CN104788365A - Isonicotinamide derivative, preparation method and applications thereof - Google Patents

Isonicotinamide derivative, preparation method and applications thereof Download PDF

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CN104788365A
CN104788365A CN201410019845.5A CN201410019845A CN104788365A CN 104788365 A CN104788365 A CN 104788365A CN 201410019845 A CN201410019845 A CN 201410019845A CN 104788365 A CN104788365 A CN 104788365A
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group
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alkyl
hydrogen
acceptable salt
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CN104788365B (en
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匡荣仁
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Shanghai Allist Medicine Polytron Technologies Inc
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Shanghai Allist Pharmaceuticals Inc
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D213/78—Carbon atoms having three bonds to hetero atoms, with at the most one bond to halogen, e.g. ester or nitrile radicals
    • C07D213/81—Amides; Imides
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings

Abstract

The present invention discloses an isonicotinamide derivative represented by a formula (I) and a pharmaceutically acceptable salt thereof, wherein R1, R2, R3 and A are defined in the specification. The present invention further discloses a preparation method of the compounds, a drug composition containing the compound, and applications of the compounds in treatment of mammal, especially human overproliferation diseases and in preparation of drugs for treatment of mammal, especially human overproliferation diseases. The formula (I) is defined in the specification.

Description

Isonicotinamide derivative, its preparation method and application
Technical field
The present invention relates to and can be used for treatment mammalian hyper-proliferative disease as Isonicotinamide derivative of tumour and preparation method thereof, pharmaceutical composition containing described compound, and described compound is treatment Mammals especially human hyperproliferative's disease and for the preparation of the application in the medicine for the treatment of Mammals especially human hyperproliferative's disease.
Background technology
Cell signaling pathway cell growth, propagation and differentiation play an important role.Mitogen activated protein kinase (mitogen-activated protein kinase, MAPK) family's signal transduction is signal path important in cell, comprises the protein kinase (ERK) of extracellular signal regulation and control, 4 approach such as protein kinase (SAPK), P38MAPK and ERK5/BMK1 that c-JunN holds kinases (JNK)/stress activate.Wherein ERK (extracellular regulated kinase) is important member's extracellular signal-regulated kinase of MAPK family, its signal transduction pathway (Ras-Raf-MEK-ERK) can be activated by multiple somatomedin, cytokine and short disintegrating agent, playing a significant role in the propagation of cell, differentiation and apoptosis, is multi-signal joint or co-channel.The cascade that the GTP of activation causes serine/threonine kinase in conjunction with Ras activates, then MEK is activated after Raf is activated, MEK belongs to rare dual specificity kinase, makes Tyr and Thr two regulatory site phosphorylations and activates ERK race (ERK-1, ERK-2).3 that in this path, people pay close attention to important target molecules are: Ras, Raf and MEK.
Ras gene is a proto-oncogene, and family member comprises K-Ras, H-Ras and N-Ras.Wherein the generation of K-Ras and tumour and evolving relations maximum, its mutation frequency in tumour is also the highest.Research finds: carcinoma of the pancreas (90%), colorectal carcinoma (50%), lung cancer (30%), ovarian cancer (15%), thyroid carcinoma (50%), bladder cancer (6%), lupus erythematosus (SLE), mammary cancer, liver cancer, skin carcinoma, rheumatoid arthritis (RA), kidney and the leukemia of some (Leukemia) have higher KRas sudden change level.
Raf is one of most important proto-oncogene of the mankind, and Raf albumen is a serine threonine kinase, has three hypotype A-Raf, B-Raf and C-Raf.In catalytic substrate, A-Raf can only take MEK1 as substrate; B-raf take then MEK1/2 as substrate, is also the topmost upstream activating kinases of MEK; And C-Raf is outside substrate divided by MEK1/2, also with Retinoblastoma Protein (Rb), BCL2/BCL-XL associated death promotive factor (BAD) etc. for substrate, therefore the effect of B-Raf is more prone to connect Ras and MEK in Ras-Raf-MEK-ERK signal path.There is B-Raf sudden change in the human tumor of nearly 8%.Most mutant forms of B-Raf are B-RafV600E sudden change, and this sudden change causes downstream MEK-ERK signal path sustained activation, to the growing multiplication of tumour and Invasion and Metastasis most important.B-Raf sudden change mainly betides in melanoma, colorectal carcinoma and thyroid carcinoma.In melanoma, B-Raf mutation rate is the highest, about has 40% ~ 68% pernicious (transitivity) melanoma that B-Raf sudden change occurs.This sudden change is first high frequency mutator gene be found in melanoma, has become melanomatous therapeutic targets at present.Also there is B-Raf sudden change in other 5% ~ 8% colorectal carcinoma.In addition B-Raf mutation rate in thyroid carcinoma is about 30%, and wherein in thyroid papillary carcinoma hypotype, mutation rate is the highest, and mutation rate is 30% ~ 70%.In lung cancer, B-Raf sudden change also has report, but mutation rate is lower, is about 2%.
MEK belongs to dual specificity kinase, and family member comprises MEK1/MEK2, is typically expressed as MEK1/2.Their substrate specificity only has ERK1 and ERK2.Can be tumor phenotypes by transforming mammalian cells by the MEK1 of sustained activation, and by suppressing MEK and then stoping the intensity of activation of ERK to stop the cell growth in vitro of Ras sudden change.Mek inhibitor is different from general kinase inhibitor, and its not kinase whose with ATP competitive target ATP-binding site, does not also compete MEK1/2 binding site with ERK1/2, but be combined with the hydrophobic region that MEK closes on.Because mek inhibitor is non ATP competitive inhibitor, so compare B-Raf inhibitor, selectivity is better, and the combining site of mek inhibitor does not have homologous sequence on other kinases simultaneously, is that in all kinase inhibitor, specificity is the strongest.In addition, much the effect of this type of mek inhibitor is reversible, and after stopping using, kinases can be reactivated.Decrease the generation of the irreversible side effect caused due to mek inhibitor.
Due to the high rate that K-RAS sudden change, B-Raf suddenly change in each comfortable tumour, and both often mutual exclusions, cause the abnormal high activation probability of Ras-Raf-MEK in tumour.Therefore by suppressing MEK, interrupting the proliferation signal that the activation of Ras-Raf-MEK-ERK signalling channel gives tumour, thus reaching oncotherapy effect, present a good prospect in theory.
MEK is suppressed in multinomial research, to demonstrate potential treatment benefit.International patent application WO2002/006213 discloses the oxygenate ester of the 4-iodoanilino benzohydroxamic acid of (1) formula as follows, as mek inhibitor, can be used for treating various proliferative disease state, such as, relate to the ergogenic illness of MEK and the disease by MEK Cascade control.Wherein representative compound is PD0325901, but is clinical II phase halted state at present.
International patent application WO2003/077914 discloses the N3 alkylated benzimidazole derivatives as mek inhibitor, be used for the treatment of excess proliferative disease in Mammals, as cancer and inflammation, general structure is as shown in the formula shown in (2), wherein representative compound is AZD6244, and itself and chemotherapeutics docetaxel (Docetaxel) coupling is at present treated the KRas nonsmall-cell lung cancer that suddenlys change and entered the clinical III phase.
International patent application WO2006/045514 discloses and can be used for the 3-arylamino pyridine derivatives of overmedication proliferative disease as a series of replacements of cancer and inflammation, and general structure is as shown in the formula shown in (3).
International patent application WO2007/044515 discloses the azetidine that can be used for the mek inhibitor for the treatment of proliferative disease, general structure is as shown in the formula shown in (4), representative compound GDC0973 is good mek inhibitor (Exelixis Inc., 210East Grand Avenue, South San Francisco, California94080, United States, ACS Med.Chem.Lett.2012,3,416-421), research that is current and Wei Luofeini (Vemurafenib) combination therapy one line metastatic disease has entered the clinical III phase.
GSK1120212 (Trametinib) is first by (in May, 2013) mek inhibitor that FDA ratifies in similar drugs, go on the market as single medicine, be used for the treatment of the metastasis melanin tumor with B-RafV600E/V600K mutator gene and can not the melanoma patient of row operative treatment.GSK1120212 structure is as follows.
The domestic exploitation to mek inhibitor is at present in the preliminary stage, does not still have compound to enter clinical.New inhibitor is provided, suppresses Ras-Raf-MEK-ERK intracellular signaling, thus effective antiproliferative effect, and then develop antitumor drug safely and effectively, remain required for clinical application, this will contribute to the therapeutic advance advancing cancer undoubtedly.
Summary of the invention
The invention provides a kind of new inhibitor, can effectively suppress Ras-Raf-MEK-ERK intracellular signaling, upstream kinases can not only be suppressed the phosphorylation of MEK, MEK can also be suppressed the phosphorylation of ERK, there is both inhibitory effects, can more effective antiproliferative effect, treatment mammalian hyper-proliferative disease is as tumour.
At this point, the invention provides a kind of as shown in the formula (I) compound, or its pharmacy acceptable salt:
In formula, R 1and R 2be selected from hydrogen, C independently of one another 1-C 4alkyl ,-O (CH 2) 2oH, or R 1and R 2formed together with the nitrogen-atoms that they connect
R 3be selected from halogen ,-S-(C 1-C 4alkyl) or C 1-C 4alkoxyl group;
A is with 1 ~ 3 substituent aryl being selected from B group group, is selected from the substituent heteroaryl of B group group with 1 ~ 3, is selected from the substituent C of B group group with 1 ~ 3 3-C 8cycloalkyl, be selected from the substituent Heterocyclylalkyl of B group group with 1 ~ 3,
Wherein, B group group is selected from following one group: hydrogen, halogen, alkyl, alkoxyl group, haloalkyl ,-OH ,-SH ,-NO 2,-CN ,-COH ,-COOH ,-CONH 2,-CONH-C 1-C 4alkyl ,-CONH-C 3-C 6cycloalkyl ,-C 1-C 4alkyl-OH ,-S-C 1-C 4alkyl ,-NR 4r 5,-SO 2r 6,-COR 6,-COOR 6,-aryl ,-O-aryl ,-NH-aryl ,-S-aryl ,-SO 2-aryl ,-CO-aryl ,-CO 2-aryl ,-CONH-aryl ,-cycloalkyl ,-heteroaryl ,-Heterocyclylalkyl;
R 4and R 5be hydrogen ,-COR independently of one another 6,-SO 2r 6,-COOR 6;
R 6be selected from-C 1-C 4alkyl ,-C 1-C 4haloalkyl ,-C 3-C 6cycloalkyl.
Present invention also offers the preparation method of described formula (I) compound.
Present invention also offers a kind of pharmaceutical composition, containing described formula (I) compound or its pharmacy acceptable salt, and pharmaceutically acceptable carrier, vehicle or thinner.
Present invention also offers described formula (I) compound or its pharmacy acceptable salt and the pharmaceutical composition containing described formula (I) compound or its pharmacy acceptable salt and pharmaceutically acceptable carrier, vehicle or the thinner medicine as overmedication proliferative disease.
Present invention also offers described formula (I) compound or its pharmacy acceptable salt and the application of pharmaceutical composition in the medicine preparing overmedication proliferative disease containing described formula (I) compound or its pharmacy acceptable salt and pharmaceutically acceptable carrier, vehicle or thinner.
Present invention also offers described formula (I) compound or its pharmacy acceptable salt and the pharmaceutical composition containing described formula (I) compound or its pharmacy acceptable salt and pharmaceutically acceptable carrier, vehicle or thinner and prepare the application in the medicine for the treatment of tumour.
Present invention also offers one can suppress mitogen activated protein kinase kinases (MEK) active and hinder mitogen activated protein kinase kinase kinase (Raf) to the method for the phosphorylation of MEK, comprises formula (I) compound to patient therapeuticallv's significant quantity or its pharmacy acceptable salt and pharmaceutically acceptable carrier, vehicle or thinner.
Present invention also offers the method for overmedication proliferative disease, comprise the step of described formula (I) compound or its pharmacy acceptable salt that give object significant quantity in need or the pharmaceutical composition containing described formula (I) compound or its pharmacy acceptable salt and pharmaceutically acceptable carrier, vehicle or thinner.
Present invention also offers the method for the treatment of tumour, comprise the step of described formula (I) compound or its pharmacy acceptable salt that give object significant quantity in need or the pharmaceutical composition containing described formula (I) compound or its pharmacy acceptable salt and pharmaceutically acceptable carrier, vehicle or thinner.
In a preferred embodiment of the invention, described formula (I) compound or its pharmacy acceptable salt, R in formula 1and R 2be independently of one another hydrogen, or R 1and R 2formed together with the nitrogen-atoms that they connect preferred R 1and R 2for hydrogen.
In a preferred embodiment of the invention, described formula (I) compound or its pharmacy acceptable salt, R in formula 3be selected from halogen or-S-(C 1-C 4alkyl), preferred R 3for I or-SMe.
In a preferred embodiment of the invention, described formula (I) compound or its pharmacy acceptable salt, in formula, A is with 1 ~ 3 substituent aryl being selected from B group group, is selected from the substituent heteroaryl of B group group with 1 ~ 3, is selected from the substituent C of B group group with 1 ~ 3 3-C 8cycloalkyl, be selected from the substituent Heterocyclylalkyl of B group group with 1 ~ 3,
Wherein, B group group is selected from following one group: hydrogen, halogen, alkyl, alkoxyl group, haloalkyl ,-OH ,-SH ,-NO 2,-CN ,-COH ,-COOH ,-CONH 2,-CONH-C 1-C 4alkyl ,-CONH-C 3-C 6cycloalkyl ,-C 1-C 4alkyl-OH ,-S-C 1-C 4alkyl ,-NR 4r 5,-SO 2r 6,-COR 6,-COOR 6; R 4and R 5be hydrogen ,-COR independently of one another 6,-SO 2r 6,-COOR 6; R 6be selected from-C 1-C 4alkyl ,-C 1-C 4haloalkyl ,-C 3-C 6cycloalkyl;
Preferred, B group group is hydrogen or-NR 4r 5, R 4for hydrogen ,-COR 6,-SO 2r 6or-COOR 6, R 5for hydrogen, R 6for-C 1-C 4alkyl ,-C 1-C 4haloalkyl or-C 3-C 6cycloalkyl.
In a preferred embodiment of the invention, described formula (I) compound or its pharmacy acceptable salt, in formula, A is the substituent aryl being selected from B group group with 1 ~ 3, wherein, B group group is selected from following one group: hydrogen, halogen, alkyl, alkoxyl group, haloalkyl ,-OH ,-SH ,-NO 2,-CN ,-COH ,-COOH ,-CONH 2,-CONH-C 1-C 4alkyl ,-CONH-C 3-C 6cycloalkyl ,-C 1-C 4alkyl-OH ,-S-C 1-C 4alkyl ,-NR 4r 5,-SO 2r 6,-COR 6,-COOR 6,-aryl ,-O-aryl ,-NH-aryl ,-S-aryl ,-SO 2-aryl ,-CO-aryl ,-CO 2-aryl ,-CONH-aryl ,-cycloalkyl ,-heteroaryl ,-Heterocyclylalkyl; R 4and R 5be hydrogen ,-COR independently of one another 6,-SO 2r 6,-COOR 6; R 6be selected from-C 1-C 4alkyl ,-C 1-C 4haloalkyl ,-C 3-C 6cycloalkyl.
In another preferred embodiment of the present invention, described formula (I) compound or its pharmacy acceptable salt, in formula, A is the substituent aryl being selected from B group group with 1 ~ 3, and wherein B group group is selected from following one group: hydrogen, halogen, alkyl, alkoxyl group, haloalkyl ,-OH ,-SH ,-NO 2,-CN ,-COH ,-COOH ,-CONH 2,-CONH-C 1-C 4alkyl ,-CONH-C 3-C 6cycloalkyl ,-C 1-C 4alkyl-OH ,-S-C 1-C 4alkyl ,-NR 4r 5,-SO 2r 6,-COR 6,-COOR 6; R 4and R 5be hydrogen ,-COR independently of one another 6,-SO 2r 6,-COOR 6; R 6be selected from-C 1-C 4alkyl ,-C 1-C 4haloalkyl ,-C 3-C 6cycloalkyl;
Preferred, B group group is hydrogen or-NR 4r 5, R 4for hydrogen ,-COR 6,-SO 2r 6or-COOR 6, R 5for hydrogen, R 6for-C 1-C 4alkyl ,-C 1-C 4haloalkyl or-C 3-C 6cycloalkyl.
In a preferred embodiment of the invention, described formula (I) compound or its pharmacy acceptable salt, in formula, A is the substituent phenyl being selected from B group group with 1 ~ 3, wherein, B group group is selected from following one group: hydrogen, halogen, alkyl, alkoxyl group, haloalkyl ,-OH ,-SH ,-NO 2,-CN ,-COH ,-COOH ,-CONH 2,-CONH-C 1-C 4alkyl ,-CONH-C 3-C 6cycloalkyl ,-C 1-C 4alkyl-OH ,-S-C 1-C 4alkyl ,-NR 4r 5,-SO 2r 6,-COR 6,-COOR 6,-aryl ,-O-aryl ,-NH-aryl ,-S-aryl ,-SO 2-aryl ,-CO-aryl ,-CO 2-aryl ,-CONH-aryl ,-cycloalkyl ,-heteroaryl ,-Heterocyclylalkyl; R 4and R 5be hydrogen ,-COR independently of one another 6,-SO 2r 6,-COOR 6; R 6be selected from-C 1-C 4alkyl ,-C 1-C 4haloalkyl ,-C 3-C 6cycloalkyl.
In another preferred embodiment of the present invention, described formula (I) compound or its pharmacy acceptable salt, in formula, A is the substituent phenyl being selected from B group group with 1 ~ 3, and wherein B group group is selected from following one group: hydrogen, halogen, alkyl, alkoxyl group, haloalkyl ,-OH ,-SH ,-NO 2,-CN ,-COH ,-COOH ,-CONH 2,-CONH-C 1-C 4alkyl ,-CONH-C 3-C 6cycloalkyl ,-C 1-C 4alkyl-OH ,-S-C 1-C 4alkyl ,-NR 4r 5,-SO 2r 6,-COR 6,-COOR 6; R 4and R 5be hydrogen ,-COR independently of one another 6,-SO 2r 6,-COOR 6; R 6be selected from-C 1-C 4alkyl ,-C 1-C 4haloalkyl ,-C 3-C 6cycloalkyl;
Preferred, B group group is hydrogen or-NR 4r 5, R 4for hydrogen ,-COR 6,-SO 2r 6or-COOR 6, R 5for hydrogen, R 6for-C 1-C 4alkyl ,-C 1-C 4haloalkyl or-C 3-C 6cycloalkyl.
In a preferred embodiment of the invention, described formula (I) compound or its pharmacy acceptable salt, in formula, A is the phenyl being selected from the replacement of B group group substitution, and wherein B group group is selected from following one group: hydrogen, halogen, alkyl, alkoxyl group, haloalkyl ,-OH ,-SH ,-NO 2,-CN ,-COH ,-COOH ,-CONH 2,-CONH-C 1-C 4alkyl ,-CONH-C 3-C 6cycloalkyl ,-C 1-C 4alkyl-OH ,-S-C 1-C 4alkyl ,-NR 4r 5,-SO 2r 6,-COR 6,-COOR 6,-aryl ,-O-aryl ,-NH-aryl ,-S-aryl ,-SO 2-aryl ,-CO-aryl ,-CO 2-aryl ,-CONH-aryl ,-cycloalkyl ,-heteroaryl ,-Heterocyclylalkyl.
In another preferred embodiment of the present invention, described formula (I) compound or its pharmacy acceptable salt, in formula, A is the phenyl being selected from the replacement of B group group substitution, and wherein B group group is selected from following one group: hydrogen, halogen, alkyl, alkoxyl group, haloalkyl ,-OH ,-SH ,-NO 2,-CN ,-COH ,-COOH ,-CONH 2,-CONH-C 1-C 4alkyl ,-CONH-C 3-C 6cycloalkyl ,-C 1-C 4alkyl-OH ,-S-C 1-C 4alkyl ,-NR 4r 5,-SO 2r 6,-COR 6,-COOR 6; R 4and R 5be hydrogen ,-COR independently of one another 6,-SO 2r 6,-COOR 6; R 6be selected from-C 1-C 4alkyl ,-C 1-C 4haloalkyl ,-C 3-C 6cycloalkyl;
Preferred, B group group is hydrogen or-NR 4r 5, R 4for hydrogen ,-COR 6,-SO 2r 6or-COOR 6, R 5for hydrogen, R 6for-C 1-C 4alkyl ,-C 1-C 4haloalkyl or-C 3-C 6cycloalkyl.
In a preferred embodiment of the invention, described formula (I) compound or its pharmacy acceptable salt, in formula, A is by-NR 4r 5the phenyl replaced, described compound has as shown in the formula (Ia) structure:
Wherein, R 4be selected from hydrogen ,-COR 6,-SO 2r 6,-COOR 6, R 5for hydrogen;
R 6be selected from-C 1-C 4alkyl ,-C 1-C 4haloalkyl ,-C 3-C 6cycloalkyl;
R 1and R 2be selected from hydrogen, C independently of one another 1-C 4alkyl ,-O (CH 2) 2oH, or R 1and R 2formed together with the nitrogen-atoms that they connect
R 3be selected from halogen ,-S-(C 1-C 4alkyl) or C 1-C 4alkoxyl group.
In a preferred embodiment of formula (Ia) compound, R 4for-COR 6, wherein R 6for-C 1-C 4alkyl or-C 1-C 4haloalkyl, preferred R 6for-C 1-C 4alkyl.
In a preferred embodiment of formula (Ia) compound, R 4for-SO 2r 6, wherein R 6for-C 1-C 4alkyl or-C 3-C 6cycloalkyl.
In a preferred embodiment of formula (Ia) compound, R 4for-COOR 6, wherein R 6for-C 1-C 4alkyl.
In a preferred embodiment of formula (Ia) compound, R 4for hydrogen.
In another preferred embodiment of formula (Ia) compound, R 1and R 2be selected from independently of one another hydrogen, or R 1and R 2formed together with the nitrogen-atoms that they connect r 3for halogen.
In another preferred embodiment of formula (Ia) compound, R 1and R 2be selected from independently of one another hydrogen or r 3for halogen or-S-(C 1-C 4alkyl).
In another preferred embodiment of formula (Ia) compound, R 1and R 2for hydrogen; R 3for halogen.
In the present invention, as the compound represented by formula (I) or its pharmacy acceptable salt, can specifically mention:
3-[(3-t-butoxycarbonyl amino) thiophenyl]-5-[(the fluoro-4-iodophenyl of 2-) is amino] Isonicotinamide;
3-(3-aminobenzene-thio)-5-[(the fluoro-4-iodophenyl of 2-) is amino] Isonicotinamide;
3-[(3-kharophen) thiophenyl]-5-[(the fluoro-4-iodophenyl of 2-) is amino] Isonicotinamide;
3-[(3-methanesulfonamido) thiophenyl]-5-[(the fluoro-4-iodophenyl of 2-) is amino] Isonicotinamide;
3-[(3-ring third sulfonamido) thiophenyl]-5-[(the fluoro-4-iodophenyl of 2-) is amino] Isonicotinamide;
3-[(3-trifluoroacetamido) thiophenyl]-5-[(the fluoro-4-iodophenyl of 2-) is amino] Isonicotinamide;
3-[(3-methanesulfonamido) thiophenyl]-5-[(2-fluoro-4-methylthio group phenyl) is amino] Isonicotinamide;
(S)-3-[(3-kharophen) thiophenyl]-5-[(the fluoro-4-iodophenyl of 2-) is amino]-N-[(2,3-dihydroxyl) propyl group] Isonicotinamide;
(S)-3-[(3-methanesulfonamido) thiophenyl]-5-[(2-fluoro-4-methylthio group phenyl) is amino]-N-[(2,3-dihydroxyl) propyl group] Isonicotinamide;
1-({ 3-[(3-kharophen) thiophenyl]-5-[(the fluoro-4-iodophenyl of 2-) is amino] pyridin-4-yl } carbonyl)-3-(piperidin-2-yl) azetidine-3-alcohol.
The present invention also provides the method for preparation formula (Ia) compound, and it comprises the following steps:
Wherein, R 4be selected from hydrogen ,-COR 6,-SO 2r 6,-COOR 6, R 5for hydrogen; R 6be selected from-C 1-C 4alkyl ,-C 1-C 4haloalkyl ,-C 3-C 6cycloalkyl; R 1and R 2be selected from hydrogen, C independently of one another 1-C 4alkyl ,-O (CH 2) 2oH, or R 1and R 2formed together with the nitrogen-atoms that they connect r 3be selected from halogen ,-S-(C 1-C 4alkyl) or C 1-C 4alkoxyl group.
With 3,5-difluoro γ-picolinic acid for starting raw material, there is substitution reaction with compound (b) in 3,5-difluoro γ-picolinic acid, obtains compound (c) under suitable conditions; Compound (c) obtains general formula compound (d) with ammoniacal liquor condensation under CDI exists; There is substitution reaction in organic solvent in compound (d) and 3-sulfydryl phenylamino t-butyl formate, obtains compound (Ie) in the presence of a base; Compound (Ie) in presence of an acid in organic solvent de-Boc protection obtain compound (If); Compound (If) in the presence of an organic base in organic solvent with acyl chlorides or acid anhydrides generation acylation reaction, obtain compound (Ig); Compound (Ig) hydrolysis obtains compound (Ih); Compound (Ih) obtains compound (Ia) through acid amide condensation in organic solvent under condensing agent, organic amine exist.
The preparation method of formula of the present invention (Ia) compound, wherein there is substitution reaction and obtain compound (Ie) in compound (d), described alkali is conventional mineral alkali, include but not limited to salt of wormwood, sodium carbonate, cesium carbonate, sodium hydroxide, potassium hydroxide etc., described organic solvent is conventional organic solvent, includes but not limited to DMF, THF, DMSO, acetone etc.; The de-Boc protection of compound (Ie) obtains compound (If), described acid is conventional organic acid or mineral acid, include but not limited to hydrochloric acid, trifluoroacetic acid etc., described organic solvent is conventional organic solvent, includes but not limited to ethyl acetate, methylene dichloride, DMF etc.; Compound (If) obtains compound (Ig) through acidylate, and described organic bases includes but not limited to pyridine, triethylamine etc., and described organic solvent is conventional organic solvent, includes but not limited to methylene dichloride, THF, DMF etc.; Compound (Ig) hydrolysis obtains compound (m), and described hydrolysising condition includes but not limited to be hydrolyzed in acetic acid under Sodium Nitrite/vitriol oil exists, be hydrolyzed in ethanol under sodium hydroxide exists; Compound (Ih) obtains compound (Ia) through acid amide condensation, described organic amine includes but not limited to DIPEA, TEA, described condensing agent includes but not limited to HOBT/EDCI, HOBT/HATU, PyBOP, and described organic solvent includes but not limited to DMF, CH 2cl 2, THF.
In another preparation method's embodiment of the present invention, the preparation method of compound V is as follows:
With the bromo-pyridine of 2-for raw material, react with 1-tertbutyloxycarbonyl-3-azetidinone under n-Butyl Lithium exists and obtain compound (i); Compound (i) obtains compound (j) through reduction; Compound (a) is obtained again with trifluoroacetic acid deprotection.
Abbreviation noun in above-mentioned each preparation process represents respectively:
THF tetrahydrofuran (THF)
CH 2cl 2methylene dichloride
DMSO dimethyl sulfoxide (DMSO)
DMF DMF
DIPEA DIPEA
TFA trifluoroacetic acid
TEA triethylamine
CDI carbonyl dimidazoles
HOBT I-hydroxybenzotriazole
EDCI 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride
HATU 2-(7-azo benzotriazole)-N, N, N ', N '-tetramethyl-urea phosphofluoric acid ester
PyBOP phosphofluoric acid benzotriazole-1-base-oxygen base tripyrrole alkyl
In the present invention, halogen refers to fluorine, chlorine, bromine, iodine etc., preferred fluorine, chlorine, bromine.
C 1-C 4alkyl refer to methyl, ethyl, propyl group, sec.-propyl, butyl, isobutyl-, sec-butyl or the tertiary butyl; Preferable methyl, ethyl, propyl group, sec.-propyl or butyl, more preferably methyl.
C 1-C 4alkoxyl group refers to methoxyl group, oxyethyl group, n-propoxy-, isopropoxy, n-butoxy, isobutoxy, sec-butoxy or tert.-butoxy; Preferred methoxyl group, oxyethyl group, n-propoxy-, isopropoxy or n-butoxy; More preferably methoxyl group.
-S-(C 1-C 4alkyl) refer to methylthio group, ethylmercapto group, rosickyite base, isopropyisulfanyl, butylthio, isobutylthio, secondary butylthio or tertiary butylthio; Preferred methylthio group, ethylmercapto group, rosickyite base, isopropyisulfanyl or butylthio, more preferably methylthio group.
C 3-C 6cycloalkyl is cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl; Preferred cyclopropyl.
C 3-C 8cycloalkyl is cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, suberyl, ring octyl group; Preferred cyclohexyl.
-C 1-C 4haloalkyl refers to by one or more halogen, the C as defined herein of preferably one to five halogen atom replacement 1-C 4alkyl, includes but not limited to chloro-2 fluoro ethyls of trifluoromethyl, trifluoroethyl, difluoromethyl, 1-etc.
Haloalkyl refers to by one or more halogen, and the alkyl as defined herein of preferably one to five halogen atom replacement, includes but not limited to chloro-2 fluoro ethyls of trifluoromethyl, trifluoroethyl, difluoromethyl, 1-etc.
Alkyl refers to the saturated monovalent hydrocarbon of side chain of the saturated monovalent hydrocarbon of the straight chain of one to eight carbon atom or three to eight carbon atoms, includes but not limited to methyl, ethyl, propyl group, sec.-propyl, butyl (comprising all isomeric form), amyl group (comprising all isomeric form) etc.
Alkoxyl group refers to-OR group, and wherein R is alkyl defined herein, includes but not limited to methoxyl group, oxyethyl group, n-propoxy-, isopropoxy, n-butoxy etc.
Cycloalkyl refer to monocycle or fused bicyclic, saturated or part is undersaturated (but non-aromatic), the monovalent hydrocarbon radical of three to ten carboatomic ring atoms.One or two ring carbon atom can by-C (O)-,-C (S)-or-C (=NH)-group substitute.Described cycloalkyl includes but not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, hexamethylene-3-thiazolinyl etc.
Heterocyclylalkyl refers to monovalent monocyclic group that is saturated or part is undersaturated (but non-aromatic) three to eight annular atomses, or monovalence fused bicyclic group that is saturated or part is undersaturated (but non-aromatic) five to ten two annular atomses, wherein one or more ring hetero atoms are independently selected from O, S, N, and all the other annular atomses are carbon.One or two ring carbon atom can by-C (O)-,-C (S)-or-C (=NH)-group substitute.Described Heterocyclylalkyl includes but not limited to that azetidine base, pyrrolidyl, piperidyl, morpholinyl, piperazinyl, 2-oxopiperazinyl, THP trtrahydropyranyl, 2-oxo-piperidine base, pyrazolidyl, imidazolinyl, imidazolidyl, dihydropyridine base, tetrahydro pyridyl, oxazolinyl, oxazolidinyl, isoxazole alkyl, thiazolinyl, thiazolidyl, octahydro indyl, octahydro pseudoindoyl, tetrahydrofuran base, tetrahydrochysene pyrrole feed base etc.
Aryl refers to aromatic ring alkyl, and preferred carbonatoms is 6 to 14, is more preferably the aryl of 6 to 10, as phenyl and naphthyl, more preferably phenyl.
Heteroaryl refers to heteroatomic 5 to the 6 yuan of bicyclic heteroaryls or itself that are selected from N, S or O containing 1 to 4 and the thick dicyclic heteroaryl of phenyl ring.Described heteroaryl includes but not limited to furyl, thienyl, pyrryl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyridyl, pyrimidyl, pyridazinyl, pyrazinyl, benzofuryl, benzothienyl, benzothiazolyl, benzimidazolyl-, indyl, pseudoindoyl, quinolyl, isoquinolyl and quinazolyl, preferred imidazolyl, thiazolyl, pyrazinyl, benzimidazolyl-and quinolyl.
The present invention is contained (I) compound pharmaceutically acceptable salt also.Term " pharmacy acceptable salt " refers to acid salt or the base addition salt of the compounds of this invention of relative nontoxic.Described acid salt is the salt that formula (I) compound and suitable mineral acid or organic acid are formed, these salt can be prepared in the last isolation andpurification process of compound, or available mistake make the formula of purifying (I) compound together free alkali form carry out reacting to prepare with suitable organic acid or mineral acid.Representative acid salt comprise hydrobromate, hydrochloride, vitriol, sulphite, acetate, oxalate, valerate, oleate, palmitate, stearate, the moon silicate, borate, benzoate, lactic acid salt, phosphoric acid salt, toluylate, Citrate trianion, maleate, fumarate, succinate, tartrate, benzoate, mesylate, tosilate, gluconate, Lactobionate and lauryl sulfonate etc.Described base addition salt is the salt that formula (I) compound and suitable mineral alkali or organic bases are formed, comprise the salt such as formed with basic metal, alkaline-earth metal, quaternary ammonium cation, such as sodium salt, lithium salts, sylvite, calcium salt, magnesium salts, tetramethyl-quaternary ammonium salt, tetraethyl-quaternary ammonium salt etc.; Amine salt, comprises and ammonia (NH 3), primary amine, secondary amine or tertiary amine formed salt, as methylamine salt, dimethylamine salt, front three amine salt, triethylamine salt, ethylamine salt etc.
Compound of the present invention can deliver medicine to Mammals and comprise people, can administration in oral, rectum, parenteral (intravenously, intramuscular or subcutaneous), topical (pulvis, ointment or drops) or knurl.Described compound can be individually dosed, or with other pharmaceutically acceptable compound Combined Preparation.It may be noted that compound of the present invention can mix administration.
The present invention also provides pharmaceutical composition, and it contains formula (I) compound or its pharmacy acceptable salt as activeconstituents, and pharmaceutically acceptable carrier, vehicle or thinner.When pharmaceutical compositions, normally by formula (I) compound or its pharmacy acceptable salt and pharmaceutically acceptable carrier, vehicle or mixing diluents.
Preparation method the described present composition can be formulated as traditional drug formulations routinely.Such as tablet, pill, capsule, powder, granule, emulsion agent, mixed floating agent, dispersion liquid, solution, syrup, elixir, ointment, drops, suppository, inhalation, propellant etc.
The solid dosage that the present composition is used for oral administration comprises such as capsule, tablet, pill, powder and granule etc.In these solid dosages, formula (I) compound mixes with at least one conventional inert excipients (or carrier) as activeconstituents, such as with Trisodium Citrate or Si Liaodengji dicalcium phosphate feed grade, or mix with following compositions: (1) filler or expanding material, such as, starch, lactose, sucrose, glucose, N.F,USP MANNITOL and silicic acid etc.; (2) tackiness agent, such as Walocel MT 20.000PV, alginate, gelatin, Polyvinylpyrolidone (PVP), sucrose and gum arabic etc.; (3) wetting Agent for Printing Inks, such as, glycerine etc.; (4) disintegrating agent, such as agar, calcium carbonate, yam starch or tapioca (flour), alginic acid, some composition silicate and sodium carbonate etc.; (5) retarding solvent, such as paraffin etc.; (6) accelerator is absorbed, such as, quaternary ammonium compound etc.; (7) wetting agent, such as hexadecanol and glyceryl monostearate etc.; (8) sorbent material, such as, kaolin etc.; (9) lubricant, such as, talcum, calcium stearate, Magnesium Stearate, solid polyethylene glycol, sodium lauryl sulphate etc., or its mixture.Also buffer reagent can be comprised in capsule, tablet and pill.
Described solid dosage such as tablet, sugar-pill, capsule, pill and granule can adopt dressing and shell material such as enteric coating and other materials well known in the art to carry out dressing or microencapsulation.They can comprise opacifying agent, and in this composition, the release of activeconstituents can discharge in certain part in a delayed fashion in digestive tube.The example of adoptable embedding component is polymeric material and Wax.If desired, activeconstituents also can form microencapsulation form with one or more in above-mentioned vehicle.
The present composition is used for the liquid dosage form of oral administration and comprises and will learn acceptable emulsion, solution, suspension, syrup and tincture etc.Except as except formula (I) compound of activeconstituents, liquid dosage form can comprise the conventional inert diluent adopted in this area, as water and other solvents, solubilizing agent and emulsifying agent, such as, the mixture etc. of ethanol, Virahol, ethyl-carbonate, ethyl acetate, propylene glycol, 1,3 butylene glycol, dimethyl formamide and oils, particularly Oleum Gossypii semen, peanut oil, maize germ, sweet oil, Viscotrol C and sesame wet goods or these materials.
Except these inert diluents, liquid dosage form of the present invention also can comprise conven-tional adjuvants, as wetting agent, emulsifying agent and suspension agent, sweeting agent, correctives and spices etc.
Described suspension agent comprises, such as, ethoxylation isooctadecane alcohol, polyoxyethylene sorbitol and anhydro sorbitol only, the mixture of Microcrystalline Cellulose, aluminum methylate and agar etc. or these materials.
The formulation that the present composition is used for parenteral injection can comprise physiologically acceptable sterile, aqueous or anhydrous solution, dispersion liquid, suspension or emulsion, and for being again dissolved into aseptic Injectable solution or the sterilized powder of dispersion liquid.Suitable carrier, thinner, solvent or vehicle comprise water, ethanol, polyvalent alcohol and suitable mixture thereof.
Preparation formulation for the inventive mixture of topical comprises ointment, powder, suppository, drops, propellant and inhalation etc.As activeconstituents formula (I) compound aseptically with physiologically acceptable carrier and optional sanitas, buffer reagent, or the propelling agent that may need if desired is mixed together.
Therefore, the present invention also provides a kind of pharmaceutical preparation, and it contains 1-1000mg formula (I) compound or its pharmacy acceptable salt, and pharmaceutically acceptable carrier, vehicle or thinner.
The present invention also provides a kind of method for the treatment of tumour.Described tumour is alleviated by suppressing MEK activity or is treated, and comprises and uses formula (I) compound of 0.1-50mg/kg body weight/day or the step of its pharmacy acceptable salt to the patient of needs treatment.
By cell experiment and experimentation on animals, prove that formula (I) compound has cancer cell multiplication restraining effect, can be used for Therapeutic cancer and the medicine for the preparation of Therapeutic cancer, described cancer includes but not limited to melanoma, mammary cancer, colon and rectum carcinoma, nonsmall-cell lung cancer or small cell lung cancer, liver cancer, kidney etc.
The drug effect using conventional procedures of the compounds of this invention anticancer propagation measures, a kind of preferred evaluation method is Sulforhodamine B (SulforhodamIne B, SRB) protein staining method, calculates the inhibiting rate of medicine to cancer cell multiplication by the change measuring the absorbance value that drug effect produces after cancer cells.
Inhibiting rate (%)=[(blank OD-dosing OD)/blank OD] × 100%
Blank OD: the OD value referring to the hole of the cell not having drug effect normal growth.
Dosing OD: the OD value referring to the hole of the cell adding compound effects to be screened.
Half inhibitor concentration (IC 50) value employing GraphPad company PrIsm software 5.0 version, four parameter fitness methods calculate.Each experiment repetition 3 times, obtains the average IC of 3 experiments 50value is the final index of rejection ability.
Tested by protein immunoblotting, also prove that the compounds of this invention has MEK inhibit activities, effectively can suppress the kinase activity of MEK.
Accompanying drawing explanation
Accompanying drawing 1 is the compounds of this invention If-2 and Compound Ig per-3 pairs of Raf-MEK-ERK signal path MEK phosphorylations and ERK phosphorylation restraining effect lab diagram.
Accompanying drawing 2 is the human colon carcinoma HT-29 mice with tumor tumor propagation curves under the compounds of this invention Ig-3 and positive control AZD6244 and GSK1120212 administration.
Accompanying drawing 3 is the human colon carcinoma HCT116 mice with tumor tumor propagation curves under the compounds of this invention Ig-3 and positive control GSK1120212 administration.
Below in conjunction with specific embodiment, set forth the present invention further.Should be understood that these embodiments are only not used in for illustrating the present invention to limit the scope of the invention.The experimental technique of unreceipted actual conditions in the following example, usually conveniently condition, or according to the condition that manufacturer advises.Unless otherwise indicated, otherwise number and per-cent are respectively weight part and weight percent.
Embodiment
I. the compounds of this invention prepares embodiment
Intermediate (a): 3-hydroxyl-3-(piperidin-2-yl) azetidine trifluoroacetate (a)
N-Boc-3-hydroxyl-3-(pyridine-2-base) azetidine (i)
Be dissolved in by 2-bromopyridine (1.57g, 10mmol) in anhydrous THF, nitrogen protection, is cooled to-78 DEG C, slowly drips 4.8mL2.5M n-butyllithium solution.Stir 1 hour at-78 DEG C, the 10mLTHF solution of 1-Boc-azetidinone (1.71g, 10mmol) is slowly dropped in reaction solution, stir at-78 DEG C and namely react completely for 1 hour.Use saturated aqueous ammonium chloride cancellation, extraction into ethyl acetate, anhydrous sodium sulfate drying, filter, concentrate to obtain crude product, (methylene dichloride: methyl alcohol=100: 1) obtain 1.8g product, yield is 72% to cross column purification.
1HNMR(400MHz，CDCl 3)δ8.55-8.50(m，1H)，7.84(td，J=7.8，1.7Hz，1H)，7.70(d，J＝8.0Hz，1H)，7.31(ddd，J=7.4，5.0，0.9Hz，1H)，5.97(br s，1H)，4.32(d，J=9.5Hz，2H)，4.13(d，J=9.5Hz，2H)，1.50(s，9H)。
N-Boc-3-hydroxyl-3-(piperidin-2-yl) azetidine (j)
2g N-Boc-3-hydroxyl-3-(pyridine-2-base) azetidine (i) is dissolved in 100mL ethanol, add 20mg platinum dioxide, at 60atm, 60 DEG C are reacted 40 hours, cross post (methylene dichloride: methyl alcohol=100: 1-methyl alcohol: triethylamine=100: 1) obtain 1g product.
1H NMR(400MHz，CDCl 3)δ3.91(dd，J=9.2，4.1Hz，2H)，3.82(t，J=9.5Hz，2H)，3.19-3.10(m，1H)，2.79-2.77(m，1H)，2.72-2.66(m，1H)，1.98-1.89(m，1H)，1.71-1.64(m，2H)，1.44(s，9H)，1.40-1.34(m，3H)。
3-hydroxyl-3-(piperidin-2-yl) azetidine trifluoroacetate (a)
4ml methylene dichloride is added in 500mg N-Boc-3-hydroxyl-3-(piperidin-2-yl) azetidine (j), cool under ice bath, slow dropping 2ml trifluoroacetic acid, naturally be warming up to room temperature reaction to spend the night, concentrated, crude product is dissolved in toluene, and the concentrated solid becoming viscosity, is directly used in subsequent reactions again.
Embodiment 1:3-[(3-t-butoxycarbonyl amino) thiophenyl]-5-[(the fluoro-4-iodophenyl of 2-) is amino] Isonicotinamide (Ie-1)
Preparation method's synthesis with reference to PCT patent WO2012/055953 intermediate 2.A obtains compound (d-1).Compound (d-1) (0.35mmol), (3-sulfydryl benzene) ammonia t-butyl formate (0.455mmol), cesium carbonate (1.05mmol) in DMF (7mL) 60 DEG C reaction 4 hours, add 20ml water, 3 times are repeatedly extracted by ethyl acetate, merge organic phase, anhydrous sodium sulfate drying after saturated common salt water washing 2 times.Preparative separation purifying obtains solid 3-[(3-t-butoxycarbonyl amino) thiophenyl]-5-[(the fluoro-4-iodophenyl of 2-) is amino] Isonicotinamide (Ie-1) 0.25mmol, and yield is 71.4%.
ESI(+)m/z：581
Embodiment 2:3-(3-aminobenzene-thio)-5-[(the fluoro-4-iodophenyl of 2-) is amino] Isonicotinamide (If-2)
Compound (Ie-1) (0.5mmol) is dissolved in 20ml ethyl acetate, 5ml trifluoracetic acid is added at O DEG C, stirred overnight at room temperature, add the neutralization of 40ml saturated sodium bicarbonate solution, be separated organic phase, organic phase anhydrous sodium sulfate drying, preparative separation purifying obtains solid 3-(3-aminobenzene-thio)-5-[(the fluoro-4-iodophenyl of 2-) is amino] Isonicotinamide (If-2) 0.41mmol, and yield is 82%.
ESI(+)m/z：481
H 1-NMR (deuterated DMSO): δ 8.13 (s, 1H), δ 8.04 (s, 1H), δ 7.88 (d, 2H), δ 7.61 (d, 1H), δ 7.47 (s, 1H), δ 7.41 (d, 1H), δ 7.04 (t, 1H), δ 6.93 (t, 1H), δ 6.63 (s, 1H), δ 6.54 (dd, 2H), δ 5.30 (s, 2H).
Embodiment 3:3-[(3-kharophen) thiophenyl]-5-[(the fluoro-4-iodophenyl of 2-) is amino] Isonicotinamide (Ig-3)
Compound (If-2) (0.26mmol) is dissolved in the mixed solvent of 3ml pyridine and 3ml methylene dichloride, adds Acetyl Chloride 98Min. (0.52mmol) under ice bath.Stirring at room temperature 5 hours, adds 1Oml water and 10ml methylene dichloride, is separated organic phase, organic phase anhydrous sodium sulfate drying.Preparative separation purifying obtains solid 3-[(3-kharophen) thiophenyl]-5-[(the fluoro-4-iodophenyl of 2-) is amino] Isonicotinamide (Ig-3) 0.21mmol, and yield is 81%.
ESI(+)m/z：523
H 1-NMR (deuterated DMSO): δ 10.05 (s, 1H), δ 8.18 (s, 1H), δ 8.07 (s, 1H), δ 7.90-7.93 (d, 2H), δ 7.68 (s, 1H), δ 7.59 (d, 1H), δ 7.51 (m, 2H), δ 7.42 (d, 1H), δ 7.32 (t, 1H), δ 7.05 (d, 1H), δ 6.96 (t, 1H), δ 7.05 (s, 3H).
Embodiment 4:3-[(3-methanesulfonamido) thiophenyl]-5-[(the fluoro-4-iodophenyl of 2-) is amino] Isonicotinamide (Ig-4)
Compound (If-2) (0.26mmol) is dissolved in the mixed solvent of 3ml pyridine and 3ml methylene dichloride, adds methylsulfonyl chloride (0.52mmol) under ice bath.Stirring at room temperature 5 hours, adds 10ml water and 10ml methylene dichloride, is separated organic phase, organic phase anhydrous sodium sulfate drying.Preparative separation purifying obtains solid 3-[(3-methanesulfonamido) thiophenyl]-5-[(the fluoro-4-iodophenyl of 2-) is amino] Isonicotinamide (Ig-4) 0.16mmol, and yield is 61.5%.
ESI(+)m/z：559
Embodiment 5:3-[(3-ring third sulfonamido) thiophenyl]-5-[(the fluoro-4-iodophenyl of 2-) is amino] Isonicotinamide (Ig-5)
Compound (If-2) (0.26mmol) is dissolved in the mixed solvent of 3ml pyridine and 3ml methylene dichloride, adds ring third SULPHURYL CHLORIDE (0.52mmol) under ice bath.Stirring at room temperature 5 hours, adds 10ml water and 10ml methylene dichloride, is separated organic phase, organic phase anhydrous sodium sulfate drying.Preparative separation purifying obtains solid 3-[(3-ring third sulfonamido) thiophenyl]-5-[(the fluoro-4-iodophenyl of 2-) is amino] Isonicotinamide (Ig-5) 0.23mmol, and yield is 88%.
ESI(+)m/z：585
Embodiment 6:3-[(3-trifluoroacetamido) thiophenyl]-5-[(the fluoro-4-iodophenyl of 2-) is amino] Isonicotinamide (Ig-6)
Compound (If-2) (0.26mmol) is dissolved in the mixed solvent of 3ml pyridine and 3ml methylene dichloride, adds trifluoro-acetic anhydride (0.52mmol) under ice bath.Stirring at room temperature 5 hours, adds 10ml water and 10ml methylene dichloride, is separated organic phase, organic phase anhydrous sodium sulfate drying.Preparative separation purifying obtains solid 3-[(3-trifluoroacetamido) thiophenyl]-5-[(the fluoro-4-iodophenyl of 2-) is amino] Isonicotinamide (Ig-6) 0.14mmol, and yield is 53.8%.
ESI(+)m/z：577
Embodiment 7:3-[(3-methanesulfonamido) thiophenyl]-5-[(2-fluoro-4-methylthio group phenyl) is amino] Isonicotinamide (Ig-7)
The synthetic method of reference example 2 compound (If-2), synthesis obtains compound (If-7).Compound (If-7) (0.26mmol) is dissolved in the mixed solvent of 3ml pyridine and 3ml methylene dichloride, adds methylsulfonyl chloride (0.52mmol) under ice bath.Stirring at room temperature 5 hours, adds 10ml water and 10ml methylene dichloride, is separated organic phase, organic phase anhydrous sodium sulfate drying.Preparative separation purifying obtains solid 3-[(3-methanesulfonamido) thiophenyl]-5-[(2-fluoro-4-methylthio group phenyl) is amino] Isonicotinamide (Ig-7) 0.19mmol, and yield is 73%.
ESI(+)m/z：479
Embodiment 8:(S)-3-[(3-kharophen) thiophenyl]-5-[(the fluoro-4-iodophenyl of 2-) is amino]-N-[(2,3-dihydroxyl) propyl group] Isonicotinamide (Ia-8)
Compound (Ig-3) (0.38mmol) is suspended in 5ml acetic acid, slowly add the concentrated sulfuric acid solution (0.25ml) of Sodium Nitrite (0.5mmol), stirring at room temperature reaction is spent the night and is obtained compound (Ih-8).Compound (Ih-8) (0.09mmol), (S)-3-amino-1,2-propylene glycol (0.18mmol), PyBop (0.105mmol), DIPEA (0.6mmol) room temperature reaction in DMF (3mL) spends the night.Add 10ml water, repeatedly extract 3 times by ethyl acetate, merge organic phase, anhydrous sodium sulfate drying after saturated common salt washes 2 times.Preparative separation purifying obtains solid (S)-3-[(3-kharophen) thiophenyl]-5-[(the fluoro-4-iodophenyl of 2-) is amino]-N-[(2,3-dihydroxyl) propyl group] Isonicotinamide (Ia-8) 0.045mmol, yield is 50%.
ESI(+)m/z：597
Embodiment 9:(S)-3-[(3-methanesulfonamido) thiophenyl]-5-[(2-fluoro-4-methylthio group phenyl) is amino]-N-[(2,3-dihydroxyl) propyl group] Isonicotinamide (Ia-9)
The synthetic method of reference example 4 compound (Ig-4), synthesis obtains compound (Ig-9).Compound (Ig-9) (0.31mmol) is dissolved in 10ml ethanol, adds sodium hydroxide (0.45mmol), refluxes and within 3 hours, obtains compound (Ih-9).Compound (Ih-9) (0.20mmol), (S)-3-amino-1,2-propylene glycol (0.40mmol), PyBop (0.26mmol), DIPEA (1.0mmol) room temperature reaction in DMF (5mL) spends the night, and adds 10ml water, repeatedly extracts 3 times by ethyl acetate, merge organic phase, anhydrous sodium sulfate drying after saturated common salt washes 2 times.Preparative separation purifying obtains solid (S)-3-[(3-methanesulfonamido) thiophenyl]-5-[(2-fluoro-4-methylthio group phenyl) is amino]-N-[(2,3-dihydroxyl) propyl group] Isonicotinamide (Ia-9) 0.12mmol, yield is 60%.
ESI(+)m/z：553
Embodiment 10:1-({ 3-[(3-kharophen) thiophenyl]-5-[(the fluoro-4-iodophenyl of 2-) is amino] pyridin-4-yl } carbonyl)-3-(piperidin-2-yl) azetidine-3-alcohol (Ia-10)
Compound (Ih-8) (0.09mmol), intermediate (a) (0.18mmol), PyBop (0.11mmol), DIPEA (0.92mmol) room temperature reaction in DMF (3mL) spends the night.Add 10ml water, repeatedly extract 3 times by ethyl acetate, merge organic phase, anhydrous sodium sulfate drying after saturated common salt washes 2 times.Preparative separation purifying obtains solid 1-({ 3-[(3-kharophen) thiophenyl]-5-[(the fluoro-4-iodophenyl of 2-) is amino] pyridin-4-yl } carbonyl)-3-(piperidin-2-yl) azetidine-3-alcohol (Ia-10) 0.035mmol, and yield is 38.8%.
ESI(+)m/z：662
(II). the compounds of this invention active testing embodiment
Testing example 1: the compounds of this invention is to human skin cancer cells (A375), human colon cancer cell (HT-29), human breast cancer cell (MDA-MB-231) inhibited proliferation
Get the cell being in logarithmic phase and be seeded in 96 orifice plates that (cell concn is 5000/hole; Cell suspension 180 μ l/ hole), 37 DEG C, 5%CO 2cultivate and make cell attachment in 24 hours.Each compound is dissolved in DMSO the storage liquid being mixed with 10mM in advance, in another 96 orifice plate, be diluted to 10 times of object concentration again with perfect medium when detecting, then in 96 orifice plates of inoculating cell, add compound 20 μ l/ hole, namely arrive object concentration.Each concentration establishes 3 multiple holes, and establishes blank.Continue at 37 DEG C, 5%CO 2middle continuation cultivation 72 hours.Stop cultivating, every hole adds 50% trichoroacetic acid(TCA) and the TCA (final concentration 10%) of 50 μ l precoolings (4 DEG C), is placed on 4 DEG C and fixes 1 hour, with purified water washing at least 5 times, and seasoning or 60 DEG C of oven dryings in air.With Sulforhodamine B and SRB of the purified water preparation 4mg/ml containing 1% glacial acetic acid, every hole adds 100 μ l, and room temperature dyes 1 hour, abandons supernatant, with 1% glacial acetic acid washing at least 5 removing non-specific bindings, and dried for standby.Every hole adds the Tris-HCl solubilize of the 10mM of 150 μ l, surveys OD value at 510nm wavelength place, and carries out data preparation calculating inhibiting rate.The results are shown in Table 1:
Table 1
Note: AZD6244 preparation method is with reference to WO2003/077914 embodiment 10; GDC0973 preparation method is with reference to WO2007/044515 embodiment 22
Test result shows: the compounds of this invention has good inhibited proliferation to human skin cancer cells (A375), human colon cancer cell (HT-29), human breast cancer cell (MDA-MB-231).Wherein Compound Ig per-3 pairs of human skin cancer cells (A375), human colon cancer cell (HT-29), human breast cancer cell (MDA-MB-231) inhibited proliferation are obviously better than AZD-6244 and GDC-0973.
By same method, test I g-3 compound on tumor cell growth inhibition IC50 value in B-RafV600 abrupt junction colon-cancer cell Colon205, K-Ras mutant human lung cell A549, colon cancer cell HCT116, pancreatic cancer cell MIA-Paca2 and nonsmall-cell lung cancer H2122, the results are shown in Table 2.
Table 2
Note: *: K-Ras mutation; #:B-Raf V600E mutation
Test result shows: the compounds of this invention Ig-3 has good inhibited proliferation to B-RafV600 abrupt junction colon-cancer cell Colon205, K-Ras mutant human lung cell A549, colon cancer cell HCT116, pancreatic cancer cell MIA-Paca2 and nonsmall-cell lung cancer H2122, and is better than AZD6244.
Testing example 2: Compound I f-2 and Compound Ig per-3 pairs of Raf-MEK-ERK intracellular signaling restraining effect
Get the human colon cancer cell (HT-29) being in logarithmic phase and be seeded in 6 orifice plates that (cell concn is 1 × 10 6individual/hole; Cell suspension 2ml perfect medium/hole), 37 DEG C, 5%CO 2cultivate and make cell attachment in 24 hours.Positive compound AZD6244, GSK1120212, test compounds If-2 and Ig-3 is dissolved in DMSO the storage liquid being mixed with 10mM respectively in advance, in another 6 orifice plate, be diluted to 1000 times of object concentration again with perfect medium when detecting, then in 6 orifice plates of inoculating cell, add compound respectively, achieve the goal concentration:
AZD624410nM，50nM，250nM；
GSK1120212 10nM，50nM；
If-2 2nM，10nM，50nM；
Ig-3 2nM，10nM，50nM
Administration timing of drug is decided to be 2 hours.After cell administration stimulates 2 hours, Aspirate medium, every hole adds the 1*SDS cell pyrolysis liquid of 100 μ l, and cell scraper is collected, and is transferred in centrifuge tube with pipettor, 95 DEG C, centrifugal 5 minutes of 5min, whizzer 10000g, carry out electrophoretic analysis, constant voltage 100V turns 2-3 hour, is gone to by albumen on PVDF (polyvinylidinedifluoride) film.Take out pvdf membrane, rinsing 1-2min in purified water, puts into confining liquid room temperature and closes 1H.(confining liquid is 5% skim-milk with TBST preparation.) using 5%BSA (TBST preparation) as in primary antibodie diluent in advance hybridization bag, add primary antibodie (p-ERK antibody, ERK antibody, P-MEK, MEK antibody) Dilution ratio is 1: 1000 (Actin is 1: 10000), again by close after pvdf membrane carry out cutting by object band, put into corresponding hybridization bag, sealing, 4 DEG C of overnight incubation.Reclaim primary antibodie, Hybond membrane puts into TBST, washes film 6 × 5min.Using 5% skim-milk (TBST preparation) as two anti-diluents, adding primary antibodie Dilution ratio is 1: 200-1: 500, then Hybond membrane is put into corresponding two anti-(coupling HRP-goat against murine/rabbit monoclonal antibodies), incubated at room 1H.Reclaim primary antibodie, Hybond membrane puts into TBST, washes film 6 × 5min.With the development of ECL luminescence reagent box, and compressing tablet.Test-results is shown in accompanying drawing 1.
Test result shows: the compounds of this invention has good restraining effect to Raf-MEK-ERK intracellular signaling, has restraining effect in various degree to the phosphorylation of MEK and ERK.Wherein namely Ig-3 compound has suppression to ERK protein phosphorylation under 2nM concentration, suitable with AZD6244 inhibition under 250nM concentration, also have certain suppression, and AZD6244 is without this restraining effect to MEK phosphorylation simultaneously.Compound Ig per-3 and the phosphorylation of GSK1120212 to MEK and ERK have the restraining effect of equal extent.
Testing example 3: to the growth-inhibiting effect of human colon carcinoma HT-29 nude mouse subcutaneous transplantation knurl
Evaluation test medicine Ig-3 treats antitumor action and the safety research of human colon carcinoma HT-29 cell strain animal xenografts model.
Method: HT-29 cell is cultivated in the Mcloy's5A substratum being added with 10%, is placed on 37 DEG C and cultivates containing in the constant incubator of 5%CO2.Collect the cell of exponential phase of growth and count, resuspended with physiological saline, be inoculated in the right dorsal scapular position of Balb/c-nu mouse, set up human colon carcinoma xenotransplantation model in situ.Back inoculation 6 × 10 on the right side of every mouse 6the HT-29 cell of number.Treat tumor average volume 128mm 3time, according to tumor size random packet.Experiment is divided into 25mg/kgBID (the one day twice) group of test medicine Ig-3, and positive controls AZD624450mg/kg BID and GSK11202123mg/kg QD group often organize 6, oral administration gavage administration 14day.Carry out safety evaluation according to the weight of animals change and death condition, carry out therapeutic evaluation time of lag according to Relative tumor appreciation rate (T/C%) and tumour.After tumor inoculation, routine monitoring includes tumor growth and the impact for the treatment of on animal normal behaviour, and particular content has the reactivity of laboratory animal, ingests and drinking-water situation, body weight increases or (body weight measures the weekly 2 times) situation of reduction, eyes, by hair and other abnormal conditions.
Gross tumor volume calculation formula is: major diameter × minor axis 2/ 2.
Relative tumor proliferation rate T/C (%): at point sometime, the percent value for the treatment of group and control group relative tumour volume.T and C is respectively treatment group and the control group relative tumour volume (RTV) at a certain particular point in time.Calculation formula is: T/C%=T rTV/ C rTV× 100%.(T rTV: treatment group RTV; C rTV: negative control group RTV).Judgement criteria is: T/C (%) >40% is invalid; T/C (%)≤40%, and be effective through statistical procedures P<0.05.
Result: testing drug Ig-3 (25mg/kg BID) treatment group is in administration the last day, all show tumor inhibition effect, T/C (%) is 13.2% respectively, and comparing negative control group statistically has significant difference P<0.001; Positive control drug AZD624450mg/kg BID group and GSK1120212mg/kg QD administration T/C the last day (%) are 29.5% and 8.9%, P value <0.001 respectively, and positive control is effectively reliable.Four experimental group tumor growth curves are shown in accompanying drawing 2,
Test result shows: the growth of the compounds of this invention to human colon carcinoma HT-29 nude mouse subcutaneous transplantation knurl has good restraining effect, and shows good security.Namely testing drug Ig-3 shows good drug action under 25mg/kg BID, and toxicity is also at tolerance interval.
Testing example 4: to the growth-inhibiting effect of human colon carcinoma HCT116 nude mouse subcutaneous transplantation knurl
Evaluation test medicine Ig-3 treats antitumor action and the safety research of KRas mutant human colorectal carcinoma HCT116 cell strain animal xenografts model.
Method: HCT116 cell is cultivated in the Mcloy's5A substratum being added with 10%, is placed on 37 DEG C and cultivates containing in the constant incubator of 5%CO2.Collect the cell of exponential phase of growth and count, resuspended with physiological saline, be inoculated in the right dorsal scapular position of Balb/c-nu mouse, set up human colon carcinoma xenotransplantation model in situ.Back inoculation 2.5 × 10 on the right side of every mouse 6the HCT-116 cell of number.Treat tumor average volume 200mm 3time, according to tumor size random packet.Experiment is divided into the 25mg/kg BID (a day twice) of test medicine Ig-3 and positive control drug GSK11202123mg/kg QD (once a day) to organize and Vehicle controls group, often organizes 6, oral administration gavage administration 14day.Carry out safety evaluation according to the weight of animals change and death condition, carry out therapeutic evaluation time of lag according to Relative tumor appreciation rate (T/C%) and tumour.After tumor inoculation, routine monitoring includes tumor growth and the impact for the treatment of on animal normal behaviour, and particular content has the reactivity of laboratory animal, ingests and drinking-water situation, body weight increases or (body weight measures the weekly 2 times) situation of reduction, eyes, by hair and other abnormal conditions.
Gross tumor volume calculation formula is: major diameter × minor axis 2/ 2.
Relative tumor proliferation rate T/C (%): at point sometime, the percent value for the treatment of group and control group relative tumour volume.T and C is respectively treatment group and the control group relative tumour volume (RTV) at a certain particular point in time.Calculation formula is: T/C%=T rTV/ C rTV× 100%.(T rTV: treatment group RTV; C rTV: negative control group RTV).Judgement criteria is: T/C (%) >40% is invalid; T/C (%)≤40%, and be effective through statistical procedures P<0.05.
Result: testing drug Ig-3 (25mg/kg BID) treatment group is in administration the last day, show obvious tumor inhibition effect, T/C (%) is 12.6%, and comparing negative control group statistically has significant difference P<0.001; Positive control drug GSK11202123mg/kg QD group administration T/C the last day (%) is 7.8%, P value <0.001, and positive control is effectively reliable.Three experimental group tumor growth curves are shown in accompanying drawing 3.
Test result shows: the growth of the compounds of this invention to human colon carcinoma HCT116 nude mouse subcutaneous transplantation knurl has good restraining effect, and shows good security.Namely testing drug Ig-3 shows good drug action under 25mg/kg BID, and toxicity is also at tolerance interval.
The all documents mentioned in this article are merged in the application all by reference.What should indicate in addition is, after the above-mentioned disclosure of having read the application, those skilled in the art can without the need to deviating from the spirit and scope of the present invention, various modification, change or amendment are made to the present invention, but these versions equally all should fall within the scope described in the application's appended claims.

Claims (23)

1. following formula (I) compound, or its pharmacy acceptable salt:
In formula, R 1and R 2be selected from hydrogen, C independently of one another 1-C 4alkyl ,-O (CH 2) 2oH, or R 1and R 2formed together with the nitrogen-atoms that they connect
R 3be selected from halogen ,-S-(C 1-C 4alkyl) or C 1-C 4alkoxyl group;
A is with 1 ~ 3 substituent aryl being selected from B group group, is selected from the substituent heteroaryl of B group group with 1 ~ 3, is selected from the substituent C of B group group with 1 ~ 3 3-C 8cycloalkyl or be selected from the substituent Heterocyclylalkyl of B group group with 1 ~ 3,
Wherein, B group group is selected from following one group: hydrogen, halogen, alkyl, alkoxyl group, haloalkyl ,-OH ,-SH ,-NO 2,-CN ,-COH ,-COOH ,-CONH 2,-CONH-C 1-C 4alkyl ,-CONH-C 3-C 6cycloalkyl ,-C 1-C 4alkyl-OH ,-S-C 1-C 4alkyl ,-NR 4r 5,-SO 2r 6,-COR 6,-COOR 6,-aryl ,-O-aryl ,-NH-aryl ,-S-aryl ,-SO 2-aryl ,-CO-aryl ,-CO 2-aryl ,-CONH-aryl ,-cycloalkyl ,-heteroaryl ,-Heterocyclylalkyl;
R 4and R 5be hydrogen ,-COR independently of one another 6,-SO 2r 6,-COOR 6;
R 6be selected from-C 1-C 4alkyl ,-C 1-C 4haloalkyl ,-C 3-C 6cycloalkyl.
2. compound as claimed in claim 1 or its pharmacy acceptable salt, is characterized in that A is with 1 ~ 3 substituent aryl being selected from B group group, is selected from the substituent heteroaryl of B group group with 1 ~ 3, is selected from the substituent C of B group group with 1 ~ 3 3-C 8cycloalkyl or be selected from the substituent Heterocyclylalkyl of B group group with 1 ~ 3,
Wherein, B group group is selected from following one group: hydrogen, halogen, alkyl, alkoxyl group, haloalkyl ,-OH ,-SH ,-NO 2,-CN ,-COH ,-COOH ,-CONH 2,-CONH-C 1-C 4alkyl ,-CONH-C 3-C 6cycloalkyl ,-C 1-C 4alkyl-OH ,-S-C 1-C 4alkyl ,-NR 4r 5,-SO 2r 6,-COR 6,-COOR 6;
R 4and R 5be hydrogen ,-COR independently of one another 6,-SO 2r 6,-COOR 6;
R 6be selected from-C 1-C 4alkyl ,-C 1-C 4haloalkyl ,-C 3-C 6cycloalkyl.
3. compound as claimed in claim 2 or its pharmacy acceptable salt, it is characterized in that A is the substituent aryl being selected from B group group with 1 ~ 3, wherein B group group is selected from following one group: hydrogen, halogen, alkyl, alkoxyl group, haloalkyl ,-OH ,-SH ,-NO 2,-CN ,-COH ,-COOH ,-CONH 2,-CONH-C 1-C 4alkyl ,-CONH-C 3-C 6cycloalkyl ,-C 1-C 4alkyl-OH ,-S-C 1-C 4alkyl ,-NR 4r 5,-SO 2r 6,-COR 6,-COOR 6;
R 4and R 5be hydrogen ,-COR independently of one another 6,-SO 2r 6,-COOR 6;
R 6be selected from-C 1-C 4alkyl ,-C 1-C 4haloalkyl ,-C 3-C 6cycloalkyl.
4. compound as claimed in claim 3 or its pharmacy acceptable salt, it is characterized in that A is the substituent phenyl being selected from B group group with 1 ~ 3, wherein B group group is selected from following one group: hydrogen, halogen, alkyl, alkoxyl group, haloalkyl ,-OH ,-SH ,-NO 2,-CN ,-COH ,-COOH ,-CONH 2,-CONH-C 1-C 4alkyl ,-CONH-C 3-C 6cycloalkyl ,-C 1-C 4alkyl-OH ,-S-C 1-C 4alkyl ,-NR 4r 5,-SO 2r 6,-COR 6,-COOR 6;
R 4and R 5be hydrogen ,-COR independently of one another 6,-SO 2r 6,-COOR 6;
R 6be selected from-C 1-C 4alkyl ,-C 1-C 4haloalkyl ,-C 3-C 6cycloalkyl.
5. compound as claimed in claim 4 or its pharmacy acceptable salt, it is characterized in that A is the phenyl being selected from the replacement of B group group substitution, wherein B group group is selected from following one group: hydrogen, halogen, alkyl, alkoxyl group, haloalkyl ,-OH ,-SH ,-NO 2,-CN ,-COH ,-COOH ,-CONH 2,-CONH-C 1-C 4alkyl ,-CONH-C 3-C 6cycloalkyl ,-C 1-C 4alkyl-OH ,-S-C 1-C 4alkyl ,-NR 4r 5,-SO 2r 6,-COR 6,-COOR 6;
R 4and R 5be hydrogen ,-COR independently of one another 6,-SO 2r 6,-COOR 6;
R 6be selected from-C 1-C 4alkyl ,-C 1-C 4haloalkyl ,-C 3-C 6cycloalkyl.
6. the compound as described in any one of Claims 1 to 5 or its pharmacy acceptable salt, is characterized in that A is by-NR 4r 5replace phenyl, its structural formula as shown in the formula (Ia),
Wherein, R 4be selected from hydrogen ,-COR 6,-SO 2r 6,-COOR 6, R 5for hydrogen;
R 6be selected from-C 1-C 4alkyl ,-C 1-C 4haloalkyl ,-C 3-C 6cycloalkyl;
R 1and R 2be selected from hydrogen, C independently of one another 1-C 4alkyl ,-O (CH 2) 2oH, or R 1and R 2formed together with the nitrogen-atoms that they connect
R 3be selected from halogen ,-S-(C 1-C 4alkyl) or C 1-C 4alkoxyl group.
7. compound as claimed in claim 6 or its pharmacy acceptable salt, is characterized in that R 4for-COR 6, wherein R 6for-C 1-C 4alkyl or-C 1-C 4haloalkyl.
8. compound as claimed in claim 7 or its pharmacy acceptable salt, is characterized in that R 4for-COR 6, wherein R 6for-C 1-C 4alkyl.
9. compound as claimed in claim 6 or its pharmacy acceptable salt, is characterized in that R 4for-SO 2r 6, wherein R 6for-C 1-C 4alkyl or-C 3-C 6cycloalkyl.
10. compound as claimed in claim 6 or its pharmacy acceptable salt, is characterized in that R 4for-COOR 6, wherein R 6for-C 1-C 4alkyl.
11. compound as claimed in claim 6 or its pharmacy acceptable salts, is characterized in that R 4for hydrogen.
12. compounds as described in claim 1 or 7 or 8 or its pharmacy acceptable salt, is characterized in that R 1and R 2be selected from independently of one another hydrogen, or R 1and R 2formed together with the nitrogen-atoms that they connect r 3for halogen.
13. compounds as described in claim 1 or 9 or its pharmacy acceptable salt, is characterized in that R 1and R 2be selected from independently of one another hydrogen or r 3for halogen or-S-(C 1-C 4alkyl).
14. compounds as described in claim 1 or 10 or 11 or its pharmacy acceptable salt, is characterized in that R 1and R 2for hydrogen; R 3for halogen.
15. compound as claimed in claim 1 or its pharmacy acceptable salts, is characterized in that described compound is selected from lower group:
3-[(3-t-butoxycarbonyl amino) thiophenyl]-5-[(the fluoro-4-iodophenyl of 2-) is amino] Isonicotinamide;
3-(3-aminobenzene-thio)-5-[(the fluoro-4-iodophenyl of 2-) is amino] Isonicotinamide;
3-[(3-kharophen) thiophenyl]-5-[(the fluoro-4-iodophenyl of 2-) is amino] Isonicotinamide;
3-[(3-methanesulfonamido) thiophenyl]-5-[(the fluoro-4-iodophenyl of 2-) is amino] Isonicotinamide;
3-[(3-ring third sulfonamido) thiophenyl]-5-[(the fluoro-4-iodophenyl of 2-) is amino] Isonicotinamide;
3-[(3-trifluoroacetamido) thiophenyl]-5-[(the fluoro-4-iodophenyl of 2-) is amino] Isonicotinamide;
3-[(3-methanesulfonamido) thiophenyl]-5-[(2-fluoro-4-methylthio group phenyl) is amino] Isonicotinamide;
(S)-3-[(3-kharophen) thiophenyl]-5-[(the fluoro-4-iodophenyl of 2-) is amino]-N-[(2,3-dihydroxyl) propyl group] Isonicotinamide;
(S)-3-[(3-methanesulfonamido) thiophenyl]-5-[(2-fluoro-4-methylthio group phenyl) is amino]-N-[(2,3-dihydroxyl) propyl group] Isonicotinamide;
1-({ 3-[(3-kharophen) thiophenyl]-5-[(the fluoro-4-iodophenyl of 2-) is amino] pyridin-4-yl } carbonyl)-3-(piperidin-2-yl) azetidine-3-alcohol.
16. methods preparing formula (Ia) compound as claimed in claim 6, it comprises the following steps:
Wherein R 1, R 2, R 3, R 4, R 5with describe in claim 6 identical;
With 3,5-difluoro γ-picolinic acid for starting raw material, there is substitution reaction with compound (b) in 3,5-difluoro γ-picolinic acid, obtains compound (c) under suitable conditions; Compound (c) obtains general formula compound (d) with ammoniacal liquor condensation under CDI exists; There is substitution reaction in organic solvent in compound (d) and 3-sulfydryl phenylamino t-butyl formate, obtains compound (Ie) in the presence of a base; Compound (Ie) in presence of an acid in organic solvent de-Boc protection obtain compound (If); Compound (If) in the presence of an organic base in organic solvent with acyl chlorides or acid anhydrides generation acylation reaction, obtain compound (Ig); Compound (Ig) hydrolysis obtains compound (Ih); Compound (Ih) obtains compound (Ia) through acid amide condensation in organic solvent under condensing agent, organic amine exist.
17. 1 kinds of pharmaceutical compositions, comprise formula (I) compound or its pharmacy acceptable salt and drug acceptable carrier, vehicle or thinner.
The application of compound described in 18. any one of claim 1 ~ 15 in the medicine of preparation treatment Mammals especially human hyperproliferative's disease.
The application of compound described in 19. any one of claim 1 ~ 15 in the medicine of preparation treatment tumour.
20. suppress mitogen activated protein kinase kinases (MEK) active and hinder mitogen activated protein kinase kinase kinase (Raf) to a method for the phosphorylation of MEK, comprise formula (I) compound to patient therapeuticallv's significant quantity or its pharmacy acceptable salt and pharmaceutically acceptable carrier, vehicle or thinner.
21. treat a method for proliferative disease, described method comprises to be used formula (I) compound or its pharmacy acceptable salt to patient or comprises treatment formula (I) compound of significant quantity and the pharmaceutical composition of drug acceptable carrier, vehicle or thinner.
22. treat a method for tumour, described method comprises to be used formula (I) compound or its pharmacy acceptable salt to patient or comprises treatment formula (I) compound of significant quantity and the pharmaceutical composition of drug acceptable carrier, vehicle or thinner.
23. methods as claimed in claim 22, wherein said tumour is melanoma, mammary cancer, colon and rectum carcinoma, nonsmall-cell lung cancer or small cell lung cancer, liver cancer, kidney.
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Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1652776A (en) * 2002-03-13 2005-08-10 阵列生物制药公司 N3 alkylated benzimidazole derivatives as MEK inhibitors
CN1219753C (en) * 2000-07-19 2005-09-21 沃尼尔·朗伯公司 Oxygenated esters of 4-iodo phenylamino benzhydroxamic acids
CN101065358A (en) * 2004-10-20 2007-10-31 应用研究***Ars股份公司 3-arylamino pyridine derivatives
US20090012091A1 (en) * 2007-07-02 2009-01-08 Kinagen, Inc. Oximide derivatives and their therapeutical application
CN101365676A (en) * 2005-10-07 2009-02-11 埃克塞利希斯股份有限公司 Azetidines as mek inhibitors for the treatment of proliferative diseases
WO2012000598A1 (en) * 2010-07-01 2012-01-05 Merck Patent Gmbh Method for the preparation of cis-1,2-diols in the kilogram scale
CN103282351A (en) * 2010-10-29 2013-09-04 拜耳知识产权有限责任公司 Substituted phenoxypyridines

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1219753C (en) * 2000-07-19 2005-09-21 沃尼尔·朗伯公司 Oxygenated esters of 4-iodo phenylamino benzhydroxamic acids
CN1652776A (en) * 2002-03-13 2005-08-10 阵列生物制药公司 N3 alkylated benzimidazole derivatives as MEK inhibitors
CN101065358A (en) * 2004-10-20 2007-10-31 应用研究***Ars股份公司 3-arylamino pyridine derivatives
CN101365676A (en) * 2005-10-07 2009-02-11 埃克塞利希斯股份有限公司 Azetidines as mek inhibitors for the treatment of proliferative diseases
US20090012091A1 (en) * 2007-07-02 2009-01-08 Kinagen, Inc. Oximide derivatives and their therapeutical application
WO2012000598A1 (en) * 2010-07-01 2012-01-05 Merck Patent Gmbh Method for the preparation of cis-1,2-diols in the kilogram scale
CN103282351A (en) * 2010-10-29 2013-09-04 拜耳知识产权有限责任公司 Substituted phenoxypyridines

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
郭宗儒: "《药物化学总论》", 31 August 2003 *

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