CN101659657B

CN101659657B - Quinoline substituted by cyan and preparation method and applications thereof

Info

Publication number: CN101659657B
Application number: CN200910152098.1A
Authority: CN
Inventors: 易崇勤; 赵鸿莲
Original assignee: Peking University Founder Group Co Ltd; PKU International Healthcare Group Co Ltd; PKUCare Pharmaceutical R&D Center
Current assignee: New Founder Holdings Development Co ltd; Peking University Medical Management Co ltd; Peking University Founder Group Co Ltd; PKU Healthcare Industry Group; PKUCare Pharmaceutical R&D Center
Priority date: 2008-08-29
Filing date: 2009-07-28
Publication date: 2014-05-14
Anticipated expiration: 2029-07-28
Also published as: CN101659657A

Abstract

The invention relates to a quinoline substituted by cyan shown in formula I, wherein R<1>, X<1>, R<2>, R<5>, n and m are defined in a specification; and the preparation method thereof comprises the medicine compounds thereof and applications in preparation of medicine which is used as an anti-proliferation agent and used for preventing or treating tumor which is sensitive to receptor tyrosine kinase inhibiting EGF and erbB. The formula I is shown in the description.

Description

Quinoline that cyano group replaces and its production and use

Technical field

The present invention relates to some novel quinoline or its pharmacy acceptable salt, the methods for the treatment of that it has anti-tumor activity and is therefore applicable to human or animal body.The invention still further relates to the preparation method of described quinoline, the pharmaceutical composition that comprises them and their application in methods for the treatment of, for example for the preparation of prevention or treatment warm-blooded animal as human body in application in the medicine of solid tumor disease.

Background technology

Disease that current many treatments are caused extremely by regulation of cell proliferation has been utilized the compound that suppresses the synthetic and cell proliferation of DNA as the scheme of psoriasis and cancer.So far, conventionally have cytotoxicity for the compound of this treatment, but they pretend use for quick noble cells as cancer cells has, this effect is favourable.Develop at present the method for replaceable these cell toxicant antitumour drugs, the selective depressant of for example Cell signal transduction pathway.The inhibitor of these types can show and strengthens selectively acting in the potentiality of antitumor cell, and therefore can reduce the probability of the adverse side effect that treatment has.

Eukaryotic cell constantly, to many different extracellular signals reactions, makes can carry out communication between the cell in organism like this.The intracellular many physical responses of these Signal Regulation, comprise propagation, differentiation, apoptosis and motion.Extracellular signal shows as various water-soluble factor form, comprises somatomedin and paracrine and endocrine factor.By binding specificity transmembrane receptor, extracellular signal is attached to intracellular signal transduction approach by these parts, therefore, signal transduction replied its extracellular signal through plasma membrane and permission cell individual.Many signal transduction processes are utilized the reverse process of protein phosphorylation, and described albumen participates in promoting above various cell responses.The phosphorylation state of target protein regulates by specificity kinases and phosphoesterase, and approximately 1/3 of all albumen of these enzymes to mammalian genes group coding regulate.Because phosphorylation is the very important regulation mechanism in signal transduction process, therefore being understood that in these cells that approach is not normal will cause Growth of Cells and prosoplasia, and (summary is referring to Cohen et al. therefore to promote transformation, Curr OpinChem Biol, 1999,3,459-465).

Well-known many Tyrosylprotein kinases sport constitutive activity form and/or after being overexpressed, cause various human cells to transform.In the human tumor that accounts for significant proportion, all there is (summarizing referring to Kolibaba et al. Biochimica et BiophysicaActa, 1997,133, F217-F248) in kinase whose these sudden changes or overexpression form.Due to the vital role that Tyrosylprotein kinase has in the propagation of various tissues and differentiation, there is the research of a lot of development of new anti-cancer therapies all to concentrate on these enzymes.This family's enzyme is divided into two groups: receptor tyrosine kinase and nonreceptor tyrosine kinase, be respectively for example EGF acceptor and SRC family. from the large quantity research result of (comprising the Human Genome Project), approximately 90 kinds of Tyrosylprotein kinases in human genome, are identified, wherein 58 kinds is receptor type, and 32 kinds for non-receptor type. they can be divided into 20 kinds of receptor tyrosine kinases and 10 kinds of nonreceptor tyrosine kinase subfamily (Robinson et al., Oncogene, 2000,19,5548-5557).

Receptor tyrosine kinase is to causing the mitogenesis signal transduction particularly important of cellular replication.These large glycoprotein of crossing over cytoplasmic membrane have for example, integrated structure city, extracellular for its ligands specific (Urogastron of EGF acceptor (EGF)).Binding partner causes the kinase whose enzymic activity of activated receptor, and kinase activity is by the cell interior Coded of acceptor.Crux tyrosine amino acid in this active phosphorus acidifying target protein, transduces through cytoplasmic membrane proliferation signal.

The erbB family of known receptor Tyrosylprotein kinase (comprises EGFR, erbB2, erbB3 and erbB4) usually participate in promoting the propagation of tumour cell and survival (to summarize referring to Olayioye et al, EMBO J., 2000, 19, 3159). the one mechanism that can complete this effect is overexpression receptor protein, overexpression is the result of gene amplification normally. and this observes in a lot of common human cancers, and (summary is referring to Mapper et al., Adv.Cancer Res., 2000, 77, 25), for example mammary cancer (Sainsbury et al., Brit.J.Cancer, 1988, 58, 458, Guerin et al., OncogeneRes., 1988,3,21, Slamon et al., Science, 1989,244,707, Kliin et al., BreastCancer Res..Treat., 1994,29,73, summary is referring to Salomon et al., Crit.Rev.Oncol.Hemato., 1995,19,183), nonsmall-cell lung cancer (NSCLC), comprises gland cancer Cernyet al., Brit.J.Cancer, 1986,54,265, Reubi et al., Int.J.Cancer, 1990,45,269, Rusch et al., Cancer Research, 1993,53,2379, Brabender et al., Clin.Cancer Res., 2001,7,1850) and other lung cancer (Hendler et al., Cancer Cells, 1989,7,347, Ohsaki et al., Oncol, Rep., 2000,7,603), bladder cancer (Neal et al., Lancet, 1985,366, Chow et al., Clin.Cancer Res., 2001,7,1957, Zhau etal., Mol Carcinog., 3,254), esophagus cancer (Mukaida et al., Cancer, 1991,68,142), for example colon and rectum carcinoma of gastrointestinal cancer or cancer of the stomach (Bolen et al., Oncogene Res., 1987,1,149, Kapitanovic et al., Gastroenterology.2000,112,1103, Ross et al., Cancer Invest., 2001,19,554), prostate cancer (Visakorpi et al., Histochem.J., 1992,24,481, Kumar et al., 2000,32,73, Scher et al., J.Natl.Cancer Inst., 2000,92,1866), leukemia (Konaka et al., Cell, 1984,37,1035, Martin-Suberoet al., Cancer Genet Cytoggenet., 2001,127,174), ovarian cancer (Hellstrom et al., Cancer Res., 2001,61,2420), head and neck cancer (Shiga et al., Head Neck, 2000,22,599) or carcinoma of the pancreas (Ovotny et al., Neoplasm, 2001,48,188).Along with more mankind tumor tissue being tested to the expression of the erbB family of its receptor tyrosine kinase, expection future their ubiquity and importance will further strengthen.

The result regulating as one or more above-mentioned acceptors (particularly erbB2) mistake, generally believes many tumours richer aggressiveness clinically, and therefore patient's poorer (the Brabender etal. of prognosis, Clin.Cancer Res., 2001,7,1850, Ross et al., Cancer Investigation, 2001,19,554, Yu et al., Bioessays, 2000,22.7,673),, except above clinical discovery, a large amount of clinical front information illustrates that receptor tyrosine kinase erbB family participates in cell transformation. comprise following observed result: one or more erbB acceptors of many tumor cell line overexpressions, EGFR or erbB2 are being transfected into after non-tumor cell, they can transform these cells. and this carciongenic potency is further confirmed: the transgenic mice of overexpression erbB2 becomes tumour in mammary gland organic growth. in addition, the verified micromolecular inhibitor that passes through of a large amount of preclinical studies, dominant negative or inhibiting antibody knock out one or more erbB activity, (summary is referring to Mendelsohn et al. can to produce antiproliferative effect, Oncogene, 2000, 19, 6550). therefore, the inhibitor that can recognize these receptor tyrosine enzymes will have important value (Yaish et al.Science as the selective depressant of mammalian cancer cells propagation, 1988, 242, 933, Kolibaba et al., Biochimicaet Biophysica Acta, 1997, 133, F217-F248, Al-Obeidi et al, 2000, Oncogenen, 19,5690-5701, Mendelsohn et al., 2000, Oncogene, 19,6550-6565). except these preclinical datas, (summary is referring to Mendelsohn et al. also to have confirmed to use the inhibition antibody (being respectively c-225 and trastuzumab) of EGFR and erbB2 to be of value to the clinical treatment of selected solid tumor, 2000, Oncogene, 19,6550-6565).

After testing amplification and/or the activity of erbB receptor Tyrosylprotein kinase, and therefore infer this amplification and/or active in a large amount of non-malignant proliferative disease, there is certain effect, for example following disease: psoriasis (Ben-Bassat, Curr.Pharm.Des., 2000,6,933; Elder et al., Science, 1989,243,811), benign prostate propagation (BPH) (Kumar et al., Int.Urol.Nephrol., 2000,32,73), pulse atherosclerosis and restenosis (Bokemeyer et al., Kidney Int, 2000,58,549).Therefore, expection erbB receptor tyrosine is gargled enzyme inhibitors and will be can be used for treating above disease and excessive other nonmalignant disease of cell proliferation.

European patent application EP 566226 discloses some 4-phenylamino quinazoline, and it is receptor tyrosine kinase inhibitors.

International Patent Application WO 96/33977, WO 96/33978, WO 96/33979, WO96/33980, WO 96/33981, WO 97/30034 and WO 97/38994 disclose some and have contained phenylamino substituting group and the quinazoline derivant at 6-and/or 7-substd in 4-position, and they have receptor tyrosine kinase and suppress active.

European patent application EP 837063 discloses the 4-amido quinazoline derivatives that aryl replaces, and its 6-at quinazoline ring or 7-position have the part that comprises aryl or heteroaryl.It is said that these compounds are applicable to overmedication proliferative disorders.

International Patent Application WO 97/30035 and WO 98/13354 disclose the 4-phenylamino quinazoline that some 7-replaces, and it is vascular endothelial growth factor receptor receptor tyrosine kinase inhibitors.

WO 00/55141 discloses the 4-phenylamino quinazoline compound that 6,7-replaces, and it is characterized in that the substituting group of 6-and/or 7-position has ester connection portion (RO-CO).

WO 00/56720 is openly used for the treatment of 6 of cancer or atopic reaction, 7-dialkoxy-4-phenylamino quinazoline compound.

WO 02/41882 discloses the 4-phenylamino quinazoline compound that 6-and/or 7-position are replaced by pyrrolidyl-alkoxyl group or piperidyl-alkoxyl group.

WO 03/082290 disclose some 6, the 4-phenylamino quinazoline compound that 7-replaces has receptor tyrosine kinase and suppresses active.So chloro-4-fluorophenyl of the concrete example 4-[(3-of this compound) amino]-6-[1-(uncle-butoxy carbonyl)-piperidin-4-yl-oxygen base]-7-methoxyl group-quinazoline.

Summary of the invention

Applicant is now surprisingly found out that, modify side chain and optionally increase other substituting group to phenylamino and produced the selected compounds group with enhanced activity, wherein compound has good erbB2 kinase inhibitory activity and EGF restraining effect, makes them be particularly suitable for being clinically applied to treatment and relating to this two kinds of kinase whose tumours.

Only be not intended to imply compound disclosed by the invention because a kind of biological procedures effect is had to pharmaceutical active, but believe that the compounds of this invention provides antitumor action by suppressing one or more receptor tyrosine kinases erbB family, described receptor tyrosine kinase erbB family participates in causing the signal transduction step of tumor cell proliferation.Especially, think that the compounds of this invention provides antitumor action by suppressing EGFR and/or erbB2 receptor tyrosine kinase.

[summary of the invention]

1. the quinoline of formula I

Wherein n is 0,1,2 or 3,

Each R ⁵independently be selected from halogen, cyano group, nitro, hydroxyl, amino, carboxyl, sulfamyl, trifluoromethyl, (1-6C) alkyl, (2-8C) alkenyl, (2-8C) alkynyl group, (1-6C) alkoxyl group, (2-6C) alkenyl oxy, (2-6C) alkynyl group oxygen base, (1-6C) alkylthio, (1-6C) alkyl sulphinyl, (1-6C) alkyl sulphonyl, (1-6C) alkylamino, two-[(1-6C) alkyl] amino, (1-6C) alkoxy carbonyl, N-(1-6C) alkylsulfamoyl group and N, N-bis--[(1-6C) alkyl] sulfamyl, C (O) NR ⁶r ⁷, wherein R ⁶and R ⁷independently be selected from hydrogen, optional (1-6C) alkyl replacing, optional (3-8C) cycloalkyl replacing or the optional aryl replacing, or R ⁶and R ⁷together with the nitrogen connecting with them, form can comprise other assorted former in optional substituted heterocycle,

X ¹directly to be good for or O;

R ¹be selected from nitrogen and (1-6C) alkyl, wherein (1-6C) alkyl is optionally replaced by one or more substituting groups, and it can be identical or different, is selected from hydroxyl and halogen, and/or be selected from amino, nitro, carboxyl, cyano group, halogen, (1-6C) alkoxyl group, hydroxyl (1-6C) alkoxyl group, (2-8C) alkenyl, (2-8C) alkynyl group, (1-6C) alkylthio, (1-6C) alkyl sulfenyl tomb, (1-6C) alkyl sulphonyl, (1-6C) alkylamino, two-[(1-6C) alkyl] amino, formamyl, N-(1-6C) alkyl-carbamoyl, N, N-bis--[(1-6C) alkyl] formamyl, (2-6C) alkyloyl, (2-6C) alkyloyl oxygen base, (2-6C) alkanoylamino, N-(1-6C) alkyl-(2-6C) alkanoylamino, (1-6C) alkoxy carbonyl, sulfamyl, N-(1-6C) alkylsulfamoyl group, N, N-bis--[(1-6C) alkyl] sulfamyl, (1-6C) alkane sulfuryl amino and N-(1-6C) alkyl-(1-6C) substituting group of alkane sulfuryl amino,

R ²be (1-6C) alkyl, (2-6C) alkenyl or (2-6C) alkynyl group, any above-mentioned group can be optionally by following replacement: fluorine, (1-6C) alkoxyl group, (1-6C) alkylthio, (1-6C) alkyl sulphinyl, (1-6C) alkyl sulphonyl or sub-formula (i):

Wherein m is 0,1,2 or 3;

R ³and R ⁴independently be selected from hydrogen or (1-6C) alkyl,

Or R ³and R ⁴together with the nitrogen-atoms connecting with them, form 5 and 6 yuan of saturated heterocycles, it optionally comprises other and is selected from oxygen, S, SO or S (O) ₂or NR ⁸heteroatoms, wherein R ⁸hydrogen, (1-6C) alkyl, (2-6C) alkenyl, (2-6C) alkynyl group, (1-6C) alkyl sulphonyl or (1-6C) alkyl-carbonyl.

2. according to the quinoline of project 1, wherein

Wherein m is 1,2 or 3;

R ³and R ⁴independently be selected from hydrogen or (1-6C) alkyl,

3. according to the quinoline of project 1 or 2, n is 1,2 or 3.

4. according to the quinoline of project 3, wherein n is 2 or 3.

5. according to the quinoline of project 4, wherein n is 2.

6. according to the quinoline of project 4, wherein n is 3.

7. according to the quinoline of aforementioned project any one, wherein each R ⁵group is halogen group.

8. according to the quinoline of aforementioned project any one, wherein each R ⁵group is selected from chlorine and fluorine.

9. according to the quinoline of aforementioned project any one, it comprises R ⁵group, this R ⁵group is positioned at its phenyl ring connecting o-(2-) position place.

10. according to the quinoline of project 9, be wherein positioned at the R at o-(2-) position place ⁵group is fluorine.

11. according to the quinoline of aforementioned project any one, sub-formula (ii) group in its Chinese style I:

Sub-formula (iii) group:

Wherein, (a) R ¹⁰or R ¹²that hydrogen and another are halogens, and R ¹¹halogen, or (b) R ¹⁰halogen, R ¹¹halogen, and R ¹²hydrogen or halogen, or (c) R ¹⁰fluorine, R ¹¹chlorine, and R ¹²be selected from hydrogen or fluorine.

12. according to the quinoline of project 11, wherein R ¹⁰or R ¹²one of be that nitrogen and another are fluorine, and R ¹¹chlorine.

13. according to the quinoline of project 11, wherein R ¹⁰fluorine, R ¹¹chlorine, and R ¹²hydrogen.

14. according to the quinoline of project 11, wherein R ¹⁰fluorine, R ¹¹chlorine, and R ¹²it is fluorine.

15. according to the quinoline of aforementioned project any one, wherein X ¹oxygen.

16. according to the quinoline of aforementioned project any one, wherein R ¹be selected from hydrogen, (1-6C) alkyl and (1-6C) alkoxyl group (1-6C) alkyl, wherein R ¹in any (1-6C) alkyl optionally contain one or more hydroxyls or halogenic substituent.

17. according to the quinoline of project 16, wherein R ¹be selected from (1-6C) alkyl, it optionally contains one or more hydroxyls or halogenic substituent.

18. according to the quinoline of aforementioned project any one, wherein R ¹-X ¹-be selected from hydrogen, methoxyl group, oxyethyl group and 2-methoxy ethoxy.

19. according to the quinoline of project 18, wherein R ¹-X ¹-be methoxyl group.

20. according to the formula IA quinoline of project 1:

Wherein R ²as project 1 defines, R ¹⁰, R ¹¹and R ¹²as the definition of project 11-14 any one, and R ¹³be selected from hydrogen, methoxyl group, oxyethyl group and 2-methoxy ethoxy.

21. according to the formula IB quinoline of project 1:

Wherein R ²as project 1 defines, and R ¹³be selected from hydrogen, methoxyl group, oxyethyl group and 2-methoxy ethoxy.

22. according to the formula IC quinoline of project 1:

23. according to the quinoline of project 20-22 any one, wherein R ¹³it is methoxyl group.

24. according to the quinoline of aforementioned project any one, wherein R ²be (1-6C) alkyl, it is optionally by following replacement: fluorine, (1-6C) alkoxyl group, (1-6C) alkylthio, (1-6C) alkyl sulphinyl, (1-6C) alkyl sulphonyl or sub-formula (i) group defining as project 1 or project 2.

25. according to the quinoline of aforementioned project any one, wherein R ²be (1-3C) alkyl, it is optionally by following replacement: fluorine, (1-6C) alkoxyl group, (1-6C) alkylthio, (1-6C) alkyl sulphinyl, (1-6C) alkyl sulphonyl or sub-formula (i) group defining as project 1 or project 2.

26. according to the quinoline of aforementioned project any one, wherein R ²be (1-6C) alkyl, its sub-formula (i) group optionally being defined as project 1 or project 2 replaces.

27. according to the quinoline of aforementioned project any one, wherein R ²be (1-3C) alkyl, its sub-formula (i) group optionally being defined as project 1 or project 2 replaces.

28. according to the quinoline of aforementioned project any one, wherein R ²it is methyl.

29. according to the quinoline of aforementioned project any one, wherein R ²comprise sub-formula (i) substituting group defining as project 1 or project 2.

30. according to the quinoline of project 29, and wherein m is 1 or 2.

31. according to the quinoline of project 30, and wherein m is 2.

32. according to the quinoline of project 30, and wherein m is 1.

33. according to the quinoline of project 29-32, wherein R ³and R ⁴together with the nitrogen-atoms connecting with them, form pyrrolidine ring, morpholine ring, piperidine ring or piperazine ring, it is optionally replaced by (1-3C) alkyl at feasible nitrogen-atoms place.

34. according to the quinoline of project 29-32, wherein R ³and R ⁴together with the nitrogen-atoms connecting with them, form pyrrolidine ring, morpholine ring, piperidine ring or piperazine ring, it is optional those methyl substituted at feasible nitrogen-atoms place.

35. according to the quinoline of project 33 or project 34, wherein R ³and R ⁴together with the nitrogen-atoms connecting with them, form pyrrolidine ring.

36. according to the quinoline of project 29-32, wherein R ³and R ⁴independently be selected from (1-3C) alkyl.

37. according to the quinoline of aforementioned project any one, wherein R ²be selected from methyl, 2-(pyrrolidin-1-yl) ethyl, 2-(dimethylamino) ethyl, 2-(diethylamino) ethyl, 2-(piperidyl) ethyl, 2-(morpholine-4-yl) ethyl and 2-(4-methylpiperazine-1-yl) ethyl.

38. according to the quinoline of project 37, wherein R ²it is 2-(pyrrolidin-1-yl) ethyl.

39. according to the quinoline of project 1, and it is selected from following one or more:

4-(the chloro-2-fluoroanilino of 3-)-7-methoxyl group-6-{[1-(methoxycarbonyl) piperidin-4-yl] oxygen base }-3-nitrile yl-quinoline;

4-(the chloro-2-fluoroanilino of 3-)-7-methoxyl group-6-{[1-{2-(pyrrolidin-1-yl) ethoxy carbonyl) piperidin-4-yl] oxygen base }-3-nitrile yl-quinoline;

4-(the chloro-2-fluoroanilino of 3-)-7-methoxyl group-6-{[1-2-(N, N-dimethylamino) ethoxy carbonyl) piperidin-4-yl] oxygen base }-3-nitrile yl-quinoline;

4-(the chloro-4-fluoroanilino of 3-)-7-methoxyl group-6-{[1-{2-(pyrrolidin-1-yl) ethoxy carbonyl } piperidin-4-yl] oxygen base }-3-nitrile yl-quinoline;

4-(3-chlorobenzene amino)-7-methoxyl group-6-{[1-{2-(pyrrolidin-1-yl) ethoxy carbonyl } piperidin-4-yl] oxygen base }-3-nitrile yl-quinoline;

4-(the chloro-2,4 difluorobenzene amino of 3-)-7-methoxyl group-6-{[1-(methoxycarbonyl) piperidin-4-yl] oxygen base }-3-nitrile yl-quinoline;

4-(the chloro-2,4 difluorobenzene amino of 3-)-7-methoxyl group-6-{[1-{2-(pyrrolidin-1-yl) ethoxy carbonyl } piperidin-4-yl] oxygen base }-3-nitrile yl-quinoline;

4-(the chloro-2,4 difluorobenzene amino of 3-)-7-methoxyl group-6-{[1-{2-(piperidin-1-yl) ethoxy carbonyl } piperidin-4-yl] oxygen base }-3-nitrile yl-quinoline;

4-(the chloro-2-fluoroanilino of 3-)-7-methoxyl group-6-{[1-{2-(piperidin-1-yl) ethoxy carbonyl } piperidin-4-yl] oxygen base }-3-nitrile yl-quinoline;

4-(the chloro-2-fluoroanilino of 3-)-6-{[1-{2-(diethylamino) ethoxy carbonyl } piperidin-4-yl] oxygen base }-7-methoxyl group-3-nitrile yl-quinoline;

4-(the chloro-2-fluoroanilino of 3-)-7-methoxyl group-6-{[1-{2-(morpholine-4-yl) ethoxy carbonyl } piperidin-4-yl] oxygen base }-3-nitrile yl-quinoline; And

4-(the chloro-2-fluoroanilino of 3-)-7-methoxyl group-6-{[1-{2-(4-methyl piperidine-1-yl) ethoxy carbonyl } piperidin-4-yl] oxygen base }-3-nitrile yl-quinoline;

Or its pharmacy acceptable salt.

40. according to the preparation method of the quinoline of aforementioned project any one, and it comprises, or

Method (a) formula II compound:

When wherein R1, X2, R5 and n have any implication defining as project 1 except needs, protect any functional group,

React with formula III compound:

Wherein R ²while having any implication as previously defined except needs with m, protect any functional group, and Lg is replaceable group,

Method (b) is modified the substituting group of another kind of formula I quinoline or its pharmaceutically-acceptable salts or is introduced substituting group, and compound is protected any functional group during as previously defined except needs,

Method (c) formula IV compound:

Wherein R ¹, X ¹, R ⁵with the related definition of n suc as formula I, react with formula V compound:

Wherein R ²as above definition and Lg are replaceable group (for example halogen are as chlorine or bromine);

The protecting group that method (d) is removed formula I quinoline or its pharmaceutically-acceptable salts;

Method (e) formula II compound defined above reacts under Mitsunobu condition with the formula III compound that is OH except Lg defined above;

Method (f) is in order to prepare wherein R ¹-X ¹be the formula I compound of hydroxyl, cracking wherein

R ¹-X ¹it is the formula I quinoline of (1-6C) alkoxyl group;

Method (g) is in order to prepare wherein X ¹be those compounds of O, make formula VI compound:

Wherein R ², R ⁵while having with n any implication defining as project 1 except needs, protect any functional group, with formula R ¹the reaction of-Lg compound, wherein R ¹while having any implication as previously defined except needs, protecting any functional group and Lg is replaceable group;

Method (h) is in order to prepare wherein R ¹the formula I compound of (1-6C) alkylamino of (1-6C) alkoxyl group that comprises (1-6C) alkoxyl group or replacement or (1-6C) alkylamino or replacement, wherein R of alkylation suitably time ¹the formula I quinoline that comprises hydroxyl or primary amine or secondary amine group;

Method (i) is in order to prepare wherein R ¹the formula I compound being replaced by group T; wherein T is selected from (1-6C) alkylamino, two-[(1-6C) alkyl] amino, (2-6C) alkanoylamino, (1-6C) alkylthio, (1-6C) alkyl sulphinyl and (1-6C) alkyl sulphonyl, makes formula VII compound:

Wherein R ², R ⁵, X ¹, n and m protect any functional group, R while having the implication defining as project 1 except needs ¹' be the R defining as herein ¹group is replaced by Lg except any T group, and Lg is replaceable group (for example chlorine or bromine), reacts with formula TH compound, protects any functional group when wherein T as above defines except needs;

Method (j) formula VIII compound:

Wherein R ¹, R ², X ¹while having with m any implication defining as project 1 except needs, protecting any functional group and Lg is replaceable group as previously mentioned,

Aniline reaction with formula IX:

Wherein R ⁵while having any implication as previously defined except needs with n, protect any functional group; Relevant to the method (j) above-mentioned those;

Method (k) makes formula X compound:

Wherein R ⁵, X ¹, R ¹while defining except needs with n as project 1, protect any functional group, and wherein Lg is leavings group, with formula R ²the reaction of-OH alcohol, wherein R ²as project 1 defines;

Method (l) is for R wherein ²comprise the compound of sub-formula (i) group, make formula XI compound

Wherein R ¹, X ¹, R ⁵with n as previously defined, R ¹⁵be (1-6C) alkyl, and Lg is leavings group,

With formula R ³r ⁴the reaction of NH compound, wherein R ³and R ⁴as the related definition of front sub-formula (i);

And after any described method, remove any blocking group of existence.

41. 1 kinds of pharmaceutical compositions, it contains just like formula I quinoline or its pharmacy acceptable salt of project 1-39 any one definition and combines by pharmaceutically acceptable diluent or carrier.

42. if the formula I quinoline of project 1-39 any one definition or its pharmacy acceptable salt are as the application of medicine.

43. if the formula I quinoline of project 1-39 any one definition or its pharmacy acceptable salt are in the purposes for the preparation of produce the medicine of antiproliferative effect in warm-blooded animal body.

44. 1 kinds produce the method for antiproliferative effect in the warm-blooded animal body of this treatment of needs, comprise and give described animal as formula I quinoline or its pharmacy acceptable salt of the definition of project 1-39 any one.

45. formula VI, VII, X or the XI compounds that define as project 40.

[detailed Description Of The Invention]

First aspect present invention provides following formula I quinoline:

Wherein n is 0,1,2 or 3,

Each R ⁵independently be selected from halogen, cyano group, nitro, hydroxyl, amino, carboxyl, sulfamyl, trifluoromethyl, (1-6C) alkyl, (2-8C) alkenyl, (2-8C) alkynyl group, (1-6C) alkoxyl group, (2-6C) alkenyl oxy, (2-6C) alkynyl group oxygen base, (1-6C) alkylthio, (1-6C) alkyl sulphinyl, (1-6C) alkyl sulphonyl, (1-6C) alkylamino, two-[(1-6C) alkyl] amino, (1-6C) alkoxy carbonyl, N-(1-6C) alkylsulfamoyl group and N, N-bis--[(1-6C) alkyl] sulfamyl, C (O) NR ⁶r ⁷, wherein R ⁶and R ⁷independently be selected from hydrogen, optional (1-6C) alkyl replacing, optional (3-8C) cycloalkyl replacing or the optional aryl replacing, or R ⁶and R ⁷together with the nitrogen connecting with them, form and can comprise other heteroatomic optional substituted heterocycle,

X ¹direct key or O;

R ¹be selected from hydrogen and (1-6C) alkyl, wherein (1-6C) alkyl is optionally replaced by one or more substituting groups, and it can be identical or different, is selected from hydroxyl and halogen, and/or be selected from amino, nitro, carboxyl, cyanogen tomb, halogen, (1-6C) alkoxyl group, hydroxyl (1-6C) alkoxyl group, (2-8C) alkenyl, (2-8C) alkynyl group, (1-6C) alkylthio, (1-6C) alkyl sulphinyl, (1-6C) alkyl sulphonyl, (1-6C) alkylamino, two-[(1-6C) alkyl] amino, formamyl, N-(1-6C) alkyl-carbamoyl, N, N-bis--[(1-6C) alkyl] formamyl, (2-6C) alkyloyl, (2-6C) alkyloyl oxygen base, (2-6C) alkanoylamino, N-(1-6C) alkyl-(2-6C) alkanoylamino, (1-6C) alkoxy carbonyl, sulfamyl, N-(1-6C) alkylsulfamoyl group, N, N-bis--[(1-6C) alkyl] sulfamyl, (1-6C) alkane sulfuryl amino and N-(1-6C) alkyl-(1-6C) substituting group of alkane sulfuryl amino,

Wherein m is 0,1,2 or 3;

R ³and R ⁴independently be selected from hydrogen or (1-6C) alkyl,

Or R ³and R ⁴together with the nitrogen-atoms connecting with them, form 5 and 6 yuan of saturated heterocycles, it optionally comprises other and is selected from oxygen, S, SO or S (O) ₂or NR ⁸assorted former in, wherein R ⁸hydrogen, (1-6C) alkyl, (2-6C) alkenyl, (2-6C) alkynyl group, (1-6C) alkyl sulphonyl or (1-6C) alkyl-carbonyl;

The present invention provides following formula I quinoline on the other hand:

Wherein n is 0,1,2 or 3,

X ¹direct key or O;

R ¹be selected from hydrogen and (1-6C) alkyl, wherein (1-6C) alkyl is optionally replaced by one or more substituting groups, and it can be identical or different, is selected from hydroxyl and halogen, and/or be selected from amino, nitro, hydroxyl, cyano group, halogen, (1-6C) alkoxyl group, hydroxyl (1-6C) alkoxyl group, (2-8C) alkenyl, (2-8C) alkynyl group, (1-6C) alkylthio, (1-6C) alkyl sulphinyl, (1-6C) alkyl sulphonyl, (1-6C) alkylamino, two-[(1-6C) alkyl] amino, formamyl, N-(1-6C) alkyl-carbamoyl, N, N bis--[(1-6C) alkyl] formamyl, (2-6C) alkyloyl, (2-6C) alkyloyl oxygen base, (2-6C) alkanoylamino, N-(1-6C) alkyl-(2-6C) alkanoylamino, (1-6C) alkoxy carbonyl, sulfamyl, N-(1-6C) alkylsulfamoyl group, N, N-bis--[(1-6C) alkyl] sulfamyl, (1-6C) alkane sulfuryl amino and N-(1-6C) alkyl-(1-6C) substituting group of alkane sulfuryl amino,

Wherein m is 1,2 or 3;

R ³and R ⁴independently be selected from hydrogen or (1-6C) alkyl,

Or R ³and R ⁴together with the nitrogen-atoms connecting with them, form 5 and 6 yuan of saturated heterocycles, it optionally comprises other and is selected from oxygen, S, SO or S (O) ₂or NR ⁸heteroatoms, wherein R ⁸hydrogen, (1-6C) alkyl, (2-6C) alkenyl, (2-6C) alkynyl group, (1-6C) alkyl sulphonyl or (1-6C) alkyl-carbonyl;

In this manual, general term " alkyl " comprises straight chain and branched-chain alkyl for example propyl group, sec.-propyl and the tertiary butyl and (3-7C) cycloalkyl (for example cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and suberyl).But while mentioning the concrete alkyl such as " propyl group ", only refer to straight chain type group, while mentioning the concrete branched-chain alkyl such as " sec.-propyl ", only refer to branched chain type group, mention concrete cycloalkyl time, for example " cyclopentyl " only refers to 5-ring, similarly agreement is applied to other general term, for example (1-6C) alkoxyl group comprises methoxyl group, oxyethyl group, ring propoxy-and cyclopentyloxy, (1-6C) alkylamino comprises methylamino, ethylamino, cyclobutyl amino and cyclohexyl amino, two [(1-6C) alkyl)] amino comprises dimethylamino, diethylamino, N-cyclobutyl-N-methylamino and N-cyclohexyl-N-ethylamino.

Term " aryl " refers to that aromatic hydrocarbon is such as phenyl and naphthyl.Term " heterocycle " and " heterocycle " comprise it can being single-and dicyclo and ring structure of comprising 3-15 atom, so wherein at least one atom and the suitably individual former oxygen of 1-4, sulphur and nitrogen heteroatom.Ring can be fragrant, non-aromatic and part fragrance, a ring that condenses ring system can be fragrance and another is non-aromatic.The object lesson of ring system comprises furyl like this, benzofuryl, tetrahydrofuran base, chromanyl, thienyl, benzothienyl, pyridyl, piperidyl, quinolyl, 1,2,3,4-tetrahydric quinoline group, isoquinolyl, 1,2,3,4-tetrahydro isoquinolyl, pyrazinyl, piperazine, pyrimidyl, pyridazinyl, quinoxalinyl, quinazolyl, cinnolines base, pyrryl, pyrrolidyl, indyl, indolinyl, imidazolyl, benzimidazolyl-, pyrazolyl, indazolyl, oxazolyl, benzoxazolyl, isoxazolyl, thiazolyl, benzothiazolyl, isothiazolyl, morpholinyl, 4H-1,4-benzoxazinyl, 4H-1,4-benzothiazine base, 1,2,3-triazoles base, 1,2,4-triazolyl, oxadiazolyl, furazanyl, thiadiazolyl group, tetrazyl, dibenzofuran group, dibenzothiophene basic ring oxirane group, oxetanyl, azetidinyl, THP trtrahydropyranyl, oxa-cyclic group in heptan, oxazepanyl, four nitrogen-Isosorbide-5-Nitrae-thiazinyl, 1,1-dioxo tetrahydrochysene Isosorbide-5-Nitrae-thiazinyl, homopiperidinyl, homopiperazine base, dihydropyridine base, tetrahydro pyridyl, dihydro-pyrimidin base, tetrahydro-pyrimidine base, tetrahydro-thienyl, tetrahydrochysene sulfo-pyranyl and thio-morpholinyl.

The object lesson of heterocyclic radical comprises THP trtrahydropyranyl, tetrahydrofuran base or N-(1-6C) alkyl pyrrolidine or N-alkyl (1-6C) piperidines.

In the time that ring comprises nitrogen-atoms, if can to have hydrogen atom or substituting group be (1-6C) alkyl when needing to meet the valence link needs of nitrogen or they can be connected with other structure by nitrogen-atoms for it.Nitrogen in heterocyclic group is former in being oxidized to N-oxide compound.

Conventionally, this compound shows that desirable physical properties is such as high resolution keeps high antiproliferative activity simultaneously. in addition, chemical compound lot of the present invention does not have activity or only has weak activity in hERG measures.

Should be understood that opticity or the racemic form that within the scope of some formula I compound defined above, may have one or more unsymmetrical carbons, present invention resides in any such opticity or the racemic form with above-mentioned activity in its definition.Can lead the well-known standard technique in city to synthesize opticity form by organic chemistry, for example, utilize opticity initial feed or prepare by resolution of racemic form.Similarly, above-mentioned activity can adopt standard laboratory technological assessment hereinafter.

It is to be further understood that the tautomeric forms that present invention resides in all formula I compounds with antiproliferative activity in its definition.

It is to be further understood that some formula I compound may exist solvate forms and non-solvent compound form, for example hydrate forms, the present invention includes all such solvate forms with antiproliferative activity.

It is to be further understood that some formula I compound may exist multiple crystal formation, the present invention includes all such forms with antiproliferative activity.

The desired value that more than relates to general group comprises those following values.

Before this or after this defined arbitrary R in this specification sheets ¹, R ², R ³or R ⁵the appropriate value of group comprises:

For halogen: fluorine, chlorine, bromine and iodine;

For (1-6C) alkyl: methyl, ethyl, propyl group, sec.-propyl, uncle

Butyl, amyl group and hexyl;

For (1-4C) alkyl: methyl, ethyl, propyl group, sec.-propyl and uncle

Butyl;

For (1-6C) alkoxyl group: methoxyl group, oxyethyl group, propoxy-, isopropyl

Oxygen base and butoxy;

For (2-8C) alkenyl: vinyl, pseudoallyl, allyl group and fourth

Thiazolinyl;

For (2-8C) alkynyl group: ethynyl, 2-propynyl and fourth-2-alkynyl;

For (2-6C) alkenyl oxygen: vinyloxy group and allyloxy;

For (2-6C) alkynyl group oxygen: second alkynyloxy group and 2-propargyl alcoholate;

For (1-6C) alkylthio: methylthio group, ethylmercapto group and rosickyite base;

For (1-6C) alkyl sulphinyl: methylsulfinyl and ethyl sulfinyl;

For (1-6C) alkyl sulphonyl: methyl sulphonyl and ethylsulfonyl;

For (1-6C) alkylamino: methylamino-, ethylamino, the third amino, isopropyl

Amino and fourth amino;

For two-[(1-6C) alkyl] amino: dimethylamino, diethylin, N-ethyl-N-

Methylamino and diisopropylaminoethyl;

For (1-6C) alkoxy carbonyl: methoxycarbonyl, ethoxy carbonyl, the third oxygen

Base carbonyl and uncle-butoxy carbonyl;

For N-(1-6C) alkyl carbamoyl N-methylamino formyl radical, N-ethylamino

Base: formyl radical, N-propyl group formamyl and N-

Isopropylamino formyl radical;

For N, N-bis--[(1-6C) alkyl] N, N-formyl-dimethylamino, N-ethyl

Formamyl :-N-methylamino formyl radical and N, N-bis-

Base formamyl;

For (2-6C) alkyloyl: ethanoyl, propionyl and isobutyryl;

For (2-6C) alkyloyl oxygen: acetoxyl group and propionyloxy;

For (2-6C) alkanoylamino: kharophen and propionamido;

For N-(1-6C) alkyl-(2-6C) alkane N-methyl kharophen and N-methyl propionyl ammonia

Acyl amino: base;

For N-(1-6C) alkyl sulphonamide N-methyl sulfamyl, N-ethyl sulphonamide

Base: base and N-sec.-propyl sulfamyl;

For N, N-bis--[(1-6C) alkyl] ammonia N, N-dimethylamino alkylsulfonyl and N-methyl

Alkylsulfonyl :-N-ethyl sulfamyl;

For (1-6C) alkane sulfuryl amino: methylsulfonyl amino and ethylsulfonylamino;

For N-(1-6C) alkyl-(1-6C) alkane N-methyl methylsulfonyl amino and N-methyl second

Sulfuryl amino: sulfuryl amino;

For hydroxyl-(1-6C) alkoxyl group: hydroxyl methoxyl group, 2-hydroxyl-oxethyl, 1-

Hydroxyl-oxethyl and 3-hydroxyl propoxy-.

Be interpreted as, work as R ¹by for example amino when replacing (1-6C) alkyl and being for example 2-amino-ethyl, be (1-6C) alkyl and radicals X ¹be connected and (or work as X ¹while being direct key, be connected with quinoline ring).

While using (1-4C) alkyl in this manual, be construed as such group and refer to the alkyl that comprises maximum 4 carbon atoms.The representative example that those of skill in the art can understand such group is the example that comprises maximum 4 carbon atoms in above-mentioned (1-4C) alkyl, for example methyl, ethyl, propyl group, sec.-propyl, butyl and the tertiary butyl.Similarly; (1-3C) alkyl refers to that the alkyl that comprises maximum 3 carbon atoms is such as methyl, ethyl, propyl group and sec.-propyl. similarly agreement is applicable to above-mentioned other group, for example (1-4C) alkoxyl group, (2-4C) alkenyl, (2-4C) alkynyl group and (2-4C) alkyloyl.

In formula I compound, hydrogen atom is present in 2,5 and 8 of quinoline ring.

The suitable pharmacy acceptable salt of formula I compound is for example acid salt of formula I compound, for example, with the inorganic or organic acid sour addition basin such as hydrochloric acid, Hydrogen bromide, sulfuric acid, trifluoroacetic acid, citric acid or toxilic acid; Or for example have the salt of the formula I compound of enough acidity, for example basic metal or alkaline earth salt such as calcium salt or magnesium salts, ammonium salt or with the salt of organic bases such as methylamine, dimethylamine, Trimethylamine 99, piperidines, morpholine or three-(2-hydroxyethyl) amine.

The object lesson of n is 1,2 or 3, suitably 2 or 3.

Suitably, each R ⁵independently be selected from halogen, trifluoromethyl, (1-C) alkyl, (2-8C) alkenyl, (2-8C) alkynyl group or C (O) NR ⁶r ⁷base, wherein R ⁶and R ⁷as above definition.

Especially, each R ⁵independently be selected from halogen such as chlorine or fluorine.

Work as R ⁶and R ⁷while being not hydrogen, its concrete substituting group comprises halogen, nitro, cyano group, hydroxyl, amino, carboxyl, formamyl, sulfamyl, trifluoromethyl, (2-8C) alkenyl, (2-8C) alkynyl group, (1-6C) alkoxyl group, (2-6C) alkenyl oxy, (2-6C) alkynyl group oxygen base, (1-6C) alkylthio, (1-6C) alkyl sulphinyl, (1-6C) alkyl sulphonyl, (1-6C) alkylamino, two-[(1-6C) alkyl] amino, (1-6C) alkoxy carbonyl, N-(1-6C) alkyl-carbamoyl, N, N-bis--[(1-6C) alkyl] formamyl, N-(1-6C) alkylsulfamoyl group, N, N-bis--[(1-6C) alkyl] sulfamyl, (3-8C) cycloalkyl, aryl and heterocyclic radical.

R ⁶and R ⁷the concrete example of aryl substituent in comprising phenyl or naphthyl, particularly phenyl.

R ⁶and R ⁷the object lesson of heterocyclic substituent comprise that 5 or 6 yuan of heterocycles are such as furyl, tetrahydrofuran base, thienyl, pyridyl, piperidyl, pyrazinyl, piperazinyl, pyrimidyl, pyridazinyl, pyrryl, pyrrolidyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, morpholinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, oxadiazolyl, furazanyl, thiadiazolyl group or tetrazyl.

Work as R ⁶and R ⁷while forming the heterocycle of optional replacement together with the nitrogen-atoms connecting with them, it is for example 5 or 6 rings, and it is saturated or unsaturated. concrete example comprises piperidyl, pyrrolidyl, morpholinyl or thio-morpholinyl.Alternative, R ⁶and R ⁷form together (3-6C) alkenyl.

R ⁶and R ⁷the nitrogen that connects with them former in together with the heterocycle that forms can be by above-mentioned and R ⁶and R ⁷relevant arbitrary group replaces.In addition, these rings can be replaced by one or more (1-6C) alkyl, himself can optionally be replaced by one or more tomb groups that are selected from below: halogen, nitro, cyano group, hydroxyl, amino, carboxyl, formamyl, sulfamyl, trifluoromethyl, (2-8C) alkenyl, (2-8C) alkynyl group, (1-6C) alkoxyl group, (2-6C) alkenyl oxy, (2-6C) alkynyl group oxygen base, (1-6C) alkylthio, (1-6C) alkyl sulphinyl, (1-6C) alkyl sulphonyl, (1-6C) alkylamino, two-[(1-6C) alkyl] amino, (1-6C) alkoxy carbonyl, N-(1-6C) alkyl-carbamoyl, N, N-bis--[(1-6C) alkyl] formamyl, N-(1-6C) alkylsulfamoyl group or N, N-bis--[(1-6C) alkyl] sulfamyl.

Work as R ⁶or R ⁷while being not hydrogen, its exemplary substituting group is cyano group, hydroxyl, (2-8C) alkenyl, (2-8C) alkynyl group, (1-6C) alkoxyl group, (1-6C) alkylthio, (1-6C) alkylamino, aryl such as phenyl or heterocyclic radical are such as furyl, in addition, work as R ⁶and R ⁷the nitrogen that connects with them former in together with while forming ring, (1-6C) alkyl is such as methyl.

In the time that n is 1,2 or 3, a R ⁵group is suitably in phenyl ring o-(2-) position.

In the time that n is 1,2 or 3, a R ⁵group is suitably in phenyl ring m-(3-) position.

Therefore, in the time that n is 1, R ⁵group is suitably at phenyl ring o-(2-) position or m-(3-).

In the present invention on the other hand, in the time that n is 2, first R ⁵group is suitably at the m-position of phenyl ring, second R ⁵group is suitably in the o-position of phenyl ring or p-position, and this ring has substituting group at the 2-of phenyl ring and 3-or 3-and 4-position thus.

In the present invention on the other hand, in the time that n is 2 or 3, first R ⁵group is suitably at the o-position of phenyl ring, second R ⁵group is suitably in m-position, and optionally (in the time that n is 3), the 3rd R ⁵group is suitably in the p-position of phenyl ring.Thus, in the time that n is 2, this ring suitably has substituting group in 2-and the 3-position of phenyl ring, and in the time that n is 3, this ring suitably has substituting group in 2-, 3-and the 4-position of phenyl ring.

The 2-that applicant has been surprised to find at phenyl ring and 3-position or 2-, the quinoline that 3-and 4-position have a substituting group (for example halogenic substituent) has substituent quinazoline derivant compared with the 3-of phenyl ring and 4-position and has given the activity that compound strengthens, these compounds have improved the effect of anti-erbB 2 and/or EGFR (particularly erbB2) in raji cell assay Raji. the 2-it is believed that at phenyl ring and 3-position or 2-, the quinoline that 3-and 4-position have substituting group (for example halogenic substituent) also will improve the effect of anti-erbB 2 and/or EGFR (particularly erbB2) in vivo.

Suitably, in the time that n is 2 or 3, each R ⁵group is that identical or different halogen atom is such as chlorine or fluorine.Suitably, at least one R ⁵group is fluorine, and at least one fluorine is suitably positioned at o-(2-) position of phenyl ring.

Suitably, in the time that n is 2, each R ⁵group is identical or different halogen atom.Especially, a R ⁵group is chlorine, and be preferably placed at its connected phenyl ring m-(3-) position, another R ⁵group is fluorine, and it is preferably placed at o-(2-) position or p-(4-) position (preferably o-(2-) position) of phenyl ring.

Suitably, in the time that n is 3, each R ⁵group is identical or different halogen atom.Especially, a R ⁵group is chlorine, and be preferably placed at its connected phenyl ring m-(3-) position, other two R ⁵respectively fluorine naturally of group, it preferably lays respectively at o-(2-) position and p-(4-) of phenyl ring.

Formula I Central Asia formula (ii) group thus:

Object lesson be sub-formula (iii) group:

Wherein, (a) R ¹¹or R ¹²be hydrogen and another be halogen such as chlorine or fluorine, particularly fluorine, and R ¹¹be halogen such as chlorine or fluorine, particularly chlorine, or (b) R ¹⁰that halogen is such as chlorine or fluorine, particularly fluorine, R ¹¹be halogen such as chlorine or fluorine, particularly chlorine, and R ¹²be hydrogen or halogen as chlorine or fluorine, particularly fluorine, or (c) R ¹⁰fluorine, R ¹¹chlorine, and R ¹²hydrogen or fluorine.Especially, R ¹⁰, R ¹¹and R ¹²define as (b) and/or (c).

In one embodiment, in the time that n is 2, each R ⁵group is identical or different halogen atom (such as fluorine and/or chlorine) and first R ⁵group is positioned at the o-position of phenyl ring and second R ⁵be positioned at m-position, now R ²not (optional replacement) (1-6C) alkyl. especially, R ²(1-6C) alkyl optionally being replaced by fluorine, (1-6C) alkoxyl group or sub-formula (i):

Wherein, m is 0 and R ³and R ⁴independently be selected from hydrogen or (1-4C) alkyl.

Suitably, X ¹oxygen.

Especially, R ¹be selected from hydrogen, (1-6C) alkyl and (1-6C) alkoxyl group (1-6C) alkyl, wherein R ¹in arbitrary (1-6C) alkyl optionally contain one or more (being suitably 1 or 2) hydroxyl or halogenic substituent.More particularly, R ¹be selected from (1-6C) alkyl, preferably (1-4C) alkyl and more preferably (1-2C) alkyl.For example, R ¹it can be methyl.

For example, R ¹-X ¹-be selected from methoxyl group, oxyethyl group, sec.-propyl oxygen base, cyclo propyl methoxy, 2-hydroxyl-oxethyl, 2-fluorine oxyethyl group, 2-methoxy ethoxy, 2,2-difluoroethoxy, 2,2,2-trifluoro ethoxy or 3-hydroxy-3-methyl butoxy.

Especially, R ¹-X-is selected from hydrogen, (1-4C) alkoxyl group and (1-4C) alkoxyl group (1-4C) alkoxyl group.For example, R ¹-X-is selected from hydrogen, methoxyl group, oxyethyl group and 2-methoxy ethoxy.

Suitably, R ²(1-6C) alkyl (particularly (1-3C) alkyl, more particularly (1-2C) alkyl), its optionally by fluorine, (1-6C) alkoxyl group, (1-6C) alkylthio, (1-6C) alkyl sulphinyl, (1-6C) alkyl sulphonyl or as defined above sub-formula (i) replace.R ²substituent object lesson be sub-formula (i) as defined above.

Especially, R ²be (1-3C) alkyl such as methyl or ethyl, it is optionally replaced by sub-formula (i) as defined above.Work as R ²while comprising sub-formula (i) substituting group, m is suitably 0,1 or 2.

Work as R ²while comprising sub-formula (i) substituting group, m is suitably 1 or 2, and particularly 2.On the other hand, m is 0 or 1.

Work as R ³and R ⁴while forming and optionally comprise that other is heteroatomic saturated 5 or 6 (preferably 6) first heterocycle together with the nitrogen-atoms connecting with them, it suitably comprises and is selected from O and NR ⁸other heteroatoms, wherein R ⁸suc as formula the related definition of I.

Work as R ³and R ⁴together with the nitrogen-atoms connecting with them, form while optionally comprising other heteroatomic saturated 5 or 6 yuan of heterocycle, it suitably contains pyridine ring, morpholine ring, piperidine ring or piperazine ring, optional former in locating by the R suc as formula I related definition at possible nitrogen ⁸replace.R ⁸the object lesson of group comprises that (1-3C) alkyl is such as methyl; (1-3C) alkyl sulphonyl is such as methyl sulphonyl; (1-3C) alkyl-carbonyl is such as ethanoyl; (2-4C) alkenyl is such as allyl group; Or (2-4C) alkynyl group such as proyl.Especially, R ⁸that (1-3C) alkyl is such as methyl.

Alternatively, R ³and R ⁴group can suitably independently be selected from (1-6C) alkyl, and particularly (1-3C) alkyl is such as methyl and ethyl.For example, on the one hand, each R ³and R ⁴group can be suitably that (1-3C) alkyl is such as R ³and R ⁴group can be ethyl separately.

R ²the object lesson of group comprises methyl, 2-(pyrrolidin-1-yl) ethyl, 2-(dimethylamino) ethyl, 2-(diethylamino) ethyl, 2-(piperidyl) ethyl, 2-(morpholine-4-yl) ethyl or 2-(4-methyl piperidine-1-yl) ethyl.More specifically, R ²the example of group comprises methyl, 2-(pyrrolidin-1-yl ethyl, 2-(diethylamino) ethyl, 2-(piperidin-1-yl) ethyl, 2-(morpholine-4-yl) ethyl or 2-(4-methylpiperazine-1-yl) ethyl.

In a specific embodiment, R ²it is methyl.In an alternative embodiment, R ²be selected from 2-(piperidin-1-yl) ethyl, 2-(4-methylpiperazine-1-yl) ethyl and 2-(pyrrolidin-1-yl) ethyl, R particularly ²it is 2-(pyrrolidin-1-yl) ethyl.In another alternative embodiment, R ²be selected from 2-(dimethylamino) ethyl and 2-(diethylamino) ethyl.In another alternative embodiment, R ²it is 2-(morpholine-4-yl) ethyl.

The object lesson of formula I compound is formula IA compound:

Wherein R ²as the related definition of above formula I, R ¹⁰, R ¹¹and R ¹²as above the related definition of sub-formula (iii), and R ¹³be selected from hydrogen, methoxyl group, oxyethyl group and 2-methoxy ethoxy, particularly methoxyl group.

Other special example of formula I compound is formula IB and IC compound:

Wherein R ²as the related definition of above formula I, and R ¹³be selected from hydrogen, methoxyl group, oxyethyl group and 2-methoxy ethoxy, particularly methoxyl group.

Other special example of formula I compound is formula ID compound:

Wherein:

R ^5aand R ^5bindependently be selected from halogen (for example fluorine and/or chlorine);

X ¹direct key or O;

R ¹be selected from hydrogen and (1-6C) alkyl, wherein (1-6C) alkyl is optionally replaced by one or more substituting groups, and it can be identical or different, is selected from hydroxyl and halogen, and/or be selected from following substituting group: amino, nitro, carboxyl, cyano group, halogen, (1-6C) alkoxyl group, hydroxyl (1-6C) alkoxyl group, (2-8C) alkenyl, (2-8C) alkynyl group, (1-6C) alkylthio, (1-6C) alkyl sulphinyl, (1-6C) alkyl sulphonyl, (1-6C) alkylamino, two-[(1-6C) alkyl] amino, formamyl, N-(1-6C) alkyl-carbamoyl, N, N bis--[(1-6C) alkyl] formamyl, (2-6C) alkyloyl, (2-6C) alkyloyl oxygen base, (2-6C) alkanoylamino, N-(1-6C) alkyl-(2-6C) alkanoylamino, (1-6C) alkoxy carbonyl, sulfamyl, N-(1-6C) alkylsulfamoyl group, N, N-bis--[(1-6C) alkyl] sulfamyl, (1-6C) alkyl sulfonyl amino and N-(1-6C) alkyl-(1-6C) alkyl sulfonyl amino,

R ²be (1-6C) alkyl, wherein should be optionally replaced by fluorine, (1-6C) alkoxyl group or sub-formula (iv) group by (1-6C) alkyl:

Wherein R ³and R ⁴group is independently selected from hydrogen or (1-4C) alkyl, or R ³and R ⁴the nitrogen that connects with them former in together with form 5 and 6 yuan of saturated heterocycles, it optionally comprises other and is selected from oxygen, S, SO or S (O) ₂or NR ⁸heteroatoms, wherein R ⁸hydrogen, (1-4C) alkyl or (1-4C) alkyl sulphonyl; Or its pharmacy acceptable salt.

In formula ID compound, R ²group is suitably (1-6C) alkyl, particularly unsubstituted (1-6C) alkyl.For example, R ²group can be methyl or ethyl, particularly methyl.

In formula ID compound, X ¹suitably oxygen.R ¹suitably be selected from hydrogen and (1-6C) alkyl, wherein R ¹in arbitrary (1-6C) alkyl optionally contain one or more (suitably 1 or 2) hydroxyl or halogenic substituent.More particularly, R ¹be selected from (1-6C) alkyl, particularly more especially (1-2C) alkyl of (1-4C) alkyl.For example, R ¹it can be methyl.R in formula ID compound ¹-X ¹object lesson be methoxyl group.

Obviously the novel especially compound of the present invention comprises formula I compound (comprising IA, IB, IC and ID) to those skilled in the art, except as otherwise noted, and wherein each R ¹, R ², R ³, R ⁵, X ¹, m and n have aforementioned arbitrary implication.

Formula I quinoline for example comprises one or more:

4-(the chloro-2,4 difluorobenzene amino of 3-)-7-methoxyl group-6-{[1-(methoxycarbonyl) piperazine smack one's lips-4-yl] oxygen base }-3-nitrile yl-quinoline;

4-(the chloro-2-fluoroanilino of 3-)-7-methoxyl group-6-{[1-{2-(morpholine-4-yl) ethoxy carbonyl } piperidin-4-yl] oxygen base }-3-itrile group-quinoline beautiful jade; And

Or its pharmacy acceptable salt.

The preferred example of formula I compound for example comprises one or more:

4-(the chloro-2-fluoroanilino of 3-)-7-methoxyl group-6-{[1-{2-(pyrrolidin-1-yl) ethoxy carbonyl } piperidin-4-yl] oxygen base }-3-nitrile yl-quinoline;

4-(the chloro-2,4 difluorobenzene amino of 3-)-7-methoxyl group-6-{[1-{2-piperidin-1-yl) ethoxy carbonyl } piperidines-4 base] oxygen base }-3-nitrile yl-quinoline

4-(the chloro-2-fluoroanilino of 3-)-7-methoxyl group-6-{[1-{2-(piperidin-1-yl) ethoxy carbonyl } piperidin-4-yl] oxygen base }-3-nitrile yl-quinoline

Or its pharmacy acceptable salt.

The special example of formula IA quinoline comprises one or more:

4-(the chloro-2-fluoroanilino of 3-)-7-methoxyl group-6-{ (1-{2-(pyrrolidin-1-yl) ethoxy carbonyl } piperidin-4-yl] oxygen base }-3-nitrile yl-quinoline;

4-(the chloro-2,4 difluorobenzene amino of 3-)-7-methoxyl group-6-{[1-{2-pyrrolidin-1-yl) ethoxy carbonyl } piperidin-4-yl] oxygen base }-3-nitrile yl-quinoline;

4-(the chloro-2,4 difluorobenzene amino of 3-)-7-methoxyl group-6-{[1-{2-piperidin-1-yl) ethoxy carbonyl } piperidin-4-yl] oxygen base }-3-nitrile yl-quinoline;

4-(the chloro-2-fluoroanilino of 3-)-6-{[1-(2-(diethylamino) ethoxy carbonyl } piperidin-4-yl] oxygen base }-7-methoxyl group-3-nitrile yl-quinoline;

4-(the chloro-2-fluoroanilino of 3-)-7-methoxyl group-6-{[1-{2-(morpholine-4-yl) ethoxy carbonyl) piperidin-4-yl] oxygen base }-3-nitrile yl-quinoline; And

Or its pharmacy acceptable salt.

The special example of formula IB quinoline comprises one or more:

4-(the chloro-2-fluoroanilino of 3-)-7-methoxyl group-6-{[1-{2-(morpholine-4-yl) ethoxy carbonyl } piperidin-4-yl] oxygen base }-3-nitrile yl-quinoline and 4-(the chloro-2-fluoroanilino of 3-)-7-methoxyl group-6-{[1-{2-(4-methyl piperidine-1-yl) ethoxy carbonyl } piperidin-4-yl] oxygen base }-3-nitrile yl-quinoline;

Or its pharmacy acceptable salt.

The special example of formula IC quinoline comprises one or more:

4-(the chloro-2,4 difluorobenzene amino of 3-)-7-methoxyl group-6-{[1-{2-(pyrrolidin-1-yl) ethoxy carbonyl } piperidin-4-yl] oxygen base }-3-nitrile yl-quinoline; And

4-(the chloro-2,4 difluorobenzene amino of 3-)-7-methoxyl group-6-{[1-{2-(piperidin-1-yl) ethoxy carbonyl } piperidin-4-yl] oxygen base }-3-nitrile yl-quinoline

Or its pharmacy acceptable salt.

The special example of formula ID quinoline comprises one or more:

4-(the chloro-2-fluoroanilino of 3-)-7-methoxyl group-6-{[1-{2-(piperidin-1-yl) ethoxy carbonyl piperidin-4-yl] oxygen base }-3-nitrile yl-quinoline;

4-(the chloro-2-benzene of 3-atmosphere base)-7-methoxyl group-6-{[1-{2-(morpholine-4-yl) ethoxy carbonyl } piperidin-4-yl] oxygen base }-3-nitrile yl-quinoline; And

4-(the chloro-2-fluoroanilino of 3-)-7-methoxyl group-6-{[1-(2-(4-methyl piperidine-1-yl) ethoxy carbonyl } piperidin-4-yl] oxygen base }-3-nitrile yl-quinoline;

Or its pharmacy acceptable salt.

Synthesizing of formula I quinoline

The present invention provides the preparation method of formula I quinoline or its pharmaceutically-acceptable salts on the other hand.The reaction that they do not need with method that may need protection of some substituting group will be interpreted as in some following method.When skilled chemist needs this protection by understanding, how such protecting group is placed in to correct position and amiable how sloughing.

About the example of protecting group can be referring to many common teaching materials about this content, ' the Protective Groups in Organic Synthesis ' (John Wiley & Sons publication) that for example TheodoraGreen shows.Removing of protecting group can be undertaken by the applicable method of removing discussed protecting group known to any ordinary method described in document or the chemist that is skilled in technique; selected these class methods should be able to be removed protecting group, and it is minimum that the interference of other group to molecule simultaneously reduces to.

Therefore,, if reactant comprises the groups such as for example amino, carboxyl or hydroxyl, described group may need protection in some reaction mentioned above.

Be for example acyl group to the applicable blocking group of amino or alkylamino, for example alkyloyl is (if ethanoyl, alkoxy carbonyl are (if methoxycarbonyl, ethoxy carbonyl or tert-butoxycarbonyl, aryl methoxy carbonyl are (for example, as benzyloxycarbonyl or aroyl (benzyl acyl group).The deprotection condition of above-mentioned protecting group must change with selected protecting group.Therefore, for example can as being hydrolyzed, remove alkali metal hydroxide (as lithium hydroxide or sodium) by the alkali with suitable such as the acyl group of alkyloyl or alkoxy carbonyl or aroyl.Or; for example can process and remove by the acid with suitable (example hydrochloric acid, sulfuric acid or phosphoric acid or trifluoroacetic acid) such as the acyl group of uncle-butoxy carbonyl, for example can be by through catalyzer (as by palladium carbon) hydrogenation or by removing with Lewis acid (as three (trifluoroacetic acid) boron) processing such as the fragrant methoxycarbonyl of benzyloxycarbonyl.Other applicable protecting group of primary amino is for example phthaloyl, and it can be by processing with alkylamine (as dimethylaminopropylamine) or removing with hydrazine processing.

The appropriate protection group of hydroxyl is for example acyl group, and for example alkane acyl for example, such as ethanoyl, aroyl (benzyl acyl group) or arylmethyl (as benzyl).The deprotection condition of above-mentioned protecting group must change with selected protecting group.Therefore, for example can remove as basic metal oxynitride (as lithium hydroxide or sodium) or ammoniacal liquor solution by the alkali with suitable such as the acyl group of alkyloyl or aroyl.Or, for example can be by removing through catalyzer (as by palladium carbon) hydrogenation such as the arylmethyl of benzyl.

The appropriate protection group of carboxyl is the group that for example becomes ester; for example methyl or ethyl; it can be by removing as sodium hydroxide hydrolysis with alkali; or for example tertiary butyl; it can be by processing and remove as organic acid (as trifluoroacetic acid) with acid; or for example benzyl, it can be by removing through catalyzer (as by palladium carbon) hydrogenation.

Also usable resins is as protecting group.

The routine techniques that can adopt chemistry neck city to know is removed protecting group in described synthetic any step easily.

Can be by being applicable to quinoline or its pharmacy acceptable salt of the known any method preparation formula I for preparing chemofacies related compounds.Provide another feature of the present invention for the preparation of the quinoline of formula I or these methods of its pharmacy acceptable salt, these methods are explained by following exemplary embodiment.Necessary raw material can obtain by organic chemistry standard method (referring to, for example Advanced Organic Chemistry (Wiley-Interscience), Jerry March).The preparation of these raw materials is described in subsidiary non-limiting example.In addition, can be by obtaining necessary raw material with the similar approach of those explanations, these methods are within organic chemist's ordinary skill ability.About the information of the essential raw material of preparation or related compound (can adopt it to form essential raw material) also can discovery in following patent and application, the content of its methods involving part is attached in the present invention by introducing at this: WO 94/27965, WO 95/03283, WO 96/33977, WO 96/33978, WO96/33979, WO 96/33980, WO 96/33981, WO 97/30034, WO 97/38994, WO 01/66099, US 5,252,586, EP 520722, EP 566226, EP 602851 and EP635507.

The present invention also provides by the following method (a) to (m) preparation formula I quinoline or its pharmaceutically-acceptable salts (wherein except as otherwise noted, each variable-definition is as above):

Step (a) through type II compound:

Wherein R ¹, X ¹, R ⁵have when any implication is except needs as defined above and protect any functional group with n, react with formula III compound:

Wherein R ²have when any implication is except needs as defined above and protect any functional group, and Lg is replaceable group, wherein this reaction is being carried out under existing easily at suitable alkali,

And after this remove existing protecting group by ordinary method.

The group L g example that is easy to replace knows it is halogen, alkane alkylsulfonyl oxygen base or arylsulfonyl oxygen groups, for example chlorine, bromine, methylsulfonyl oxygen base, 4-oil of mirbane alkylsulfonyl oxygen base or toluene-4-alkylsulfonyl oxygen base (suitable is methylsulfonyl oxygen base, 4-oil of mirbane alkylsulfonyl oxygen or toluene-4-alkylsulfonyl oxygen base).

This reaction has advantage under alkali exists.Suitable alkali is for example that organic amine alkali is as diisopropyl ethyl amine, pyridine, 2,6-lutidine, trimethylpyridine, 4-dimethylaminopyridine, triethylamine, N-methylmorpholine or diazabicyclo [5.4.0] 11-7-alkene, or for example basic metal or alkaline earth metal carbonate or oxyhydroxide, for example sodium carbonate, salt of wormwood, cesium carbonate, calcium carbonate, sodium hydroxide or potassium hydroxide. alternatively, this alkali is for example that alkalimetal hydride is as sodium hydride, basic metal or alkaline-earth metal acid amides, the alkali metal halide of for example acid amide sodium or two (trimethyl silyl) acid amides sodium or enough alkalescence is as cesium fluoride and sodium iodide. and this reaction is suitably carried out under inert solvent or thinner existence, and for example alcohol of solvent or thinner or ester are as methyl alcohol, ethanol, 2-propyl alcohol or ethyl acetate, halogenated solvent is as methylene dichloride, trichloromethane or tetracol phenixin, ether is as tetrahydrofuran (THF) or Isosorbide-5-Nitrae-dioxane, and aromatic hydrocarbon solvent is as toluene, or suitable dipolar aprotic solvent is as DMF, N,N-dimethylacetamide, N-picoline alkane-2-ketone or dimethyl sulfoxide (DMSO). this reaction in a certain temperature range as 10-150 ℃ (or solvent boiling point) carries out easily, suitably at 20-90 ℃.

Suitable especially alkali is cesium fluoride. this reaction is suitably carried out in as N,N-dimethylacetamide or DMF in solvent in the non-matter of inertia dipole.

This reaction is suitably carried out at 25-85 ℃ of temperature.

The substituting group of method (b) by modification formula I quinoline or its pharmaceutically-acceptable salts or introduce substituting group, compound is protected any functional group during as previously defined except needs, and is after this removing any protecting group by ordinary method.

Known in the art substituting group is converted into other substituent method.For example alkylthio can be oxidized to alkyl sulphinyl or alkyl sulphonyl; cyano reduction is amino; nitroreduction is amino; hydroxyl turns to methoxyl group through alkyl; bromo is converted into alkylthio, and amino can generate alkanoylamino (example is known and suitable acyl chlorides or anhydride reaction) or alkyloyl oxygen based hydrolysable is hydroxyl (for example acetoxyl group ethanoyl can be converted into hydroxyacetyl) through acidylate.Easily, can be by a R as the final step of preparation I compound ¹groups converted is another R ¹group.

Method (c) through type IV compound:

Wherein R ¹, X ¹, R ⁵relevant with n suc as formula I, react with formula V or V ' compound:

Wherein R ²as above definition, and Lg is replaceable group (for example halogen is as chlorine or bromine or 1-imidazolyl).Above-mentioned reaction is at suitable alkali (those that describe in such as method (a), for example salt of wormwood or two-sec.-propyl ethamine) exist lower and under inert solvent or thinner (inert solvent of for example, describing in method (a) and thinner are as acetonitrile, N,N-dimethylacetamide, methyl alcohol, ethanol or methylene dichloride) existence, carry out easily easily.

Method (d) is by removing the protecting group of formula I quinoline or its pharmaceutically-acceptable salts.

The known proper method of removing protecting group is also discussed in this article.For example, in order to generate wherein R ¹the formula I compound that comprises primary amine or secondary amine, cracking is R wherein ¹the corresponding formula I compound that comprises protected primary amine or secondary amine group.

Suitable chloro protecting group is for example disclosed arbitrary amino protecting group before this.The proper method of these amino protecting groups of cracking is disclosed before this.Especially, suitable protecting group be compared with lower alkoxycarbonyl such as tert-butoxycarbonyl, it can be in popular response condition such as cracking under acid-catalyzed hydrolysis, for example, under trifluoroacetic acid exists.

Method (e) is reacted with formula III compound defined above by formula II compound defined above under Mitsunobu condition, except Lg is OH, after this removes the protecting group of any existence by ordinary method.

Suitable Mitsunobu condition comprises, for example, under suitable three grades of phosphines and two-alkyl azodicarboxylate exist, organic solvent as THF or suitable methylene dichloride and 0 ℃ of-60 ℃ of temperature range in suitably reaction at ambient temperature.Three grades of suitable phosphines for example comprise three-n-butyl phosphine or are suitably three-Phenylphosphine. suitable two-alkyl azodicarboxylate for example comprises diethylazodicarboxylate (DEAD) or is suitably azo-2-carboxylic acid's two-tert-butyl ester.The details of Mitsunobu reaction is included in Tet.Letts., and 31,699, (1990); The Mitsunobu Reaction, D.L.Hughes, Organic Reactions, 1992, Vol.42,335-656 and Progress in the MitsunobuReaction, D.L.Hughes, Organic Preparations and ProceduresInternational, 1996, Vol.28, in 127-164.

Method (f) is by wherein R of cracking ¹-X ¹the formula I quinoline that is (1-6C) alkoxyl group is prepared wherein R ¹-X ¹it is the formula I compound of hydroxyl.

This scission reaction can complete by one of known many methods that realize this conversion.Wherein R ¹that the scission reaction of formula I compound of (1-6C) alkyl is for example by being used basic metal (1-6C) alkyl sulfur compounds process or use basic metal diaryl phosphides processing can complete as diphenylphosphine lithium as sulfur alcohol sodium.Alternatively, this scission reaction is for example by being used boron or aluminium trihalid to process quinazoline derivant or by having reacted as trifluoroacetic acid with mineral acid as boron tribromide.These reactions are suitably carried out under above-mentioned suitable inert solvent or thinner existence.Preferred scission reaction is to use pyridine hydrochloride to process the quinoline of formula I.This scission reaction is for example suitably completing in 10-200 ℃ of temperature range.Sometimes can for example at 25-80 ℃ of temperature, complete, but while sometimes for example using pyrrole to fill the air with noses hydrochlorate deprotection, be suitable 160-200 ℃ of fusing conventionally.

Wherein R ², R ⁵have when any implication is except needs as defined above and protect any functional group with n, with formula R ¹the reaction of-Lg compound, wherein R ¹have that when any implication is except needs as defined above, to protect any functional group, Lg be replaceable group, wherein this is instead carrying out under existing easily at suitable alkali;

Remove subsequently any protecting group of existence by any ordinary method.

Suitable replaceable group, Lg, as definition in front method (a), for example, is chlorine or bromine.This reaction is suitably suitably being carried out under alkali existence.Appropriate solvent, thinner or alkali for example comprise those that preceding method (a) is relevant.Alternatively, Lg is hydroxyl, and this reaction can be carried out under Mitsunobu condition, as front method (e) associated description.

Method (h) is in order to prepare wherein R ¹the formula I compound of (1-6C) alkylamino of (1-6C) alkoxyl group that comprises (1-6C) alkoxyl group or replacement or (1-6C) alkylamino or replacement, suitably time, wherein R of alkylation under the suitable alkali as described in front method (a) exists ¹the formula I quinoline that comprises hydroxyl or primary amine or secondary amine group.

For example alkylate hydroxyl known in the art of suitable alkylating reagent forms the alkoxyl group of alkoxyl group or replacement or any reagent of the amino alkylamino that forms alkylamino or replacement of alkylation, the halogenide of for example alkyl or substituted alkyl, for example (1-6C) alkyl chloride, bromide, iodide, easily under suitably alkali exists as defined above, as defined above suitably natural instincts solvent or thinner and in 10-140 ℃ of temperature range for example easily in envrionment temperature or approach under envrionment temperature.Can use similar approach by (2-6C) alkyloyl oxygen base, (2-6C) alkanoylamino that optionally replace and (1-6C) alkane sulfuryl amino introduce R ¹.

In order to prepare easily wherein R ¹the formula I compound of (1-6C) alkylamino that comprises (1-6C) alkylamino or replacement, can use formaldehyde or (2-6C) alkyl aldehydes (for example acetaldehyde or propionic aldehyde) implement reduction amination.For example,, in order to prepare wherein R ¹those formulas I compound that comprises N-methyl, the respective compound that comprises N-H group can suitably reacted under reductive agent existence with formaldehyde.Suitable reductive agent is for example that nitride reductive agent for example formic acid, aluminum hydride basic metal compound are as lithium aluminum hydride or suitably for alkali metal borohydride is as sodium borohydride, trimethoxy sodium borohydride and sodium triacetoxy borohydride.This reaction suitably natural instincts solvent or thinner for example for compared with strong reductant as the tetrahydrofuran (THF) of lithium aluminum hydride and ether or for carrying out easily as methyl alcohol or ethanol as the methylene dichloride of sodium triacetoxy borohydride and sodium cyanoborohydride or protic solvent compared with weak reductant.In the time that reductive agent is formic acid, this reaction is used aqueous formic acid to carry out easily.This reaction 10-100 ℃ such as 70-90 ℃ of temperature range in or approaching under envrionment temperature and carrying out.Easily, in the time that reductive agent is formic acid, on NH group, prevent that alkylating protecting group is such as tert-butoxycarbonyl (for example, from synthetic starting raw material time just exist) can be removed on the spot reaction process.

Wherein R ², R ⁵, X ¹, n and m protect any functional group, R while having implication as previously defined except needs ¹' be R as defined herein ¹group is replaced by Lg except any T group, and Lg is replaceable group (for example chlorine or bromine, or the alkyl sulfonic ester of aryl/(1-6C) is as methanesulfonates), react with formula TH compound, when wherein T as above defines except needs, protect any functional group;

Remove subsequently any blocking group of existence by ordinary method.

This reaction is suitably being carried out under alkali existence easily.This reaction can be carried out easily in suitable inert solvent or thinner.Suitably alkali, solvent and thinner are for example described in method (a).This reaction is suitably carried out at the temperature of for example 10-150 ℃, for example 30-60 ℃.

To be interpreted as, before aforesaid method or can introduce by the fragrant substitution reaction of standard following closely or can generate some different ring substituents in the compounds of this invention by conventional modified with functional group, these methods are also included within method of the present invention aspect.These reactions and modification for example comprise by fragrant substitution reaction introduces substituting group, reduction substituting group, alkylation substituting group and oxidation substituting group.The reagent of these methods and reaction are that chemical field is known.The concrete ion of fragrance substitution reaction comprises use concentrated nitric acid introducing nitro, for example uses carboxylic acid halides and lewis' acid (as aluminum chloride) to introduce acyl group under Friedel Crafts condition; Use haloalkane and lewis' acid (as aluminum chloride) to introduce alkyl under Friedel Crafts condition; And introducing halogen group.

Method (j) through type VIII compound:

Wherein R ¹, R ², X ¹while having foregoing implication except needs with m, protecting any functional group and Lg is interchangeable group as previously mentioned, and the aniline reaction of formula IX:

Wherein R ⁵while having foregoing any implication except needs with n, protect any functional group, and wherein this reaction is suitably being carried out under acid existence easily,

Remove subsequently any blocking group of existence by ordinary method.

As previously mentioned, particularly halogen is as chlorine for the suitable replaceable group of Lg representative.

This reaction at suitable inert solvent or for example alcohol of thinner or ester as methyl alcohol, ethanol, Virahol or ethyl acetate, halogenated solvent is as methylene dichloride, chloroform or tetracol phenixin, ether is as tetrahydrofuran (THF) or 1,4-diox, aromatic solvent as toluene or the non-matter of dipole in solvent as N, dinethylformamide, N, in N-N,N-DIMETHYLACETAMIDE, NMP acetonitrile or dimethyl sulfoxide (DMSO), carry out easily, this reaction for example 10-250 ℃ in 40-120 ℃ of temperature range or under the reflux temperature of solvent or thinner, carry out easily easily.Easily, formula VIII compound and formula IX compound at polar solvent as Virahol, easily in acid under for example catalytic amount acid exists, react under these conditions.Suitable acid comprises ether Huo dioxane solution and the spirit of salt of hydrogen chloride gas, the hydrogenchloride dioxane solution of for example 4M.Alternative, this reaction at non-polar solvent as diox or dipolar aprotic solvent as N, in N-methylacetamide or acetonitrile, under the ether Huo dioxane solution of for example hydrogen chloride gas of acid or spirit of salt exist, can carry out easily.

Lg is the formula VIII compound of halogen, can under acid exists, react with formula IX compound.In this reaction, replace halogen leavings group Lg and cause forming on the spot HLg acid this reaction of autocatalysis.Easily, this reaction is carried out in suitable for example Virahol, diox of inert organic solvents or N,N-dimethylacetamide.The suitable condition of this reaction is described above.

Alternative, formula VIII compound can suitably react under alkali existence with formula IX compound.Be applicable to the alkali of this reaction described in front method (a).For example, suitable alkali is that alkaline-earth metal acid amides is as two (trimethyl silyl) acid amides sodium.This reaction is for example carried out in above-mentioned those relevant with method (j) easily at natural instincts solvent or thinner;

Method (k) is those formulas I compound of 1,2 or 3 in order to prepare m wherein, makes formula X compound:

Wherein R ⁵, X ¹, R ¹with n as previously defined, and wherein Lg is that leavings group is such as halogen especially chlorine or 1-imidazolyl, with formula R ²the alcohol reaction of-OH, wherein R ²as previously defined.This reaction is suitably carried out in as DCM at aprotic solvent under alkali exists as triamine/pyridine.Skilled chemical technology personnel obviously will know suitable temperature.

Method (1) is for R wherein ²comprise the compound of sub-formula (i) group, make formula XI compound

With formula R ³r ⁴the reaction of H compound, wherein R ³and R ⁴as the related definition of front sub-formula (i).Now suitable leavings group Lg comprises that halogen is if chlorine or alkyl/aryl sulphonate are as methanesulfonates.This reaction is suitably carried out in as N,N-DIMETHYLACETAMIDE, N-Methyl pyrrolidone or dimethyl formamide at organic solvent under propiodal exists as potassiumiodide or tetrabutylammonium iodide.Suitably, use excessive R ³r ⁴nH amine.For example, in the time that Lg is chlorine, this can advantageously remove the hydrogenchloride forming in reaction process.Suitably, can use for example 50-120 ℃ of high temperature, for example approximately 80 ℃.

It will be appreciated by those skilled in the art that, for obtaining the compounds of this invention in other situation and some occasion in mode more easily, order that can be different carries out above-described each process step, and/or carries out each reaction (adopting the intermediate relevant with being different from above-mentioned concrete reaction to carry out each chemical conversion) with the different stages in whole route.

For example, in the time needing the pharmacy acceptable salt (acid salt) of quinoline of formula I, can adopt conventional method, for example, by described quinoline and suitable acid-respons are obtained.For be easy to separate described compound in preparation process, can prepare the compound of the form of non-pharmacy acceptable salt.Then modify obtained salt by routine techniques, generate the pharmacy acceptable salt of described compound.These technology comprise for example ion exchange technique or pharmaceutically acceptable counter ion there is the technology of compound described in lower redeposition.For example, redeposition under applicable acid (as HCl) exists, obtains salt acid salt.

In this part, above-mentioned wording " inert solvent " refers to the solvent that can not react with starting raw material, reagent, intermediate or product adversely to affect the mode of desired product yield.

The preparation of starting raw material

Formula II compound be commercially available or can adopt routine techniques or with similar method preparation described in the prior.Especially above listed those patents and application, as WO96/15118, WO 01/66099 and EP 566226.For example, can be according to reaction process 1 preparation formula II compound:

Reaction process 1

Wherein R ¹, X ¹, R ⁵, Lg and n as previously defined and Pg be hydroxyl protecting group.

(i) reaction is suitable in inertia matter in solvent (as alkanol, for example Virahol), aprotic solvent (as dioxane) or the non-matter of dipole in solvent (as N, N-N,N-DIMETHYLACETAMIDE) in, under acid (as the hydrogen chloride gas in ether or dioxane or hydrochloric acid) exists, under the conditions of similarity described in aforesaid method (i), carry out.

Or, described reaction can above-mentioned inertia molten one of not in, under alkali (as salt of wormwood) exists, carry out expediently.More than react in scope for for example in 0-150 ℃, suitably or approach under the reflux temperature of reaction solvent and carry out expediently.

(ii) Pg of these reactions of cracking under standard conditions.For example, in the time that Pg is alkyloyl (as ethanoyl), can be by heating under existing at Methanol ammonia solution by its cracking.

Formula XII compound is known compound or can adopts the currently known methods preparation of preparing similar compound.If can not provide commercially available, the method that formula XII compound can be by being selected from standard chemical technology, be similar to the technology of synthetic known structure analogue compounds or be similar to described in an embodiment step is prepared.For example, the standard chemical technology of describing in Houben Weyl.For example, adopt the method for explanation in reaction process 2 can preparation formula XII compound, wherein R ¹-X ¹for methoxyl group, Lg is that chlorine and Pg are ethanoyl:

Reaction process 2

Compound (for example introducing the substituting group of non-methoxyl group on the 7-position of described quinoline ring) in this specification sheets that technician can be applied to reaction process 2 not illustrate.

Formula III compound be commercially available or can adopt standard technique preparation, for example, in US5252586 and WO 94/27965 technology of explanation.

Formula IV compound can react and prepare with XVa or XVb compound by through type II compound:

Wherein Lg is that replaceable group and Pg are as previously defined suitable blocking groups.

Formula II compound carries out with the condition that can describe in method (a) above being similar to of reacting of formula XV compound, removes protecting group subsequently under standard conditions.

Formula II compound can carry out with reacting of formula XVb compound under as the Mitsunobu condition of aforesaid method (a), removes protecting group subsequently under standard conditions.

Formula IV compound also can through type IX compound and formula XVc compound:

Reaction preparation, wherein Lg, X ¹and R ¹as previously defined and Pg be suitable protecting group.

Formula IX compound carries out with the condition that can describe in method (j) above being similar to of reacting of formula XVc compound, removes protecting group subsequently under standard conditions.

Formula V and IX compound can be buied maybe can use standard technique preparation.

Formula VI compound can through method above, (e0 preparation be used the compound of for example preparing through method (a) as starting raw material.

Formula VII compound can be through for example method (a) or method (d) method (e) preparation, wherein R ¹the group of representative suitably by suitable replaceable group L g as chlorine or bromine functionalization.

Formula VIII compound can be through ordinary method preparation well known in the art.For example remove the hydroxyl protecting group in above-mentioned reaction process 1 formula XII compound, Pg, production XIV compound:

Pg protecting group can be removed through routine techniques from formula XII compound.

Then, formula XIV compound can with the coupling of aforementioned formula III compound, use the condition of describing in method (a) or method (e) that is similar to.

Formula X can be through method preparation well known in the art.For example, formula IV compound is reacted with formula Lg-CO-Lg compound, wherein Lg is that interchangeable group for example, as halogen (chlorine or bromine) or imidazolyl.Formula IV compound and formula Lg-CO-Lg compound react can be through being similar to aforesaid method (c) condition under weak base exists as pyridine or lutidine and for example, carry out under natural instincts solvent (acetonitrile or methylene dichloride). the suitable example of formula Lg-CO-Lg compound is phosgene and 1,1 '-carbonyl dimidazoles.

In method, some novel intermediates used is provided as further feature of the present invention with together with their preparation method above.

According to a further general feature of the present invention, provide formula VI described above, VII, X and XI compound or its salt, (comprising its pharmacy acceptable salt), as previously mentioned.

biological assay

Not the protein tyrosine kinase mensuration based on cell and the inhibition activity of having measured compound in the proliferation assay based on cell, in xenotransplantation transplanting research, evaluating afterwards their activity in vivo.

A) protein tyrosine kinase phosphorylation assay

This test determination test compound suppresses the ability of the peptide substrate phosphorylation that contains tyrosine by EGFR or ErbB2 Tyrosylprotein kinase.

Reconstitution cell internal fragment EGFR, erbB2 and erbB4 (catalog number (Cat.No.) is respectively X00588, X03363 and L07868) are cloned and expressed in baculovirus/Sf21 system.By by ice-cooled molten born of the same parents' damping fluid (20mM HEPES (HEPES) pH7.5,150mM NaCl, 10% glycerine, 1%TritonX-100,1.5mM MgCl for these cells ₂, 1mM ethylene glycol-bis-(beta-amino ether) N ', N ', N ', N '-tetraacethyl (EGTA)) add proteinase inhibitor processing, then prepare lysate by centrifugal clarification.

The ability that makes a kind of synthetic peptide (randomcopolymer of L-glutamic acid, L-Ala and the tyrosine that is 6: 3: 1 by ratio forms) that phosphorylation occur by it is measured the formation kinase activity of described recombinant protein.Specifically, by Maxisorb ^tMsynthetic peptide coating (0.2 μ g peptide is in 100Ld phosphate-buffered saline (PBS) and 4 ℃ of overnight incubation) for 96 hole immunization test boards.Under room temperature, by each for plate PBS-T (with the phosphate-buffered saline of 0.5% polysorbas20) then wash to remove any excessive unconjugated synthetic peptide with 50mM HEPES pH 7.4.By at 22 ℃, at 100mMHEPES pH 7.4, for adenosine triphosphate (ATP), 10mMMnCl under the Km concentration of each enzyme ₂, 0.1mm Na3VO ₄, in 0.2mM DL-sulphur threitol (DTT), 0.1%TritonX-100, assay plate coated peptide is hatched 20 minutes together with the DMSO of test compound solution (final concentration 2.5%), measure the tyrosine kinase activity of EGFR, ErbB2 or ErbB4.By removing the liquid ingredient of measuring in liquid, then wash each plate termination reaction with PBS-T.

By immobilized phosphoric acid-peptide (phospho-peptide) product reacting described in determination of immunological methods.First, at room temperature, each plate is hatched 90 minutes together with the anti-Tyrosine O-phosphate primary antibody (deriving from the 4G10 of Upstate Biotechnology) of cultivating in mouse.Fully after washing, under room temperature, each plate is used to (horseradish peroxidase (HRP) that derives from the NXA931 of Amersham is processed 60 minutes in conjunction with goat-anti mouse secondary antibodies.Further, after washing, adopt as 2 of substrate 2 '-azino-bis--[3-ethyl benzo thiazole phenanthroline sulfonic acid (6)] di-ammonium salts crystal (ABTS1 ^tM, purchased from Roche) and the HRP activity in every hole in colorimetric estimation plate.

By measuring absorbancy by Molecular Devices ThermoMax microplate under 405nm, to developing the color quantitatively, measure thus enzymic activity.The kinase inhibitory activity of given compound is with IC ₅₀value representation.The required compound concentration of 50% restraining effect that provides phosphorylated by calculating in this mensuration is measured IC ₅₀value.From positive (medium adds ATP) and negative (medium subtracts ATP) contrast position, calculate phosphorylated scope.

B) the KB cell proliferation test that EGFR drives

This test determination test compound suppresses the ability of KB cell (KB cell is obtained by AmericanType Culture Collection (ATCC)) propagation.

At 7.5%CO ₂in air incubator, at 37 ℃, in the DulbeccoShi improvement EagleShi substratum (DMEM) that contains 10% foetal calf serum, 2mM glutamine and non-essential amino acid, cultivate KB cell (KB cell is provided by ATCC).Adopt trypsinase/ethylenediamine tetraacetic acid (EDTA) (EDTA), harvested cell from deposit bottle.Adopt hematimeter to measure cell density, then with every hole 1.25 × 10 ³the density of individual cell, is inoculated in 7.5%CO ₂, in 96 orifice plates at 37 ℃ containing in the DMEM of 2.5% activated carbon decolorizing blood plasma (stripped serum), 1mm glutamine and non-essential amino acid, fix 4 hours, then adopt trypan blue solution to calculate vigor.

Cell adhesion on plate after, cell, with (or need not) EGF (final concentration 1ng/ml) with methyl-sulphoxide (DMSO) (final concentration 0.1%) processing of the compound within the scope of (or need not) finite concentration, is then hatched 4.Hatch after end, by adding the bromination 3-of 50 μ l, (4,5-dimethylthiazole-2-base-2,5-phenylbenzene tetrazolium (storing solution 5mg/ml) are measured cell count 2 hours.Then MTT solution is inclined to, culture plate is rapped dry, then add 100 μ l DMSO to dissolve described cell.

By adopting Molecular Devices ThermoMax microplate, the absorbancy of reading the cell of dissolving under 540nm.By the restraining effect IC of propagation ₅₀value representation.Measure IC by the required compound concentration of 50% restraining effect calculating producing propagation ₅₀value.From positive (medium adds EGP) and negative (medium subtracts EGP) control value, calculate multiplication nursery.

C) in body, xenotransplantation is measured

(i)LOVO

This test detects the ability that test compound suppresses LoVo tumour (rectum cancer is derived from ATCC) growth in female Swiss athymic mouse (AlderleyPark, nu/nu genotype).

Raise female Swiss nude mouse (nu/nu genotype), then remain in the Aldefiey Park in negative pressure Isolator (shield retaining) (PFI Systems Ltd.).Mouse is closed and supported in the barrier apparatus with 12 little time/dark circulations, aseptic food and water are freely provided.In steps all adopt at least 8 week ages mouse carry out.By giving 1 × 10 of the fresh culture of every mouse subcutaneous injection in 100 μ l serum free mediums ⁷individual cell, sets up the xenotransplantation of LoVo tumour cell (nodus hemorrhoidalis enteraden disease, is derived from ATCC) at the rear side belly of donor mice.After transplanting the 5th day, mouse is divided into 7 groups at random, then give compound 1 time every day with the amount of 0.1ml/10g body weight or control media is processed.Measure gross tumor volume by two-sided slide caliper 2 times weekly, adopt following formula to calculate: (length x width) × √ (length x width) × (Л/6), wherein length is that width is corresponding perpendicular diameter by this swollen sick longest diameter.The mean change of the gross tumor volume by compare group and treatment group is calculated from studying initial growth-inhibiting, adopts the statistical significance between two groups of Students t check assessments.

(ii) in body, BT-474 xenotransplantation is measured

This test detects test compound at female Swiss athymic mouse (Alderley Park, nu/nu genotype) middle BT-474 tumour cell xenotransplantation (the human breast cancer that suppresses, be derived from DrVaselga, Laboratorio Recerca Oncologica, Paseo Vall D ' Hebron 119-129, Barcelona 08035, Spain) ability (Baselga of growth, J.et al. (1998) CancerResearch, 58,2825-2831).

Raise female Swiss nude mouse (nu/nu genotype), then remain in the Aldefiey Park in negative pressure Isolator (shield retaining) (PFI Systems Ltd.).Mouse is closed and supported in the barrier apparatus with 12 little time/dark circulations, aseptic food and water are freely provided.In steps all adopt at least 8 week ages mouse carry out.By giving 1 × 10 of the fresh culture of every mouse subcutaneous injection in the 100 μ l serum free mediums that contain 50% matrigel ⁷individual cell, the xenotransplantation of setting up BT-474 tumour cell at the rear side belly of donor mice.After transplanting the 14th day, be divided into the most at random 10 groups by little, then give compound 1 time every day with the amount of 0.1ml/10g body weight or control media is processed.Measure gross tumor volume by two-sided slide caliper 2 times weekly, adopt following formula to calculate: (length x width) × √ (length x width) × (Л/6), wherein length is the longest diameter by this tumour, and width is corresponding perpendicular diameter.The mean change of the gross tumor volume by compare group and treatment group is calculated from studying initial growth-inhibiting, adopts the statistical significance between two groups of Students t check assessments.

D) the potassium channel inhibition test of hERG-coding

This test determination test compound suppresses the mobile ability of tail current by the potassium channel of people ether-a-go-go-genes involved (hERG)-coding.

Make to express human embryo kidney (HEK) (HEK) Growth of Cells of hERG--coding pass in adding 10% foetal calf serum (Labtech International; Production number 4-101-500), 10%M1 serum-free supplement liquid (EggTechnologies; Production number 70916) and 0.4mg/ml Geneticin G418 (Sigma-Aldrich; Catalog number (Cat.No.) G7034) Eagle minimum essential medium (EMEM; Sigma-Aldrich; Catalog number (Cat.No.) M2279) in.Before each on-test 1 or 2 day, adopt normal structure cultural method, with Accutase (TCS Biologicals) isolated cell from tissue culture flasks.Then they are placed on the glass cover slide in the 12 each holes of well culture plate, with the covering of 2ml growth medium.

For recording every porocyte, auspicious the glass that cell is housed cover glass is placed in to the bottom, the Perspex chamber of containing body lotion (seeing below) under room temperature (20 ℃). an inversion is fixed on in this chamber and differs on the platform that shows emblem mirror.Cover glass is placed in to this chamber, from a gravity supply storehouse (gravity-fed reservoir), in described chamber, pours into body lotion 2 minutes with the speed of about 2ml/min immediately.Then stop perfusion.

To pull out that device (Sutter Instrument Co.) makes from borosilicate glass tube (GC120F, Harvard Apparatus) by P-97 trace pipette membranaceous for pipette pipette solution (seeing below) be full of.By silver/silver chloride line, described pipette is connected in to the top of patch clamp augmentor (Axopatch 200B, Axon Instruments).This top/bottom part is connected to earth polar.It is made up of the side silver chloride line being inserted in 3% agar being made up of 0.85% sodium-chlor.

Whole-cell configuration with patch clamp technique records cell. and carry out " swarming into (break-in) " and carry out after the suitable debugging of series resistance and Capacity control maintaining current potential-80mV (being arranged by augmentor), adopt electric physiology software (Clampex, Axon Instruments) arrange maintain current potential (80mV), then transmit an agreement voltage (voltage protocol).The application of every 15 seconds of this agreement once, by 1 second to+40mV step and subsequently 1s extremely-50mV step forms.Under 1kHz, low by the electric current of the each application protocol voltage of filtering response by augmentor.Then pass through with similar quanxtizer similarly signal digitizing in augmentor, the signal that acquisition is filtered online. on the computer that moves Clampex software (Axon Instruments), catch digitized signal afterwards.Maintaining in voltage and extremely+40mV step process, under 1kHz, survey sample electric current.Then the sample rate of remaining voltage protocol is made as to 5kHz.

In following table, list the penetration degree of described composition, pH and described body lotion and pipette solution.

Salt	Suction pipe (mM)	Body lotion (mM)
			NaCl	-	137
KCl	130	4
			MgCl ₂	1	1
CaCl ₂	-	1.8
			HEPES	10	10
Glucose	-	10
			Na ₂ATP	5	-
EGTA	5	-

Parameter	Suction pipe	Body lotion
			pH	7.18-7.22	7.40
PH adjusting agent	1M KOH	1M NaOH
			Penetration degree (mOsm)	275-285	285-295

By Clampex software (Axonlnstruments) online record voltage from+40mV to-50mV step the amplitude of potassium channel tail current of hERG-coding.After the amplitude stability of this tail current, to the body lotion that adds the medium that contains tested substance in cell.Add medium to have no significant effect the amplitude of tail current, then set up the accumulative total mass action curve to compound.

By the lower amplitude of expressing tail current of the given concentration at test compound (percentage concentration existing in medium), the effect of the each concentration to test compound is quantitative.

By adopting normal data matching software package, the percentage restraining effect value being made up of is fitted to the usefulness (IC that deletes equation and come determination test compound of quaternary parameter concentration-effect ₅₀).If being seen inhibition level is no more than 50% under the highest experimental concentration, does not produce efficiency value and quote the percent inhibition under this concentration.

E) clone 24 phosphoric acid-erbB2 (phospho-erbB2) measures

This immunofluorescence endpoint determination has been measured the ability that test compound suppresses erbB2 phosphorylation in the clone of source MCF7 (mammary cancer), and clone is wherein by being used total length erbB2 gene to express the clone (being after this called " clone 24 " cell) of whole audience wild-type erbB2 albumen with the mistake producing through standard method transfection MCF7 cell.

At 37 ℃ in 7.5%CO ₂in air incubator, will clone 24 cell cultures in growth medium (, containing the phenol red improved Eagle ' of Dulbecco ' s s substratum (DMEM), not comprising 10% foetal calf serum, 2mM L-glutamic acid and 1.2mg/ml G418).By in PVS (phosphate-buffered saline, pH7.4, Gibco No.10010-015) in washing once and use 2ml trypsin 1.25mg/ml) (0.8mg/ml) solution results of/ethylenediamine tetraacetic acid (EDTA) (EDTA), thereby from T75 deposit bottle harvested cell.Cell is suspended in growth medium again.Further in growth medium, before dilution, use hemocytometer to measure cell density and also use trypan blue solution to calculate vitality, by cell with 1 × 10 ⁴the density in the every hole of individual cell (in 100 μ l) is inoculated in to be clarified in 96 orifice plates that have the end (Packard, No.6005182).

After 3 days, from each hole, remove growth medium and replace with 100 μ l and measure substratum (not containing phenol red DMEM, 2mM L-glutamic acid, 1.2mg/ml G418), containing or do not contain erbB inhibitor compound.This plate is sent back in incubator and placed and then in each hole, add the PBS solution of 20 μ l 20% formaldehyde and each plate is placed in to room temperature 30 minutes for 4 hours.Use multichannel pipettor to remove this fixative solution, in each hole, add 100 μ l PBS then to use multichannel pipettor to remove, and then add 50 μ l PBS to each hole.Seal each plate and reach 2 weeks in 4 ℃ of storages.

Under room temperature, carry out immunostaining.Then use 200 μ l PBS/ polysorbas20s (preparing to adding 1 bag PBS/ tween dry powder (Sigma, No.P3563) in 1L distilled water) to wash each hole through plate washing device adds 200 μ l retardance liquid (Blocking Solution) (the PBS/ polysorbas20 solution of the skimming milk (Nestle) that 5%Marvel is dry) and hatches 10 minutes.Use plate washing device to remove retardance liquid, add 200 μ l0.5%Triton X-100/PBS permeation cells.After 10 minutes, use 200 μ l PBS/ polysorbas20s to wash this plate and again add 200 μ l retardance liquid to hatch 15 minutes.Use plate washing device to remove after retardance liquid, to adding in each hole through the 30 μ l rabbit polyclonal anti-phosphoric acid ErbB2IgG antibody (epitope phosphoric acid-Tyr 1248, SantaCruz, No.SC-12352-R) of 1: 250 retardance liquid dilution and hatching 2 hours.Then use plate washing device from each hole, to remove this trie primary antibody solutions, with after use 200 μ lPBS/ polysorbas20 washed twice through plate washing device.Then add the 30 μ lAlexa-Fluor 488 goat anti-rabbit igg secondary antibody (Molecular Probes, No.A-11008) through the dilution of 1: 750 retardance liquid to each hole.From now on, as possible, the plate that keeps in Dark Place, now by sealing with black tape base.Hatch these plates and within 45 minutes, then from each hole, remove secondary antibody solution, with after use 200 μ l PBS/ polysorbas20 washed twice through plate washing device.Then remove through plate washing device to adding in each hole 100 μ l PBS to hatch after 10 minutes.And then add 100 μ l PBS in each plate, without hatching for a long time, remove through plate washing device.Then in each hole, add 50 μ l PBS, again seal these plates and reached 2 days in 4 ℃ of storages before analyzing with black tape base.

Use Acumen Explorer Instrument (Acumen Bioscience Ltd.) to measure the fluorescent signal in each hole, this instrument is the plate count device that can be used for fast quantification laser scanning image feature.Thereby set this apparatus measures and provide the measurement for erbB2 phosphorylation state of protein higher than the fluorescence number of predetermined threshold.The fluorescence dose response data that each compound is obtained is applied to suitable software package (such as Origin) to carry out curve the matching analysis.The inhibition of erbB2 phosphorylation is expressed as IC ₅₀value.It suppresses 50% the required compound concentration of erbB2 phosphorylation signal by calculating and determines.

Although the pharmacological properties of formula I compound is as changing with structural changes of estimating, generally, more than one or more of, in test, under following concentration or dosage, susceptible of proof formula I compound has activity:

Test (a) :-IC50 is within the scope of 0.001-10 μ M for example;

Test (b) :-IC ₅₀within the scope of 0.001-10 μ M for example:

Test (e) :-IC ₅₀within the scope of 0.001-10 μ M for example;

Test (c) :-for example thering is activity in the dosage range of 1-200mg/kg/ days;

By the mode of embodiment, Table A for example understands the activity of representative compound of the present invention.Table A the 2nd hurdle shows that test (a) suppresses the IC of EGFR Tyrosylprotein kinase protein phosphorylation ₅₀; The 3rd hurdle shows that test (a) suppresses the IC of erbB2 Tyrosylprotein kinase protein phosphorylation ₅₀value; The 4th hurdle shows to suppress in above-mentioned test (b) IC of KB cell proliferation ₅₀value; The 5th hurdle shows the IC of erbB2 Tyrosylprotein kinase protein phosphorylation in the middle clone that suppresses to derive from MCF7 of above-mentioned test (e) ₅₀value:

Table A

No. embodiment	IC ₅₀(μ M) tests (a): suppress EGFR Tyrosylprotein kinase protein phosphorylation	IC ₅₀(μ M) tests (a): suppress erbB2 Tyrosylprotein kinase protein phosphorylation	IC ₅₀(μ M) tests (b): suppress KB cell proliferation	IC ₅₀(μ M) tests (e): suppress erbB2 Tyrosylprotein kinase protein phosphorylation
					1	0.011	0.052	0.038	0.013
2	0.007	0.016	0.0021	0.035
					4	0.005	0.009	0.017	0.027
6	0.004	0.007	0.011	0.053

two kinds of compounds suppress specific activity to protein tyrosine kinase phosphorylation

Preferred compound embodiment 3 and document 7-Alkoxy-4-phenylamino-3-quinolinecarbonitriles as Dual Inhibitors ofSrc and Abl Kinases.Journal of Medicinal Chemistry (2004) in comparative study this patent, 47 (7), 1599-1601. in disclosed compound 4-[(2, the chloro-5-p-methoxy-phenyl of 4-bis-) amino]-6-methoxyl group-7-[(1-methyl-4-piperidyl) oxygen base]-3-quinolinecarbonitriles (4-[(2, 4-dichloro-5-methoxyphenyl) amino]-6-methoxy-7-[(1-methyl-4-piperidinyl) oxy]-3-Quinolinecarbonitrile) (CAS:681821-34-1) protein tyrosine kinase phosphorylation is suppressed to active.

Test method refers to " a) protein tyrosine kinase phosphorylation assay " under biological assay item, test adopts reconstitution cell internal fragment EGFR and erbB2 to clone and express in baculovirus/sf21 system, the inhibition ability of the peptide substrate that mensuration compound contains tyrosine to protein tyrosine kinase phosphorylation.Test-results shows, this patent embodiment 3 medicines are significantly better than documentation compound (p < 0.05) to EGFR and erbB2 protein tyrosine kinase phosphorylation inhibition, the results are shown in Table B.

Table B this patent embodiment 3 compounds and documentation compound suppress specific activity to protein tyrosine kinase phosphorylation

Invention a kind of medicinal compositions is provided on the other hand, this medicinal compositions comprises as the quinoline of formula I defined above or its pharmacy acceptable salt and pharmaceutically acceptable diluent or carrier.

Composition of the present invention can be form (for example tablet that is applicable to orally use, lozenge, hard or soft capsule, water-based or oily suspensions, emulsion, dispersible pulvis or granule, sugar dress agent or elixir), be applicable to the local form using (as creme, paste, jelling agent, water-based or oily solution or suspension), be applicable to the form (as finely divided powder or liquid aerosol) of inhalation, be applicable to be blown into the form (as finely divided powder agent) of administration or for example, for being applicable to form (sterile aqueous or the oily solution of parenterai administration, can be for vein, subcutaneous, intramuscular administration, or be the suppository for rectal administration).

Can adopt various conventional pharmaceutical excipient well known in the art, prepare composition of the present invention by ordinary method.Therefore, being intended for use the composition of oral administration can be containing just like one or more tinting materials, sweeting agent, correctives and/or sanitas.

The amount that one or more vehicle combine the active ingredient that obtains single formulation must change according to treated host and concrete route of administration.For example, the preparation that is intended for use people's oral administration is applicable to conventionally containing (being more suitable for as 0.5-100mg just like 0.5mg-0.5g active medicine, as be 1-30mg), and in said preparation, also contain appropriate vehicle, the amount of vehicle can be the approximately 5%-98% of composition total weight.

Certainly,, according to the known principle of medical field, in the time that formula I compound is used for the treatment of or prevents object, its dosage size should change according to animal or patient's disease character and severity, age and sex and route of administration.

In the time that formula I compound is used for the treatment of or prevents object, be generally, if desired can be with several divided dose administrations as the scope administration of 0.1mg/kg-75mg/kg body weight take for example per daily dose.Generally, in the time adopting parenteral administration, should give lower dosage.Therefore, for example, in the time of intravenously administrable, the dosage range conventionally adopting is as 0.1mg/kg-30mg/kg body weight.Similarly, in the time of inhalation, the dosage range conventionally adopting is for example 0.05mg/kg-25mg/kg body weight.But preferred oral administration, particularly with the form oral administration of tablet.Generally speaking, in unit dosage, contain the 0.5mg-0.5g compound of the present invention of having an appointment.

We find that the compounds of this invention has anti proliferative properties such as anticancer character, and the erbB2/EGF character that this character is considered to particularly to be mixed by its erbB family receptors tyrosine-kinase enzyme inhibition activity causes.

Therefore, expect that the compounds of this invention can be used for treatment by erbB receptor tyrosine kinase separately or disease or the illness of part mediation, described compound is used in and in the warm-blooded animal that needs this kind for the treatment of, produces erbB receptor tyrosine kinase restraining effect.Therefore, the compounds of this invention provides a kind of method for the treatment of malignant cell, the method is characterised in that one or more erbB family that suppresses receptor tyrosine kinase. specifically, the compounds of this invention can be used for producing by suppressing erbB receptor tyrosine kinase separately or antiproliferative and/or proapoptosis and/or the anti-erosion effect of part mediation.Especially expect that the compounds of this invention can be used for prevention or treats suppressing those tumours of one or more erbB receptor tyrosine kinase sensitivity, these receptor tyrosine kinases are relevant with the signal conduction step of these tumor cell proliferations of driving and existence.Therefore, expect that the compounds of this invention is by providing antiproliferative effect, can be used for treating psoriasis, benign prostate (BPH), atherosclerosis and restenosis and/or cancer, in particular for the cancer for the treatment of erbB receptor tyrosine kinase sensitivity. these optimum or malignant tumours can affect any tissue, comprise non-solid tumour, as leukemia, multiple myeloma or lymphatic cancer, and solid tumor, as bile duct, bone, bladder, brain/CNS, breast, colorectum, uterine endometrium, stomach, head and neck, liver, lung, neurone, oesophagus, ovary, pancreas, prostate gland, kidney, skin, testis, Tiroidina, uterus and the carcinoma of vulva.

One aspect of the present invention provides as the formula I compound of medicine or its pharmacy acceptable salt.

Another aspect of the present invention provides formula I compound or its pharmacy acceptable salt to produce the purposes of antiproliferative effect in such as people warm-blooded animal.

Therefore, one aspect of the present invention provides formula I quinoline as previously defined or its pharmacy acceptable salt in the purposes for the preparation of producing in such as people warm-blooded animal in the medicine of antiproliferative effect.

According to another feature of this aspect of the present invention, the warm-blooded animal the invention provides in this type for the treatment of of needs produces the method for antiproliferative effect in such as people, and the method comprises quinoline or its pharmacy acceptable salt of the formula I of the aforementioned definitions that gives described animal effective dose.

Another aspect of the present invention provide the quinoline of formula I of aforementioned definitions or its pharmacy acceptable salt for the preparation of prevention or treatment to suppressing erbB receptor tyrosine kinase such as the purposes in the medicine of those tumours of the associating sensitivity of EGFR and erbB2, it is relevant that described receptor tyrosine kinase and the signal that causes tumor cell proliferation conduct step.

This another feature on the one hand of the present invention provides erbB family to suppressing one or more receptor tyrosine kinase one prevention or methods for the treatment of such as those tumours of the associating sensitivity of EGFR and erbB2, described receptor tyrosine kinase with cause the signal conduction step of tumor cell proliferation and/or existence relevant, the method comprises quinoline or its pharmacy acceptable salt of the formula I of the aforementioned definitions that gives described animal effective dose.

According to another feature of this aspect of invention, the invention provides formula I compound or its pharmacy acceptable salt in prevention or treatment to suppressing erbB receptor tyrosine kinase such as the purposes in those tumours of the associating sensitivity of EGFR and erbB2, it is relevant that described receptor tyrosine kinase and the signal that causes tumor cell proliferation conduct step.

According to another aspect of the present invention, the invention provides the quinoline of formula I of aforementioned definitions or its pharmacy acceptable salt in the purposes for the preparation of providing in the EGFR of associating and the medicine of erbB2 tyrosine kinase inhibitory activity.

According to another feature of this aspect of the present invention, the invention provides a kind of method of EGFR and erbB2 tyrosine kinase inhibitory activity that associating is provided, the method comprises quinoline or its pharmacy acceptable salt of the formula I of the aforementioned definitions that gives described animal effective dose.

According to another feature of this aspect of the present invention, the invention provides EGFR for associating is provided and formula I compound or its pharmacy acceptable salt of erbB2 tyrosine kinase inhibitory activity.

According to another feature of this aspect of the present invention, the invention provides the quinoline of formula I of aforementioned definitions or its pharmacy acceptable salt purposes in for example, medicine for the preparation for the treatment of cancer (be selected from following cancer: leukemia, multiple myeloma, lymphatic cancer, bile duct, bone, bladder, brain/CNS, breast, colorectum, uterine endometrium, stomach, head and neck, liver, lung, neurone, oesophagus, ovary, pancreas, prostate gland, kidney, skin, testis, Tiroidina, uterus and the carcinoma of vulva).

According to another feature of this aspect of the present invention, the invention provides a kind of warm-blooded animal in this kind for the treatment of of needs treats cancer in such as people and (is for example selected from following cancer: leukemia, multiple myeloma, lymphatic cancer, bile duct, bone, bladder, brain/CNS, breast, colorectum, uterine endometrium, stomach, head and neck, liver, lung, neurone, oesophagus, ovary, pancreas, prostate gland, kidney, skin, testis, Tiroidina, uterus and the carcinoma of vulva) method, the method comprises quinoline or its pharmacy acceptable salt of the formula I of the aforementioned definitions that gives described animal effective dose.

According to another feature of this aspect of the present invention, the invention provides formula I compound or its pharmacy acceptable salt that one is used for the treatment of cancer (be for example selected from following cancer: leukemia, multiple myeloma, lymphatic cancer, bile duct, bone, bladder, brain/CNS, breast, colorectum, uterine endometrium, stomach, head and neck, liver, lung, neurone, oesophagus, ovary, pancreas, prostate gland, kidney, skin, testis, Tiroidina, in palace and the carcinoma of vulva).

As mentioned above, treatment or the large young pathbreaker of the needed dosage of a kind of disease specific of prophylactic treatment must change according to the severity of other factors, the host who treats, route of administration and the disease for the treatment of.

Antiproliferative treatment defined above can be used as single therapy method and uses separately, or except quinoline derivatives beyond the region of objective existence of the present invention, use can also combine with conventional therapeutic method of surgery, radiotherapy or chemical therapeutic method. this type of chemical therapeutic method can comprise one or more following all kinds of Hangzhoupro tumour medicines:

(i) antiproliferative/antitumour drug adopting in medical oncology, for example, as alkylating agent (cis-platinum, carboplatin, endoxan, mustargen, melphalan, Chlorambucil, busulfan and Nitrosourea); Antimetabolite (as anti-folic acid class (antifolates), for example fluoropyrimidines (as 5 FU 5 fluorouracil and Tegafur), Raltitrexed, the cry of certain animals of first ammonia butterfly, the Arabic grain glycosides of cytosine(Cyt) and hydroxyurea; Antitumor antibiotics (as anthracyclines, as Zorubicin, bleomycin, Dx, daunorubicin, epirubicin, darubicin, Mitomycin-C, actinomycin and duomycin); Antimitotic agent (as vinca alkaloids, as vincristin, vinealeucoblastine(VLB), vindesine and vinorelbine, and taxanes (taxoids), as taxol and taxotere); And topoisomerase enzyme inhibitor (as Etoposide class, as Etoposide and teniposide, amsacrine, Topotecan and camptothecine);

(ii) born of the same parents' inhibitor, if anti-estrogens is (as tamoxifen, Toremitene, raloxifene, droloxifene and iodoxyfene), estrogen receptor reduces conditioning agent (as fulvestrant), anti-androgens is (as bicalutamide, flutamide, Nilutamide and CPA), LHRT antagonist or LHRT agonist are (as goserelin, Leuprolide and buserelin), progestogens (as acetic acid megestrol), arimedex is (as Anastrozole, letrozole, fluorine chlorazol (vorazole) and Exemestane) and 5a-reductase inhibitor, as finasteride,

(iii) anticancer is invaded agent (as inhibitors of metalloproteinase (as Marimastat) and black kinases Profibrinolysin activator function of receptors inhibitor);

(iv) somatomedin depressant of functions, as this type of inhibitor packages draw together growth factor antibodies, growth because of in receptor antibody (as anti-erbb2 antibody trastuzumab [Herceptin ^tM] and anti-erbbl antibody Cetuximab [C225]), farnesyl transferase inhibitor, tyrosine kinase inhibitor and silk ammonia are expected threonine kinase enzyme inhibitors, the inhibitor of for example epidermal growth factor family is (as EGIR family Tyrosylprotein kinase suppresses other dose N-(the chloro-4-fluorophenyl of 3-)-7-methoxyl group-6-(3-morpholino propoxy-) quinazoline-4-amine (gefitinib, AZD1839), N-(3-ethynyl phenyl-6, two (2-methoxy ethoxy) quinazoline-4-amine (erlotinib of 7-, and 6-acrylamido-N-(the chloro-4-fluorophenyl-7-of 3-(3-morpholino propoxy-) quinazoline-4-amine (CI1033)) OSI-774), the inhibitor of for example inhibitor of thrombocyte class growth factor family and pHGF family,

(v) anti-angiogenic agent, as those suppress vascular endothelial growth factors in compound, (for example anti-angiogenic endothelium is carefully executed growth factor antibodies rhuMAb-VEGF [Avastin ^tM], disclosed compound in International Patent Application WO 97/22596, WO 97/30035, WO 97/32856 and WO 98/13354) and those compounds that work by other mechanism (as linomide, integral protein av β 3 depressant of functions and angiostatin);

(vi) vascular damaging agents, as disclosed compound in combretastatin A4 and International Patent Application WO 99/02166, WO00/40529, WO 00/41669, WO 01/92224, W002/04434 and WO 02/08213;

(vii) antisense therapy agent, for example, relate to the medicine (as ISIS 2503, a kind of anti-ras antisense drug) of above listed target;

(viii) gene therapy approach, comprise as replaced aberrant gene approach, as abnormal p53 or abnormal BRCA1 or BRCA2, GDEPT (relating to the therapy of the enzyme precursor medicine of gene) approach, as adopt those approach of Isocytosine deaminase, thymidine kinase or bacterium nitroreductase, and strengthen the approach of patient to chemotherapy or radiotherapeutic tolerance, as multi-medicament resistant gene therapy; With

(ix) immunotherapy approach, comprise the immunogenic in vitro and in vivo approach as strengthened patient tumors cell, as used cytokine (as interleukin II, interleukin-4 or rHuGM-CSF) transfection, reduce T-cell anergia approach, adopt transfection immunocyte (as the dendritic cell of cytokine transfection) approach, adopt the approach of the tumor cell line of cytokine transfection, and adopt the approach of anti-atopic antibody.

By simultaneously, order or each component of separately treating, can carry out this type of combination therapy.This type of joint product can use compound of the present invention in aforesaid dosage range, and other medical active agent is also used in the amount ranges of its approval.

A kind of medicinal product is provided according to an aspect of the present invention, this product comprise aforementioned definitions formula I quinoline and for other antitumour drug of the aforementioned definitions of cancer combination therapy.

Although formula I compound is mainly the medicine as warm-blooded animal (comprising people), while needs, they also can be for suppressing the effect of erbB receptor tyrosine protein kinase.Therefore, these compounds can also be as the medicinal standard product in new biological test exploitation and new pharmacology drug research.

Embodiment

Now by following non-limiting example, the present invention is further described, wherein, unless otherwise indicated, otherwise:

(i) degree be degree Celsius (℃); All operations, all in room temperature or envrionment temperature, carries out in the temperature range of 18-25 ℃;

(ii) organic solvent is through anhydrous magnesium sulfate drying; Adopt rotatory evaporator, at decompression (600-4000; 4.5-30mmHg), carry out the evaporation of solvent under not higher than the bath temperature of 60 ℃;

(iii) chromatography refers to flash chromatography on silica gel; Thin-layer chromatography (TLC) carries out on silica-gel plate;

(iv) conventionally after reaction process, carry out TLC and/or analytical LCMS, the reaction times providing is only for explanation;

(v) end product all has gratifying proton magnetic resonance (PMR) (NMR) spectrum and/or mass-spectrometric data;

(vi) productive rate of giving, only for explanation, might not be the yield obtaining by conscientiously studying flow process; If need more product, can repeat preparation;

(vii) in the time providing NMR data, unless otherwise indicated, NMR data with Main Diagnosis matter in δ value form provide, ppm (ppm) using the tetramethylsilane with respect to as internal standard substance (TMS) represents, adopting complete deuterated dimethyl (DMSO-d6) is solvent, under 400MHz, measures; Adopt following abbreviation: s, unimodal; D, bimodal; T, triplet; Q, quartet; M, multimodal; B, broad peak;

(viii) chemical symbol has common meaning; Use SI units and symbol;

(ix) the solvent ratio providing is volume: volume (v/v); With

(x) mass spectrum (MS) adopts 70 electron-volts of electron energies under chemi-ionization (CI) mode, and application directly exposes probe and detects, from being undertaken by electron spray(ES) in changing; Provide m/z value; Conventionally only the matter of report expression parent quality in; Unless otherwise indicated, mass ion is expressed as (MH) ⁺;

(xi) carbon that unless otherwise indicated, contains Asymmetrical substitute and/or the compound of sulphur atom are without fractionation;

(xii) in a certain synthetic and previous embodiment, be confused when similar when describing, the mmole ratio of institute's usage quantity is equal to the ratio using in described previous embodiment;

(xiii) adopt following abbreviation:

DCM methylene dichloride;

DMF DMF;

DMA N,N-dimethylacetamide;

THF tetrahydrofuran (THF);

(xiv) one synthetic while obtaining acid salt (as HCl salt) when describing, do not confirm the concrete stoichiometry of this salt;

(xv) in embodiment 1-12, unless otherwise indicated, all NMR data of report are free base material, and it is converted into the form of free alkali before evaluation by the salt separating.

Embodiment 1

4-(3-oxygen-2-fluoroanilino)-7-methoxyl group-6-{[1-(methoxycarbonyl) piperidin-4-yl] oxygen base }-3-itrile group quinoline

By methyl-chloroformate (23 μ l, 0.3mmol) dropwise add ice-cold 4-(the chloro-2-fluoroanilino of 3-)-7-methoxyl group-6-[(piperidin-4-yl) oxygen base]-3-itrile group quinoline (0.3mmol) and diisopropylethylamine (63 μ l, 0.36mmol) be in the mixture of methylene dichloride (5ml).Stirring the mixture 1 hour. organic solution is through water and salt water washing, dried over mgso.After vacuum is steamed and desolventized, residue generates title compound through chromatography purification (eluent: the dichloromethane solution of 2%7N ammonium formiate (methanolic ammonia)) on silica gel, is white solid (60mg). ¹h NMR (300MHz, DMSO-d ₆) δ ppm:1.90 (m, 2H), 2.00 (m, 2H), 3.44 (m, 2H), 3.72 (s, 3H), 3.84 (m, 2H), 4.01 (s, 3H), 4.64 (m, 1H); 7.17 (m, 3H), 7.42 (m, 2H), 8.54 (s, 1H), 9.77 (s, 1H); Mass spectrum: MH ⁺485.

As 4-(the chloro-2-fluoroanilino of 3-)-7-methoxyl group-6-[(piperidin-4-yl of starting raw material) oxygen base]-3-itrile group quinoline is prepared as follows:

Step 1

6-acetoxyl group-4-(the chloro-2-fluoroanilino of 3-)-7-methoxyl group-3-itrile group quinoline hydrochloride

Chloro-6-acetoxyl group-4-7-methoxyl group-3-itrile group quinoline and the chloro-2-fluoroaniline of 3-(3.46g, 23.8mmol) are suspended in Virahol (200ml).This mixture is heated to 80 ℃ to reflux 3 hours.Evaporating solvent, residue is crystallization from acetonitrile, generates hydrochloride product, is lightpink crystalline solid (7.36g):

¹h NMR (300MHz, DMSO-d ₆) δ ppm:2.37 (s, 3H), 4.00 (s, 3H), 7.34 (m, 3H), 7.61 (m, 2H), 8.62 (s, 1H), 9.76 (s, 1H); Mass spectrum: 386.4,388.4.

Step 2

4-(the chloro-2-fluoroanilino of 3-)-6-hydroxyl-7-methoxyl group-3-itrile group quinoline

6-acetoxyl group-4-from step 1 (the chloro-2-fluoroanilino of 3-)-7-methoxyl group-3-itrile group quinoline hydrochloride (21.9mmol) is dissolved in to (200ml) in methyl alcohol.Add strong aqua (15ml), under stirring, in 50 ℃ of heated solutions 2 hours, produce milk oil sample hydrochloric acid solid precipitation.Collect after filtration solid, through ether (3 × 200ml) washing, and exist the dry bath of vacuum under Vanadium Pentoxide in FLAKES to generate product in 60 ℃, be pale solid (5.26g);

¹h NMR (300MHz, DMSO-d ₆) δ ppm:3.95 (s, 3H), 7.33 (m, 3H), 7.50 (s, 1H), 7.64 (s, 1H), 8.32 (s, 1H), 9.23 (brs, 1H), 9.77 (s, 1H); Mass spectrum: 344.4,346.4.

Step 3

Uncle 6-{[(1--butoxy carbonyl) piperidin-4-yl] oxygen base }-4-(the chloro-2-fluoroanilino of 3-)-7-methoxyl group-3-itrile group quinoline

4-from step 2 (the chloro-2-fluoroanilino of 3-)-6-hydroxyl-7-methoxyl group-3-itrile group quinoline (5.85mmol) is dissolved in DMA (50ml).(as Chemical & Pharmaceutical Bulletin 2001,49 (7), prepared by 822-829 to add (4-methylsulfonyl oxygen base) piperidines-1-carboxylic acid tert-butyl ester; 490mg, 1.76mmol) and cesium fluoride (890mg, 5.85mmol), lower heating this mixture to 85 ℃ stirred.2 hours, 4 hours and 6 hours intermittently, to (4-methylsulfonyl oxygen base) piperidines-1-carboxylic acid tert-butyl ester and the cesium fluoride that adds above-mentioned amount in reaction mixture.After adding for the last time, continue again in 85 ℃ of heating 6 hours.Evaporating solvent, residue is at DCM (150ml) and H ₂in O (150ml), distributing. water layer is through DCM (4 × 100ml) extraction, combining extraction liquid and DCM layer.The DCM part merging is through MgSO ₄be dried and evaporate.Residue is through chromatography purification, through 0-2.5% (7: the dense NH of 1MeOH/ ₄the OH aqueous solution) DCM eluant solution.Merge suitable part evaporation, generate product, be light brown foam (2.25g);

¹h NMR (300MHz, DMSO-d ₆) δ ppm:1.40 (s, 9H), 1.60-1.65 (m, 2H), 1.95-2.00 (m, 2H), 3.20-3.25 (m, 2H), 3.65-3.70 (m, 2H), 3.92 (s, 3H), 4.68 (m, 1H), 7.21 (s, 1H), 7.27 (dd, 1H), 7.47 (ddd, 1H), 7.51 (dd, 1H), 7.85 (s, 1H), 8.56 (s, 1H), 9.77 (s, 1H); Mass spectrum: 527.5,529.5.

Step 4

4-(the chloro-2-fluoroanilino of 3-)-7-methoxyl group-6-[(piperidin-4-yl) oxygen base]-3-itrile group quinoline

By the uncle 6-{[(1--butoxy carbonyl from step 3) piperidin-4-yl] oxygen base }-4-(the chloro-2-fluoroanilino of 3-)-7-methoxyl group-3-itrile group quinoline (0.70mmol) is dissolved in trifluoroacetic acid (5ml), and places this solution 2 hours.Evaporate excessive trifluoroacetic acid, residue is through twice of DCM azeotropic.Residue, through chromatography purification, uses 0-4% (7: the dense NH of 1MeOH/ ₄the OH aqueous solution) DCM eluant solution.Evaporate suitable part and generate product, be pale solid (260mg):

¹h NMR (300MHz, DMSO-d ₆) δ ppm:1.53-1.64 (m, 2H), 2.00-2.05 (m, 2H), 2.64-2.72 (m, 2H), 3.00-3.07 (m, 2H), 3.92 (s, 3H), 4.60 (m, 1H), 7.20 (s, 1H), 7.26 (ddd, 1H), 7.47 (dd, 1H), 7.50 (dd, 1H), 7.82 (s, 1H), 8.43 (s, 1H), 9.77 (s, 1H); Mass spectrum: 427.2,429.2.

Embodiment 2

4-(the chloro-2-fluoroanilino of 3-)-7-methoxyl group-6-{[1-{2-(pyrrolidin-1-yl) ethoxy carbonyl } piperidin-4-yl] oxygen base }-3-itrile group quinoline

By 6-{[1-(2-chloroethoxy carbonyl) piperidin-4-yl] oxygen base }-4-(the chloro-2-fluoroanilino of 3-)-7-methoxyl group-3-itrile group quinoline (1mmol), tetramethyleneimine (0.33ml, 4mmol) and potassiumiodide (330mg, 2mmol) in N,N-DIMETHYLACETAMIDE in 80 ℃ heating 4 hours.After cooling, high vacuum evaporation solvent.Residue distributes then through dichloromethane extraction in water and methylene dichloride.

Organic layer is through water and salt water washing and through dried over mgso.After evaporating solvent, residue on silica gel through chromatography purification (eluent: the dichloromethane solution of 2-3%7N ammonium formiate) and further at the HPLC post (C18 of preparative HPLC-MS system, 5 microns, 19mm diameter, 100mm length) upper purifying, make water (comprising 5% methyl alcohol and 1% acetic acid) and acetonitrile wash-out (gradient).After evaporating solvent, residue is dissolved in methylene dichloride and wet chemical in.Organic layer is through dried over mgso.Evaporating solvent also grinds residue in pentane, obtains title compound, is white solid (228mg). ¹h NMR (300MHz, DMSO-d ₆) δ ppm:1.80 (m, 4H), 1.91 (m, 2H), 2.00 (m, 2H), 2.60 (m, 4H), 2.77 (t, 2H), 3.45 (m, 2H), 3.83 (m, 2H), 4.01 (s, 3H), 4.26 (t, 2H), 4.64 (m, 1H), 7.16 (m, 2H), 7.20 (s, 1H), 7.30 (m, 2H), 8.52 (m, 1H), 9.77 (s, 1H); Mass spectrum: MH ⁺568.

As 6-{[1-(the 2-chloroethoxy carbonyl) piperidin-4-yl of starting raw material] oxygen base }-4-(the chloro-2-fluoroanilino of 3-)-7-methoxyl group-3-itrile group quinoline is prepared as follows:

Mono Chloro Acetic Acid 2-chloroethene ester is dropwise added to ice-cold 4-(the chloro-2-fluoroanilino of 3-)-7-methoxyl group-6-[(piperidin-4-yl) oxygen base]-3-itrile group quinoline (5mmol) is in the mixture of the methylene dichloride (100ml) of diisopropylethylamine (1.05ml, 6mmol).Stir this mixture 1 hour in 0 ℃.Organic solution is through water and salt water washing, and magnesium sulfate is done bath.After vacuum is steamed and desolventized, residue generates title compound through chromatography purification (eluent: the dichloromethane solution of 2%7N ammonium formiate) on silica gel, is white solid (1.5g). ¹h NMR (300MHz, DMSO-d ₆) δ ppm:1.90 (m, 2H), 2.00 (m, 2H), 3.47 (m, 2H), 3.71 (t, 2H), 3.83 (m, 2H), 4.01 (s, 3H), 4.36 (t, 2H), 4.65 (m, 1H), 7.15 (m, 2H), 7.21 (s, 1H), 7.41 (m, 2H), 8.46 (s, 1H), 9.77 (s, 1H); Mass spectrum: MH ⁺533,535.

Embodiment 3

4-(the chloro-2,4 difluorobenzene amino of 3-)-7-methoxyl group-6-{[1-(methoxycarbonyl) piperidin-4-yl] oxygen base }-3-itrile group quinoline

By methyl-chloroformate (40 μ l, 0.48mmol) dropwise add ice-cold 4-(3-chloro-2,4-difluorobenzene amino)-7-methoxyl group-6-[(piperidin-4-yl) oxygen base] in methylene dichloride (2ml) mixture of-3-itrile group quinoline (0.48mmol) and diisopropylethylamine (170 μ l, 0.95mmol).Stir this mixture 90 minutes in 0 ℃.After evaporating solvent, residue is dissolved in to purifying in DMSO and on the HPLC post (C18,5 microns, 19mm diameter, 100mm length) of preparative HPLC-MS system, uses the water and the acetonitrile wash-out (gradient) that comprise 2g/l ammonium formiate.After vacuum evaporating solvent, residue on silica gel through chromatography purifying (eluent: the dichloromethane solution of 0-3%7N ammonium formiate) again.After evaporating solvent, residue grinds and generates title compound in acetonitrile, is white solid (35mg). ¹h NMR (300MHz, DMSO-d ₆) δ ppm:1.67 (m, 2H), 2.02 (m, 2H), 3.35 (m, 2H), 3.61 (s, 3H), 3.73 (m, 2H), 3.94 (s, 3H), 4.72 (m, 1H), 7.23 (s, 1H), 7.42 (m, 1H), 7.58 (m, 1H), 7.86 (s, 1H), 8.45 (s, 1H), 9.75 (s, 1H); Mass spectrum: MH ⁺503.

As 4-(the chloro-2,4 difluorobenzene amino of 3-)-7-methoxyl group-6-[(piperidin-4-yl of starting raw material) oxygen base]-3-itrile group quinoline is prepared as follows:

By chloro-3-2, Virahol (2ml) solution of 4-difluoroaniline (1.7g, 10.1mmol) and 5N hydrogenchloride adds the chloro-7-methoxyl group-3-of 4-[(4-itrile group quinoline-6-yl) oxygen base] piperidines-1-carboxylic acid tert-butyl ester (10.1mmol) is in the suspension of Virahol (50ml).Stir this mixture 3 hours in 80 ℃.After evaporating solvent, residue generates 4-(3-chloro-2 through chromatography purification (eluent: the dichloromethane solution of 5-10%7N ammonium formiate) on silica gel, 4-difluorobenzene amino)-7-methoxyl group-6-[(piperidin-4-yl) oxygen base]-3-itrile group quinoline (3.65g) is white solid. ¹h NMR (300MHz, DMSO-d ₆) δ ppm:2.15 (m, 2H), 2.30 (m, 2H), 3.34 (m, 2H), 3.47 (m, 2H), 4.01 (s, 3H), 4.91 (m, 1H), 7.03 (m, 2H), 7.58 (m, 2H), 8.32 (s, 1H), 9.54 (s, 1H); Mass spectrum: MH ⁺445.

Embodiment 4 and 5

By 4-(3-chloro-2,4-difluorobenzene amino)-6-{[1-(2-chloroethoxy carbonyl) piperidin-4-yl] oxygen base }-7-methoxyl group-3-itrile group quinoline (0.66mmol) and suitable amine (2.6mmol) and the mixture of potassiumiodide (220mg, 1.33mmol) in N,N-DIMETHYLACETAMIDE (5ml) be in 95 ℃ of heating 2 hours.Residue dilutes and mistake filter solid in methylene dichloride.Evaporation filtrate after, residue on silica gel through chromatography purification (eluent: the dichloromethane solution of 2%7N ammonium formiate).Evaporating solvent obtains title compound.

As the 4-(3-chloro-2 of starting raw material, 4-difluorobenzene amino)-6-{[1-(2-chloroethoxy carbonyl) piperidin-4-yl] oxygen base }-7-methoxyl group-3-itrile group quinoline is by chloroformic acid 2-chloroethene ester and 4-(3-chloro-2,4-difluorobenzene amino)-7-methoxyl group-6-[(piperidin-4-yl) oxygen base]-3-itrile group quinoline warp and same procedure preparation described in embodiment 2, obtain 465mg. ¹H NMR(300MHz，DMSO-d ₆)δppm：1.70(m，2H)，2.03(m，2H)，3.30(m，2H)，3.74(m，2H)，3.83(t，2H)，3.94(s，3H)，4.28(t，2H)，4.73(m，1H)，7.21(s，1H)，7.40(m，1H)，7.58(m，1H)，7.87(s，1H)，8.37(s，1H)，9.71(s，1H).

Embodiment 4

4-(3-oxygen-2,4 difluorobenzene amino)-7-methoxyl group-6-{[1-2-(pyrrolidin-1-yl) ethoxy carbonyl } piperidin-4-yl] oxygen base }-3-itrile group quinoline

Amine used is tetramethyleneimine (0.22ml, 2.6mmol).

Yield: 182mg. ¹h NMR (300MHz, DMSO-d ₆) δ ppm:1.80 (m, 4H), 1.90 (m, 2H), 2.00 (m, 2H), 2.57 (m, 4H), 2.76 (t, 2H), 3.43 (m, 2H), 3.83 (m, 2H), 4.01 (s, 3H), 4.25 (t, 2H), 4.64 (m, 1H), 7.07 (m, 1H), 7.21 (m, 2H), 7.30 (s, 1H), 8.36 (s, 1H), 9.77 (s, 1H); Mass spectrum: MH ⁺586.

Embodiment 5

4-(the chloro-2,4 difluorobenzene amino of 3-)-7-methoxyl group-6-{ (1-{2-(piperidin-1-yl) ethoxy carbonyl piperidin-4-yl] oxygen base }-3-itrile group quinoline

Amine used is piperidines.

Yield: 52mg. ¹h NMR (300MHz, DMSO-d ₆) δ ppm:1.45 (m, 2H), 1.60 (m, 4H), 1.90 (m, 2H), 2.00 (m, 2H), 2.46 (m, 4H), 2.63 (t, 2H), 3.43 (m, 2H), 3.83 (m, 2H), 4.01 (s, 3H), 4.24 (t, 2H), 4.64 (m, 1H), 7.07 (m, 1H), 7.18 (m, 1H), 7.21 (s, 1H), 7.30 (s, 1H), 8.35 (s, 1H), 9.71 (s, 1H); Mass spectrum: MH ⁺598.

Embodiment 6-9

By 6-{[1-(2-chloroethoxy carbonyl) piperidin-4-yl] oxygen base-4-(the chloro-2-fluoroanilino of 3-)-7-methoxyl group-3-itrile group quinoline (0.4mmol), potassiumiodide (134mg, 0.8mmol) and suitably the mixture of amine (1.6mmol) in N,N-DIMETHYLACETAMIDE (4ml) in 80 ℃ of heating 4 hours.After cooling, high vacuum evaporation solvent.Residue distributes in water and methylene dichloride, and through dichloromethane extraction. and organic layer is through water and salt water washing and through dried over mgso.After evaporating solvent, residue residue grinds and obtains title compound through chromatography purification (eluent: the dichloromethane solution of 2-3%7N ammonium formiate) and in pentane on silica gel.

Embodiment 6

4-(3-chlorine 2-fluoroanilino)-7-methoxyl group-6-{[1-{2-(piperidin-1-yl) ethoxy carbonyl } piperidin-4-yl] oxygen base }-3-itrile group quinoline

Amine used is piperidines.

Yield: 142mg, (reacting with 0.5mmol scale). ¹h NMR (300MHz, DMSO-d ₆) δ ppm:1.44 (m, 2H), 1.60 (m, 4H), 1.88 (m, 2H), 2.00 (m, 2H), 2.46 (m, 4H), 2.63 (t, 2H), 3.43 (m, 2H), 3.82 (m, 2H), 4.01 (s, 3H), 4.24 (t, 2H), 4.64 (m, 1H), 7.16 (m, 2H), 7.20 (s, 1H), 7.39 (m, 2H), 8.48 (s, 1H), 9.77 (s, 1H): mass spectrum: MH ⁺580.

Embodiment 7

4-(the chloro-2-fluoroanilino of 3-)-6-{[1-{2-(diethylamino) ethoxy carbonyl } piperidin-4-yl] oxygen base }-7-methoxyl group-3-itrile group quinoline

Amine used is diethylamine.

Yield: 100mg (reacting with excessive diethylamine far away in tube sealing). ¹h NMR (300MHz, DMSO-d ₆) δ ppm:1.04 (m, 6H), 1.90 (m, 2H), 1.99 (m, 2H), 2.59 (m, 4H), 2.73 (t, 2H), 3.72 (m, 2H), 3.82 (m, 2H), 4.01 (s, 3H), 4.18 (t, 2H), 4.64 (m, 1H), 7.16 (m, 2H), 7.20 (s, 1H), 7.35 (m, 2H), 8.48 (s, 1H), 9.77 (s, 1H); Mass spectrum: MH ⁺570.

Embodiment 8

4-(the chloro-2-chlorobenzene of 3-amino)-7-methoxyl group-6-{[1-{2-(morpholine-4-yl) ethoxy carbonyl } piperidin-4-yl] oxygen base }-3-itrile group quinoline

Amine used is beautiful jade.

Yield: 126mg. ¹h NMR (300MHz, DMSO-d ₆) δ ppm:1.91 (m, 2H), 2.00 (m, 2H), 2.53 (m, 4H), 2.65 (t, 2H), 3.44 (m, 2H), 3.72 (m, 4H), 3.83 (m, 2H), 4.01 (s, 3H), 4.25 (t, 2H), 4.64 (m, 1H), 7.16 (m, 2H), 7.20 (s, 1H), 7.33 (m, 2H), 8.46 (s, 1H), 9.77 (s, 1H); Mass spectrum: MH ⁺584.

Embodiment 9

4-(the chloro-2-fluoroanilino of 3-)-7-methoxyl group-6-{[1-{2-(4-methyl piperidine-1-yl) ethoxy carbonyl piperidin-4-yl] oxygen base }-3-itrile group quinoline

Amine used is 4-methyl piperidine.

Yield: 112mg. ¹h NMR (300MHz, DMSO-d ₆) δ ppm:1.89 (m, 2H), 2.00 (m, 2H), 2.28 (s, 3H), 2.6-2.3 (m, 8H), 2.68 (t, 2H), 3.44 (m, 2H), 3.82 (m, 2H), 4.01 (s, 3H), 4.25 (t, 2H), 4.64 (m, 1H), 7.16 (m, 2H), 7.20 (s, 1H), 7.32 (m, 2H), 8.48 (m, 1H), 9.77 (s, 1H); Mass spectrum: MH ⁺598.

Embodiment 10

Pharmaceutical composition

For example understand exemplary pharmaceutical dosage form of the present invention (activeconstituents is called as " compounds X ") as described herein below, for the treatment in human body or prophylactic applications:

(a) tablet I mg/ sheet

Compounds X 100

Lactose Ph.Eur. 182.75

Croscarmellose sodium 12.0

Corn starch paste (5%w/v paste) 2.25

Magnesium Stearate 3.0

(b) injection I (50mg/ml)

Compounds X 5.0%w/v

1M sodium hydroxide solution 15.0%v/v

0.1m hydrochloric acid (regulating pH to 7.6)

Poly(oxyethylene glycol) 400 4.5%w/v

Water for injection to 100%

Can obtain above-mentioned preparation by the known ordinary method of pharmaceutical field.For example tablet can be prepared in flakes by being mixed together each component pressing mixt.

Claims

1. the quinoline of formula I

Wherein n is 2 or 3,

Sub-formula (ii) group in formula I:

Sub-formula (iii) group:

Wherein, R ¹⁰halogen, R ¹¹halogen, and R ¹²it is hydrogen or halogen;

X ¹o;

R ¹be selected from (1-6C) alkyl, it optionally contains one or more hydroxyls or halogenic substituent;

R ²be (1-6C) alkyl, above-mentioned group can optionally be replaced by sub-formula (i):

Wherein m is 0,1,2 or 3;

R ³and R ⁴together with the nitrogen-atoms connecting with them, form pyrrolidine ring, morpholine ring, piperidine ring or piperazine ring, it is optionally replaced by (1-3C) alkyl at feasible nitrogen-atoms place.

2. according to the quinoline of claim 1, wherein m is 1,2 or 3.

3. according to the quinoline of claim 1, wherein R ¹⁰fluorine, R ¹¹chlorine, and R ¹²be selected from hydrogen or fluorine.

4. according to the quinoline of claim 3, wherein R ¹⁰fluorine, R ¹¹chlorine, and R ¹²hydrogen.

5. according to the quinoline of claim 3, wherein R ¹⁰fluorine, R ¹¹chlorine, and R ¹²it is fluorine.

6. according to the quinoline of claim 1, wherein R ¹-X ¹-be selected from methoxyl group, oxyethyl group.

7. according to the quinoline of claim 1, it is the compound of formula IA:

Wherein R ²as claim 1 defines, R ¹⁰, R ¹¹and R ¹²as the definition of claim 3-5 any one, and R ¹³be selected from methoxyl group, oxyethyl group.

8. according to the quinoline of claim 1, it is the compound of formula IB:

Wherein R ²as claim 1 defines, and R ¹³be selected from methoxyl group, oxyethyl group.

9. according to the quinoline of claim 1, it is the compound of formula IC:

10. according to the quinoline of claim 1, wherein R ²be (1-3C) alkyl, its sub-formula (i) group optionally being defined by claim 1 or claim 2 replaces.

11. quinoline according to claim 1, wherein R ²comprise sub-formula (i) substituting group defining as claim 1 or claim 2.

12. according to the quinoline of claim 1, wherein R ³and R ⁴together with the nitrogen-atoms connecting with them, form pyrrolidine ring.

13. according to the quinoline of claim 1, wherein R ²be selected from methyl, 2-(pyrrolidin-1-yl) ethyl, 2-(piperidyl) ethyl, 2-(morpholine-4-yl) ethyl and 2-(4-methylpiperazine-1-yl) ethyl.

14. quinoline, it is selected from following one or more:

Or its pharmacy acceptable salt.

The preparation method of the quinoline of 15. claims 1, it is selected from:

Method (a) formula II compound:

Wherein R ¹, X ¹, R ⁵define as claim 1 with n,

React with formula III compound:

Wherein R ²as claim 1 defines, and Lg is replaceable group,

Method (c) formula IV compound:

Wherein R ¹, X ¹, R ⁵, as defining, claim 1 reacts with formula V compound with n:

Wherein R ²as above definition and Lg are replaceable groups;

Method (g) makes formula VI compound:

Wherein R ², R ⁵define as claim 1 with n, with formula R ¹the reaction of-Lg compound, wherein R ¹as claim 1 defines and Lg is replaceable group;

Method (j) formula VIII compound:

Wherein R ¹, R ², X ¹as claim 1 defines and Lg is replaceable group,

Aniline reaction with formula IX:

Wherein R ⁵define as claim 1 with n;

Method (k) makes formula X compound:

Wherein R ⁵, X ¹, R ¹define as claim 1 with n, and wherein Lg is leavings group, with formula R ²the reaction of-OH alcohol, wherein R ²as claim 1 defines;

Wherein R ¹, X ¹, R ⁵define R as claim 1 with n ¹⁵be (1-6C) alkyl, and Lg is leavings group,

With formula R ³r ⁴the reaction of NH compound, wherein R ³and R ⁴as claim 1 defines.

16. methods according to claim 15, wherein, in method (c), Lg is halogen.

17. methods according to claim 16, wherein, in method (c), Lg is chlorine or bromine.

18. methods according to claim 15, wherein, described quinoline prepares by following method:

The described formula II compound of method (e) reacts under Mitsunobu condition with the described formula III compound that is OH except Lg.

19. 1 kinds of pharmaceutical compositions, it comprises as quinoline or its pharmacy acceptable salt of the definition of claim 1-14 any one and combines by pharmaceutically acceptable diluent or carrier.

20. if the quinoline of claim 1-14 any one definition or its pharmacy acceptable salt are in the purposes of medicine for the preparation of suppressing erbB2 receptor tyrosine kinase.

21. according to the purposes of claim 20, and wherein said medicine is also for suppressing EGFR receptor tyrosine kinase.

22. formula VI, X or the XI compounds that define as claim 15, substituting group wherein as defined in claim 15.