CN102558173B

CN102558173B - Compound inhibiting conduction of WNT signal, and composition and application thereof

Info

Publication number: CN102558173B
Application number: CN201010624876.5A
Authority: CN
Inventors: 安松柱; 李楚芳; 黄晨; 周桂生
Original assignee: Curegenix Inc
Current assignee: Curegenix Inc
Priority date: 2010-12-31
Filing date: 2010-12-31
Publication date: 2015-05-20
Anticipated expiration: 2030-12-31
Also published as: CN102558173A

Abstract

The invention relates to a compound shown in a general formula (I), a composition of the compound, and a method for inhibiting the conduction of a WNT signal by using the compound and the composition of the compound. The compound and the composition of the compound disclosed by the invention can effectively inhibit the secretion of WNT protein and the conduction of the WNT signal, thus having a great treatment effect on WNT-mediated diseases.

Description

Suppress the compound of WNT intracellular signaling, composition and application thereof

Technical field

The present invention relates to a kind of compound of the WNT of suppression intracellular signaling and comprise the pharmaceutical composition of this compound, and described compound or pharmaceutical composition are suppressing the application in WNT intracellular signaling.

Background technology

WNT signal path has vital effect to the fetal development of adult animal and adult stable state, and this signal path mainly comprises following process: 1, the generation of WNT protein and secretion; 2, the combination of WNT protein and cell associated receptor; 3, a series of biochemical reaction (Mikels and Nusse, 2006 in the cell that WNT-receptors bind causes; MacDonald, 2009; Moon, 2005).

Classical WNT signal path is combined with Frizzled and the LRP5/6 coreceptor of cell surface by WNT protein and is activated, thus causes beta chain protein delivery in nucleus, and TCF/LEF family transcription factor interaction, promotes transcribing of specific gene.

The conduction of non-classical WNT signal path completes primarily of a series of different intracellular protein.The function of this signal paths is mainly reflected in the polarity controlling insect plane cell and other physiological development process, as the formation of vertebrates gastrula.

The function of WNT signal path is also embodied in the versatility and differentiation (Nusse, 2008) that control embryonic stem cell and adult stem cell.For example, in the growth course of gastrula, the activation of the formation of former bar and WNT signal orientation in embryo has close relationship (ten Berge, 2008).From the broad variety cell that embryonic stem cell is derivative, such as heart cell, pancreatic cell, dopamine neuronal cell, liver cell, be all subject to regulation and control (Yang, 2008 of WNT signal path; D ' Amour, 2006; Inestrosa and Arenas, 2010; Sullivan, 2010).WNT signal path, in the growth of bone tissue, comprises the generation of osteogenic tissue and the generation of cartilaginous tissue, all plays very important role (Hoeppner, 2009; Chun, 2008).In addition, WNT signal path also has important effect (Lie, 2005) in the neuron regeneration of adult central nervous system.

The exception of WNT signal path activity can cause a series of relevant disease.Such as, the activation of classical WNT signal path can cause the misgrowth (Reya and Clevers, 2005) of cell.It is worth noting most, 90% colorectal carcinoma is the main supressor due to WNT/ beta chain protein signal path---the disappearance of tumprigenicity polyp of colon (APC) gene function and causing (Kinzler and Vogelstein, 1996).The high expression level of WNT protein, or the disappearance of the extracellular supressor of suppression WNT protein function, also can cause producing the tumour (Polakis, 2007) relevant to WNT signal path.Nearest research shows, non-classical WNT signal path many cancers occur and development process in work (Camilli and Weeraratna, 2010).Up-to-date achievement in research also shows the generation (Takahashi-Yanaga and Kahn, 2010) that WNT signal path relates to tumor stem cell.

A large amount of evidence display WNT signal transduction pathway magnetic target therapies is widely used (Barker and Clevers, 2006) in the treatment of numerous disease.The sudden change of the APC, beta chain albumen or the axle albumen-1 that cause the homoeostasis of classical WNT path to activate is most important in many development of cancer of people, comprise (Polakis, 2007) such as straight colorectal carcinoma, malignant melanoma, hepatocellular carcinoma, cancer of the stomach, ovarian cancers.In kinds cancer, use genetic method or chemical process to suppress WNT path can limit abnormal Growth of Cells (Herbst and Kolligs, 2007) significantly.Further, suppress this path may to support growth of cancer cells and cause cancer metastasis because have direct effect, these factors are considered to the main resistance factor to traditional embolic chemotherapy.

Except the WNT activity caused because the gene product of WNT receptor downstream is undergone mutation raises, the abnormal activity of the WNT path that other mechanism causes also with many cancers there is close contacting.These cancers mainly comprise: the cancer such as lung (minicell, non-small cell) cancer, mastocarcinoma, prostate cancer, carcinoid tumor, bladder cancer, cancer of the stomach, carcinoma of the pancreas, liver cancer (liver cell, liver protoblast), colorectal cancer, neck tesselated epithelium, esophagus cancer, ovarian cancer, cervical cancer, carcinoma of endometrium, lung carcinoma mesothelial, malignant melanoma, sarcoma, osteosarcoma, thyroid carcinoma, fibroma durum, acute myeloblastic leukemia, chronic myelocytic leukemia.Cancer cells depends on the autocrine of WNT signal path or the rise of paracrine signal to have had many examples to confirm.From osteosarcoma, colorectal cancer, mammary cancer, head and neck cancer and ovarian cancer cancerous cell line confirmed that the WNT signal of autocrine or paracrine can protect cancer cells to enter apoptosis (Kansara, 2009; Bafico, 2004; Akiri, 2009; DeAlmeida, 2007; Chan, 2007; Chen, 2009; And Rhee, 2002).

Further, abnormal WNT path has been presented in Fibrotic generating process has effect, mainly comprise: pulmonary fibrosis (idiopathic pulmonary fibrosis, radioactive fibrosis), renal fibrosis and hepatic fibrosis (Morrisey, 2003; Hwang, 2009; Cheng, 2008).

The Other diseases relevant to abnormal WNT intracellular signaling mainly includes but not limited to: bone or cartilage disease, such as osteoporosis and osteoarthritis; The obesity that type-II diabetes is relevant; Nerve degenerative diseases is as alzheimer's disease (Hoeppner, 2009; Ouchi, 2010; Blom, 2010 and Boonen, 2009).The self of WNT intracellular signaling and hemopoietic stem cell and remain relevant, therefore the functional disorder of WNT intracellular signaling can cause the various diseases relevant to hemopoietic stem cell, as leukemia and multiple other cancer relevant to blood (Reya, 2005).

Therefore, find the method and compound that can regulate and control the cell response that WNT path is correlated with, by for providing an effective approach with the treatment of this signal paths relative disease.

Summary of the invention

The object of this invention is to provide a kind of compound that can be used on the inhibitor of WNT intracellular signaling and the method suppressing WNT intracellular signaling with it.

" the WNT signal transduction pathway " or " WNT path " mentioned in this article refers to the path that the combination of WNT protein and cell receptor causes cell behavior to change.WNT path comprises much albumen, comprise FZ (Frizzled), disheveled protein (Disheveled), axle albumen (Axin), APC, GSK3 β, beta chain albumen, LEF/TCF transcription factor, and to the synthesis of WNT protein with secrete the relevant factor.To the synthesis of WNT protein with secrete the relevant factor and comprise, the albumen such as WNTless/evenness interrupted (Wls/Evi), porcupine (Porcn), Vps35p.Wherein Wls/Evi is a kind of albumen with seven transmembrane structure, mainly concentrates on golgi body, can secrete Wg (fruit bat), MOM-2 (nematode) and WNT3A.It comprises the structural motif of a high conservative, and the structure and function of this die body is also in unknown state at present.Porcn is film a member in conjunction with O-acyltransferase (MBOAT) family of palmitoyl transferase.The modification of lipid acid is extremely important for the function of WNT.WNT protein carries out palmitoylation modification on the position of one or two high conservative.The supressor of Porcn can stop the synthesis of all WNT, thus affects the function of WNT signal path.Vps35p is the subunit being referred to as retromer albumen composition, and the function of this albumen composition relates generally to the transhipment of intracellular protein.The main wherein effect of Vps35p is combining target albumen, such as WNT protein, forms transport vesica.

Term " WNT intracellular signaling supressor " or " WNT path supressor " normally micromolecular compound, molecular weight, greatly about below 800g/mol, can suppress the activity of WNT signal path.

The term method of intracellular signaling " suppress WNT " refers to the method for the biochemical reaction suppressing known, and generation or the thin intramicellar reaction of WNT protein of described biochemical reaction and functional WNT protein are relevant.As noted herein, small molecules may suppress WNT to react, and is just meeting this definition.

WNT protein is a kind of ligandin jointly combined with FZ and LRP5/6 two kinds of acceptors, and can activate classical or non-classical WNT signal path.WNT protein mainly comprises: WNT-1 (NM005430), WNT-2 (NM003391), WNT-2B/WNT-13 (NM004185), WNT-3 (NM030753), WNT3a (NM033131), WNT-4 (NM030761), WNT-5A (NM003392), WNT-5B (NM032642), WNT-6 (NM006522), WNT-7A (NM004625), WNT-7B (NM058238), WNT-8A (NM058244), WNT-8B (NM003393), WNT-9A/WNT-14 (NM003395), WNT-9B/WNT-15 (NM003396), WNT-10A (NM025216), WNT-10B (NM003394), WNT-11 (NM004626), WNT-16 (NM016087).

" WNT pathway dependent diseases " refers to the state or disease that cause when WNT intracellular signaling is in error state (ERST).On the one hand, abnormal WNT intracellular signaling refers in the cell suffering from certain disease or tissue, has the WNT intracellular signaling level exceeding normal cell or tissue.In a particular aspects, the disease that WNT path is relevant comprises cancer and fibrosis.

The growth that when term " cancer " refers to that human body is in pathological state, cell is unordered.Current cancer main Types includes but not limited to: cancer, lymphoma, blastoma and leukemia.Cancer more specifically example includes but not limited to: the cancer such as lung (minicell, non-small cell) cancer, mastocarcinoma, prostate cancer, carcinoid tumor, bladder cancer, cancer of the stomach, carcinoma of the pancreas, liver (liver cell, liver protoblast), colorectal cancer, neck squamous cell carcinoma, esophagus cancer, ovarian cancer, cervical cancer, carcinoma of endometrium, lung carcinoma mesothelial, malignant melanoma, sarcoma, osteosarcoma, thyroid carcinoma, fibroma durum, acute myeloblastic leukemia, chronic myelocytic leukemia.

Term " fibrosis " mainly refers to that people is in a kind of pathological state, the principal character of this case state be have random growth inoblast and organize stiff.Main disease comprises: pulmonary fibrosis (idiopathic pulmonary fibrosis, radioactive fibrosis), renal fibrosis, hepatic fibrosis comprise liver cirrhosis.

" suppression ", " treatment " or " methods for the treatment of " refers to the remission method of disease, methods for the treatment of and/or prevention method, and object is wherein the pathological condition of alleviating or curing relative disease.In one embodiment, the inhibitor of WNT signal transduction pathway is applied to cancer patients, the tumour in patient body likely can diminish." treatment " or " methods for the treatment of " is mainly manifested in: 1, suppress the symptom of disease to further develop; 2, improve or alleviate the disease symptoms and situation that have shown at present or experienced; 3, obvious, detectable improvement result is had to the disease symptoms shown at present or experiencing and situation.WNT path supressor can anticancer growth and/or directly kill cancer cells, this may the growth of also anticancer, or has cytotoxicity.

Term " treatment significant quantity " refers to the dosage effectively can treating the WNT path supressor needed for WNT signal transduction pathway relative disease in patient or Mammals.In cancer therapy, the treatment significant quantity of medicine effectively can reduce the quantity of cancer cells, the size reducing tumour, other tissue of inhibition tumor cell wet periphery (perimeter) and organ, the migration of inhibition tumor cell, Tumor suppression and grow to a certain degree, and/or can alleviate one or more symptoms relevant to cancer to a certain extent.

" Combined Preparation " in conjunction with one or several treatment reagent refer to different order simultaneously or continuity administration.In this article, the medicament production that term " drug regimen " refers to mixing or combines activated effective constituent and obtain, comprises the fixing of effective constituent and non-fixed combinations.Term " fixed Combination " refers to that effective constituent is as the compound of molecular formula (1) and another composition, simultaneously for patient under the condition of single formulation or dosage.Term " non-fixed combinations " refers to that effective constituent is as the compound of molecular formula (1) and another composition, with the while of different formulations or in succession for patient, do not have other special time limitation, wherein this administering mode can provide the treatment level of significance of activeconstituents in patient body.The second administering mode is also applied to cocktail type treatment, such as, combine the administration of three kinds or more effective constituent.

" chemical treatment reagent " refers to the compound being applied to cancer therapy.Main example includes but not limited to: gemcitabine, irinotecan, Zorubicin, 5 FU 5 fluorouracil, cytosine arabinoside (" Ara-C "), endoxan (Cyclophosphamide), tespamin, busulfan, endoxan (Cytoxin), taxol, methotrexate, cis-platinum, melphalan, alkali green for a long time and carboplatin.

First object of the present invention is to provide compound and the pharmacy acceptable salt thereof that one has general formula (1):

Wherein,

X ₁, X ₂, X ₃, X ₄, X ₅, X ₆, X ₇, X ₈be CR independently ₄or N;

Y ₁for hydrogen or CR ₄;

Y ₂, Y ₃be hydrogen, halogen or CR independently ₃;

R ₁for morpholinyl, piperazinyl, quinolyl, aryl, C _1-6heterocycle, to be selected from heteroatomic 5 or 6 yuan of heteroaryls of N, O and S containing 1-2, wherein each can optionally by R ₄replace;

R ₂for hydrogen, halogen, morpholinyl, piperazinyl, quinolyl, aryl, C _1-6heterocycle, to be selected from heteroatomic 5 or 6 yuan of heteroaryls of N, O and S containing 1-2, wherein each can optionally by R ₄replace;

Preferably, described 5 or 6 yuan of heteroaryls include but not limited to following radicals:

Preferably, R ₁and R ₂can independently and optionally by 1-2 R ₄group replaces;

R ₃for hydrogen, halogen, cyano group, C _1-6alkyl or C _1-6alkoxyl group, wherein C _1-6alkyl and C _1-6alkoxyl group is optionally replaced by halogen, amino, hydroxy, alkoxyl group or cyano group;

R ₄for hydrogen, halogen, C _1-6alkoxyl group ,-S (O) ₂oR ₅,-C (O) OR ₅,-C (O) R ₅,-C (O) NR ₆r ₇, C _1-6alkyl, C _2-6thiazolinyl or C _2-6alkynyl, wherein each can optionally be replaced by halogen, amino, hydroxy, alkoxyl group or cyano group;

R ₅, R ₆and R ₇be hydrogen, C independently _1-6alkyl, C _2-6thiazolinyl or C _2-6alkynyl, wherein each can optionally be replaced by halogen, amino, hydroxy, alkoxyl group or cyano group;

R ₈for hydrogen or C _1-6alkyl.

As in present specification use, any substituting group (such as CH in compound ₂) in H atom comprise all suitable isotropic substance variations, such as H, ²h and ³h.

Preferably, the application's compound arbitrarily other atom substituent can be Isotopic variations, includes but not limited to ¹¹c, ¹³c, ¹⁴c, ¹⁵n, ¹⁷o, ¹⁸o, ³⁵s, ¹⁸f, ³⁶i and/or ¹²³i.

Above-mentioned isotropic substance variation will have Detectable effects in the research application of compound, effect and distribution.

Preferably, molecular formula (1) has but is not limited to following core texture:

In a preferred embodiment, compound of the present invention includes but not limited to:

6-(2-picoline-4-base)-N-(4-(2-picoline-4-base) phenmethyl)-2,7-naphthyridines-1-amine;

N-(3-methyl-4-(2-picoline-4-base) phenmethyl)-6-(2-picoline-4-base)-2,7-naphthyridines-1-amine;

6-(3-fluorophenyl)-N-((2 '-methyl-[2,4 '-dipyridyl]-5-base) methyl) isoquinoline 99.9-1-amine;

2-(2-picoline-4-base)-N-(4-(2-picoline-4-base) phenmethyl)-1,6-naphthyridines-5-amine;

N-(4-(2-picoline-4-base) phenmethyl)-2-phenylpyridine also [3,4-b] pyrazine-5-amine;

N-(4-(2-picoline-4-base) phenmethyl)-6-(pyridin-4-yl)-2,7-naphthyridines-1-amine;

N-(4-(2-picoline-4-base) phenmethyl)-6-phenyl-2,7-naphthyridines-1-amine;

6-(3-chloro-phenyl-)-N-(4-(2-picoline-4-base) phenmethyl)-2,7-naphthyridines-1-amine;

6-(3-fluorophenyl)-N-(4-(2-picoline-4-base) phenmethyl)-2,7-naphthyridines-1-amine;

6-(3-fluorophenyl)-N-((2 '-methyl-[2,4 '-dipyridyl]-5-base) methyl)-2,7-naphthyridines-1-amine;

6-(3-fluorophenyl)-N-(4-(2-(trifluoromethyl) pyridin-4-yl) phenmethyl)-2,7-naphthyridines-1-amine;

N-((2 ', 3-dimethyl-[2,4 '-dipyridyl]-5-base) methyl)-6-(3-fluorophenyl)-2,7-naphthyridines-1-amine;

N-(4-(2-picoline-4-base) phenmethyl)-6-(pyrimidine-5-base)-2,7-naphthyridines-1-amine;

6-(5-picoline-3-base)-N-(4-(2-picoline-4-base) phenmethyl)-2,7-naphthyridines-1-amine;

6-(6-picoline-3-base)-N-(4-(2-picoline-4-base) phenmethyl)-2,7-naphthyridines-1-amine;

3-(8-((4-(2-picoline-4-base) phenmethyl) is amino)-2,7-naphthyridines-3-bases) cyanobenzene;

4-(8-((4-(2-picoline-4-base) phenmethyl) is amino)-2,7-naphthyridines-3-bases) cyanobenzene;

6-(4-fluorophenyl)-N-(4-(2-picoline-4-base) phenmethyl)-2,7-naphthyridines-1-amine;

N-(4-(2-picoline-4-base) phenmethyl)-6-(m-tolyl)-2,7-naphthyridines-1-amine;

N-(4-(2-picoline-4-base) phenmethyl)-2,7-naphthyridines-1-amine;

N-(4-(2-picoline-4-base) phenmethyl)-6-(pyridine-2-base)-2,7-naphthyridines-1-amine;

6-(2-fluorine pyridin-4-yl)-N-(4-(2-picoline-4-base) phenmethyl)-2,7-naphthyridines-1-amine;

6-(2-fluorophenyl)-N-(4-(2-picoline-4-base) phenmethyl)-2,7-naphthyridines-1-amine;

N-(4-(2-picoline-4-base) phenmethyl)-6-(pyridin-3-yl)-2,7-naphthyridines-1-amine;

N-([1,1 '-xenyl]-4-ylmethyl)-6-(2-picoline-4-base)-2,7-naphthyridines-1-amine;

6-(2-picoline-4-base)-N-((5-phenylpyridine-2-base) methyl)-2,7-naphthyridines-1-amine;

6-(3-fluorophenyl)-N-((2 '-(trifluoromethyl)-[2,4 '-dipyridyl]-5-base) methyl)-2,7-naphthyridines-1-amine;

N-(the fluoro-4-of 3-(2-fluorine pyridin-4-yl) phenmethyl)-6-(2-picoline-4-base)-2,7-naphthyridines-1-amine;

6-(2-picoline-4-base)-N-((2 '-(trifluoromethyl)-[2,4 '-dipyridyl]-5-base) methyl)-2,7-naphthyridines-1-amine;

N-((fluoro-2 '-(trifluoromethyl) of 3--[2,4 '-dipyridyl]-5-base) methyl)-6-(2-picoline-4-base)-2,7-naphthyridines-1-amine;

N-(the fluoro-4-of 3-(2-picoline-4-base) phenmethyl)-6-(2-picoline-4-base)-2,7-naphthyridines-1-amine;

N-((2 '-fluoro-[2,4 '-dipyridyl]-5-base) methyl)-6-(2-picoline-4-base)-2,7-naphthyridines-1-amine;

4-(5-(((6-(2-picoline-4-base)-2,7-naphthyridines-1-bases) are amino) methyl) pyridine-2-base) thiomorpholine 1,1-dioxide;

6-(2-picoline-4-base)-N-(4-(pyridazine-4-base) phenmethyl)-2,7-naphthyridines-1-amine;

N-(4-(2-picoline-4-base) phenmethyl)-6-(pyrazine-2-base)-2,7-naphthyridines-1-amine;

N-(4-(2-picoline-4-base) phenmethyl)-6-(pyridazine-4-base)-2,7-naphthyridines-1-amine;

N-(4-(2-picoline-4-base) phenmethyl)-6-morpholinyl-2,7-naphthyridines-1-amine;

6-(4-methylpiperazine-1-yl)-N-(4-(2-picoline-4-base) phenmethyl)-2,7-naphthyridines-1-amine;

4-(8-((4-(2-picoline-4-base) phenmethyl) is amino)-2,7-naphthyridines-3-bases) thiomorpholine 1,1-dioxide;

N-(the fluoro-4-of 3-(2-fluorine pyridin-4-yl) phenmethyl)-6-(3-fluorophenyl)-2,7-naphthyridines-1-amine;

N-(the fluoro-4-of 3-(2-picoline-4-base) phenmethyl)-6-(3-fluorophenyl)-2,7-naphthyridines-1-amine;

N-((fluoro-2 '-(trifluoromethyl) of 3--[2,4 '-dipyridyl]-5-base) methyl)-6-(3-fluorophenyl)-2,7-naphthyridines-1-amine;

N-((2 '-fluoro-[2,4 '-dipyridyl]-5-base) methyl)-6-(3-fluorophenyl)-2,7-naphthyridines-1-amine;

6-(3-fluorophenyl)-N-(3-methyl-4-(2-picoline-4-base) phenmethyl)-2,7-naphthyridines-1-amine;

4-(5-(((6-(3-fluorophenyl)-2,7-naphthyridines-1-base) is amino) methyl) pyridine-2-base) thiomorpholine 1,1-dioxide;

N-(4-chlorophenylmethyl)-6-(2-picoline-4-base)-2,7-naphthyridines-1-amine;

N-(4-methylbenzyl)-6-(2-picoline-4-base)-2,7-naphthyridines-1-amine;

6-(2-picoline-4-base)-N-(pyridin-3-yl methyl)-2,7-naphthyridines-1-amine;

N-([1,1 '-xenyl]-4-ylmethyl)-6-(3-fluorophenyl) isoquinoline 99.9-1-amine;

N-((2-fluoro-[1,1 '-xenyl]-4-base) methyl)-6-(3-fluorophenyl) isoquinoline 99.9-1-amine;

N-((2 '-methyl-[2,4 '-dipyridyl]-5-base) methyl)-6-phenyl isoquinolin quinoline-1-amine;

6-(3-chloro-phenyl-)-N-((2 '-methyl-[2,4 '-dipyridyl]-5-base) methyl) isoquinoline 99.9-1-amine;

N-(4-(2-picoline-4-base) phenmethyl)-6-phenyl isoquinolin quinoline-1-amine;

6-(2-picoline-4-base)-N-(4-(2-picoline-4-base) phenmethyl) isoquinoline 99.9-1-amine;

N-(4-(2-picoline-4-base) phenmethyl)-6-(pyridin-4-yl) isoquinoline 99.9-1-amine;

6-(6-picoline-3-base)-N-(4-(2-picoline-4-base) phenmethyl) isoquinoline 99.9-1-amine;

N-(4-(2-picoline-4-base) phenmethyl)-6-(m-tolyl) isoquinoline 99.9-1-amine;

N-(4-(2-picoline-4-base) phenmethyl)-6-(pyridin-3-yl) isoquinoline 99.9-1-amine;

N-(4-(2-picoline-4-base) phenmethyl)-6-(pyrazine-2-base) isoquinoline 99.9-1-amine;

N-(4-(2-picoline-4-base) phenmethyl)-6-(pyridazine-4-base) isoquinoline 99.9-1-amine;

N-((2 '-methyl-[2,4 '-dipyridyl]-5-base) methyl)-6-(pyrazine-2-base) isoquinoline 99.9-1-amine;

N-((2 ', 3-dimethyl-[2,4 '-dipyridyl]-5-base) methyl)-6-(pyrazine-2-base) isoquinoline 99.9-1-amine;

N-((2 ', 3-dimethyl-[2,4 '-dipyridyl]-5-base) methyl)-6-(pyridine-2-base) isoquinoline 99.9-1-amine;

N-((2 ', 3-dimethyl-[2,4 '-dipyridyl]-5-base) methyl)-6-(3-fluorophenyl) isoquinoline 99.9-1-amine;

N-((2 ', 3-dimethyl-[2,4 '-dipyridyl]-5-base) methyl)-6-(5-picoline-3-base) isoquinoline 99.9-1-amine;

N-phenmethyl-2-(3-fluorophenyl)-1,6-naphthyridines-5-amine;

2-(3-fluorophenyl)-N-((2 '-methyl-[2,4 '-dipyridyl]-5-base) methyl)-1,6-naphthyridines-5-amine;

N-((2 '-methyl-[2,4 '-dipyridyl]-5-base) methyl)-2-(2-picoline-4-base)-1,6-naphthyridines-5-amine;

N-((6-(3-fluorophenyl) pyridin-3-yl) methyl)-2-(2-picoline-4-base)-1,6-naphthyridines-5 amine;

N-(4-(2-fluorine pyridin-4-yl) phenmethyl)-2-(2-picoline-4-base)-1,6-naphthyridines-5-amine;

2-(2-picoline-4-base)-N-(4-(2-(trifluoromethyl) pyridin-4-yl) phenmethyl)-1,6-naphthyridines-5-amine;

N-((2 ', 3-dimethyl-[2,4 '-dipyridyl]-5-base) methyl)-2-(2-picoline-4-base)-1,6-naphthyridines-5-amine;

N-((2 '-methyl-[2,4 '-dipyridyl]-5-base) methyl)-2-phenylpyridine also [3,4-b] pyrazine-5-amine.

Second object of the present invention is to provide a kind of pharmaceutical composition, and described pharmaceutical composition comprises compound of the present invention or the compounds of this invention pharmacy acceptable salt and the pharmaceutically acceptable carrier of at least one or thinner.Such pharmaceutical composition can be oral compositions, Injectable composition or suppository.Described pharmaceutical composition can adopt conventional mode by mixing, granulation or coating method manufacture.

In one embodiment of the invention, described pharmaceutical composition is oral compositions, can be tablet or gelatine capsule.Preferably, described oral compositions contains compound of the present invention and a) thinner, such as lactose, glucose, sucrose, N.F,USP MANNITOL, sorbyl alcohol, Mierocrystalline cellulose and/or carbohydrate gum; B) lubricant, such as silicon-dioxide, talcum, stearic acid, stearic magnesium salts or calcium salt and/or polyoxyethylene glycol; For tablet, also comprise c) tackiness agent, such as neusilin salt, starch paste, gelatin, tragamayth, methylcellulose gum, Xylo-Mucine and/or polyvinylpyrrolidone; And when needing, d) decomposition agent can also be comprised, such as, starch, agar, alginic acid or its sodium salt, or effervescent mixture; And/or e) additive, as absorption agent, tinting material, spices and/or sweetener.

In another embodiment of the invention, described pharmaceutical composition is Injectable composition, and it can be aqueous isotonic solutions or suspension.

In an embodiment more of the present invention, described pharmaceutical composition is suppository, is prepared from by fats emulsion or suspension.

Preferably, described composition can be aseptic and/or comprise auxiliary agent, such as sanitas, stablizer, wetting agent or emulsifying agent, solution promoters, for regulating the salt of seepage water pressure, buffer reagent and/or their combination.

Alternatively or additionally, described composition can also comprise the upper valuable material of other treatment in different application, as solubilizing agent, stablizer, absorption enhancer, buffer reagent and/or sanitas.

In one embodiment of the invention, described composition can be the suitable preparation for percutaneous dosing.Such preparation includes compound of the present invention and the carrier of effective amount.Preferably, described carrier can comprise absorbable physiologically acceptable solvent to help the skin by body.Such as, transdermal delivery device is the form of bandage, described bandage comprises strut member, storage vault containing described compound (alternatively containing carrier), optional speed control barrier with the controlled skin described compound being released into body with predetermined speed within for some time extended, and ensures that described device arrives the instrument of skin.In addition, matrix percutaneous drug administration preparation can also be used.

In another embodiment of the invention, described pharmaceutical composition can be the suitable preparation of topical application, such as, be applied to skin and eyes.Such preparation can be aqueous solution well-known in the art, ointment, emulsifiable paste or gel.

Another object of the present invention is to provide a kind of WNT that suppresses from the method for emiocytosis, comprises the compound of the present invention by treatment significant quantity or its pharmaceutical composition thereof cell.

Another object of the present invention is to provide the method for WNT intracellular signaling in a kind of T suppression cell, comprises the compound of the present invention by treatment significant quantity or its pharmaceutical composition thereof cell.In one embodiment, cell is any cell comprising mammalian cell, and the dosage of administration is treatment significant quantity.In another embodiment, the suppression of WNT intracellular signaling can cause the stopping of Growth of Cells further.In further embodiment, described cell is cancer cells.Again in further embodiment, described cell is brotic cells.

The detection of cell proliferation is used in the method that those skilled in the art know very much.Such as, detect Growth of Cells one easily method be CellTiter-Glo ^tMdetect, sell in Promega (Madison, WI) company.This experimental technique is mainly by CellTiter- reagent joins in porous plate cultured cells, in photometer or imaging device, detect luminous signal.The ATP quantity existed in this signal and cell is proportional, and the viable cell quantity of ATP and culturing cell is proportional.In addition, be also the conventional method in this area by the growth velocity that colony-forming test carrys out analysis of cells.

The present invention provides the method for cancer that a kind of compounds for treating of the present invention of significant quantity is relevant to WNT signal transduction pathway or fibrosis equally.Whether one or more technique means that those skilled in the art can use this area common are relevant to WNT path to analyze cancer cells.Such as, immunity can be used to detect the abnormal expression of the albumen relevant to WNT signal path or mRNA level in-site in cancer cells with nucleic acid means.

The cancer relevant to WNT path mentioned in this article or fibrosis comprise: wherein in WNT signal path, the activity of one or several components has exceeded Normocellular substrate level.In one embodiment, WNT path is suppressed to comprise the secretion suppressing WNT protein.In another kind of embodiment, WNT path is suppressed to comprise the downstream component of T suppression cell surface receptor.In another kind of embodiment, suppress WNT secretion to comprise to suppress any and functional WNT secretes the activity of relevant albumen.

On the other hand, the invention provides the method for the disease for the treatment of WNT signal path: the WNT supressor patient of this kind of disease being provided to treatment significant quantity.In one embodiment, this kind of disease is presented as the exception of cell proliferation, mainly abnormal relevant to the activity of WNT intracellular signaling, as the rising of activity.In another embodiment, the generation of this kind of disease is the rising due to WNT protein expression amount.In another embodiment, be cancer cells during uncontrolled cellular proliferation, include but not limited to: the cancers such as lung (minicell, non-small cell) cancer, mastocarcinoma, prostate cancer, carcinoid tumor, bladder cancer, cancer of the stomach, carcinoma of the pancreas, liver (liver cell, liver protoblast) cancer, colorectal cancer, neck tesselated epithelium, esophagus cancer, ovarian cancer, cervical cancer, carcinoma of endometrium, lung carcinoma mesothelial, malignant melanoma, sarcoma, osteosarcoma, thyroid carcinoma, fibroma durum, acute myeloblastic leukemia, chronic myelocytic leukemia.In another embodiment, the out of control of cell proliferation is fibrosis, includes but not limited to: systemic sclerosis, fibrosis of skin, pulmonary fibrosis comprise idiopathic pulmonary fibrosis, radioactive fibrosis, drug-induced fibrosis, renal fibrosis, hepatic fibrosis comprise liver cirrhosis.In another embodiment, described out of control be osteoarthritis, Parkinson's disease, retinopathy, macular degeneration.

Be in therepic use, can by routine well known in the art and acceptable any means by compound of the present invention individually to treat significant quantity administration.In this article, treatment significant quantity may difference very large, this depends on the severity of the disease of patient, age and relative health, the drug effect of compound used and other factors.Usually, the daily dose systemic administration display of about 0.03-2.5mg/kg body weight can obtain gratifying result.In one embodiment, required in the larger Mammals such as mankind daily dosage is about between 100mg at about 0.5mg-.Preferably, described compound reached 4 points of agent administrations or sustained release forms administration with one day.Suitable unit dosage form for oral administration comprises about 1-100mg active ingredient.

Alternatively, compound of the present invention can treat administration together with reagent with one or more, as drug regimen as activeconstituents to treat significant quantity.Synergistic effect can be produced when compound of the present invention and chemotherapeutic combination known in the art use.The dosage of the compound of Combined Preparation is different according to the type of used combination medicine, the certain drug used, treatment condition etc.

Compound of the present invention or its pharmaceutical composition can by the administrations of any conventional.In one embodiment, be digestive tube administration, such as oral, adopt the form of tablet or capsule.In another embodiment, described administration is parenteral administration, such as, adopt the form administration of Injectable solution or suspension.In another embodiment, described administration is topical, such as, adopt emulsion, gel, ointment or cream forms administration, or with nose with or the form administration of suppository.

On the other hand, present invention also offers drug regimen, such as test kit, comprise a) the first reagent, described reagent is as compound disclosed in present specification or its pharmacy acceptable salt, and b) at least one combines reagent.Described test kit can comprise the specification sheets for its administration.

Combined therapy of the present invention can be used in vivo or in vitro.These methods can comprise simultaneously or carry out administration to these reagent within for some time, and wherein said material produces desirable result for the treatment of when individually dosed.This can by making cell, tissue or organ and comprising the independent composition of two or more reagent or pharmaceutical formulation thereof realizes, or realize by making the cell composition different with two or more or preparation contact, a kind of composition wherein comprises a kind of reagent, and other compositions comprise other reagent.Compound of the present invention can before described another kind of reagent, parallel with described another kind of reagent and/or after described another kind of reagent, space-number is minute to administration several weeks.In one embodiment, when described reagent is used for cell, tissue or organ individually, reagent can guarantee that the effective time cycle can not exceed the timed interval discharged each time usually, thus it is favourable in conjunction with effect that described reagent still can be produced described cell, tissue or organ.Such as, under these circumstances, a kind of reagent as described alternative material, can with two kinds, three kinds, four kinds or more plant form basic while, namely contacted with described cell, tissue or organ within about one minute.In another embodiment, one or more reagent can administration in about 1 minute-14 days.

On the other hand, the invention provides the method preparing compound or its salt or derivatives thereof of the present invention.

In one embodiment, the compound of molecular formula (1) can be had according to any one preparation in the synthetic method described in following examples part.When wishing to comprise functional group such as hydroxy, amino, imino-, thio group or the carboxyl with reactive behavior in final product, these functional groups can be protected in described reaction unnecessarily to participate in reaction to avoid it.According to usual experience (see such as T.W.Greene and P.G.M.Wuts; " blocking group in organic chemistry " (Protecting Groups in Organic Chemistry), John Wiley and Sons.1991) conventional blocking group can be used.The suitable leavings group used in described synthetic method comprises halo leaving group, and the leavings group of those other routines in the common knowledge of those skilled in the art.Preferably, described leavings group is chlorine or bromine.

In another embodiment, compound of the present invention and salt thereof also can be obtained by the form of hydration, or their crystal can comprise such as the solvent (existing with solvate) of crystallization.Usually salt can be changed into the compound of free form, such as, adopt suitable alkaline reagents process, such as alkali-metal carbonate, alkali-metal supercarbonate or alkali-metal oxyhydroxide, such as salt of wormwood or sodium hydroxide.By adopting suitable acid (such as hydrochloric acid etc.) process, the compounds of this invention of addition salt forms can be changed into corresponding free acid.Due to the intimacy between the new compound of free form and salt (comprising the salt as intermediate) thereof such as in the purifying or qualification process of described new compound, suitably can being interpreted as, being also mentioned to its corresponding salt when mentioning described free compound.

Mode well known in the art can be adopted to prepare the salt of the compounds of this invention containing salt forming group.By can obtain the acid salt of the compound with molecular formula (1) with acid or suitable anionite process.The salt of the pharmaceutically acceptable form of compound of the present invention can be the form of the acid salt of the compound containing basic nitrogen atom such as using organic or inorganic acid and molecular formula (1).

Suitable mineral acid includes but not limited to: hydracid is hydrochloric acid, sulfuric acid or phosphoric acid such as.Suitable organic acid includes but not limited to: carboxylic acid, phosphoric acid, sulfonic acid, thionamic acid, such as acetic acid, propionic acid, sad, capric acid, dodecylic acid, oxyacetic acid, lactic acid, fumaric acid, succsinic acid, hexanodioic acid, pimelic acid, suberic acid, nonane diacid, oxysuccinic acid, tartrate, citric acid, amino acid such as L-glutamic acid or aspartic acid, toxilic acid, hydroxymaleic acid, methyl-maleic acid, hexahydrobenzoic acid, adamantanecarboxylic acid, phenylformic acid, Whitfield's ointment, 4-ASA, phthalandione, phenylacetic acid, amygdalic acid, styracin, methane-or ethane-sulfonic acid, 2-hydroxyethanesulfonic acid, ethane-1, 2-disulfonic acid, Phenylsulfonic acid, 2-naphthene sulfonic acid, 1, 5-naphthalene-disulfonic acid, 2-, 3-or 4-toluene sulfonic acide, methylsulphonic acid, ethylsulfonic acid, dodecyl sodium sulfonate, N-cyclohexylsulfamic, N-methyl-, N-ethyl-or N-propylcarbamic sulfonic acid or the acid of other organic proton, such as xitix.

Alternatively, in order to the object of abstraction and purification, pharmaceutically unacceptable salt also can be used, such as picrate or perchlorate.In order to the object for the treatment of, only adopt the compound (when the form of pharmaceutical preparation can be adopted) of pharmacy acceptable salt or free form.

In another embodiment, by using the N-oxide compound of reductive agent and the compounds of this invention to react in suitable inert organic solvents, at 0-80 DEG C, the compound of the present invention of non-oxidised form can be prepared into.Preferably, described inert organic solvents is acetonitrile, ethanol, water-based dioxane or analogue.Preferably, described reductive agent is sulphur, sulfurous gas, triphenylphosphine, lithium borohydride, sodium borohydride, phosphorus trichloride, phosphorus tribromide or analogue.

In another embodiment, method known to a person of ordinary skill in the art can be adopted (such as, the more details of described method are see the people such as Saulnier (1994), biological organic and pharmaceutical chemistry bulletin (Bioorganic and Medicinal Chemistry Letters), 4th volume, the 1985th page) prepare the prodrug derivatives of the compounds of this invention.Such as; can by the compounds of this invention of non-derived and suitable carbamoylating agent (such as 1,1-acyloxyallcyl chloro-formic ester (do not contain N in this compound, can as carbamoylating agent); p-nitrophenyl carbonic ether, or analogue).Method known to a person of ordinary skill in the art can be adopted to prepare the derivative of protected the compounds of this invention.About the generation of spendable blocking group and the detailed description of the technology of removal thereof are disclosed in (third edition, John Wiley and Sons company limited, 1999) in " blocking group in organic chemistry " of T.W.Greene.

In another embodiment, compound of the present invention can be prepared to their respective steric isomers, by the racemic mixture of described compound and optic active resolving agent being reacted the compound to generate a pair non-cubic isomer, being separated described non-cubic isomer and obtaining optic pure enantiomorph.The covalency non-cubic isomer derivative of the compounds of this invention or the segregation by using separable mixture (the non-cubic isomer salt of such as crystallization) to carry out enantiomorph can be used.Non-cubic isomer has different physicalies (such as fusing point, boiling point, solubleness, reactive behavior etc.) and utilizes the difference of these non-cubic isomer to be easily isolated.Can pass through fractional crystallisation, chromatography or by based on the separation/maceration technique of different solubility by described non-cubic isomer separation.Then by racemic empirical method can not be caused arbitrarily to obtain optic pure enantiomorph and segregation reagent.About the more detailed description of the spendable technology making the steric isomer of compound and their racemic mixture emanate is disclosed in JeanJacques, Andre Collet, Samuel H.Wilen, " enantiomorph, racemoid and segregation " (Enantiomers, Racemates and Resolutions) " John Wiley and Sons company limited, 1981.

In sum, compound of the present invention can be prepared by the method described in embodiment; And

Be optionally pharmacy acceptable salt by converting compounds of the present invention;

Optionally the non-oxidised form of compound is converted into pharmaceutically acceptable N-oxide compound;

From isomer mixture, optionally isolate the independent isomer of compound of the present invention; And

Optionally the compounds of this invention of non-derived is converted into pharmaceutically acceptable prodrug derivatives.

The not special manufacturing processed described about starting raw material, described compound is known or the method similar with the known method of this area can be adopted to prepare or adopt the method disclosed in embodiment as hereafter to prepare.It will be appreciated by those skilled in the art that above-mentioned transformation example is only the representative of the method preparing the compounds of this invention, and other well-known method can be used equally.

Compound of the present invention and composition thereof effectively can suppress the secretion of WNT protein, suppress WNT intracellular signaling, therefore have good result for the treatment of to the disease of WNT mediation.

Embodiment

By hereafter and the embodiment of preparation describing the compounds of this invention in detail illustrate the present invention further, but the present invention is not limited to this.

Abbreviation definition or lexical or textual analysis

DCM methylene dichloride

DIEA N, N '-diisopropylethylamine

DMF DMF

Eq. equivalent

TEA triethylamine

THF hydrogen furans

RT room temperature

EA ethyl acetate

Pd ₂(dba) ₃three (dibenzalacetone) two palladium (0)

S-Phos 2-dicyclohexyl phosphino--2 ', 6 '-dimethoxy-biphenyl

Pd (PPh ₃) ₄tetrakis triphenylphosphine palladium

Pd (OAc) ₂acid chloride (II)

BINAP 2,2 '-bis-(biphenyl phosphino-)-1,1 '-dinaphthalene

M-CPBA 3-Chloroperbenzoic acid

Embodiment 1:

N-(4-(2-picoline-4-base) phenmethyl)-6-(2-picoline-4-base)-2,7-naphthyridines-1-amine (compound 1)

Step 1:

2-malonamide nitrile (50g, 601.8mmol) and methyl aceto acetate (75mL, 601.8mmol) are dissolved in methyl alcohol.KOH (37.0g, 1.1eq.) is dissolved in methyl alcohol, and described KOH is dropwise added in mixture, occur some white solids.By described mixture reflux 8h in oil bath, be then cooled to room temperature.Filtering solids and described solid is dissolved in the hot water again, and then filter.Add in filtrate in 6N HCl and until pH < 7.Described white solid again occurs and is filtered.Use solid described in methyl alcohol, water and methanol wash further, then by vacuum-drying to obtain final product 3-ethynyl-4-picoline-2,6-glycol (productive rate ~ 41%)

Step 2:

3-ethynyl-4-picoline-2,6-glycol (28.0g, 195.2mmol) is dissolved in POCl ₃(60.0mL) in.Described reaction mixture to be sealed in penstock and to be heated to 180 DEG C of heating 6h.After described reactant is cooled to room temperature, remove excessive POCl under vacuo ₃.Trash ice is added in described mixture lentamente, has then occurred solid.Solid filtering is also dry not needed the final product 2,6-bis-chloro-4-picoline-3-formonitrile HCN (productive rate ~ 92%) be further purified under vacuo.

Step 3:

At 2 of 200mL, DMF dimethylacetal (12.82g is added in the chloro-4-picoline of 6-bis--3-formonitrile HCN (20.0g, 107.5mmol) aqueous isopropanol, 107.5mmol), and at 65 DEG C, described reactant is stirred 18h.After described reactant is cooled to RT, also 50mL washed with isopropyl alcohol is used by collected by filtration thing, then do not need the chloro-4-of product 2,6-bis-((E)-2-(dimethylamino) vinyl) pyridine-3-formonitrile HCN (productive rate ~ 26%) be further purified with acquisition by dry air.

Step 4:

Chloro-for 2,6-bis-4-((E)-2-(dimethylamino) vinyl) the dense HCl of pyridine-3-formonitrile HCN (4.0g, 16.6mmol) and 20mL is added in sealed tube.At 45 DEG C, described reactant is stirred 18h.After described reactant is cooled to RT, frozen water is added into the slip forming deep yellow in described solution.By collected by filtration thing and with the washing of cold water, ether and ethyl acetate, then dry to obtain chloro-2,7-naphthyridines-1 (2H)-one (productive rate ~ 80%) of flaxen solid 6,8-bis-under vacuo.MS m/z 215.0(M+1)。 ¹HNMR(300MHz，DMSO-d6)：δ11.75(s，1H)，7.76(s，1H)，7.50(t，J＝6.6Hz，1H)，6.52(d，J＝6.6Hz，1H)。

Step 5:

Chloro-for 6,8-bis-2,7-naphthyridines-1 (2H)-one (3.0g, 13.96mmol) are dissolved in iPrOH (120mL) and form a kind of suspension.In ice bath, described solution is cooled to 0 DEG C, then dropwise adds hydrazine solution (5.6g, 80%, 10eq.).Described mixture is stirred 15 minutes under RT, then heated overnight in the oil bath of 55 DEG C.After described reactant is cooled to RT, filter directly to obtain solid, then use solid described in 70mL methanol wash and drying under vacuo.In step reaction below, direct use does not need the product 6-chloro-8-diazanyl-2,7-naphthyridines-1 (2H)-one (productive rate ~ 98%) be further purified.

Step 6:

Chloro-for 6-8-diazanyl-2,7-naphthyridines-1 (2H)-one (1.50g, 7.12mmol) is dissolved in acetonitrile (90mL) to generate a kind of suspension.Add 1N NaOH (17.80mL, 2.5eq), then the water (107.80mL) of the cumulative volume equivalent with acetonitrile and NaOH is added in described mixture.Heat at 50 DEG C and stir described reaction-ure mixture until described mixture becomes clear soln.Described solution is cooled to 0 DEG C again, and dropwise adds NaOCl (11.05g, 12% solution, 2.5eq), then at room temperature described reactant is stirred and spend the night.After completion of reaction, described solution is cooled to 0 DEG C, then adds 1N HCl and neutralize (pH ~ 6).Collecting precipitation thing and extract described filtrate with 100mLx2EA.Organic layer is mixed and passes through Na ₂sO ₄dry also evaporation is to obtain extra crude product.Directly mixed chloro-2,7-naphthyridines-1 (2H)-one (productive rate ~ 93%) of the solid materials 6-not needing to be further purified are used in reaction below.MS m/z 181.1(M+1)。

Step 7:

In penstock, chloro-for 6-2,7-naphthyridines-1 (2H)-one (400mg, 2.2mmol) are added into POCl ₃(20.0mL) in.Described reaction-ure mixture is heated to 160 DEG C of heating 4h to obtain clear soln.Described solution be cooled to room temperature and pour in DCM, then adding trash ice lentamente.By saturated NaHCO ₃be added in described mixture with the HCl produced in neutralization reaction.Remove DCM under vacuo and the aqueous solution be left with 100mLx2EA extraction.With the organic layer of saturated common salt water washing mixed once, then use Na ₂sO ₄drying, then vaporising under vacuum does not need chloro-2, the 7-naphthyridines (productive rate ~ 73%) of solid 1,6-bis-be further purified for step reaction below to obtain.MS m/z 199.0(M+1)。

Step 8:

(4-bromophenyl) methylamine (1.00g, 5.37mmol) and 2-picoline-4-base-4-boric acid (883.30mg, 6.45mmol) are dissolved in butanols (10.0mL) and water (2.0mL).At N ₂lower interpolation K ₃pO ₄(2.28g, 10.75mmol), Pd ₂(dba) ₃(120.20mg, 0.27mmol) and S-phos (220.70mg, 0.54mmol).Described reaction-ure mixture to be sealed in penstock and to be heated to 125 DEG C of heating 1h.After described reactant is cooled to RT, described mixture is poured into water and extracts with 100mLx3EA.With the organic layer of saturated common salt water washing mixing, Na ₂sO ₄dry and concentrated to obtain crude product under vacuo.Purify described solid to obtain pure (4-(2-picoline-4-base) phenyl) methylamine (productive rate ~ 89%) with being dissolved in containing the DCM of 10% methyl alcohol (comprising ~ 2N ammonia) by Silica hydrogel post.MS m/z 199.1(M+1)。

Step 9:

By 1, chloro-2, the 7-naphthyridines (160mg, 0.80mmol) of 6-bis-and (4-(2-picoline-4-base) phenyl) methylamine (239.10mg, 1.21mmol) be dissolved in butanols (5.0mL), be then heated to 115 DEG C and spend the night.After described reactant is cooled to RT, remove described organic solvent under vacuo.By crude product described in Silica hydrogel flash chromatography EA/ hexane (1: 1) purifying to obtain solid N-(4-(2-picoline-4-base) phenmethyl) chloro-2, the 7-naphthyridines-1-amine (productive rate ~ 90%) of-6-.MS m/z 361.1(M+1)。

Step 10:

By N-(4-(2-picoline-4-base) phenmethyl)-6-chloro-2,7-naphthyridines-1-amine (50.00mg, 0.14mmol) be dissolved in butanols (3.0mL) and water (0.6mL) with 2-picoline-4-base-4-boric acid (56.90mg, 0.42mmol).At N ₂lower to K ₃pO ₄(88.20mg, 0.028mmol), Pd ₂(dba) ₃(6.20mg, 0.014mmol) and S-phos (11.40mg, 0.011mmol) are added in described mixture.Described mixture is sealed in penstock and is then heated to 105 DEG C and spends the night.After described reactant is cooled to RT, described mixture is poured into water and extracts three times with EA.With the organic layer of saturated common salt water washing mixing, use Na ₂sO ₄dry and concentrate under vacuo.Crude product is further purified to obtain final product N-(4-(2-picoline-4-base) phenmethyl)-6-(2-picoline-4-base)-2,7-naphthyridines-1-amine (productive rate ~ 70%) by the DCM of preparative thin layer chromatography containing 5% methyl alcohol.MS m/z 418.2(M+1)。 ¹HNMR(300MHz，CDCl ₃)：δ2.46(s，3H)，2.63(s，3H)，4.94(d，J＝5.10Hz，2H)，5.94(br，1H)，6.97(d，J＝5.70Hz，1H)，7.31(d，J＝4.20Hz，1H)，7.36(s，1H)，7.54(d，J＝8.10Hz，2H)，7.63(d，J＝8.40Hz，2H)，7.90(s，1H)，8.19(d，J＝6.00Hz，1H)，8.22(s，1H)，8.51(m，2H)，9.08(s，1H)，9.30(s，1H)。

Embodiment 2:

N-(3-methyl-4-(2-picoline-4-base) phenmethyl)-6-(2-picoline-4-base)-2,7-naphthyridines-1-amine (compound 2)

Step 1:

By chloro-for 6-2,7-naphthyridines-1 (2H)-one (200mg, 1.10mmol) be dissolved in butanols (5.0mL) and water (1.0mL) with 2-picoline-4-base-4-boric acid (227.60mg, 1.66mmol).At N ₂under add K ₃pO ₄(705.20g, 3.32mmol), Pd ₂(dba) ₃(49.60mg, 0.22mmol) and S-phos (91.00mg, 0.11mmol).Reaction-ure mixture in penstock is heated to 130 DEG C of heating 1h.After described reactant is cooled to RT, described mixture is poured into water, extracts three times with EA.With the organic layer of saturated common salt water washing mixing, Na ₂sO ₄drying, concentrated to obtain crude product under vacuo.Purify described crude product to obtain finalization compound 6-(2-picoline-4-base)-2,7-naphthyridines-1 (2H)-one (productive rate ~ 61%) by the DCM of column chromatography containing 5% methyl alcohol.MS m/z 238.1(M+1)。

Step 2:

6-(2-picoline-4-base)-2,7-naphthyridines-1 (2H)-one (150mg, 0.63mmol) are dissolved in POCl ₃(15.0mL), in, seal described penstock and be heated to 160 DEG C of heating 4h.After described reactant is cooled to RT, remove excessive POCl under vacuo ₃.Trash ice is added in described mixture lentamente, then adds NaHCO ₃neutralization is until pH ~ 7.5.With EA by described solution extraction three times, with the organic layer of saturated common salt water washing mixing, Na ₂sO ₄drying, concentrates under vacuo.By column chromatography EA/ hexane (1: 1) purification of crude product to obtain the chloro-6-of compound 1-(2-picoline-4-base)-2,7-naphthyridines (productive rate ~ 55%).MS m/z 256.1(M+1)。

Step 3:

By chloro-for 1-6-(2-picoline-4-base)-2,7-naphthyridines (10.00mg, 0.039mmol) be dissolved in toluene (1.0mL) with (3-methyl-4-(2-picoline-4-base) phenyl) methylamine (10.00mg, 0.047mmol).At N ₂lower to KO ^tbu (8.80mg, 0.078mmol), Pd (OAc) ₂(0.90mg, 0.0039mmol) and BINAP (4.90mg, 0.0078mmol) are added in mixture.Described reactant is heated to 100 DEG C spend the night.After described reactant is cooled to RT, described mixture is poured into water, extracts three times with EA.With the organic layer of saturated common salt water washing mixing, Na ₂sO ₄drying, then concentrates under vacuo.By Preparative TLC chromatographic grade EA/ hexane (4: 1) purification of crude product to obtain N-(3-methyl-4-(2-picoline-4-base) phenmethyl)-6-(2-picoline-4-base)-2,7-naphthyridines-1-amine (8.8mg, productive rate ~ 52%).1H NMR(300MHz，CDCl3)：δ2.31(s，3H)，2.63(s，3H)，2.70(s，3H)，4.91(d，J＝5.10Hz，2H)，5.88(br，1H)，7.00(d，J＝5.40Hz，1H)，7.08(d，J＝5.10Hz，1H)，7.12(s，1H)，7.22(d，J＝7.50Hz，1H)，7.36(m，2H)，7.77(d，J＝4.50Hz，1H)，7.88(s，1H)，7.98(s，1H)，8.24(d，J＝6.00Hz，1H)，8.53(d，J＝4.80Hz，1H)，8.64(d，J＝5.40Hz，1H)，9.31(s，1H)。MS m/z 432.2(M+1)。

Embodiment 3:

6-(3-fluorophenyl)-N-((6-(2-picoline-4-base) pyridin-3-yl) methyl) isoquinoline 99.9-1-amine (compound 3)

Step 1:

6-bromo-isoquinoline (1.80g, 8.66mmol) is dissolved in DCM (40mL), after described reactant is cooled to 0 DEG C, adds m-CPBA (2.30g, 1.3eq, maximum 77%) on a small quantity, lentamente.Described reactant is warming up to RT to generate a kind of white suspension.In 4 hours, 100mLDCM is added in described solution, then uses saturated Na ₂cO ₃solution, water and saturated common salt water washing.Use Na ₂sO ₄the dry organic layer be separated also is removed to obtain yellow solid N-oxide compound 6-bromo-isoquinoline (1.82g, productive rate ~ 93%) not needing to be further purified under vacuo.

Step 2:

N-oxide compound 6-bromo-isoquinoline (1.82g, 8.12mmol) is dissolved in dry DCM (80mL), interpolation POCl dropwise under RT ₃(1.12ml, 1.5eq).Described reactant is heated to 45 DEG C 2 hours.After described reactant is cooled to RT, remove DCM and excessive POCl under vacuo ₃.Crude product is dissolved in 100mLDCM again, and uses saturated Na ₂cO ₃, water and saturated common salt water washing.Na ₂sO ₄the dry organic layer be separated, then concentrates and obtains brown solid.By rapid column chromatography with the DCM purification of crude product be dissolved in containing 2% methyl alcohol to obtain faint yellow solid 6-bromine 1-chlorine isoquinoline 99.9 (1.27g, productive rate ~ 65%).MS m/z 242.0(M+1)。

Step 3:

Propyl carbinol (10mL) and water (2mL) is used (6-chloropyridine-3-base) methylamine (300mg, 2.1mmol) and 2-picoline-4-ylboronic acid (345mg, 2.52mmol) to be dissolved in penstock.Add K under nitrogen protection ₃pO ₄(893mg, 4.2mmol), Pd ₂(dba) ₃(96.3mg, 0.105mmol) and S-phos (86.4mg, 0.21mmol).Described reactant is heated to 125 DEG C of heating 30 minutes, is then cooled to room temperature.Described solution is poured into water and extracts three times with EA.With the organic layer of saturated common salt water washing mixing, and use Na ₂sO ₄drying, then concentrates under vacuo.Crude product is further purified to obtain pure (6-(2-picoline-4-base) pyridin-3-yl) methylamine (0.19g, productive rate ~ 45%) by the DCM of flash chromatography containing 10% methyl alcohol (containing ~ 2N ammonia).MS m/z 200.1(M+1)。

Step 4:

In sealed tube, bromo-for 6-1-chlorine isoquinoline 99.9 (100mg, 0.41mmol) and (6-(2-picoline-4-base) pyridin-3-yl) methylamine (165mg, 0.82mmol) are dissolved in 0.5mL propyl carbinol.Described reactant is heated to 160 DEG C of heating 6h, is then cooled to RT.Crude product is further purified to obtain the bromo-N-of pure 6-(6-(2-picoline-4-base) pyridin-3-yl) methyl by the DCM of flash chromatography containing 8% methyl alcohol (containing ~ 2N ammonia)) isoquinoline 99.9-1-amine (116mg, productive rate ~ 70%).MS m/z405.2(M+1)。

Step 5:

By bromo-for 6-N-(6-(2-picoline-4-base) pyridin-3-yl) methyl) isoquinoline 99.9-1-amine (20mg, 0.05mmol), 3-flurophenyl boronic acid (10.5mg, 0.075mmol), Na ₂cO ₃(21mg, 0.2mmol) and tetrakis triphenylphosphine palladium (5.8mg, 0.005mmol) are added in penstock.Dioxane/water (3: 1,2mL) is added in pipe, and is heated to 125 DEG C of heating 10 minutes.After described reactant is cooled to RT, dilutes described solution with 50mL water and extract 3 times with EA.Use Na ₂sO ₄the organic layer of dry mixed, and concentrate under vacuo.Crude product is further purified to obtain pure 6-(3-fluorophenyl)-N-((6-(2-picoline-4-base) pyridin-3-yl) methyl) isoquinoline 99.9-1-amine (15.8mg, ~ 75%) by the DCM of flash chromatography containing 10% methyl alcohol (containing ~ 2N ammonia).1H NMR(400MHz，CDCl3)：δ2.71(s，3H)，5.00(d，J＝5.6Hz，2H)，7.32-7.38(m，2H)，7.59-7.65(m，1H)，7.75-7.83(m，3H)，8.10(d，J＝8.4Hz，1H)，8.21(d，J＝8.8Hz，1H)，8.27-8.31(m，2H)，8.39(s，2H)，8.72(d，J＝8.8Hz，1H)，8.79(d，J＝6.0Hz，1H)，8.91(d，J＝1.6Hz，1H)，10.02(s，1H)。MS m/z 421.2(M+1)。

Embodiment 4:

N-(4-(2-picoline-4-base) phenmethyl)-2-(2-picoline-4-base)-1,6-naphthyridines-5-amine (compound 4)

Step 1:

1,6-naphthyridines-5 (6H)-one (2.9g, 19.84mmol) is dissolved in POCl ₃(40mL), in, be then heated to 100 DEG C of heating 24h. after described reactant is cooled to room temperature, remove excessive POCl under vacuo ₃.Add a small amount of saturated Na lentamente ₂cO ₃the trash ice of solution, and produce a large amount of bubble and solid.Filtering solids, then extracts 3 times by described solution EA.Use Na ₂sO ₄the organic layer of dry mixed, and concentrate under vacuo.The solid of further dry mixed is to obtain chloro-1, the 6-naphthyridines of 5-(2.6g, productive rate ~ 80%) not needing to be further purified under vacuo.MS m/z 165.1(M+1)。

Step 2:

Chloro-for 5-1,6-naphthyridines (1.5g, 9.11mmol) is dissolved in DCM (45mL), is then cooled by ice bath, add m-CPBA (3.7g, 2eq, maximum 77%) on a small quantity, lentamente.Described reactant is warming up to RT and keeps 3 hours.The DCM being greater than 100mL is added in described solution, then uses saturated Na ₂cO ₃solution, water and saturated common salt water washing.Na ₂sO ₄dry organic layer, and concentrated to obtain chloro-1, the 6-naphthyridines of the yellow solid N-oxide compound 5-(1.25g, productive rate ~ 76%) not needing to be further purified under vacuo.

Step 3:

Chloro-for N-oxide compound 5-1,6-naphthyridines (1.2g, 6.64mmol) is dissolved in dry DCM (30mL), adds Et3N (1.85mL, 13.29mmol), then dropwise add the POCl be dissolved in the DCM of 5mL drying ₃(0.93mL, 9.97mmol).Described reactant is heated to 48 DEG C of heating 2h.Na ₂sO ₄dry organic layer, and concentrated to obtain yellow solid under vacuo.Crude product is further purified to obtain chloro-1, the 6-naphthyridines of white solid 2,5-bis-(0.6g, productive rate ~ 45%) by silicon column chromatography EA/ hexane (1: 4).MS m/z 199.0(M+1)。

Step 4:

By chloro-for 2,5-bis-1,6-naphthyridines (200mg, 1.0mmol), 2-picoline-4-base 4-boric acid (137mg, 1.0mmol), Na ₂cO ₃(424mg, 4.0mmol) and tetrakis triphenylphosphine palladium (116mg, 0.1mmol) add in flask, then add 16mL dioxane and 4mL water.Described reactant is fully stirred and is heated to 90 DEG C of heating 4h.After described reactant is cooled to RT, dilutes described solution with 100mL water and extract 3 times with EA.Na ₂sO ₄the organic layer of dry mixed, and concentrate under vacuo.Crude product is further purified to obtain the chloro-2-of solid 5-(2-picoline-4-base)-1,6-naphthyridines (143mg, productive rate ~ 56%) by flash chromatography EA/ hexane (1: 1).MS m/z256.1(M+1)。

Step 5:

By chloro-for 5-2-(2-picoline-4-base)-1,6-naphthyridines (20.00mg, 0.078mmol) be dissolved in toluene (2.0mL) with (4-(2-picoline-4-base) phenyl) methylamine (25mg, 0.118mmol).At N ₂lower to KOtBu (13.2mg, 0.118mmol), Pd (OAc) ₂(2.7mg, 0.012mmol) and BINAP (15.0mg, 0.024mmol) are added in described mixture.Described reactant is heated to 100 DEG C spend the night.After described reactant is cooled to RT, described mixture is poured into water, extracts three times with EA.With the organic layer of saturated common salt water washing mixing, Na ₂sO ₄drying, then concentrates under vacuo.The DCM purification of crude product of 8% methyl alcohol is contained to obtain N-(4-(2-picoline-4-base) phenmethyl)-2-(2-picoline-4-base)-1 by Preparative TLC chromatographic grade, 6-naphthyridines-5-amine (31mg, productive rate ~ 61%). ¹H NMR(400MHz，DMSO-d6)：δ9.12(d，J＝8.8Hz，1H)，8.77-8.83(m，2H)，8.49(d，J＝8.4Hz，1H)，8.40(s，1H)，8.31(d，J＝6.4Hz，1H)，8.21(s，1H)，8.11(d，J＝5.6Hz，1H)，8.06(d，J＝6.4Hz，1H)，7.99(d，J＝8.4Hz，2H)，7.65(d，J＝8.4Hz，2H)，7.23(d，J＝6.4Hz，1H)，5.76(s，1H)，4.93(d，J＝5.6Hz，2H)，2.72(s，6H)。MS m/z 432.2(M+1)。

Embodiment 5:

N-(4-(2-picoline-4-base) phenmethyl)-2-phenylpyridine also [4,3-b] pyrazine-5-amine (compound 5)

Step 1:

Phenyl oxalic dialdehyde monohydrate (940mg, 6.99mmol) and chloro-3, the 4-diamino-pyridines (1000mg, 6.99mmol) of 2-are added to 20mL ethanol.Described mixture is refluxed spend the night.After described reactant cooling, thick precipitated product is filtered and uses 15mL washing with alcohol, then dry to obtain the chloro-2-phenylpyridine of 5-also [3, the 4-b] pyrazine (1.28g not needing to be further purified under vacuo, productive rate ~ 76%), MS m/z 241.0 (M+1); 1H NMR (300MHz, DMSO-d6): δ 9.82 (s, 1H), 8.64 (d, J=6.0Hz, 1H), 8.38-8.43 (m, 2H), 8.07 (d, J=6.0Hz, 1H), 7.64-7.68 (m, 3H).

Step 2:

By N-(4-(2-picoline-4-base) phenmethyl)-2-phenylpyridine also [3,4-b] pyrazine-5-amine (50mg, 0.21mmol) be dissolved in toluene (4.0mL) with (4-(2-picoline-4-base) phenyl) methylamine (42mg, 0.21mmol).At N ₂lower to KO ^tbu (24mg, 0.21mmol), Pd (OAc) ₂(4.5mg, 0.021mmol) and BINAP (26.4mg, 0.042mmol) are added in described mixture.Described reactant heat is spent the night to 100 DEG C.After described reactant is cooled to room temperature, by mixture down in water, extract 3 times with EA.With the organic layer of saturated common salt water washing mixing, Na ₂sO ₄drying, then concentrates under vacuo.By flash chromatography with containing the DCM purification of crude product of 7% methyl alcohol to obtain N-(4-(2-picoline-4-base) phenmethyl)-2-phenylpyridine also [4,3-b] pyrazine-5-amine (61mg, productive rate ~ 72%).MS m/z＝404.2(M+1)； ¹H NMR(400MHz，DMSO-d6)δ9.53(s，1H)，8.77(d，J＝6.4Hz，1H)，8.35-8.39(m，2H)，8.21(s，1H)，8.11(d，J＝6.0Hz，1H)，8.07(d，J＝6.4Hz，1H)，7.96(d，J＝8.4Hz，2H)，7.60-7.65(m，5H)，7.14(d，J＝6.0Hz，1H)，5.76(s，1H)，4.90(d，J＝6.4Hz，2H)，2.71(s，3H)。

In the present invention, other all compound is by according to method similar in above-described embodiment and this area routine techniques Preparation and identification.

Other compound list:

Experimental example 1:WNT signal path reporter gene assays

Materials and methods: l cell NIH3T3 (American type culture collection, Manassas, Virginia) transfection comprises the plasmid of the luciferase gene of 5 copy TCF unit and driving thereof, the cell obtained 37 DEG C, containing 5%CO ₂condition under cultivate, substratum comprises Eagle ' s substratum (DMEM that Dulbecco ' s revises, Invitrogen, Carlsbad, Canada), 10%FBS (Invitrogen), 50 units/mL penicillin, 50 μ g/mL Streptomycin sulphates (Invitrogen), add 1 μ g/mL bleomycin (Invitrogen) simultaneously and screen cell.HEK293 cell (ATCC) transfection suspended contains the plasmid of the cDNA sequence of the total length WNT3a of people, is driven by CMV promoter.Cell screening in FreeStyle 293 substratum (Invitrogen) that with the addition of 100 μ g/mL G418 of stably express.

NIH3T3TCF-Luc cell and 293WNT3a cell co-cultivation in 96 orifice plates, substratum is DMEM substratum and 0.5%FBS.After 16 hours, at Steady-Glo ^tMthe activity of Photinus pyralis LUC is detected in Luciferase Assay System (Promega).The compound obtained in this invention is with different concns and these cell co-cultures.Reduce 50% of fluorescence intensity by compound and calculate the IC of compound ₅₀.In order to quantity and the activity of standardized cell, CellTiter Glo detects will repeat twice.

The IC of all compounds in the present invention ₅₀value is all less than 5 micromoles per liter, selects compound to be listed in the table below:

Compound number:	IC ₅₀(μM)
		1	＜0.003
3	0.010
		5	0.070
23	0.020
		28	＜0.003
35	＜0.003
		37	0.020
50	0.035
		68	0.025
70	＜0.003

The study mechanism of experimental example 2:WNT signal pathway inhibitor

In our preliminary study, the compound obtained can effectively suppress WNT-3a cell to the induction of TCF reporter gene expression activity.In ensuing study mechanism, main object finds the target spot of these compound effects.Wherein the agonist of two main Water demand comprises, one is WNT-3a recombinant protein (the StemRD Inc. of purifying, Burlingame, CA), another is the supressor 6-bromoindirubin-3 '-oxime (StemRD Inc.) of GSK-3b.

The result of study mechanism is presented at some activated compounds in this invention and can suppresses the activity of WNT signal path, but the activity of the protein induced TCF reporter gene of restructuring WNT-3a can not be suppressed, illustrate that the action target spot of compound should be the upstream protein of WNT-3a and receptors bind.The target spot material standed for that may act on mainly comprises WNTless/evenness interrupted (Wls/Evi), Porcn, Vps35p.

The function of experimental example 3:WNT path inhibition in cancer cells

Suppress the secretion of WNT protein and the compound of Cellular Signaling Transduction Mediated, may the propagation of anticancer, because the growth of some cancer cells depends on the WNT signal of autocrine.WNT path inhibition can need to play a role in the Growth of Cells of WNT autocrine signal at these, comprises the growth of 2-D cultured cells, does not rely on the cell proliferation of upholder, anti-the dying property of tune etc. of clone.To the analysis that the evaluation of these compounds will use the WNT dependent form cell mentioned in the current works delivered to carry out standard.These WNT dependent form cells comprise: PA-1 (ovary teratoblastoma), MDA-MB-157 (mammary cancer), Saos-2 (osteosarcoma), SNU1076 (nasopharyngeal carcinoma).In these clones, observe the function of inhibition, confirm the activity desired by these compounds further.

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Claims

1. one kind has compound and the pharmacy acceptable salt thereof of general formula (1):

Wherein, described compound is following listed compound 1, compound 3, compound 5, compound 23, compound 28, compound 35, compound 37, compound 50, compound 68 or compound 70.

2. a pharmaceutical composition, comprises as the compound according to claim 1 of activeconstituents or its pharmacy acceptable salt and the pharmaceutically acceptable carrier of at least one or thinner.

3. compound according to claim 1 or pharmaceutical composition according to claim 2 are preparing the application in the medicine suppressing WNT to secrete from cell.

4. compound according to claim 1 or the application of pharmaceutical composition according to claim 2 in the medicine preparing WNT intracellular signaling in T suppression cell.

5. compound according to claim 1 or pharmaceutical composition according to claim 2 application in the medicine out of control of preparation treatment WNT mediation, wherein, described out of control abnormal to WNT signaling activity relevant, be describedly out of controlly: cancer, fibrosis, osteoarthritis, Parkinson's disease, retinopathy or macular degeneration.

6. apply as claimed in claim 5, wherein, described cancer is: small cell lung cancer, nonsmall-cell lung cancer, mastocarcinoma, prostate cancer, carcinoid tumor, bladder cancer, cancer of the stomach, carcinoma of the pancreas, hepatocellular carcinoma, liver protoblast cancer, colorectal cancer, neck squamous cell carcinoma, esophagus cancer, ovarian cancer, cervical cancer, carcinoma of endometrium, lung carcinoma mesothelial, malignant melanoma, sarcoma, osteosarcoma, thyroid carcinoma, fibroma durum, acute myeloblastic leukemia, chronic myelocytic leukemia.

7. apply as claimed in claim 5, wherein, described fibrosis is: systemic sclerosis, fibrosis of skin, pulmonary fibrosis, radioactive fibrosis, drug-induced fibrosis, renal fibrosis, hepatic fibrosis.

8. apply as claimed in claim 7, wherein, described pulmonary fibrosis is idiopathic pulmonary fibrosis.

9. apply as claimed in claim 7, wherein, described hepatic fibrosis is liver cirrhosis.