention also provides the purposes of compound of the present invention or its each optical isomer, each crystal formation, pharmaceutically acceptable inorganic or organic salt, hydrate, solvate or prodrug, and it is used to prepare tyrosine kinase inhibitor.
The present invention also provides the purposes of compound of the present invention or its each optical isomer, each crystal formation, pharmaceutically acceptable inorganic or organic salt, hydrate, solvate or prodrug, and it is used to that preparation suppresses the medicine of tyrosine kinase activity or for the preparation of the medicine of the too high relevant disease for the treatment of, prevention and alleviation and tyrosine kinase activity.
In a preference of the present invention, described is selected from tumour with the too high relative disease of tyrosine kinase activity, includes but not limited to liver cancer, lung cancer, brain tumor, cancer of the stomach, kidney, colorectal carcinoma, mammary cancer, ovarian cancer, prostate cancer, osteocarcinoma, leukemia, skin carcinoma.
Simultaneously, vegf receptor family is the closest with vasculogenesis (angiogenesis) relation in current known family tyrosine kinase member, those skilled in the art should anticipate, vegf receptor family member is had to good inhibiting compound, to tumor-blood-vessel growth, should also there is good restraining effect.
Given this, the present invention also provides the purposes of compound of the present invention or its each optical isomer, each crystal formation, pharmaceutically acceptable inorganic or organic salt, hydrate, solvate or prodrug, and it is used to prepare angiogenesis inhibitor.
The present invention also provides the purposes of compound of the present invention or its each optical isomer, each crystal formation, pharmaceutically acceptable inorganic or organic salt, hydrate, solvate or prodrug, and it is used to the medicine of preparation prevention or treatment tumour.
For evaluating the synthetic compound of the present invention and the anti-tumor activity of pharmaceutical composition thereof, compound in the present invention has been carried out to the pharmacologically active test of molecular level, result shows that this compounds generally has good vitro inhibition tyrosine kinase activity, particularly VEGFR2 and VEGFR3 is had to good inhibition activity.
Accompanying drawing explanation
Fig. 1 is the linear lower-most point concentration chromatogram of quantitative limit in the experiment of compound K L-11 pharmacokinetics.
Fig. 2 is the linear vertex concentration chromatogram of quantitative limit in the experiment of compound K L-11 pharmacokinetics.
Fig. 3 is the typical color spectrogram of compound K L-11 and interior mark KL-13 in gavage group blood plasma in pharmacokinetics experiment.
Fig. 4 is gavage group rat 1N-11 Plasma Concentration-time data deployment conditions in the experiment of compound K L-11 pharmacokinetics.
Fig. 5 is average Plasma Concentration-time curve of gavage group rat in the experiment of compound K L-11 pharmacokinetics.
Embodiment
Below in conjunction with specific embodiment, the invention will be further described.Should be understood that following examples are only for the present invention is described but not for limiting scope of the present invention.
Embodiment 1:
The preparation of 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline (II)
In 250ml round-bottomed bottle, add 4-chloro-6,7-dimethoxy-quinoline 22.5g(100.6mmol), 6-hydroxyl-1,2,3,4-tetrahydroquinoline 15g(100.5mmol), N-Methyl pyrrolidone (NMP) 100ml, uniform stirring dissolves, ice bath adds tertiary butyl potassium alcoholate 33.9g(301.8mmol lower minute three times), be heated to 100 ℃, stirring reaction approximately 16 hours, TLC follows the tracks of reaction, and (developping agent is sherwood oil: ethyl acetate=2.5:2.5), react the complete aftertreatment of carrying out.Pressure reducing and steaming NMP, residue washing, obtains faint yellow solid, and methyl alcohol-sherwood oil recrystallization filters, dry, obtains sterling 19g.Productive rate: 56.2%.
1HNMR(600MHz,DMSO)δ8.43(d,J=5.2Hz,1H),7.49(s,1H),7.36(s,1H),6.77(d,J=2.7Hz,1H),6.76(s,1H),6.53(d,J=8.2Hz,1H),6.40(d,J=5.2Hz,1H),5.74(s,1H),3.94(s,3H),3.93(s,3H),3.22-3.19(m,2H),2.69(t,J=6.2Hz,2H),1.83-1.78(m,2H).HRMS[M+H]337.48
Embodiment 2's is synthetic)
In 100ml round-bottomed bottle, add 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline 0.5g(1.5mmol), chloroform 10ml, dissolve, stirring at room, separately gets tertiary octyl group isocyanic ester 0.47g(3.0mmol) in chloroform 2ml, extremely dissolve, after dissolving, slowly add in round-bottomed bottle, after room temperature reaction 1 hour, TLC tracking reaction (developping agent is methylene dichloride: methyl alcohol=4.5:0.5), and room temperature reaction 70 hours, aftertreatment.
Reaction solution moves in separating funnel, and chloroform layer adds water 10ml vibration washing 2 times, and chloroform layer is again with saturated sodium bicarbonate solution 5ml washing 1 time, add again water 10ml vibration washing 2 times, chloroform layer is dried 1 hour with anhydrous magnesium sulfate 1.5g, filters, and evaporate to dryness chloroform obtains crude product.Crude product adds normal hexane to micro-muddiness after dissolving with a small amount of methylene dichloride, reheats to clarification, places, and crystallization, filters, and solid obtains fine work 0.18g with Preparative TLC silica-gel plate purifying again.Productive rate: 24.3%.
1HNMR(600MHz,DMSO)δ8.48(d,J=5.2Hz,1H),7.54(d,J=8.8Hz,1H),7.49(s,1H),7.39(s,1H),7.02-6.96(m,2H),6.52(d,J=5.2Hz,1H),6.43(d,J=7.8Hz,1H),3.95(s,3H),3.93(s,3H),3.61-3.58(m,2H),3.40-3.37(m,2H),2.71(t,J=6.5Hz,2H),1.87-1.08(m,17H).HRMS[M+H]492.56。
Embodiment 3's is synthetic)
In 100ml round-bottomed bottle, add 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline 0.5g(1.5mmol), chloroform 10ml, dissolve, stirring at room, separately gets 2-ethylphenyl isocyanic ester 0.44g(3.0mmol) in chloroform 2ml, extremely dissolve, after dissolving, slowly add in round-bottomed bottle, after room temperature reaction 1 hour, TLC follows the tracks of reaction, and (developping agent is ethyl acetate: methyl alcohol=4.7:0.3), room temperature reaction is 65 hours altogether, aftertreatment.
Reaction solution moves in separating funnel, and chloroform layer adds water 10ml vibration washing 2 times, and chloroform layer is again with saturated sodium bicarbonate solution 5ml washing 1 time, add again water 10ml vibration washing 2 times, chloroform layer is dried 1 hour with anhydrous magnesium sulfate 1.5g, filters, and evaporate to dryness chloroform obtains crude product.Crude product adds normal hexane to micro-muddiness after dissolving with a small amount of methylene dichloride, reheats to clarification, places, and crystallization, filters, and solid obtains fine work 0.25g with Preparative TLC silica-gel plate purifying again.Productive rate: 34.5%.
1HNMR(600MHz,DMSO)δ8.48(d,J=5.2Hz,1H),8.26(s,1H),7.72(dd,J=11.4,4.4Hz,1H),7.61(d,J=8.9Hz,1H),7.50(s,1H),7.39(s,1H),7.34-7.31(m,1H),7.24(dd,J=7.5,1.5Hz,1H),7.08(d,J=2.6Hz,1H),7.04(dd,J=8.5,3.0Hz,1H),7.01(td,J=7.6,0.8Hz,1H),6.52(d,J=5.2Hz,1H),3.94(s,3H),3.93(s,3H),3.80-3.76(m,2H),2.79(t,J=6.5Hz,2H),2.63(dd,J=15.1,7.6Hz,2H),1.97–1.92(m,2H),1.15(t,J=7.5Hz,3H).HRMS[M+H]484.40。
Embodiment 4's is synthetic)
In 100ml round-bottomed bottle, add 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline 0.5g(1.5mmol), chloroform 10ml, dissolve, stirring at room, separately gets 2,4-dimethylphenyl isocyanate 0.44g(3.0mmol) in chloroform 2ml, extremely dissolve, after dissolving, slowly add in round-bottomed bottle, room temperature reaction is after 1 hour, and TLC follows the tracks of reaction, and (developping agent is ethyl acetate: methyl alcohol=4.7:0.3), room temperature reaction is 65 hours altogether, aftertreatment.
Reaction solution moves in separating funnel, and chloroform layer adds water 10ml vibration washing 2 times, and chloroform layer is again with saturated sodium bicarbonate solution 5ml washing 1 time, add again water 10ml vibration washing 2 times, chloroform layer is dried 1 hour with anhydrous magnesium sulfate 1.5g, filters, and evaporate to dryness chloroform obtains crude product.Crude product adds normal hexane to micro-muddiness after dissolving with a small amount of methylene dichloride, reheats to clarification, places, and crystallization, filters, and solid obtains fine work 0.20g with Preparative TLC silica-gel plate purifying again.Productive rate: 27.6%.
1HNMR(600MHz,DMSO)δ8.48(d,J=5.2Hz,1H),8.23(s,1H),7.62(d,J=8.9Hz,1H),7.50(s,1H),7.39(s,1H),7.19(d,J=8.1Hz,1H),7.07(d,J=2.6Hz,1H),7.04–7.01(m,2H),6.96(d,J=8.4Hz,1H),6.53(d,J=5.2Hz,1H),3.95(s,3H),3.93(s,3H),3.78-3.75(m,2H),2.78(t,J=6.5Hz,2H),2.25(s,3H),2.18(s,3H),1.93(dt,J=13.8,6.7Hz,2H).HRMS[M+H]484.45。
Embodiment 5's is synthetic)
In 100ml round-bottomed bottle, add 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline 0.5g(1.5mmol), chloroform 10ml, dissolve, stirring at room, separately gets 2-isopropyl benzene isocyanic ester 0.48g(3.0mmol) in chloroform 2ml, extremely dissolve, after dissolving, slowly add in round-bottomed bottle, after room temperature reaction 1 hour, TLC follows the tracks of reaction, and (developping agent is ethyl acetate: methyl alcohol=4.7:0.3), room temperature reaction is 65 hours altogether, aftertreatment.
Reaction solution moves in separating funnel, and chloroform layer adds water 10ml vibration washing 2 times, and chloroform layer is again with saturated sodium bicarbonate solution 5ml washing 1 time, add again water 10ml vibration washing 2 times, chloroform layer is dried 1 hour with anhydrous magnesium sulfate 1.5g, filters, and evaporate to dryness chloroform obtains crude product.Crude product adds normal hexane to micro-muddiness after dissolving with a small amount of methylene dichloride, reheats to clarification, places, and crystallization, filters, and solid obtains fine work 0.31g with Preparative TLC silica-gel plate purifying again.Productive rate: 41.6%.
1HNMR(600MHz,DMSO)δ8.48(d,J=5.2Hz,1H),8.29(s,1H),7.61(d,J=8.9Hz,1H),7.50(s,1H),7.39(s,1H),7.32-7.30(m,1H),7.27-7.24(m,1H),7.21-7.14(m,2H),7.07(d,J=3.3Hz,1H),7.03(dd,J=8.8,2.8Hz,1H),6.52(d,J=5.3Hz,1H),3.94(s,3H),3.93(s,3H),3.80-3.77(m,2H),3.19-3.13(m,1H),2.79(t,J=6.6Hz,2H),1.97-1.91(m,2H),1.17(d,J=6.8Hz,6H).HRMS[M+H]498.41。
Embodiment 6's is synthetic)
In 100ml round-bottomed bottle, add 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline 0.5g(1.5mmol), chloroform 10ml, dissolve, stirring at room, separately gets 4-normal-butyl phenol isocyanic ester 0.53g(3.0mmol) in chloroform 2ml, extremely dissolve, after dissolving, slowly add in round-bottomed bottle, after room temperature reaction 1 hour, TLC follows the tracks of reaction, and (developping agent is ethyl acetate: methyl alcohol=4.7:0.3), room temperature reaction is 65 hours altogether, aftertreatment.
Reaction solution moves in separating funnel, and chloroform layer adds water 10ml vibration washing 2 times, and chloroform layer is again with saturated sodium bicarbonate solution 5ml washing 1 time, add again water 10ml vibration washing 2 times, chloroform layer is dried 1 hour with anhydrous magnesium sulfate 1.5g, filters, and evaporate to dryness chloroform obtains crude product.Crude product adds normal hexane to micro-muddiness after dissolving with a small amount of methylene dichloride, reheats to clarification, places, and crystallization, filters, and solid obtains fine work 0.38g with Preparative TLC silica-gel plate purifying again.Productive rate: 49.5%.
1HNMR(600MHz,DMSO)δ8.77(s,1H),8.50(d,J=5.2Hz,1H),7.50-7.48(m,2H),7.40-7.38(m,3H),7.10-7.07(m,3H),7.02(dd,J=9.0,2.8Hz,1H),6.56(d,J=5.2Hz,1H),3.95(s,3H),3.93(s,3H),3.76-3.73(m,2H),2.78(t,J=6.3Hz,2H),2.53-2.50(m,2H),1.94-1.90(m,2H),1.55-1.50(m,2H),1.29(dt,J=15.1,7.6Hz,2H),0.89(t,J=7.4Hz,3H).HRMS[M+H]512.44。
Embodiment 7's is synthetic)
In 100ml round-bottomed bottle, add 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline 0.5g(1.5mmol), chloroform 10ml, dissolve, stirring at room, separately gets p-Methoxyphenyl isocyanate 0.45g(3.0mmol) in chloroform 2ml, extremely dissolve, after dissolving, slowly add in round-bottomed bottle, after room temperature reaction 1 hour, TLC follows the tracks of reaction, and (developping agent is ethyl acetate: methyl alcohol=4.7:0.3), room temperature reaction is 65 hours altogether, aftertreatment.
Reaction solution moves in separating funnel, and chloroform layer adds water 10ml vibration washing 2 times, and chloroform layer is again with saturated sodium bicarbonate solution 5ml washing 1 time, add again water 10ml vibration washing 2 times, chloroform layer is dried 1 hour with anhydrous magnesium sulfate 1.5g, filters, and evaporate to dryness chloroform obtains crude product.Crude product adds normal hexane to micro-muddiness after dissolving with a small amount of methylene dichloride, reheats to clarification, places, and crystallization, filters, and solid obtains fine work 0.29g with Preparative TLC silica-gel plate purifying again.Productive rate: 39.8%.
1HNMR(600MHz,DMSO)δ8.70(s,1H),8.49(d,J=5.3Hz,1H),7.51(d,J=8.8Hz,2H),7.41-7.37(m,3H),7.07(s,1H),7.02(d,J=7.6Hz,1H),6.86(d,J=8.0Hz,2H),6.55(d,J=4.7Hz,1H),3.95(s,3H),3.93(s,3H),3.76-3.73(m,2H),3.72(s,3H),2.77(t,J=6.2Hz,2H),1.94-1.89(m,2H).HRMS[M+H]486.35。
Embodiment 8's is synthetic)
In 100ml round-bottomed bottle, add 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline 0.5g(1.5mmol), chloroform 10ml, dissolve, stirring at room, separately gets m-methoxy phenyl isocyanate 0.45g(3.0mmol) in chloroform 2ml, extremely dissolve, after dissolving, slowly add in round-bottomed bottle, after room temperature reaction 1 hour, TLC follows the tracks of reaction, and (developping agent is ethyl acetate: methyl alcohol=4.7:0.3), room temperature reaction is 65 hours altogether, aftertreatment.
Reaction solution moves in separating funnel, and chloroform layer adds water 10ml vibration washing 2 times, and chloroform layer is again with saturated sodium bicarbonate solution 5ml washing 1 time, add again water 10ml vibration washing 2 times, chloroform layer is dried 1 hour with anhydrous magnesium sulfate 1.5g, filters, and evaporate to dryness chloroform obtains crude product.Crude product obtains fine work 0.37g with ether purifying.Productive rate: 50.8%.
1HNMR(600MHz,CDCl 3)δ8.52(d,J=5.6Hz,1H),7.67(s,1H),7.56(s,1H),7.48(d,J=9.0Hz,1H),7.21-7.17(m,2H),7.09-7.06(m,2H),6.93(s,1H),6.84(dd,J=7.9,1.6Hz,1H),6.64-6.63(m,1H),6.62(d,J=2.4Hz,1H),4.09(s,3H),4.07(s,3H),3.87(t,J=6.3Hz,2H),3.81(s,3H),2.84(t,J=6.6Hz,2H),2.07-2.03(m,2H).HRMS[M+H]486.40。
Embodiment 9's is synthetic)
In 100ml round-bottomed bottle, add 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline 0.5g(1.5mmol), chloroform 10ml, dissolve, stirring at room, separately gets O-methoxy phenylisocyanate 0.45g(3.0mmol) in chloroform 2ml, extremely dissolve, after dissolving, slowly add in round-bottomed bottle, after room temperature reaction 1 hour, TLC follows the tracks of reaction, and (developping agent is ethyl acetate: methyl alcohol=4.7:0.3), room temperature reaction is 72 hours altogether, aftertreatment.
Reaction solution moves in separating funnel, and chloroform layer adds water 10ml vibration washing 2 times, and chloroform layer is again with saturated sodium bicarbonate solution 5ml washing 1 time, add again water 10ml vibration washing 2 times, chloroform layer is dried 1 hour with anhydrous magnesium sulfate 1.5g, filters, and evaporate to dryness chloroform obtains crude product.Crude product obtains fine work 0.26g with ether purifying.Productive rate: 35.7%.
1HNMR(600MHz,CDCl 3)δ8.51(d,J=5.5Hz,1H),8.29-8.27(m,1H),7.81(s,1H),7.57(d,J=9.0Hz,3H),7.08-7.05(m,2H),7.00-6.97(m,2H),6.86–6.84(m,1H),6.57(d,J=5.3Hz,1H),4.08(s,3H),4.07(s,3H),3.91-3.88(m,2H),3.80(s,3H),2.84(t,J=6.6Hz,2H),2.07-2.02(m,2H).HRMS[M+H]486.40。
Embodiment 10's is synthetic)
In 100ml round-bottomed bottle, add 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline 0.5g(1.5mmol), chloroform 10ml, dissolve, stirring at room, separately gets 2-methoxyl group-5-methylbenzene isocyanic ester 0.49g(3.0mmol) in chloroform 2ml, extremely dissolve, after dissolving, slowly add in round-bottomed bottle, after room temperature reaction 1 hour, TLC follows the tracks of reaction, and (developping agent is ethyl acetate: methyl alcohol=4.7:0.3), room temperature reaction is 55 hours altogether, aftertreatment.
Reaction solution moves in separating funnel, and chloroform layer adds water 10ml vibration washing 2 times, and chloroform layer is again with saturated sodium bicarbonate solution 5ml washing 1 time, add again water 10ml vibration washing 2 times, chloroform layer is dried 1 hour with anhydrous magnesium sulfate 1.5g, filters, and evaporate to dryness chloroform obtains crude product.Crude product adds normal hexane to micro-muddiness after dissolving with a small amount of methylene dichloride, reheats to clarification, places, and crystallization, filters, and solid adds normal hexane to micro-muddiness after dissolving with methylene dichloride again, reheats to clarification, places, and crystallization obtains fine work 0.40g.Productive rate: 53.3%.
1HNMR(600MHz,CDCl 3)δ8.51(d,J=5.4Hz,1H),8.13(d,J=1.7Hz,1H),7.77(s,1H),7.58-7.53(m,3H),7.08-7.04(m,2H),6.78(dd,J=8.2,1.3Hz,1H),6.73(d,J=8.2Hz,1H),6.56(d,J=5.4Hz,1H),4.08(s,3H),4.07(s,3H),3.90-3.87(m,2H),3.77(s,3H),2.83(t,J=6.6Hz,2H),2.32(s,3H),2.05(p,J=6.5Hz,2H).HRMS[M+H]500.46。
Embodiment 11's is synthetic)
In 100ml round-bottomed bottle, add 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline 0.5g(1.5mmol), chloroform 10ml, dissolve, stirring at room, separately gets 4-phenyl methyl ketone based isocyanate 0.48g(3.0mmol) in chloroform 2ml, extremely dissolve, after dissolving, slowly add in round-bottomed bottle, after room temperature reaction 1 hour, TLC follows the tracks of reaction, and (developping agent is ethyl acetate: methyl alcohol=4.7:0.3), room temperature reaction is 70 hours altogether, aftertreatment.
Reaction solution moves in separating funnel, and chloroform layer adds water 10ml vibration washing 2 times, and chloroform layer is again with saturated sodium bicarbonate solution 5ml washing 1 time, add again water 10ml vibration washing 2 times, chloroform layer is dried 1 hour with anhydrous magnesium sulfate 1.5g, filters, and evaporate to dryness chloroform obtains crude product.Crude product, with methylene dichloride 15ml reflux, has insoluble solids, filters, the concentrated major part of going of filtrate, add normal hexane to micro-muddiness, reheat to clarification, place, crystallization, filter, solid adds normal hexane to micro-muddiness after dissolving with methylene dichloride again, reheats to clarification, place, crystallization obtains fine work 0.37g.Productive rate: 49.6%.
1HNMR(600MHz,CDCl 3)δ8.53(d,J=5.5Hz,1H),7.93(d,J=8.7Hz,2H),7.61(d,J=8.5Hz,1H),7.55(s,1H),7.51(d,J=8.7Hz,2H),7.46(d,J=8.4Hz,1H),7.16(s,1H),7.12-7.09(m,2H),6.63(d,J=5.3Hz,1H),4.08(s,3H),4.07(s,3H),3.89(t,J=6.3Hz,2H),2.84(t,J=6.6Hz,2H),2.57(s,3H),2.06(p,J=6.5Hz,2H).HRMS[M+H]498.38。
Embodiment 12's is synthetic)
In 100ml round-bottomed bottle, add 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline 0.5g(1.5mmol), chloroform 10ml, dissolve, stirring at room, separately gets 4-fluorophenyl isocyanic ester 0.41g(3.0mmol) in chloroform 2ml, extremely dissolve, after dissolving, slowly add in round-bottomed bottle, after room temperature reaction 1 hour, TLC follows the tracks of reaction, and (developping agent is ethyl acetate: methyl alcohol=20:1), room temperature reaction is 65 hours altogether, aftertreatment.
Reaction solution moves in separating funnel, and chloroform layer adds water 10ml vibration washing 2 times, and chloroform layer is again with saturated sodium bicarbonate solution 5ml washing 1 time, add again water 10ml vibration washing 2 times, chloroform layer is dried 1 hour with anhydrous magnesium sulfate 1.5g, filters, and evaporate to dryness chloroform obtains crude product.Crude product, with appropriate methylene dichloride reflux, has insoluble solids, filters, the concentrated part of going of filtrate, add sherwood oil to micro-muddiness, reheat to clarification, place, crystallization, filter, solid adds sherwood oil to micro-muddiness after dissolving with methylene dichloride again, reheats to clarification, place, crystallization obtains fine work 0.18g.Productive rate: 25.4%.
1HNMR(600MHz,CDCl 3)δ8.52(d,J=5.6Hz,1H),8.29(d,J=8.7Hz,1H),7.65(s,1H),7.59-7.54(m,3H),7.50(d,J=8.6Hz,1H),7.43(s,1H),7.17(t,J=7.6Hz,1H),7.12-7.08(m,2H),6.56(d,J=5.5Hz,1H),4.10(s,3H),4.08(s,3H),3.90(t,J=6.3Hz,2H),2.85(t,J=6.7Hz,2H),2.06(p,J=6.6Hz,2H).HRMS[M+H]474.38。
Embodiment 13's is synthetic)
In 100ml round-bottomed bottle, add 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline 0.5g(1.5mmol), chloroform 10ml, dissolve, stirring at room, separately gets 3-fluorophenyl isocyanic ester 0.41g(3.0mmol) in chloroform 2ml, extremely dissolve, after dissolving, slowly add in round-bottomed bottle, after room temperature reaction 1 hour, TLC follows the tracks of reaction, and (developping agent is ethyl acetate: methyl alcohol=20:1), room temperature reaction is 65 hours altogether, aftertreatment.
Reaction solution moves in separating funnel, and chloroform layer adds water 10ml vibration washing 2 times, and chloroform layer is again with saturated sodium bicarbonate solution 5ml washing 1 time, add again water 10ml vibration washing 2 times, chloroform layer is dried 1 hour with anhydrous magnesium sulfate 1.5g, filters, and evaporate to dryness chloroform obtains crude product.Crude product, with appropriate methylene dichloride reflux, has insoluble solids, filters, the concentrated part of going of filtrate, add sherwood oil to micro-muddiness, reheat to clarification, place, crystallization, filter, solid adds sherwood oil to micro-muddiness after dissolving with methylene dichloride again, reheats to clarification, place, crystallization obtains fine work 0.20g.Productive rate: 28.2%.
1HNMR(600MHz,CDCl 3)δ8.52(s,1H),7.98(s,2H),7.60(s,1H),7.54(d,J=8.7Hz,1H),7.39(d,J=11.2Hz,1H),7.11(d,J=7.7Hz,2H),7.03(d,J=9.7Hz,1H),6.97(s,1H),6.78(dd,J=8.1,2.0Hz,1H),6.76(d,J=1.7Hz,1H),4.14(s,3H),4.09(s,3H),3.89(t,J=6.2Hz,2H),2.87(t,J=6.6Hz,2H),2.10-2.05(m,2H).HRMS[M+H]474.36。
Embodiment 14's is synthetic)
In 100ml round-bottomed bottle, add 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline 0.5g(1.5mmol), chloroform 10ml, dissolve, stirring at room, separately gets 2-fluorophenyl isocyanic ester 0.41g(3.0mmol) in chloroform 2ml, extremely dissolve, after dissolving, slowly add in round-bottomed bottle, after room temperature reaction 1 hour, TLC follows the tracks of reaction, and (developping agent is ethyl acetate: methyl alcohol=4.5:0.5), room temperature reaction is 70 hours altogether, aftertreatment.
Reaction solution moves in separating funnel, and chloroform layer adds water 10ml vibration washing 2 times, and chloroform layer is again with saturated sodium bicarbonate solution 5ml washing 1 time, add again water 10ml vibration washing 2 times, chloroform layer is dried 1 hour with anhydrous magnesium sulfate 1.5g, filters, and evaporate to dryness chloroform obtains crude product.Crude product, with appropriate methylene dichloride reflux, has insoluble solids, filters, the concentrated part of going of filtrate, add sherwood oil to micro-muddiness, reheat to clarification, place, crystallization, filter, solid adds sherwood oil to micro-muddiness after dissolving with methylene dichloride again, reheats to clarification, place, crystallization obtains fine work 0.31g.Productive rate: 43.7%.
1HNMR(600MHz,CDCl 3)δ8.53(d,J=5.5Hz,1H),8.24(td,J=8.2,1.5Hz,1H),7.57-7.50(m,3H),7.29(d,J=3.2Hz,1H),7.15(t,J=8.0Hz,1H),7.10(dd,J=8.6,2.8Hz,1H),7.08-7.03(m,2H),7.02-6.97(m,1H),6.59(d,J=5.4Hz,1H),4.08(s,3H),4.06(s,3H),3.89(t,J=6.3Hz,2H),2.84(t,J=6.6Hz,2H),2.06(p,J=6.6Hz,2H).HRMS[M+H]474.35。
Embodiment 15's is synthetic)
In 100ml round-bottomed bottle, add 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline 0.5g(1.5mmol), chloroform 10ml, dissolve, stirring at room, separately gets 2,4-difluorophenyl isocyanate 0.47g(3.0mmol) in chloroform 2ml, extremely dissolve, after dissolving, slowly add in round-bottomed bottle, room temperature reaction is after 1 hour, and TLC follows the tracks of reaction, and (developping agent is methylene dichloride: methyl alcohol=6.5:0.5), room temperature reaction is 70 hours altogether, aftertreatment.
Reaction solution moves in separating funnel, and chloroform layer adds water 10ml vibration washing 2 times, and chloroform layer is again with saturated sodium bicarbonate solution 5ml washing 1 time, add again water 10ml vibration washing 2 times, chloroform layer is dried 1 hour with anhydrous magnesium sulfate 1.5g, filters, and evaporate to dryness chloroform obtains crude product.Crude product, with appropriate methylene dichloride reflux, has insoluble solids, filters, the concentrated part of going of filtrate, add sherwood oil to micro-muddiness, reheat to clarification, place, crystallization, filter, solid adds sherwood oil to micro-muddiness after dissolving with methylene dichloride again, reheats to clarification, place, crystallization obtains fine work 0.25g.Productive rate: 33.8%.
1HNMR(600MHz,CDCl 3)δ8.53(d,J=5.4Hz,1H),8.15(td,J=9.1,5.9Hz,1H),7.55(s,1H),7.53(s,1H),7.51(d,J=8.6Hz,1H),7.14(d,J=2.7Hz,1H),7.11-7.06(m,2H),6.91-6.81(m,2H),6.58(d,J=5.4Hz,1H),4.07(s,3H),4.06(s,3H),3.88(t,J=6.3Hz,2H),2.83(t,J=6.6Hz,2H),2.05(p,J=6.6Hz,2H).HRMS[M+H]492.36。
Embodiment 16's is synthetic)
In 100ml round-bottomed bottle, add 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline 0.5g(1.5mmol), chloroform 10ml, dissolve, stirring at room, separately gets 4-chloro-phenyl-isocyanic ester 0.46g(3.0mmol) in chloroform 2ml, extremely dissolve, after dissolving, slowly add in round-bottomed bottle, after room temperature reaction 1 hour, TLC follows the tracks of reaction, and (developping agent is methylene dichloride: methyl alcohol=4.5:0.5), room temperature reaction is 72 hours altogether, complete reaction, aftertreatment.
Reaction solution moves in separating funnel, and chloroform layer adds water 10ml vibration washing 2 times, and chloroform layer is again with saturated sodium bicarbonate solution 5ml washing 1 time, add again water 10ml vibration washing 2 times, chloroform layer is dried 1 hour with anhydrous magnesium sulfate 1.5g, filters, and evaporate to dryness chloroform obtains crude product.Crude product, with appropriate methylene dichloride reflux, has insoluble solids, filters, the concentrated part of going of filtrate, add sherwood oil to micro-muddiness, reheat to clarification, place, crystallization, filter, solid adds sherwood oil to micro-muddiness after dissolving with methylene dichloride again, reheats to clarification, place, crystallization obtains fine work 0.39g.Productive rate: 52.7%.
1HNMR(600MHz,CDCl 3)δ8.53(d,J=5.5Hz,1H),7.64(s,1H),7.56(s,1H),7.47(d,J=8.2Hz,1H),7.36(d,J=8.8Hz,2H),7.28(s,2H),7.10-7.07(m,2H),6.94(s,1H),6.63(d,J=4.8Hz,1H),4.09(s,3H),4.06(s,3H),3.87(t,J=6.3Hz,2H),2.84(t,J=6.6Hz,2H),2.07-2.01(m,2H).HRMS[M+H]490.35。
Embodiment 17's is synthetic)
In 100ml round-bottomed bottle, add 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline 0.5g(1.5mmol), chloroform 10ml, dissolve, stirring at room, separately gets 3-chloro-phenyl-isocyanic ester 0.46g(3.0mmol) in chloroform 2ml, extremely dissolve, after dissolving, slowly add in round-bottomed bottle, after room temperature reaction 1 hour, TLC follows the tracks of reaction, and (developping agent is methylene dichloride: methyl alcohol=4.5:0.5), room temperature reaction is 72 hours altogether, aftertreatment.
Reaction solution moves in separating funnel, and chloroform layer adds water 10ml vibration washing 2 times, and chloroform layer is again with saturated sodium bicarbonate solution 5ml washing 1 time, add again water 10ml vibration washing 2 times, chloroform layer is dried 1 hour with anhydrous magnesium sulfate 1.5g, filters, and evaporate to dryness chloroform obtains crude product.Crude product, with appropriate methylene dichloride reflux, has insoluble solids, filters, the concentrated part of going of filtrate, add sherwood oil to micro-muddiness, reheat to clarification, place, crystallization, filter, solid adds sherwood oil to micro-muddiness after dissolving with methylene dichloride again, reheats to clarification, place, crystallization obtains fine work 0.32g.Productive rate: 43.2%.
1HNMR(600MHz,CDCl3)δ8.53(d,J=5.6Hz,1H),7.61(s,1H),7.56(s,1H),7.50(t,J=1.9Hz,1H),7.46(d,J=8.4Hz,1H),7.22(t,J=8.0Hz,2H),7.11-7.07(m,2H),7.04(d,J=8.4Hz,1H),6.95(s,1H),6.63(d,J=5.5Hz,1H),4.08(s,3H),4.06(s,3H),3.87(t,J=6.3Hz,2H),2.84(t,J=6.6Hz,2H),2.07-2.03(m,2H).HRMS[M+H]490.32。
Embodiment 18's is synthetic)
In 100ml round-bottomed bottle, add 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline 0.5g(1.5mmol), chloroform 10ml, dissolve, stirring at room, separately gets 2-chloro-phenyl-isocyanic ester 0.46g(3.0mmol) in chloroform 2ml, extremely dissolve, after dissolving, slowly add in round-bottomed bottle, after room temperature reaction 1 hour, TLC follows the tracks of reaction, and (developping agent is methylene dichloride: methyl alcohol=4.5:0.5), room temperature reaction is 65 hours altogether, aftertreatment.
Reaction solution moves in separating funnel, and chloroform layer adds water 10ml vibration washing 2 times, and chloroform layer is again with saturated sodium bicarbonate solution 5ml washing 1 time, add again water 10ml vibration washing 2 times, chloroform layer is dried 1 hour with anhydrous magnesium sulfate 1.5g, filters, and evaporate to dryness chloroform obtains crude product.Crude product, with appropriate methylene dichloride reflux, has insoluble solids, filters, the concentrated part of going of filtrate, add sherwood oil to micro-muddiness, reheat to clarification, place, crystallization, filter, solid adds sherwood oil to micro-muddiness after dissolving with methylene dichloride again, reheats to clarification, place, crystallization obtains fine work 0.37g.Productive rate: 50%.
1HNMR(600MHz,CDCl 3)δ8.52(d,J=5.4Hz,1H),8.38(dd,J=8.3,1.4Hz,1H),7.73(s,1H),7.57(d,J=9.6Hz,2H),7.54(s,1H),7.33(dd,J=8.0,1.4Hz,1H),7.30–7.27(m,1H),7.11(dd,J=8.6,2.7Hz,1H),7.08(d,J=2.7Hz,1H),6.98(td,J=7.8,1.5Hz,1H),6.54(d,J=5.4Hz,1H),4.08(s,3H),4.07(s,3H),3.91(t,J=6.3Hz,2H),2.85(t,J=6.6Hz,2H),2.06(p,J=6.6Hz,2H).HRMS[M+H]490.43。
Embodiment 19's is synthetic)
In 100ml round-bottomed bottle, add 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline 0.5g(1.5mmol), chloroform 10ml, dissolve, stirring at room, separately gets 2-methyl-4-chloro-phenyl-isocyanic ester 0.5g(3.0mmol) in chloroform 2ml, extremely dissolve, after dissolving, slowly add in round-bottomed bottle, after room temperature reaction 1 hour, TLC follows the tracks of reaction, and (developping agent is methylene dichloride: methyl alcohol=4.5:0.5), room temperature reaction is 75 hours altogether, aftertreatment.
Reaction solution moves in separating funnel, and chloroform layer adds water 10ml vibration washing 2 times, and chloroform layer is again with saturated sodium bicarbonate solution 5ml washing 1 time, add again water 10ml vibration washing 2 times, chloroform layer is dried 1 hour with anhydrous magnesium sulfate 1.5g, filters, and evaporate to dryness chloroform obtains crude product.Crude product, with appropriate methylene dichloride reflux, has insoluble solids, filters, the concentrated part of going of filtrate, add sherwood oil to micro-muddiness, reheat to clarification, place, crystallization, filter, solid adds sherwood oil to micro-muddiness after dissolving with methylene dichloride again, reheats to clarification, place, crystallization obtains fine work 0.22g.Productive rate: 28.9%.
1HNMR(600MHz,CDCl 3)δ8.51(d,J=5.4Hz,1H),7.84(d,J=8.7Hz,1H),7.56-7.51(m,3H),7.19(dd,J=8.7,2.4Hz,1H),7.14(d,J=2.2Hz,1H),7.08(dd,J=6.1,3.0Hz,2H),6.84(s,1H),6.54(d,J=5.4Hz,1H),4.08(s,3H),4.06(s,3H),3.88(t,J=6.3Hz,2H),2.84(t,J=6.6Hz,2H),2.14(s,3H),2.08-2.03(m,2H).HRMS[M+H]504.43。
Embodiment 20's is synthetic)
In 100ml round-bottomed bottle, add 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline 0.5g(1.5mmol), chloroform 10ml, dissolve, stirring at room, separately gets 4-trifluoromethylbenzene based isocyanate 0.56g(3.0mmol) in chloroform 2ml, extremely dissolve, after dissolving, slowly add in round-bottomed bottle, after room temperature reaction 1 hour, TLC follows the tracks of reaction, and (developping agent is methylene dichloride: methyl alcohol=4.5:0.5), room temperature reaction is 75 hours altogether, aftertreatment.
Reaction solution moves in separating funnel, and chloroform layer adds water 10ml vibration washing 2 times, and chloroform layer is again with saturated sodium bicarbonate solution 5ml washing 1 time, add again water 10ml vibration washing 2 times, chloroform layer is dried 1 hour with anhydrous magnesium sulfate 1.5g, filters, and evaporate to dryness chloroform obtains crude product.Crude product, with appropriate methylene dichloride reflux, has insoluble solids, filters, the concentrated part of going of filtrate, add sherwood oil to micro-muddiness, reheat to clarification, place, crystallization, filter, solid adds sherwood oil to micro-muddiness after dissolving with methylene dichloride again, reheats to clarification, place, crystallization obtains fine work 0.25g.Productive rate: 31.6%.
1HNMR(600MHz,CDCl 3)δ8.54(d,J=5.3Hz,1H),7.58-7.51(m,6H),7.45(t,J=7.7Hz,1H),7.12-7.08(m,3H),6.62(d,J=4.7Hz,1H),4.09(s,3H),4.06(s,3H),3.89(t,J=6.3Hz,2H),2.84(t,J=6.6Hz,2H),2.09-2.03(m,2H).HRMS[M+H]524.32。
Embodiment 21's is synthetic)
In 100ml round-bottomed bottle, add 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline 0.5g(1.5mmol), chloroform 10ml, dissolve, stirring at room, separately gets 2-trifluoromethylbenzene based isocyanate 0.56g(3.0mmol) in chloroform 2ml, extremely dissolve, after dissolving, slowly add in round-bottomed bottle, after room temperature reaction 1 hour, TLC follows the tracks of reaction, and (developping agent is methylene dichloride: methyl alcohol=4.5:0.5), room temperature reaction is 70 hours altogether, aftertreatment.
Reaction solution moves in separating funnel, and chloroform layer adds water 10ml vibration washing 2 times, and chloroform layer is again with saturated sodium bicarbonate solution 5ml washing 1 time, add again water 10ml vibration washing 2 times, chloroform layer is dried 1 hour with anhydrous magnesium sulfate 1.5g, filters, and evaporate to dryness chloroform obtains crude product.Crude product, with appropriate methylene dichloride reflux, has insoluble solids, filters, the concentrated part of going of filtrate, add sherwood oil to micro-muddiness, reheat to clarification, place, crystallization, filter, solid adds sherwood oil to micro-muddiness after dissolving with methylene dichloride again, reheats to clarification, place, crystallization obtains fine work 0.17g.Productive rate: 21.5%.
1HNMR(600MHz,CDCl3)δ8.53(d,J=5.5Hz,1H),7.61-7.53(m,6H),7.47(t,J=8.2Hz,1H),7.15-7.07(m,3H),6.56(d,J=5.5Hz,1H),4.10(s,3H),4.08(s,3H),3.90(t,J=6.3Hz,2H),2.85(t,J=6.7Hz,2H),2.10-2.04(m,2H).HRMS[M+H]524.37。
Embodiment 22's is synthetic)
In 100ml round-bottomed bottle, add 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline 0.5g(1.5mmol), chloroform 10ml, dissolve, stirring at room, separately gets 4-cyano-phenyl isocyanic ester 0.43g(3.0mmol) in chloroform 2ml, extremely dissolve, after dissolving, slowly add in round-bottomed bottle, after room temperature reaction 1 hour, TLC follows the tracks of reaction, and (developping agent is methylene dichloride: methyl alcohol=4.5:0.5), room temperature reaction is 75 hours altogether, aftertreatment.
Reaction solution moves in separating funnel, and chloroform layer adds water 10ml vibration washing 2 times, and chloroform layer is again with saturated sodium bicarbonate solution 5ml washing 1 time, add again water 10ml vibration washing 2 times, chloroform layer is dried 1 hour with anhydrous magnesium sulfate 1.5g, filters, and evaporate to dryness chloroform obtains crude product.Crude product, with appropriate methylene dichloride reflux, has insoluble solids, filters, the concentrated part of going of filtrate, add sherwood oil to micro-muddiness, reheat to clarification, place, crystallization, filter, solid adds sherwood oil to micro-muddiness after dissolving with methylene dichloride again, reheats to clarification, place, crystallization obtains fine work 0.30g.Productive rate: 41.7%.
1HNMR(600MHz,CDCl 3)δ8.53(d,J=5.3Hz,1H),7.60-7.58(m,2H),7.55-7.52(m,3H),7.50(s,1H),7.41(d,J=8.7Hz,1H),7.21(s,1H),7.12-7.09(m,2H),6.59(d,J=5.3Hz,1H),4.07(s,3H),4.06(s,3H),3.88(t,J=6.4Hz,2H),2.83(t,J=6.6Hz,2H),2.05(m,J=6.6Hz,2H).HRMS[M+H]481.33。
Embodiment 23's is synthetic)
In 100ml round-bottomed bottle, add 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline 0.5g(1.5mmol), chloroform 10ml, dissolve, stirring at room, separately gets 2-chloro-4 nitrophenyl isocyanic ester 0.6g(3.0mmol) in chloroform 2ml, extremely dissolve, after dissolving, slowly add in round-bottomed bottle, after room temperature reaction 1 hour, TLC follows the tracks of reaction, and (developping agent is methylene dichloride: methyl alcohol=4.5:0.5), room temperature reaction is 70 hours altogether, aftertreatment.
Reaction solution moves in separating funnel, and chloroform layer adds water 10ml vibration washing 2 times, and chloroform layer is again with saturated sodium bicarbonate solution 5ml washing 1 time, add again water 10ml vibration washing 2 times, chloroform layer is dried 1 hour with anhydrous magnesium sulfate 1.5g, filters, and evaporate to dryness chloroform obtains crude product.Crude product, with appropriate methylene dichloride reflux, has insoluble solids, filters, the concentrated part of going of filtrate, add sherwood oil to micro-muddiness, reheat to clarification, place, crystallization, filter, solid adds sherwood oil to micro-muddiness after dissolving with methylene dichloride again, reheats to clarification, place, crystallization obtains fine work 0.27g.Productive rate: 33.8%.
1HNMR(600MHz,CDCl 3)δ8.69(d,J=9.3Hz,1H),8.53(d,J=5.6Hz,1H),8.28(dd,J=13.0,2.6Hz,1H),8.19(dd,J=9.3,2.5Hz,1H),8.09(s,1H),7.67(s,1H),7.58-7.53(m,2H),7.17-7.11(m,2H),6.57(d,J=4.6Hz,1H),4.10(s,3H),4.07(s,3H),3.93(t,J=6.4Hz,2H),2.87(t,J=6.6Hz,2H),2.10(p,J=6.5Hz,2H).HRMS[M+H]535.27。
Embodiment 24's is synthetic)
In 100ml round-bottomed bottle, add 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline 0.5g(1.5mmol), chloroform 10ml, dissolve, stirring at room, separately gets 3-nitro-4-chloro-phenyl-isocyanic ester 0.6g(3.0mmol) in chloroform 2ml, extremely dissolve, after dissolving, slowly add in round-bottomed bottle, after room temperature reaction 1 hour, TLC follows the tracks of reaction, and (developping agent is methylene dichloride: methyl alcohol=4.5:0.5), room temperature reaction is 70 hours altogether, aftertreatment.
Reaction solution moves in separating funnel, and chloroform layer adds water 10ml vibration washing 2 times, and chloroform layer is again with saturated sodium bicarbonate solution 5ml washing 1 time, add again water 10ml vibration washing 2 times, chloroform layer is dried 1 hour with anhydrous magnesium sulfate 1.5g, filters, and evaporate to dryness chloroform obtains crude product.Crude product, with appropriate methylene dichloride reflux, has insoluble solids, filters, the concentrated part of going of filtrate, add sherwood oil to micro-muddiness, reheat to clarification, place, crystallization, filter, solid adds sherwood oil to micro-muddiness after dissolving with methylene dichloride again, reheats to clarification, place, crystallization obtains fine work 0.36g.Productive rate: 45%.
1HNMR(600MHz,CDCl 3)δ8.52(d,J=5.8Hz,1H),8.04(d,J=2.5Hz,1H),7.88(d,J=86.2Hz,1H),7.67(dd,J=8.8,2.5Hz,2H),7.57(s,1H),7.46(dt,J=15.6,7.6Hz,2H),7.13(dt,J=4.6,2.6Hz,2H),6.72(d,J=5.7Hz,1H),4.09(s,3H),4.08(s,3H),3.89(t,J=6.3Hz,2H),2.85(t,J=6.6Hz,2H),2.09-2.04(m,2H).HRMS[M+H]535.29。
Embodiment 25's is synthetic)
In 100ml round-bottomed bottle, add 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline 0.5g(1.5mmol), chloroform 10ml, dissolve, stirring at room, separately gets 2-methyl-4-nitro isocyanic ester 0.53g(3.0mmol) in chloroform 2ml, extremely dissolve, after dissolving, slowly add in round-bottomed bottle, after room temperature reaction 1 hour, TLC follows the tracks of reaction, and (developping agent is methylene dichloride: methyl alcohol=4.5:0.5), room temperature reaction is 75 hours altogether, aftertreatment.
Reaction solution moves in separating funnel, and chloroform layer adds water 10ml vibration washing 2 times, and chloroform layer is again with saturated sodium bicarbonate solution 5ml washing 1 time, add again water 10ml vibration washing 2 times, chloroform layer is dried 1 hour with anhydrous magnesium sulfate 1.5g, filters, and evaporate to dryness chloroform obtains crude product.Crude product, with appropriate methylene dichloride reflux, has insoluble solids, filters, the concentrated part of going of filtrate, add sherwood oil to micro-muddiness, reheat to clarification, place, crystallization, filter, solid adds sherwood oil to micro-muddiness after dissolving with methylene dichloride again, reheats to clarification, place, crystallization obtains fine work 0.31g.Productive rate: 40.3%.
1HNMR(600MHz,CDCl 3)δ8.52(d,J=5.7Hz,1H),8.40(d,J=9.1Hz,1H),8.14(d,J=2.4Hz,1H),8.04(d,J=2.3Hz,1H),7.69(s,1H),7.57-7.53(m,2H),7.28(s,1H),7.15-7.12(m,2H),6.59(d,J=4.5Hz,1H),4.10(s,3H),4.08(s,3H),3.92(t,J=6.3Hz,2H),2.87(t,J=6.2Hz,2H),2.19(s,3H),2.11-2.07(m,2H).HRMS[M+H]515.36。
Embodiment 26's is synthetic)
In 100ml round-bottomed bottle, add 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline 0.5g(1.5mmol), chloroform 10ml, dissolve, stirring at room, separately gets cyclopentyl isocyanic ester 0.33g(3.0mmol) in chloroform 2ml, extremely dissolve, after dissolving, slowly add in round-bottomed bottle, after room temperature reaction 1 hour, TLC follows the tracks of reaction, and (developping agent is methylene dichloride: methyl alcohol=4.5:0.5), room temperature reaction is 75 hours altogether, aftertreatment.
Reaction solution moves in separating funnel, and chloroform layer adds water 10ml vibration washing 2 times, and chloroform layer is again with saturated sodium bicarbonate solution 5ml washing 1 time, add again water 10ml vibration washing 2 times, chloroform layer is dried 1 hour with anhydrous magnesium sulfate 1.5g, filters, and evaporate to dryness chloroform obtains crude product.Crude product, with appropriate methylene dichloride reflux, has insoluble solids, filters, the concentrated part of going of filtrate, add sherwood oil to micro-muddiness, reheat to clarification, place, crystallization, filter, solid adds sherwood oil to micro-muddiness after dissolving with methylene dichloride again, reheats to clarification, place, crystallization obtains fine work 0.19g.Productive rate: 28.4%.
1HNMR(600MHz,CDCl 3)δ8.51(d,J=5.3Hz,1H),7.54(s,1H),7.47(s,1H),7.40(d,J=8.5Hz,1H),7.01(dd,J=9.4,3.5Hz,2H),6.55(d,J=5.3Hz,1H),4.99(d,J=6.9Hz,1H),4.22-4.16(m,1H),4.06(s,3H),4.05(s,3H),3.78-3.75(m,2H),2.77(t,J=6.6Hz,2H),2.04-1.94(m,4H),1.67-1.60(m,4H),1.42-1.35(m,2H).HRMS[M+H]448.51。
Embodiment 27's is synthetic)
In 100ml round-bottomed bottle, add 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline 0.5g(1.5mmol), chloroform 10ml, dissolve, stirring at room, separately gets cyclohexyl isocyanate 0.38g(3.0mmol) in chloroform 2ml, extremely dissolve, after dissolving, slowly add in round-bottomed bottle, after room temperature reaction 1 hour, TLC follows the tracks of reaction, and (developping agent is methylene dichloride: methyl alcohol=4.5:0.5), room temperature reaction is 75 hours altogether, aftertreatment.
Reaction solution moves in separating funnel, and chloroform layer adds water 10ml vibration washing 2 times, and chloroform layer is again with saturated sodium bicarbonate solution 5ml washing 1 time, add again water 10ml vibration washing 2 times, chloroform layer is dried 1 hour with anhydrous magnesium sulfate 1.5g, filters, and evaporate to dryness chloroform obtains crude product.Crude product, with appropriate methylene dichloride reflux, has insoluble solids, filters, the concentrated part of going of filtrate, add sherwood oil to micro-muddiness, reheat to clarification, place, crystallization, filter, solid adds sherwood oil to micro-muddiness after dissolving with methylene dichloride again, reheats to clarification, place, crystallization obtains fine work 0.33g.Productive rate: 47.8%.
1HNMR(600MHz,DMSO)δ8.51(d,J=5.3Hz,1H),7.55(d,J=8.8Hz,1H),7.52(s,1H),7.40(s,1H),7.02(d,J=2.7Hz,1H),6.99(dd,J=8.8,2.8Hz,1H),6.56(d,J=5.2Hz,1H),6.44(d,J=7.5Hz,1H),3.96(s,3H),3.94(s,3H),3.61-3.58(m,2H),3.38(q,J=7.0Hz,1H),2.72(t,J=6.5Hz,2H),1.86-1.70(m,6H),1.29-1.24(m,4H),1.10(dd,J=12.5,5.5Hz,2H).HRMS[M+H]462.50。
Test example 1: vitro inhibition tyrosine kinase activity test
(1) experimental technique
(a) preparation of damping fluid
With 50mMHEPES, pH7.5,0.0015%Brij-35,10mMMgCl 2, 2mMDTT kinases preparation damping fluid, 100mMHEPES(pH7.5), 0.015%Brij-35,0.2%CoatingReagent#3,50mMEDTA prepare stop buffer.
(b) preparation of sample solution
Survey the storage liquid that before living, given the test agent is made into 10mM, guaranteeing with damping fluid, to be diluted to desired concn under the prerequisite that DMSO concentration is 10%; As only needed, measure the inhibiting rate under a certain concentration, adopt a concentration; If need, measure IC 50value, initial concentration is 10mM, and extension rate is 3, and 10 concentration are set, and each concentration is established multiple hole.
(c) kinase reaction
In basic damping fluid, add kinases with preparation kinase buffer liquid; In basic damping fluid, add FAM-labeled peptide and ATP with preparation peptide damping fluid.The sample solution that adds 10 μ l different concns in the test hole of 384 orifice plates, adds 10 μ l kinase buffer liquid during institute is porose, at room temperature cultivates after 10min, in institute is porose, adds 10 μ l peptide damping fluids, and continues to cultivate 60min at 28 ℃.Add subsequently 25 μ l stop buffers with stopped reaction, by instrument image data and make curve, obtain IC 50value.
We have tested the inhibiting rate to VEGFR-2 (KDR) and VEGFR-3 (FLT4) under whole compound 30nM concentration, carry out preliminary screening:
Table 1: kinases kind and experiment condition
Figure BDA0000388073950000191
(1) experimental result
Inhibiting rate experiment under table 2:30nM concentration
Figure BDA0000388073950000192
Figure BDA0000388073950000201
In inhibiting rate experiment under 30nM concentration, that contrast is selected is Staurosporine (Staurosporine, STS), the IC to VEGFR-2 50for 7.2nM, the IC to VEGFR-3 50for 1.5nM.
We are chosen under 30nM VEGFR-2 (KDR) and the higher compound of VEGFR-3 (FLT4) inhibiting rate have been carried out to IC 50the mensuration of value:
Table 3: the kinase inhibition IC of part preferred compound 50the mensuration of value
Figure BDA0000388073950000202
Staurosporine (Staurosporine) is selected in contrast.VEGFR inhibition experiment is presented at tested compound all has certain restraining effect to VEGFR-2 (KDR) or VEGFR-3 (FLT4), and wherein part of compounds all has stronger restraining effect to two kinds of acceptors, active in contrast Staurosporine.
Above kinase activity test Jun Shanghai Ruizhi Chemical Study Co., Ltd. carries out, and above data are all provided by the project report being provided by wise and farsighted chemistry.
The hepatomicrosome metabolic rate research of test example 2 part preferred compounds
1. the preparation of reference substance:
Weigh: precision takes each 3mg of KL-07, KL-11, KL-13 and KL-17 in EP pipe;
Dissolve: add 1mL methyl alcohol, 4mLDMSO dissolves, and final concentration is 0.6mg/mL.
2. Preparatory work of experiment
The acetonitrile solution approximately 100 μ mol/L of 1.1 preparation KL-07, KL-11, KL-13 and KL-17 are as storing solution, A5 (chemical name: N-(5-(4-(4-((dimethylamino) methyl) phenyl) quinoline-6-yl)-2-picoline-3-yl)-2, the 4-phenyl-difluoride sulphonamide) solution of another preparation 100 μ mol/L contrasts;
1.2PBS solution;
1.3β-NADPH16.7mg/mL；
1.4 rat liver microsomes;
The preparation of 1.5 inner mark solutions
KL-07, KL-11, KL-13, KL-17 and A5 are with N-(4-acetyl the phenyl)-6-(6 of 100ng/mL, 7-dimethoxyquinazoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide (2N-27) solution is interior mark, N-(4-acetyl phenyl)-6-(6,7-dimethoxyquinazoline-4-oxygen base)-3, it is interior mark that 4-dihydroquinoline-1 (2H)-methane amide (2N-27) be take the KL-17 of 100ng/mL.
3. the analysis condition of compound
KL-07：
Chromatographic condition:
Chromatographic column: Shiseido 3.0 * 100,3 μ m; Flow velocity: 0.4mL/min; Sample size: 5 μ L; Moving phase: 5mmol ammonium acetate+0.1% formic acid water: acetonitrile=55:45; Analysis time: 10min.
Mass spectrum condition:
Positive ion mode, SIM monitors [M+H] +486.0
Dry gas volume: 10L
Spraying gun pressure: 40psig; Dry gas temperature: 350 ℃
Cracking voltage: 70eV.
KL-11：
Chromatographic condition:
Chromatographic column: Shiseido 3.0 * 100,3 μ m; Flow velocity: 0.4mL/min; Sample size: 5 μ L; Moving phase: 5mmol ammonium acetate+0.1% formic acid water: acetonitrile=55:45; Analysis time: 10min.
Mass spectrum condition:
Positive ion mode, SIM monitors [M+H] +497.90
Dry gas volume: 10L
Spraying gun pressure: 40psig; Dry gas temperature: 350 ℃
Cracking voltage: 70eV.
KL-13：
Chromatographic condition:
Chromatographic column: Shiseido 3.0 * 100,3 μ m; Flow velocity: 0.4mL/min; Sample size: 5 μ L; Moving phase: 5mmol ammonium acetate+0.1% formic acid water: acetonitrile=55:45; Analysis time: 10min.
Mass spectrum condition:
Positive ion mode, SIM monitors [M+H] +474.0
Dry gas volume: 10L
Spraying gun pressure: 40psig; Dry gas temperature: 350 ℃
Cracking voltage: 70eV.
KL-17：
Chromatographic condition:
Chromatographic column: Shiseido 3.0 * 100,3 μ m; Flow velocity: 0.4mL/min; Sample size: 5 μ L; Moving phase: 5mmol ammonium acetate+0.1% formic acid water: acetonitrile=55:45; Analysis time: 10min.
Mass spectrum condition:
Positive ion mode, SIM monitors [M+H] +490.0
Dry gas volume: 10L
Spraying gun pressure: 40psig; Dry gas temperature: 350 ℃
Cracking voltage: 70eV.
4. sample determination result:
The measurement result of table 4:KL-07
Figure BDA0000388073950000221
The measurement result of table 5:KL-11
Figure BDA0000388073950000222
The measurement result of table 6:KL-13
Figure BDA0000388073950000223
Figure BDA0000388073950000231
The measurement result of table 7:KL-17
The measurement result of table 8:A5
Figure BDA0000388073950000233
The absorption characteristic research of test example 3 part preferred compounds in Caco-2 model
1. the preparation of need testing solution
A series of concentration of preparation KL-07, KL-11, KL-13 and KL-17: 20 μ mol/L, 10 μ mol/L, 5 μ mol/L, 2 μ mol/L, 1 μ mol/L, 0.5 μ mol/L, 0.2 μ mol/L, 0.1 μ mol/L.
2. cell cultures and toxicity test
Inoculating cell: be mixed with single Caco-2 cell (deriving from ATCC) suspension with the MEM substratum containing 10% foetal calf serum, be inoculated in 96 orifice plates with 5000, every hole cell.
Be placed in incubator and cultivate, administration after cell is all adherent (at least adherent reach 80%).
After cultivating 24-36h, each hole adds 20 μ LMTT, and (5mg/mL prepares with PBS, pH=7.4) cultivate after 4h sucking-off supernatant liquor and add 150 μ LDMSO, jolting 10min, select 492nm wavelength, on enzyme linked immunological monitor, measure each hole absorbance value, record result, take the time as X-coordinate, and light absorption value is that ordinate zou is drawn cell growth curve.
Get the concentration administration Caco-2 Cell uptake model of inhibiting rate < 2% of relative Caco-2 cell, measure its apparent permeability coefficient.Result, four compound 20 μ mol/L, 10 μ mol/L, 5 μ mol/L show obvious cyto-inhibition to Caco-2 cell, final KL-11, KL-13, KL-17 choose 1 μ M, and KL-27 chooses the safe administration concentration of 0.5 μ M and carries out the transmitance experiment of Caco-2 cell.
3. transport experiment
3.1Caco-2 cell model is set up
Caco-2 cell (contains 10%FBS, 1%NEAA, 100UmL at MEM substratum -1penicillin-Streptomycin sulphate, 10mmolL -1hEPES), the 5%CO of 37 ℃ 2in constant incubator, cultivate.After cell cultures 21 days, cell model is verified, chosen cell transmembrane resistance and be greater than 600 Ω cm 2and uranine transmitance is lower than 0.6%h -1cm -2cell carry out drug transport experiment.
The transhipment of 3.2 compounds and picked-up experiment
Cell cleans three times with HBSS liquid is careful, hatches for the last time 30min in incubator, blots HBSS liquid.Blank group all adds HBSS liquid at AP and BL both sides, and administration group adds the HBSS liquid (AP side 0.5ml, BL side 1.5ml) of 0.5 μ M2N-27 in administration side, and receiver side adds blank HBSS liquid.Then culture plate is put into incubator and hatch, collect respectively BL side and AP side 0,30,60,90 and the solution 100 μ L of 120min, after sampling, add blank HBSS liquid 100 μ L, it is frozen that sample is placed in-20 ℃ of refrigerators.By the chemical composition in HPLC quantitative analysis transhipment liquid.
The preparation of 3.3 linear solution
Get KL-07, KL-11, KL-13 and KL-17 and obtain with HBSS solution preparation the HBSS liquid that concentration is 1 μ mol/mL, 0.5 μ mol/mL, 0.25 μ mol/mL, 0.125 μ mol/mL, 0.0625 μ mol/mL, 0.3125 μ mol/mL; Get 100 μ LHBSS liquid+300 μ L(containing interior mark) acetonitrile.The centrifugal 5min of vortex 20S gets supernatant and obtains linear sample; Inner mark solution is prepared with acetonitrile: 100ng/mLKL-17 and 100ng/mL2N-27.
Concentration determination adopts high performance liquid chromatography.Chromatographic column: Shiseido C 183.0 * 100mm3 μ m; Flow velocity: 0.4mL/min sample size: 5 μ L, moving phase: 5mmol ammonium acetate+0.1% formic acid water: acetonitrile=55:45; Analysis time: 10min.
Table 9
Figure BDA0000388073950000251
y=0.7707x-0.1367R 2=0.976
Table 10
Figure BDA0000388073950000252
y=0.5721x-0.11R 2=0.9038
Table 11
y=0.1692x-0.0399R 2=0.9698
From above data, can find out the apparent permeability coefficient P of compound of mensuration appall be greater than 10 -5, (two other compound does not reach quantitative limit and cannot measure) be it is generally acknowledged, apparent permeability coefficient P appbe greater than 10 -6the oral absorption that shows compound is good.This explanation, the absorption characteristic of KL-11 and KL-13 is very good, the compound providing due to the application all has identical parent nucleus, chemical structure difference is less to each other, therefore those skilled in the art should estimate, the compound that the application provides should all have good absorption, is expected to develop the oral preparations that bioavailability is high.
The pharmacokinetics experiment of test example 4 compound K L-11
1. analysis condition
Chromatographic condition
Chromatographic column: Shiseido 3.0 * 100mm, 3 μ m, C 18post; Sample size: 5 μ L; Flow velocity: shunt 1:1 after 0.8mL/min post;
Moving phase: 0.1% formic acid water: acetonitrile=64:36
Mass spectrum condition: positive ion mode, SIM=498.2 (KL-11) SIM=474.0 (KL-13)
Dry gas volume: 10L
Spraying gun pressure: 40psig; Dry gas temperature: 350 ℃
Cracking voltage: 70eV
2, sample treatment
Inside being designated as KL-13(concentration is 60.5ng/mL)
With acetonitrile compound concentration, be followed successively by the standardized solution of 13ug/ml, 6.5ug/ml, 3.25ug/ml, 1.3ug/ml, 0.65ug/ml, 0.325ug/ml, 0.013ug/ml, 0.065ug/ml, obtain typical curve 1;
In label taking directrix curve 1, each concentration 10ul adds and adds 190ul after 100ul plasma sample and contain interior target acetonitrile solution respectively, and vortex 30S, gets the analysis of supernatant liquor sample introduction after 12000rpm, centrifugal 10min.
Three, experimental result
Table 12: gavage group KL-11 typical curve
Figure BDA0000388073950000261
y=9.4248x-0.0994
R2=0.9992
Table 13:KL-11 lower limit of quantitation experimental result
Figure BDA0000388073950000262
Table 14:KL-11 withinday precision experimental result
Figure BDA0000388073950000271
Table 15:KL-11 matrix effect and extraction recovery experimental result
Figure BDA0000388073950000272
Table 16: gavage group rat KL-11 Plasma Concentration (ug/ml)
Figure BDA0000388073950000281
The pharmacokinetic parameter of 1N-11 concentration in table 17 gavage group rat plasma
Figure BDA0000388073950000282
More than show and described ultimate principle of the present invention and principal character.Those skilled in the art should understand; the present invention is not restricted to the described embodiments; that in above-described embodiment and specification sheets, describes just illustrates principle of the present invention; the present invention also has various changes and modifications without departing from the spirit and scope of the present invention, and these changes and improvements all fall in the claimed scope of the invention.The claimed scope of the present invention is defined by appending claims and equivalent thereof.

Claims (10)

1. the compound shown in formula I, or its each optical isomer, each crystal formation, pharmaceutically acceptable inorganic or organic salt, hydrate, solvate or prodrug:
Figure FDA0000388073940000011
Wherein R is selected from H, C 1-10alkyl, C 1-6thiazolinyl, C 1-6alkynyl, C 1-6alkoxyl group, C 1-6alkylthio, C 1-6carbonyl, C 1-6the carbonylamino of carbalkoxy, carbonylamino, replacement, sulfuryl amino, phenyl, substituted-phenyl, cycloalkyl, substituted cycloalkyl, Heterocyclylalkyl, substituted heterocycle alkyl, heteroaryl or substituted heteroaryl.
2. compound according to claim 1, is characterized in that, R is selected from H, C 1-10alkyl, C 1-6alkoxyl group, or be selected from following structural unit:
Figure FDA0000388073940000012
Wherein, R 1, R 2independently be selected from separately H, halogen, C 1-6alkyl, C 1-6thiazolinyl, C 1-6alkynyl, C 1-6alkoxyl group, nitro, trifluoromethyl, trifluoromethoxy or C 1-6alkyl-carbonyl.
3. compound according to claim 1, is characterized in that, described compound is selected from following compound:
N-(2,4,4-trimethylpentane-2-yl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide;
N-(2-ethylphenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide;
N-(2,4-3,5-dimethylphenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide;
N-(2-isopropyl phenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide;
N-(4-n-butylphenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide;
N-(4-p-methoxy-phenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide;
N-(3-p-methoxy-phenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide;
N-(2-p-methoxy-phenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide;
N-(2-methoxyl group-5-aminomethyl phenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide;
N-(4-acetylphenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide;
N-(4-fluorophenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide;
N-(3-fluorophenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide;
N-(2-fluorophenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide;
N-(2,4 difluorobenzene base)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide;
N-(4-chloro-phenyl-)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide;
N-(3-chloro-phenyl-)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide;
N-(2-chloro-phenyl-)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide;
N-(4-chloro-2-methyl phenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide;
N-(4-(trifluoromethyl) phenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide;
N-(3-(trifluoromethyl) phenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide;
N-(4-cyano-phenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide;
N-(2-chloro-4 nitrophenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide;
N-(the chloro-3-nitrophenyl of 4-)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide;
N-(2-methyl-4-nitrophenyl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide;
N-cyclopentyl-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide;
N-cyclohexyl-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide;
N-(benzo [d] [1,3] dioxolane-5-yl)-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-methane amide.
4. prepare a method for compound as claimed in claim 1, it is characterized in that, containing having the following steps:
Figure FDA0000388073940000031
(1) 4-is chloro-6,7-dimethoxy-quinoline and 6-hydroxyl-1, and 2,3,4-tetrahydroquinoline reacts and generates 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline (II) under the existence of alkali, catalyzer;
(2) 6,7-dimethoxy-4 '-(1,2,3,4-tetrahydroquinoline-6 oxygen base) quinoline (II) reacts with the isocyanate compound of various replacements and generates corresponding N-replacement-6-(6,7-dimethoxy-quinoline-4-oxygen base)-3,4-dihydroquinoline-1 (2H)-Carbox amide (I);
(3) according to ordinary method, be prepared as the form of pharmacy acceptable salt.
5. a pharmaceutical composition, it is characterized in that, it contains pharmaceutically acceptable vehicle or carrier, and compound claimed in claim 1 or its each optical isomer, each crystal formation, pharmaceutically acceptable inorganic or organic salt, hydrate, solvate or prodrug.
6. a purposes for compound claimed in claim 1 or its each optical isomer, each crystal formation, pharmaceutically acceptable inorganic or organic salt, hydrate, solvate or prodrug, is characterized in that, for the preparation of tyrosine kinase inhibitor.
7. the purposes of a compound claimed in claim 1 or its each optical isomer, each crystal formation, pharmaceutically acceptable inorganic or organic salt, hydrate, solvate or prodrug, it is characterized in that, for the preparation of suppressing the medicine of tyrosine kinase activity or for the preparation of the medicine for the treatment of, prevention and alleviation and the too high relative disease of tyrosine kinase activity.
8. purposes as claimed in claim 7, is characterized in that, the described and too high relative disease of tyrosine kinase activity is selected from tumour.
9. a purposes for compound claimed in claim 1 or its each optical isomer, each crystal formation, pharmaceutically acceptable inorganic or organic salt, hydrate, solvate or prodrug, is characterized in that, for the preparation of angiogenesis inhibitor.
10. a purposes for compound claimed in claim 1 or its each optical isomer, each crystal formation, pharmaceutically acceptable inorganic or organic salt, hydrate, solvate or prodrug, is characterized in that, for the preparation of the medicine of prevention or treatment tumour.
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